00610nas a2200169 4500008004100000022001400041245011400055210006900169490000700238100002500245700002600270700002000296700002500316700002600341700002500367856004800392 2024 eng d a2296-861X00aMammary epithelium permeability during established lactation: associations with cytokine levels in human milk0 aMammary epithelium permeability during established lactation ass0 v111 aKivlighan, Katie, T.1 aSchneider, Sallie, S.1 aBrowne, Eva, P.1 aPentecost, Brian, T.1 aAnderton, Douglas, L.1 aArcaro, Kathleen, F. uhttp://dx.doi.org/10.3389/fnut.2024.125890500575nas a2200157 4500008004100000022001400041245015100055210006900206300001400275490000700289100001900296700002000315700002500335700001600360856004100376 2024 eng d a1473-018900aRapid cell isolation in breastmilk in a non-clinical setting by a deterministic lateral displacement device and selective water and fat absorption0 aRapid cell isolation in breastmilk in a nonclinical setting by a a604–6140 v241 aHawkins, Jamar1 aBrowne, Eva, P.1 aArcaro, Kathleen, F.1 aSun, Yubing uhttp://dx.doi.org/10.1039/d3lc00899a00491nas a2200145 4500008004100000245010000041210006900141300000900210490000700219100001900226700001900245700002100264700002100285856003900306 2023 eng d00aEffect of Maternal Diet on Maternal Milk and Breastfed Infant Gut Microbiomes: A Scoping Review0 aEffect of Maternal Diet on Maternal Milk and Breastfed Infant Gu a14200 v151 aTaylor, Rachel1 aKeane, Deirdre1 aBorrego, Paulina1 aArcaro, Kathleen uhttps://doi.org/10.3390/nu1506142000447nas a2200121 4500008004100000245012000041210006900161300001400230490000800244100001700252700001300269856004300282 2022 eng d00aBerberine alleviates LPS-induced apoptosis, oxidation, and skewed lineages during mouse preimplantation development0 aBerberine alleviates LPSinduced apoptosis oxidation and skewed l a699–7090 v1061 aMiao, Xiaosu1 aCui, Wei uhttps://doi.org/10.1093/biolre/ioac00202330nas a2200289 4500008004100000022001400041245012400055210006900179260001500248300001200263490000800275520141000283653001201693653001401705653001401719653001701733653001101750653001101761653002401772653001201796653000901808653003001817653001401847100001701861700001301878856014901891 2022 eng d a1529-726800aBerberine alleviates LPS-induced apoptosis, oxidation, and skewed lineages during mouse preimplantation development†.0 aBerberine alleviates LPSinduced apoptosis oxidation and skewed l c2022 04 26 a699-7090 v1063 a
Female infertility is a heterogeneous disorder with a variety of complex causes, including inflammation and oxidative stress, which are also closely associated with the pathogenesis of polycystic ovary syndrome (PCOS). As a new treatment for PCOS, berberine (BER), a natural compound from Berberis, has been clinically applied recently. However, the mechanisms underlying the association between BER and embryogenesis are still largely unknown. In this study, effects of BER on preimplantation development were evaluated under both normal and inflammatory culture conditions induced by lipopolysaccharide (LPS) in mice. Our data first suggest that BER itself (25 nM) does not affect embryo quality or future developmental potency; however, it can effectively alleviate LPS-induced embryo damage by mitigating apoptosis via reactive oxygen species (ROS)-/caspase-3-dependent pathways and by suppressing proinflammatory cytokines via inhibition of the NF-κB signaling pathway during preimplantation embryonic development. In addition, skewed cell lineage specification in the inner cell mass (ICM) and primitive endoderm (PE) caused by LPS can also be successfully rescued with BER. In summary, these findings for the first time demonstrate the nontoxicity of low doses of BER and its antiapoptotic and antioxidative properties on embryonic cells during mammalian preimplantation development.
10aAnimals10aApoptosis10aBerberine10aCell Lineage10aFemale10aHumans10aLipopolysaccharides10aMammals10aMice10aPolycystic Ovary Syndrome10aPregnancy1 aMiao, Xiaosu1 aCui, Wei uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/berberine-alleviates-lps-induced-apoptosis-oxidation-and-skewed-002119nas a2200277 4500008004100000022001400041245008000055210006900135260001200204300001200216490000700228520125800235653001201493653001401505653001401519653001601533653001401549653001201563653002001575653001301595100002301608700001601631700002601647700001701673856015101690 2022 eng d a1098-279500aBorcs6 is required for endo-lysosomal degradation during early development.0 aBorcs6 is required for endolysosomal degradation during early de c2022 08 a337-3500 v893 aEarly development and differentiation require precise control of cellular functions. Lysosomal degradation is a critical component of normal cellular homeostasis, allowing for degradation of signaling molecules, proteins, and other macromolecules for cellular remodeling and signaling. Little is known about the role of lysosomal function in mammalian embryos before gastrulation. Borcs6 is a protein involved in lysosomal trafficking as well as endo-lysosomal and autophagosome fusion. Here, we show that Borcs6 is necessary for efficient endo-lysosomal degradation in the early embryo. Although embryos lacking Borcs6 are developmentally comparable to control littermates at E5.5, they are characterized by large cells containing increased levels of late endosomes and abnormal nuclei. Furthermore, these embryos display a skewed ratio of extraembryonic and embryonic cell lineages, are delayed by E6.5, and do not undergo normal gastrulation. These results demonstrate the essential functions of lysosomal positioning and fusion with endosomes during early embryonic development and indicate that the early lethality of BORCS6 mutant embryos is primarily due to defects in the HOPS-related function of BORC rather than lysosomal positioning.
10aAnimals10aAutophagy10aEndosomes10aHomeostasis10aLysosomes10aMammals10aMembrane Fusion10aProteins1 aBell, Charlotte, J1 aGupta, Neha1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/borcs6-required-endo-lysosomal-degradation-during-early-development02907nas a2200325 4500008004100000022001400041245009200055210006900147260001300216300001000229490000800239520192000247653001202167653001302179653001102192653001002203653001202213653000902225653001402234653002402248653001402272653001402286100002302300700002002323700001702343700002102360700002102381700002602402856015302428 2022 eng d a1095-564X00aDeciphering the role of retinoic acid in hepatic patterning and induction in the mouse.0 aDeciphering the role of retinoic acid in hepatic patterning and c2022 Nov a31-420 v4913 aRetinoic acid (RA), a metabolite of vitamin A, is a small molecule and morphogen that is required for embryonic development. While normal RA signals are required for hepatic development in a variety of vertebrates, a role for RA during mammalian hepatic specification has yet to be defined. To examine the requirement for RA in murine liver induction, we performed whole embryo culture with the small molecule RA inhibitor, BMS493, to attenuate RA signaling immediately prior to hepatic induction and through liver bud formation. BMS493 treated embryos demonstrated a significant loss of hepatic specification that was confined to the prospective dorsal anterior liver bud. Examination of RA attenuated embryos demonstrates that while the liver bud displays normal expression of foregut endoderm markers and the hepato-pancreatobiliary domain marker, PROX1, the dorsal/anterior liver bud excludes the critical hepatic marker, HNF4α, indicating that RA signals are required for dorsal/anterior hepatic induction. These results were confirmed and extended by careful examination of Rdh10 embryos, which carry a genetic perturbation in RA synthesis. At E9.5 Rdh10 embryos display a similar yet more significant loss of the anterior/dorsal liver bud. Notably the anterior/dorsal liver bud loss correlates with the known dorsal-ventral gradient of the RA synthesis enzyme, Aldh1a2. In addition to altered hepatic specification, the mesoderm surrounding the liver bud is disorganized in RA abrogated embryos. Analysis of E10.5 Rdh10 embryos reveals small livers that appear to lack the dorsal/caudal lobes. Finally, addition of exogenous RA prior to hepatic induction results in a liver bud that has failed to thicken and is largely unspecified. Taken together our ex vivo and in vivo evidence demonstrate that the generation of normal RA gradients is required for hepatic patterning, specification, and growth.
10aAnimals10aEndoderm10aFemale10aLiver10aMammals10aMice10aPregnancy10aProspective Studies10aTretinoin10aVitamin A1 aGuertin, Taylor, M1 aPalaria, Amrita1 aMager, Jesse1 aSandell, Lisa, L1 aTrainor, Paul, A1 aTremblay, Kimberly, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/deciphering-role-retinoic-acid-hepatic-patterning-and-induction-mouse03726nas a2200457 4500008004100000022001400041245010000055210006900155260000900224300001100233490000700244520235400251653001302605653002202618653001102640653001102651653002102662653002102683653002102704653001102725653001402736653001902750653001602769653001402785653001402799653002002813653001502833653001102848653003602859100002702895700002402922700002202946700002102968700002502989700002503014700001903039700001603058700001603074700002403090856015403114 2022 eng d a1664-322400aDurable antibody and effector memory T cell responses in breastmilk from women with SARS-CoV-2.0 aDurable antibody and effector memory T cell responses in breastm c2022 a9852260 v133 aBackground: Given that only 25% of pregnant women elect to receive a COVID-19 vaccine, maternal SARS-CoV-2 infection remains an important route of conferring protective passive immunity to breastfed infants of mothers who are not vaccinated.
Methods: We enrolled 30 lactating participants between December 2020 and March 2021 who had a positive PCR-test and their first COVID-19 symptoms within the previous 21 days. Participants were asked to provide serial bilateral milk samples at 12 timepoints (~ every 3 days) over a period of 35 days. A second set of samples was collected at least four months after the beginning of the first set. Participants also were asked to provide their dried blood spots and infant stool samples. All samples were tested for receptor-binding domain (RBD)-specific immunoglobulin (Ig)A, IgG, and IgM. Milk samples were assessed for neutralizing ability against the spike protein and four SARS-CoV-2 variants: D614G, Alpha (B.1.1.7), Beta (B.1.351), and Gamma (P.1). Permeability of the breast epithelium was assessed by measuring the sodium to potassium ions (Na:K) in milk. Using flow cytometry, memory CD4 and CD8 T cells (CD45RO and CCR7) and mucosal-homing CD4 and CD8 T cells (CD103) were determined in cells from milk expressed at 35 days and at least 4 months after their first milk donation.
Results: Milk antibodies from SARS-CoV-2 positive participants neutralized the spike complex. Milk from 73, 90, and 53% of participants had binding reactivities to RBD-specific IgA, IgG, and IgM, respectively. In contrast to blood spots, which showed increased levels of IgG, but not IgA or IgM, the COVID-19 response in milk was associated with a robust IgA response. Twenty-seven percent of participants had increased breast-epithelium permeability, as indicated by Na:K ≥ 0.6. The percentage of CD45ROCCR7 effector-memory T cells in the day ≥120 milk samples was significantly higher than day 35 samples (< 0.05).
Conclusions: Antibodies in milk from participants with recent SARS-CoV-2 infection and those who recovered can neutralize the spike complex. For the first time we show that breastmilk T cells are enriched for mucosal memory T cells, further emphasizing the passive protection against SARS-CoV-2 conferred to infants breastmilk.
10aCOVID-1910aCOVID-19 Vaccines10aFemale10aHumans10aImmunoglobulin A10aImmunoglobulin G10aImmunoglobulin M10aInfant10aLactation10aMemory T Cells10aMilk, Human10aPotassium10aPregnancy10aReceptors, CCR710aSARS-CoV-210aSodium10aSpike Glycoprotein, Coronavirus1 aNarayanaswamy, Vignesh1 aPentecost, Brian, T1 aTelfer, Janice, C1 aBurnside, Amy, S1 aSchneider, Sallie, S1 aAlfandari, Dominique1 aBaker, Ryan, L1 aSaiju, Aman1 aNodiff, Sam1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/durable-antibody-and-effector-memory-t-cell-responses-breastmilk-women00505nas a2200181 4500008004100000245005200041210005200093300001300145490000700158100001700165700001400182700001800196700002000214700002100234700001700255700001300272856003800285 2022 eng d00aEarly embryonic lethality of mice lacking POLD20 aEarly embryonic lethality of mice lacking POLD2 a98–1080 v901 aWu, Xiaoqing1 aLiu, Yong1 aWang, Wenying1 aCrimmings, Kate1 aWilliams, Andrea1 aMager, Jesse1 aCui, Wei uhttps://doi.org/10.1002/mrd.2366300455nas a2200121 4500008004100000245011100041210006900152260001000221100001900231700002400250700002000274856003900294 2022 eng d00aEffect of early or late blood sampling on thyrotropin releasing hormone stimulation test results in horses0 aEffect of early or late blood sampling on thyrotropin releasing bWiley1 aThane, Kristen1 aUricchio, Cassandra1 aFrank, Nicholas uhttps://doi.org/10.1111/jvim.1636202436nas a2200277 4500008004100000022001400041245013300055210006900188260001500257300001200272490000700284520143200291653002701723653003101750653002101781653003301802653001801835100002001853700001801873700002001891700002201911700002901933700002401962700002001986856015202006 2022 eng d a1520-481200aEvaluation of Cellular Targeting by Fab' vs Full-Length Antibodies in Antibody-Nanoparticle Conjugates (ANCs) Using CD4 T-cells.0 aEvaluation of Cellular Targeting by Fab vs FullLength Antibodies c2022 03 16 a486-4950 v333 aTargeted delivery of chemotherapeutic drugs can improve their therapeutic efficiency by localizing their toxic effects at the diseased site. This is often achieved either by direct conjugation of drugs to antibodies targeting overexpressed receptors on cancer cells (antibody-drug conjugates/ADCs) or by conjugating antibodies to nanoparticles bearing drugs (antibody-nanoparticle conjugates/ANCs). Here, we report a platform for utilizing hinge cysteines on antigen-binding fragment (Fab') of an anti-CD4 antibody for site-specific conjugation to nanoparticles giving rise to anti-CD4 Fab'-nanoparticle conjugates (Fab'-NCs). We demonstrate a convenient route for obtaining functional anti-CD4 Fab' from full-length antibody and examine the targeted delivery efficiencies of anti-CD4 Fab'-NCs vs ANCs for selective delivery to CD4 mT-ALL cells. Our results indicate that higher avidity of full-length anti-CD4 antibody, i.e., protein alone translated to higher binding ability to CD4 mT-ALL cells in comparison with anti-CD4 Fab' alone. However, the targeted delivery efficiency of anti-CD4 Fab'-NCs was comparable to ANCs indicating that the avidity of Fab' is restored in a nanoparticle-conjugate format. Fab'-NCs are equally capable of achieving targeted drug delivery to CD4 T-cells as ANCs and are a versatile alternative to ANCs by offering site-selective modification strategy while retaining their advantages.
10aAntibodies, Monoclonal10aCD4-Positive T-Lymphocytes10aImmunoconjugates10aImmunoglobulin Fab Fragments10aNanoparticles1 aSingh, Khushboo1 aCanakci, Mine1 aKanjilal, Pintu1 aWilliams, Natalie1 aShanthalingam, Sudarvili1 aOsborne, Barbara, A1 aThayumanavan, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evaluation-cellular-targeting-fab-vs-full-length-antibodies-antibody03036nas a2200421 4500008004100000022001400041245014000055210006900195260001500264300001300279490000700292520161700299653001201916653002401928653002301952653001101975653002001986653001702006653002202023653001502045653001802060653001202078653002002090653004302110653004402153653002502197653002502222653002402247653002402271100002302295700002602318700002102344700001902365700002202384700003202406700002402438856015202462 2022 eng d a1098-552200aIdentification of Leptospiral Protein Antigens Recognized by WC1 γδ T Cell Subsets as Target for Development of Recombinant Vaccines.0 aIdentification of Leptospiral Protein Antigens Recognized by WC1 c2022 01 25 ae00492210 v903 aPathogenic species cause leptospirosis, a neglected zoonotic disease recognized as a global public health problem. It is also the cause of the most common cattle infection that results in major economic losses due to reproductive problems. γδ T cells play a role in the protective immune response in livestock species against , while human γδ T cells also respond to . Thus, activation of γδ T cells has emerged as a potential component in the optimization of vaccine strategies. Bovine γδ T cells proliferate and produce gamma interferon (IFN-γ) in response to vaccination with inactivated leptospires, and this response is mediated by a specific subpopulation of the WC1-bearing γδ T cells. WC1 molecules are members of the group B scavenger receptor cysteine-rich (SRCR) superfamily and are composed of multiple SRCR domains, of which particular extracellular domains act as ligands for Since WC1 molecules function as both pattern recognition receptors and γδ TCR coreceptors, the WC1 system has been proposed as a novel target to engage γδ T cells. Here, we demonstrate the involvement of leptospiral protein antigens in the activation of WC1 γδ T cells and identify two leptospiral outer membrane proteins able to interact directly with them. Interestingly, we show that the protein-specific γδ T cell response is composed of WC1.1 and WC1.2 subsets, although a greater number of WC1.1 γδ T cells respond. Identification of protein antigens will enhance our understanding of the role γδ T cells play in the leptospiral immune response and in recombinant vaccine development.
10aAnimals10aAntigens, Bacterial10aBacterial Proteins10aCattle10aCattle Diseases10aImmunization10aImmunophenotyping10aLeptospira10aLeptospirosis10aLigands10aProtein Binding10aProtein Interaction Domains and Motifs10aReceptors, Antigen, T-Cell, gamma-delta10aRecombinant Proteins10aT-Lymphocyte Subsets10aVaccine Development10aVaccines, Synthetic1 aTeixeira, Aline, F1 aGillespie, Alexandria1 aYirsaw, Alehegne1 aBritton, Emily1 aTelfer, Janice, C1 aNascimento, Ana, Lucia Tabe1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-leptospiral-protein-antigens-recognized-wc1-gd-t-cell02782nas a2200349 4500008004100000022001400041245010500055210006900160260001200229300001200241490000700253520162400260653003701884653001201921653002101933653001601954653001401970653001101984653001101995653003002006653003602036653002802072653000902100653002502109100002102134700002502155700002302180700002802203700001802231700003002249856015302279 2022 eng d a1573-703900aInduced mammary cancer in rat models: pathogenesis, genetics, and relevance to female breast cancer.0 aInduced mammary cancer in rat models pathogenesis genetics and r c2022 06 a185-2100 v273 aMammary cancer, or breast cancer in women, is a polygenic disease with a complex etiopathogenesis. While much remains elusive regarding its origin, it is well established that chemical carcinogens and endogenous estrogens contribute significantly to the initiation and progression of this disease. Rats have been useful models to study induced mammary cancer. They develop mammary tumors with comparable histopathology to humans and exhibit differences in resistance or susceptibility to mammary cancer depending on strain. While some rat strains (e.g., Sprague-Dawley) readily form mammary tumors following treatment with the chemical carcinogen, 7,12-dimethylbenz[a]-anthracene (DMBA), other strains (e.g., Copenhagen) are resistant to DMBA-induced mammary carcinogenesis. Genetic linkage in inbred strains has identified strain-specific quantitative trait loci (QTLs) affecting mammary tumors, via mechanisms that act together to promote or attenuate, and include 24 QTLs controlling the outcome of chemical induction, 10 QTLs controlling the outcome of estrogen induction, and 4 QTLs controlling the outcome of irradiation induction. Moreover, and based on shared factors affecting mammary cancer etiopathogenesis between rats and humans, including orthologous risk regions between both species, rats have served as useful models for identifying methods for breast cancer prediction and treatment. These studies in rats, combined with alternative animal models that more closely mimic advanced stages of breast cancer and/or human lifestyles, will further improve our understanding of this complex disease.
10a9,10-Dimethyl-1,2-benzanthracene10aAnimals10aBreast Neoplasms10aCarcinogens10aEstrogens10aFemale10aHumans10aMammary Neoplasms, Animal10aMammary Neoplasms, Experimental10aQuantitative Trait Loci10aRats10aRats, Sprague-Dawley1 aMiller, James, L1 aBartlett, Arianna, P1 aHarman, Rebecca, M1 aMajhi, Prabin, Dhangada1 aJerry, Joseph1 aVan de Walle, Gerlinde, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/induced-mammary-cancer-rat-models-pathogenesis-genetics-and-relevance03161nas a2200373 4500008004100000022001400041245013800055210006900193260001300262300001100275490000800286520189800294653002802192653002102220653001102241653002002252653001102272653003002283653003202313100002402345700002802369700002302397700002202420700001902442700002202461700001702483700002802500700002202528700001902550700002502569700001802594700002502612856015002637 2022 eng d a1090-241400aInterindividual variation contributes to differential PCB 126 induced gene expression in primary breast epithelial cells and tissues.0 aInterindividual variation contributes to differential PCB 126 in c2022 Aug a1137220 v2413 aPCB 126 is a pervasive, dioxin-like chemical pollutant which can activate the aryl hydrocarbon receptor (AhR). Despite being banned from the market, PCB 126 can be detected in breast milk to this day. The extent to which interindividual variation impacts the adverse responses to this chemical in the breast tissue remains unclear. This study aimed to investigate the impact of 3 nM PCB 126 on gene expression in a panel of genetically diverse benign human breast epithelial cell (HBEC) cultures and patient derived breast tissues. Six patient derived HBEC cultures were treated with 3 nM PCB 126. RNAseq was used to interrogate the impact of exposure on differential gene expression. Gene expression changes from the top critical pathways were confirmed via qRT-PCR in a larger panel of benign patient derived HBEC cultures, as well as in patient-derived breast tissue explant cultures. RNAseq analysis of HBEC cultures revealed a signature of 144 genes significantly altered by 3 nM PCB 126 treatment. Confirmation of 8 targets using a panel of 12 HBEC cultures and commercially available breast cell lines demonstrated that while the induction of canonical downstream target gene, CYP1A1, was consistent across our primary HBECs, other genes including AREG, S100A8, IL1A, IL1B, MMP7, and CCL28 exhibited significant variability across individuals. The dependence on the activity of the aryl hydrocarbon receptor was confirmed using inhibitors. PCB 126 can induce significant and consistent changes in gene expression associated with xenobiotic metabolism in benign breast epithelial cells. Although the induction of most genes was reliant on the AhR, significant variability was noted between genes and individuals. These data suggest that there is a bifurcation of the pathway following AhR activation that contributes to the variation in interindividual responses.
10aCytochrome P-450 CYP1A110aEpithelial Cells10aFemale10aGene Expression10aHumans10aPolychlorinated Biphenyls10aReceptors, Aryl Hydrocarbon1 aMorin, Stephanie, M1 aMajhi, Prabin, Dhangada1 aCrisi, Giovanna, M1 aGregory, Kelly, J1 aFranca, Renata1 aSchalet, Benjamin1 aMason, Holly1 aCasaubon, Jesse, Thomas1 aCao, Qing, Jackie1 aHaddad, Sandra1 aMakari-Judson, Grace1 aJerry, Joseph1 aSchneider, Sallie, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/interindividual-variation-contributes-differential-pcb-126-induced02755nas a2200313 4500008004100000022001400041245007600055210006900131260001200200300001000212490000800222520171400230653002101944653003201965653001401997653004602011653002502057653001702082653001702099653002502116100002002141700002102161700002102182700002202203700002402225700002502249700002002274856014702294 2022 eng d a1095-564X00aMcrs1 is required for branchial arch and cranial cartilage development.0 aMcrs1 is required for branchial arch and cranial cartilage devel c2022 09 a62-750 v4893 aMcrs1 is a multifunctional protein that is critical for many cellular processes in a wide range of cell types. Previously, we showed that Mcrs1 binds to the Six1 transcription factor and reduces the ability of the Six1-Eya1 complex to upregulate transcription, and that Mcrs1 loss-of-function leads to the expansion of several neural plate genes, reduction of neural border and pre-placodal ectoderm (PPR) genes, and pleiotropic effects on various neural crest (NC) genes. Because the affected embryonic structures give rise to several of the cranial tissues affected in Branchio-otic/Branchio-oto-renal (BOR) syndrome, herein we tested whether these gene expression changes subsequently alter the development of the proximate precursors of BOR affected structures - the otic vesicles (OV) and branchial arches (BA). We found that Mcrs1 is required for the expression of several OV genes involved in inner ear formation, patterning and otic capsule cartilage formation. Mcrs1 knockdown also reduced the expression domains of many genes expressed in the larval BA, derived from either NC or PPR, except for emx2, which was expanded. Reduced Mcrs1 also diminished the length of the expression domain of tbx1 in BA1 and BA2 and interfered with cranial NC migration from the dorsal neural tube; this subsequently resulted in defects in the morphology of lower jaw cartilages derived from BA1 and BA2, including the infrarostral, Meckel's, and ceratohyal as well as the otic capsule. These results demonstrate that Mcrs1 plays an important role in processes that lead to the formation of craniofacial cartilages and its loss results in phenotypes consistent with reduced Six1 activity associated with BOR.
10aBranchial Region10aBranchio-Oto-Renal Syndrome10aCartilage10aGene Expression Regulation, Developmental10aHomeodomain Proteins10aNeural Crest10aNeural Plate10aRNA-Binding Proteins1 aKeer, Stephanie1 aCousin, Hélène1 aJourdeuil, Karyn1 aNeilson, Karen, M1 aTavares, Andre, L P1 aAlfandari, Dominique1 aMoody, Sally, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mcrs1-required-branchial-arch-and-cranial-cartilage-development03571nas a2200805 4500008004100000022001400041245010000055210006900155260001500224300000800239490000700247520115900254653001201413653003301425653002801458653002301486653001801509653002201527653001901549653001601568653001201584653001001596653002501606653003201631653002001663653001801683653003501701653001101736653000901747653002301756653002201779653002001801653004001821653003801861653004301899653002501942653001401967100001801981700002201999700002602021700001802047700001902065700001602084700002902100700002402129700002102153700002402174700002102198700002302219700002302242700002102265700001402286700001702300700002102317700002002338700001802358700002102376700001602397700001602413700001502429700002702444700002602471700002002497700002202517700002102539700001702560700002402577700001902601856014502620 2022 eng d a2041-172300aMicrobial enzymes induce colitis by reactivating triclosan in the mouse gastrointestinal tract.0 aMicrobial enzymes induce colitis by reactivating triclosan in th c2022 01 10 a1360 v133 aEmerging research supports that triclosan (TCS), an antimicrobial agent found in thousands of consumer products, exacerbates colitis and colitis-associated colorectal tumorigenesis in animal models. While the intestinal toxicities of TCS require the presence of gut microbiota, the molecular mechanisms involved have not been defined. Here we show that intestinal commensal microbes mediate metabolic activation of TCS in the colon and drive its gut toxicology. Using a range of in vitro, ex vivo, and in vivo approaches, we identify specific microbial β-glucuronidase (GUS) enzymes involved and pinpoint molecular motifs required to metabolically activate TCS in the gut. Finally, we show that targeted inhibition of bacterial GUS enzymes abolishes the colitis-promoting effects of TCS, supporting an essential role of specific microbial proteins in TCS toxicity. Together, our results define a mechanism by which intestinal microbes contribute to the metabolic activation and gut toxicity of TCS, and highlight the importance of considering the contributions of the gut microbiota in evaluating the toxic potential of environmental chemicals.
10aAnimals10aAnti-Infective Agents, Local10aAnticarcinogenic Agents10aBacterial Proteins10aBinding Sites10aBiotransformation10aCarcinogenesis10aCarcinogens10aColitis10aColon10aColorectal Neoplasms10aGastrointestinal Microbiome10aGene Expression10aGlucuronidase10aGlycoside Hydrolase Inhibitors10aHumans10aMice10aMice, Inbred C57BL10aModels, Molecular10aProtein Binding10aProtein Conformation, alpha-Helical10aProtein Conformation, beta-Strand10aProtein Interaction Domains and Motifs10aRecombinant Proteins10aTriclosan1 aZhang, Jianan1 aWalker, Morgan, E1 aSanidad, Katherine, Z1 aZhang, Hongna1 aLiang, Yanshan1 aZhao, Ermin1 aChacon-Vargas, Katherine1 aYeliseyev, Vladimir1 aParsonnet, Julie1 aHaggerty, Thomas, D1 aWang, Guangqiang1 aSimpson, Joshua, B1 aJariwala, Parth, B1 aBeaty, Violet, V1 aYang, Jun1 aYang, Haixia1 aPanigrahy, Anand1 aMinter, Lisa, M1 aKim, Daeyoung1 aGibbons, John, G1 aLiu, LinShu1 aLi, Zhengze1 aXiao, Hang1 aBorlandelli, Valentina1 aOverkleeft, Hermen, S1 aCloer, Erica, W1 aMajor, Michael, B1 aGoldfarb, Dennis1 aCai, Zongwei1 aRedinbo, Matthew, R1 aZhang, Guodong uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/microbial-enzymes-induce-colitis-reactivating-triclosan-mouse02329nas a2200181 4500008004100000022001400041245010100055210006900156260001600225520163700241100001801878700001701896700001601913700001801929700002301947700002601970856015101996 2022 eng d a1741-789900aMultidrug resistance transporter-1 dysfunction perturbs meiosis and Ca2+ homeostasis in oocytes.0 aMultidrug resistance transporter1 dysfunction perturbs meiosis a c2022 Oct 013 aMDR-1 is a transmembrane ATP-dependent effluxer present in organs that transport a variety of xenobiotics and byproducts. Previous findings by our group demonstrated that this transporter is also present in the oocyte mitochondrial membrane and that its mutation led to abnormal mitochondrial homeostasis. Considering the importance of these organelles in the female gamete, we assessed the impact of MDR-1 dysfunction on mouse oocyte quality, with a particular focus on the meiotic spindle organization, aneuploidies, Ca2+ homeostasis, ATP production and mtDNA mutations. Our results demonstrate that young Mdr1 mutant mice produce oocytes characterized by lower quality, with a significant delay in the germinal vesicle (GV) to germinal vesicle breakdown (GVBD) transition, an increased percentage of symmetric divisions, chromosome mis-alignments and a severely altered meiotic spindle shape compared to the wild types. Mutant oocytes exhibit 7000 more single nucleotide polymorphisms (SNPs) in the exomic DNA and twice the amount of mitochondrial DNA SNPs compared to the wild-type ones. Ca2+ analysis revealed the inability of MDR-1 mutant oocytes to manage Ca2+ storage content and oscillations in response to several stimuli and ATP quantification shows that mutant oocytes trend towards lower ATP levels compared to wild types. Finally, 1-year-old mutant ovaries express a lower amount of Sirt1, Sirt3, Sirt5, Sirt6 and Sirt7 compared to wild type levels. These results, together emphasize the importance of MDR-1 in mitochondrial physiology and highlight the influence of MDR-1 on oocyte quality and ovarian aging.
1 aNabi, Dalileh1 aBosi, Davide1 aGupta, Neha1 aThaker, Nidhi1 aFissore, Rafael, A1 aBrayboy, Lynae, Maria uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/multidrug-resistance-transporter-1-dysfunction-perturbs-meiosis-and03550nas a2200421 4500008004100000022001400041245011700055210006900172260001500241300001200256490000800268520224500276653001002521653002902531653002202560653001902582653001902601653002202620653001402642653001102656653001102667653002102678653002102699653001102720653002002731653001602751653001602767653001502783653001602798100002702814700002402841700002202865700002502887700002502912700001602937700002402953856015102977 2022 eng d a1873-233X00aNeutralizing Antibodies and Cytokines in Breast Milk After Coronavirus Disease 2019 (COVID-19) mRNA Vaccination.0 aNeutralizing Antibodies and Cytokines in Breast Milk After Coron c2022 02 01 a181-1910 v1393 aOBJECTIVE: To evaluate immune responses to coronavirus disease 2019 (COVID-19) mRNA-based vaccines present in breast milk and transfer of the immune responses to breastfeeding infants.
METHODS: We enrolled 30 lactating women who received mRNA-based COVID-19 vaccines from January through April 2021 in this cohort study. Women provided serial milk samples, including milk expressed before vaccination, across 2-3 weeks after the first dose, and across 3 weeks after the second dose. Women provided their blood, spotted on cards (dried blood spots), 19 days after the first dose and 21 days after the second dose. Stool samples from the breastfed infants were collected 21 days after mothers' second vaccination. Prepandemic samples of milk, dried blood spots, and infant stool were used as controls. Milk, dried blood spots, and infant stool were tested by enzyme-linked immunosorbent assay for receptor-binding domain (RBD)-specific immunoglobulin (Ig)A and IgG. Milk samples were tested for the presence of neutralizing antibodies against the spike and four variants of concern: D614G, Alpha (B.1.1.7), Beta (B.1.351), and Gamma (P.1). Levels of 10 cytokines were measured in milk samples.
RESULTS: Milk from COVID-19-immunized women neutralized the spike and four variants of concern, primarily driven by anti-RBD IgG. The immune response in milk also included significant elevation of interferon-γ. The immune response to maternal vaccination was reflected in breastfed infants: anti-RBD IgG and anti-RBD IgA were detected in 33% and 30% of infant stool samples, respectively. Levels of anti-RBD antibodies in infant stool correlated with maternal vaccine side effects. Median antibody levels against RBD were below the positive cutoffs in prepandemic milk and infant stool samples.
CONCLUSION: Humoral and cellular immune responses to mRNA-based COVID-19 vaccination are present in most women's breast milk. The milk anti-RBD antibodies can neutralize severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike and variants of concern. Anti-RBD antibodies are transferred to breastfed infants, with the potential to confer passive immunity against SARS-CoV-2.
10aAdult10aAntibodies, Neutralizing10aAntibodies, Viral10aBreast Feeding10aCohort Studies10aCOVID-19 Vaccines10aCytokines10aFemale10aHumans10aImmunoglobulin A10aImmunoglobulin G10aInfant10aInfant, Newborn10aMiddle Aged10aMilk, Human10aSARS-CoV-210aVaccination1 aNarayanaswamy, Vignesh1 aPentecost, Brian, T1 aSchoen, Corina, N1 aAlfandari, Dominique1 aSchneider, Sallie, S1 aBaker, Ryan1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/neutralizing-antibodies-and-cytokines-breast-milk-after-coronavirus02001nas a2200205 4500008004100000022001400041245009800055210006900153260001300222300001100235490000800246520129200254653001201546653001101558653001701569653002201586653001401608100001901622856015401641 2022 eng d a1095-993900aNew chemistries for the control of human head lice, Pediculus humanus capitis: A mini-review.0 aNew chemistries for the control of human head lice Pediculus hum c2022 Feb a1050130 v1813 aPediculus lice represent one of the longest and most prevalent parasitic infestations of humans. Head lice are an economic and social concern whereas body lice pose a more serious public health threat. Significant progress has been made in the study of human lice over the last 10 years, allowing for new approaches in their control. An in vitro rearing system has made it possible to maintain insecticide-susceptible and -resistant reference strains, which allowed an in depth study of pediculicide resistance, including its underlying molecular mechanisms and the detection and monitoring of resistance. The generation of inbreed strains facilitated the efficient sequencing, assembly and annotation of the genomes and transcriptomes of both lice. The use of functional genomics and reverse genetics elucidated the genetics involved in the evolution of resistance and the discovery of novel target sites for the development of new pediculicides. In this review, four new effective pediculicide products, each with different mode of action and unique chemistries, will be presented. They have been found to be safe and selective, and control resistant lice. As such, they meet the criteria necessary to be used in rotations as a sustainable resistance management strategy.
10aAnimals10aHumans10aInsecticides10aLice Infestations10aPediculus1 aClark, John, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/new-chemistries-control-human-head-lice-pediculus-humanus-capitis-mini03458nas a2200325 4500008004100000022001400041245024400055210006900299260001500368300000800383490000700391520230500398653001002703653001502713653001902728653002102747653001502768653000902783653001102792653001002803653001102813653001102824653004202835653001502877100002302892700002002915700002602935700002402961856014702985 2022 eng d a1472-687400aNew Moms Wellness Study: trial study protocol for an intervention study to increase fruit and vegetable intake and lower breast cancer risk through weekly counseling and supplemental fruit and vegetable box delivery in breastfeeding women.0 aNew Moms Wellness Study trial study protocol for an intervention c2022 09 24 a3890 v223 aBACKGROUND: Laboratory studies indicate that chemicals in fruits and vegetables have anti-carcinogenic and anti-inflammatory activities that can lower breast cancer risk. However, epidemiologic studies of the association between fruit and vegetable intake and breast cancer risk have produced mixed results. Measurement error, confounding, and an emphasis on diet in later adulthood may contribute to weak associations. This paper describes a randomized controlled diet intervention trial in breastfeeding women to examine the effect of high fruit and vegetable intake on breast cancer risk factors, including weight, DNA methylation and inflammatory markers.
METHODS: Eligible breastfeeding women who reside within a 35-mile radius of Amherst, MA are enrolled at five to six weeks postpartum and randomly assigned to a Fruit and Vegetable Intervention Arm (target n = 200) or to a USDA MyPlate Control Arm (target n = 200). The Fruit and Vegetable Intervention group receives weekly telephone or video-based counseling to encourage intake of at least eight to ten daily servings of fruits and vegetables and a weekly delivery of a supplemental box of fruits and vegetables for 20 weeks, and less intensive counseling for up to one year. Breastmilk and infant fecal specimens are collected at baseline, 10 and 20 weeks. Anthropometric measurements are obtained at these timepoints and at the 1-year follow-up. The primary outcomes are change in DNA methylation in breast epithelial cells and change in inflammatory markers in breastmilk from randomization to 20 weeks; and change in weight, waist circumference, and fruit and vegetable intake for the period from randomization to 20 weeks and 1 year.
DISCUSSION: This 1-year randomized diet intervention trial in breastfeeding women will assess whether intake of at least eight to ten daily servings of fruits and vegetables per day improves biomarkers of breast cancer risk directly in the breast (i.e., DNA methylation and inflammatory markers) and helps women maintain a healthy weight.
TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04374747. Registered May 5, 2020. https://www.
CLINICALTRIALS: gov/ct2/show/NCT04374747 .
10aAdult10aBiomarkers10aBreast Feeding10aBreast Neoplasms10aCounseling10aDiet10aFemale10aFruit10aHumans10aInfant10aRandomized Controlled Trials as Topic10aVegetables1 aSturgeon, Susan, R1 aSibeko, Lindiwe1 aBalasubramanian, Raji1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/new-moms-wellness-study-trial-study-protocol-intervention-study02419nas a2200301 4500008004100000022001400041245009200055210006900147260001500216300001100231490000800242520146100250653001201711653001801723653001101741653000901752653001201761653000901773653001201782653003701794653003901831653001601870100001601886700002001902700002001922700002301942856015201965 2022 eng d a1741-789900aSPERM FACTORS AND EGG ACTIVATION: ICSI and the discovery of the sperm factor and PLCZ1.0 aSPERM FACTORS AND EGG ACTIVATION ICSI and the discovery of the s c2022 05 23 aF9-F200 v1643 aThe discovery of PLCZ1 nearly 20 years ago as the primary Ca2+ oscillation-inducing factor in the sperm of mammals represented a significant breakthrough in our quest to elucidate the molecules and pathways that promote egg activation during fertilization. The advent of the intracytoplasmic sperm injection (ICSI) technique, which made fertilization possible without sperm capacitation, acrosome reaction, and gamete fusion, strengthened the research that led to the discovery of PLCZ1 and became an essential clinical tool for humans. The use of ICSI combined with the detection of PLCZ1 expression and mutations in infertile patients established the fundamental role of PLCZ1 in human fertility while leading to the discovery of novel components of the perinuclear theca, the site of the residence of PLCZ1 in sperm before fertilization. Remarkably, the more extensive use of ICSI in species other than humans and mice revealed poor success and exposed gaps in our understanding of PLCZ1 release and/or activation. Similarly, fertilization using sperm from mouse models lacking Plcz1 has produced striking results whose true implications are yet to be determined. Nevertheless, answers to these unresolved questions will produce a complete picture of the adaptations and molecular players that mammalian species employ to ensure the success of the triggering event of embryo development that has linked generations since the beginning of times.
10aAnimals10aFertilization10aHumans10aMale10aMammals10aMice10aOocytes10aPhosphoinositide Phospholipase C10aSperm Injections, Intracytoplasmic10aSpermatozoa1 aGupta, Neha1 aAkizawa, Hiroki1 aLee, Hoi, Chang1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sperm-factors-and-egg-activation-icsi-and-discovery-sperm-factor-and02506nas a2200265 4500008004100000022001400041245015900055210006900214260001200283300001300295490000700308520158500315653001501900653001201915653002601927653002201953653002601975653001202001653000902013100001902022700001702041700001302058700001602071856015302087 2022 eng d a1742-566200aSurface functionalization of poly(dimethylsiloxane) substrates facilitates culture of pre-implantation mouse embryos by blocking non-selective adsorption.0 aSurface functionalization of polydimethylsiloxane substrates fac c2022 04 a202109290 v193 aPoly(dimethylsiloxane) (PDMS) is widely used in biomedical settings such as microfluidics for its optical transparency, castability, gas permeability and relative biocompatibility. While PDMS devices with certain modifications or treatments have been used for mammalian pre-implantation embryo culture, it is unclear why native PDMS leads to significant embryo death. In this study, we employ Nile Red as a model hydrophobic small molecule to demonstrate that significant hydrophobic sequestration occurs on native PDMS substrates even with a bovine serum albumin-containing KSOM pre-equilibration. Our results suggest that this small molecule sequestration has detrimental effects on mouse embryo development in PDMS static culture wells, with 0% blastocyst development rates from embryos cultured on native PDMS. We found that prior saturation of the PDMS culture well with water vapour only rescues about 10% of blastocyst development rates, indicating osmolality alone is not responsible for the high rates of embryo arrest. We also present a safe and simple Pluronic F127 pretreatment for PDMS substrates that successfully circumvented the harmful effects of native PDMS, achieving a blastocyst and implantation rate akin to that of our polystyrene controls. Our results call into question how researchers and clinicians can account for the alterations in medium composition and embryo secretions when using hydrophobic substrates, especially in the mammalian embryo culture setting where minimum effective concentrations of peptides and amino acids are commonplace.
10aAdsorption10aAnimals10aDimethylpolysiloxanes10aEmbryo, Mammalian10aEmbryonic Development10aMammals10aMice1 aHawkins, Jamar1 aMiao, Xiaosu1 aCui, Wei1 aSun, Yubing uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/surface-functionalization-polydimethylsiloxane-substrates-facilitates02152nas a2200421 4500008004100000022001400041245010600055210006900161260000900230300001100239490000700250520086000257653001401117653003101131653001101162653001801173653001301191653001401204653002101218100002201239700001501261700001601276700001801292700001901310700002501329700002401354700001901378700002101397700002001418700002001438700002201458700001301480700002301493700002401516700001901540700001701559856015401576 2022 eng d a1664-322400aTargeting the Cbl-b-Notch1 axis as a novel immunotherapeutic strategy to boost CD8+ T-cell responses.0 aTargeting the CblbNotch1 axis as a novel immunotherapeutic strat c2022 a9872980 v133 aA critical feature of cancer is the ability to induce immunosuppression and evade immune responses. Tumor-induced immunosuppression diminishes the effectiveness of endogenous immune responses and decreases the efficacy of cancer immunotherapy. In this study, we describe a new immunosuppressive pathway in which adenosine promotes Casitas B-lineage lymphoma b (Cbl-b)-mediated Notch1 degradation, causing suppression of CD8+ T-cells effector functions. Genetic knockout and pharmacological inhibition of Cbl-b prevents Notch1 degradation in response to adenosine and reactivates its signaling. Reactivation of Notch1 results in enhanced CD8+ T-cell effector functions, anti-cancer response and resistance to immunosuppression. Our work provides evidence that targeting the Cbl-b-Notch1 axis is a novel promising strategy for cancer immunotherapy.
10aAdenosine10aCD8-Positive T-Lymphocytes10aHumans10aImmunotherapy10aLymphoma10aNeoplasms10aReceptor, Notch11 aMonticone, Giulia1 aHuang, Zhi1 aCsibi, Fred1 aLeit, Silvana1 aCiccone, David1 aChamphekar, Ameya, S1 aAustin, Jermaine, E1 aUcar, Deniz, A1 aHossain, Fokhrul1 aIbba, Salome, V1 aBoulares, Hamid1 aCarpino, Nicholas1 aXu, Keli1 aMajumder, Samarpan1 aOsborne, Barbara, A1 aLoh, Christine1 aMiele, Lucio uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/targeting-cbl-b-notch1-axis-novel-immunotherapeutic-strategy-boost-cd802165nas a2200325 4500008004100000022001400041245012500055210006900180260001500249300001400264490000800278520109900286653001201385653001501397653001101412653000901423653003101432653001801463653001701481100002101498700002201519700002301541700001401564700001701578700002301595700002501618700002101643700002101664856015401685 2022 eng d a1537-661300aUse of a Contained Mycobacterium tuberculosis Mouse Infection Model to Predict Active Disease and Containment in Humans.0 aUse of a Contained Mycobacterium tuberculosis Mouse Infection Mo c2022 05 16 a1832-18400 v2253 aPrevious studies have identified whole-blood transcriptional risk and disease signatures for tuberculosis; however, several lines of evidence suggest that these signatures primarily reflect bacterial burden, which increases before symptomatic disease. We found that the peripheral blood transcriptome of mice with contained Mycobacterium tuberculosis infection (CMTI) has striking similarities to that of humans with active tuberculosis and that a signature derived from these mice predicts human disease with accuracy comparable to that of signatures derived directly from humans. A set of genes associated with immune defense are up-regulated in mice with CMTI but not in humans with active tuberculosis, suggesting that their up-regulation is associated with bacterial containment. A signature comprising these genes predicts both protection from tuberculosis disease and successful treatment at early time points where current signatures are not predictive. These results suggest that detailed study of the CMTI model may enable identification of biomarkers for human tuberculosis.
10aAnimals10aBiomarkers10aHumans10aMice10aMycobacterium tuberculosis10aTranscriptome10aTuberculosis1 aDuffy, Fergal, J1 aOlson, Gregory, S1 aGold, Elizabeth, S1 aJahn, Ana1 aAderem, Alan1 aAitchison, John, D1 aRothchild, Alissa, C1 aDiercks, Alan, H1 aNemeth, Johannes uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/use-contained-mycobacterium-tuberculosis-mouse-infection-model-predict00512nas a2200145 4500008004100000245007700041210006900118260003900187490000700226100002100233700002700254700001900281700002600300856004000326 2021 eng d00aAlveolar macrophages: novel therapeutic targets for respiratory diseases0 aAlveolar macrophages novel therapeutic targets for respiratory d bCambridge University Press ({CUP})0 v231 aLim, Pamelia, N.1 aCervantes, Maritza, M.1 aPham, Linh, K.1 aRothchild, Alissa, C. uhttps://doi.org/10.1017/erm.2021.2102134nas a2200253 4500008004100000022001400041245007800055210006900133260001500202300000800217490000700225520132000232653001301552653001101565653000901576653002601585653001401611653001501625100002001640700002601660700001801686700002501704856015101729 2021 eng d a1462-399400aAlveolar macrophages: novel therapeutic targets for respiratory diseases.0 aAlveolar macrophages novel therapeutic targets for respiratory d c2021 11 26 ae180 v233 aAlveolar macrophages (AMs) are lung-resident myeloid cells that sit at the interface of the airway and lung tissue. Under homeostatic conditions, their primary function is to clear debris, dead cells and excess surfactant from the airways. They also serve as innate pulmonary sentinels for respiratory pathogens and environmental airborne particles and as regulators of pulmonary inflammation. However, they have not typically been viewed as primary therapeutic targets for respiratory diseases. Here, we discuss the role of AMs in various lung diseases, explore the potential therapeutic strategies to target these innate cells and weigh the potential risks and challenges of such therapies. Additionally, in the context of the COVID-19 pandemic, we examine the role AMs play in severe disease and the therapeutic strategies that have been harnessed to modulate their function and protect against severe lung damage. There are many novel approaches in development to target AMs, such as inhaled antibiotics, liposomal and microparticle delivery systems, and host-directed therapies, which have the potential to provide critical treatment to patients suffering from severe respiratory diseases, yet there is still much work to be done to fully understand the possible benefits and risks of such approaches.
10aCOVID-1910aHumans10aLung10aMacrophages, Alveolar10aPandemics10aSARS-CoV-21 aLim, Pamelia, N1 aCervantes, Maritza, M1 aPham, Linh, K1 aRothchild, Alissa, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/alveolar-macrophages-novel-therapeutic-targets-respiratory-diseases01621nas a2200277 4500008004100000022001400041245009400055210006900149260001500218490000700233520069400240653001200934653002700946653002600973653001500999653003001014653002001044653001101064653001101075653003801086100001901124700001701143700001301160700001601173856015401189 2021 eng d a1460-240700aBiophysical optimization of preimplantation embryo culture: what mechanics can offer ART.0 aBiophysical optimization of preimplantation embryo culture what c2021 01 220 v273 aOwing to the rise of ART and mounting reports of epigenetic modification associated with them, an understanding of optimal embryo culture conditions and reliable indicators of embryo quality are highly sought after. There is a growing body of evidence that mechanical biomarkers can rival embryo morphology as an early indicator of developmental potential and that biomimetic mechanical cues can promote healthy development in preimplantation embryos. This review will summarize studies that investigate the role of mechanics as both indicators and promoters of mammalian preimplantation embryo development and evaluate their potential for improving future embryo culture systems.
10aAnimals10aBiomedical Engineering10aBiophysical Phenomena10aBlastocyst10aEmbryo Culture Techniques10aFallopian Tubes10aFemale10aHumans10aReproductive Techniques, Assisted1 aHawkins, Jamar1 aMiao, Xiaosu1 aCui, Wei1 aSun, Yubing uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/biophysical-optimization-preimplantation-embryo-culture-what-mechanics00612nas a2200181 4500008004100000245010900041210006900150260001600219300001000235490000700245100002400252700002500276700002200301700001900323700002300342700002300365856004200388 2021 eng d00aCaput Ligation Renders Immature Mouse Sperm Motile and Capable to Undergo cAMP-Dependent Phosphorylation0 aCaput Ligation Renders Immature Mouse Sperm Motile and Capable t b{MDPI} {AG} a102410 v221 aTourzani, Darya, A.1 aBattistone, Maria, A1 aSalicioni, Ana, M1 aBreton, Sylvie1 aVisconti, Pablo, E1 aGervasi, Maria, G. uhttps://doi.org/10.3390/ijms22191024100526nas a2200169 4500008004100000245009100041210006900132300001400201490000700215100001500222700001500237700001500252700001500267700001900282700001700301856003800318 2021 eng d00aThe contraceptive efficacy of a self-assembling intra-uterine device in domestic mares0 acontraceptive efficacy of a selfassembling intrauterine device i a130–1360 v991 aJoonè, CJ1 aGradil, CM1 aPicard, JA1 aTaylor, JD1 ade Tonnerre, D1 aCavalieri, J uhttps://doi.org/10.1111/avj.1305501973nas a2200385 4500008004100000022001400041245004700055210004500102260001500147490000700162520087700169653001201046653002001058653000901078653000901087653001901096653001801115653002001133653003001153100001501183700002701198700002001225700001701245700002101262700001201283700002501295700002301320700001701343700002201360700002101382700002301403700002301426700001701449856012101466 2021 eng d a2050-084X00aDeficient spermiogenesis in mice lacking .0 aDeficient spermiogenesis in mice lacking c2021 02 230 v103 aThe X-linked gene plays major roles in female mouse development and reproduction, where it is crucial for the maintenance of imprinted X chromosome inactivation in extraembryonic tissues of embryos. However, while females carrying a systemic knockout (KO) die around implantation, male KO mice appear healthy and are fertile. Here, we report an important role for in testis where it is highly expressed in post-meiotic round spermatids as well as in Sertoli cells. Systemic deletion of the gene results in lower numbers of mature sperm that contains excess cytoplasm, leading to decreased sperm motility and in vitro fertilization rates. Targeting the conditional cKO specifically to the spermatogenic cell lineage largely recapitulates this phenotype. These results reveal functions of in male reproduction specifically in round spermatids during spermiogenesis.
10aAnimals10aGenes, X-Linked10aMale10aMice10aMice, Knockout10aSertoli Cells10aSpermatogenesis10aUbiquitin-Protein Ligases1 aWang, Feng1 aGervasi, Maria, Gracia1 aBošković, Ana1 aSun, Fengyun1 aRinaldi, Vera, D1 aYu, Jun1 aWallingford, Mary, C1 aTourzani, Darya, A1 aMager, Jesse1 aZhu, Lihua, Julie1 aRando, Oliver, J1 aVisconti, Pablo, E1 aStrittmatter, Lara1 aBach, Ingolf uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/deficient-spermiogenesis-mice-lacking02340nas a2200337 4500008004100000022001400041245012000055210006900175260001500244490000800259520126000267653001201527653001201539653002901551653001101580653001401591653001801605653001801623653001601641653000901657653001901666653001201685653002501697653002501722100002001747700002001767700002001787700002101807700002001828856015401848 2021 eng d a1477-913700aDeletion of TRPV3 and CaV3.2 T-type channels in mice undermines fertility and Ca2+ homeostasis in oocytes and eggs.0 aDeletion of TRPV3 and CaV32 Ttype channels in mice undermines fe c2021 07 010 v1343 aCa2+ influx during oocyte maturation and after sperm entry is necessary to fill the internal Ca2+ stores and for complete egg activation. We knocked out the transient receptor potential vanilloid member 3 (TRPV3) and the T-type channel, CaV3.2, to determine their necessity for maintaining these functions in mammalian oocytes/eggs. Double-knockout (dKO) females were subfertile, their oocytes and eggs showed reduced internal Ca2+ stores, and, following sperm entry or Plcz (also known as Plcz1) cRNA injection, fewer dKO eggs displayed Ca2+ responses compared to wild-type eggs, which were also of lower frequency. These parameters were rescued and/or enhanced by removing extracellular Mg2+, suggesting that the residual Ca2+ influx could be mediated by the TRPM7 channel, consistent with the termination of divalent-cation oscillations in dKO eggs by a TRPM7 inhibitor. In total, we demonstrated that TRPV3 and CaV3.2 mediate the complete filling of the Ca2+ stores in mouse oocytes and eggs. We also showed that they are required for initiating and maintaining regularly spaced-out oscillations, suggesting that Ca2+ influx through PM ion channels dictates the periodicity and persistence of Ca2+ oscillations during mammalian fertilization.
10aAnimals10aCalcium10aCalcium Channels, T-Type10aFemale10aFertility10aFertilization10aGene Deletion10aHomeostasis10aMice10aMice, Knockout10aOocytes10aTRPM Cation Channels10aTRPV Cation Channels1 aMehregan, Aujan1 aArdestani, Goli1 aAkizawa, Hiroki1 aCarvacho, Ingrid1 aFissore, Rafael uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/deletion-trpv3-and-cav32-t-type-channels-mice-undermines-fertility-and00602nas a2200157 4500008004100000022001400041245007700055210006900132260000900201300001100210490000600221100002400227700002000251700002200271856015100293 2021 eng d a2296-634X00aEditorial: Targeting Developmental Pathways in Inflammation and Disease.0 aEditorial Targeting Developmental Pathways in Inflammation and D c2021 a7911150 v91 aAnastasiadou, Eleni1 aMinter, Lisa, M1 aFelli, Maria, Pia uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/editorial-targeting-developmental-pathways-inflammation-and-disease00592nas a2200157 4500008004100000022001400041245007200055210006900127260000900196300001100205490000600216100002400222700003000246700002300276856013500299 2021 eng d a2296-634X00aEditorial: The Fertilization Success From the Oocyte's Perspective.0 aEditorial The Fertilization Success From the Oocytes Perspective c2021 a8104200 v91 aMichaut, Marcela, A1 aSouza-Fabjan, Joanna, M G1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/editorial-fertilization-success-oocytes-perspective02931nas a2200421 4500008004100000022001400041245012100055210006900176260001500245490000800260520158600268653001201854653002001866653002501886653001101911653001401922653000901936653002701945653002201972653000901994653002402003653001302027653001402040653003802054100002102092700002702113700001702140700002202157700002702179700002302206700002202229700001902251700002102270700001802291700002502309700002502334856015002359 2021 eng d a1945-717000aExposure to Propylparaben During Pregnancy and Lactation Induces Long-Term Alterations to the Mammary Gland in Mice.0 aExposure to Propylparaben During Pregnancy and Lactation Induces c2021 06 010 v1623 aThe mammary gland is a hormone sensitive organ that is susceptible to endocrine-disrupting chemicals (EDCs) during the vulnerable periods of parous reorganization (ie, pregnancy, lactation, and involution). Pregnancy is believed to have long-term protective effects against breast cancer development; however, it is unknown if EDCs can alter this effect. We examined the long-term effects of propylparaben, a common preservative used in personal care products and foods, with estrogenic properties, on the parous mouse mammary gland. Pregnant BALB/c mice were treated with 0, 20, 100, or 10 000 µg/kg/day propylparaben throughout pregnancy and lactation. Unexposed nulliparous females were also evaluated. Five weeks post-involution, mammary glands were collected and assessed for changes in histomorphology, hormone receptor expression, immune cell number, and gene expression. For several parameters of mammary gland morphology, propylparaben reduced the effects of parity. Propylparaben also increased proliferation, but not stem cell number, and induced modest alterations to expression of ERα-mediated genes. Finally, propylparaben altered the effect of parity on the number of several immune cell types in the mammary gland. These results suggest that propylparaben, at levels relevant to human exposure, can interfere with the effects of parity on the mouse mammary gland and induce long-term alterations to mammary gland structure. Future studies should address if propylparaben exposures negate the protective effects of pregnancy on mammary cancer development.
10aAnimals10aCells, Cultured10aEndocrine Disruptors10aFemale10aLactation10aMale10aMammary Glands, Animal10aMaternal Exposure10aMice10aMice, Inbred BALB C10aParabens10aPregnancy10aPrenatal Exposure Delayed Effects1 aMogus, Joshua, P1 aLaPlante, Charlotte, D1 aBansal, Ruby1 aMatouskova, Klara1 aSchneider, Benjamin, R1 aDaniele, Elizabeth1 aSilva, Shannon, J1 aHagen, Mary, J1 aDunphy, Karen, A1 aJerry, Joseph1 aSchneider, Sallie, S1 aVandenberg, Laura, N uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/exposure-propylparaben-during-pregnancy-and-lactation-induces-long03299nas a2200541 4500008004100000022001400041245017100055210006900226260001200295300001400307490000700321520151500328653001201843653002101855653002301876653002701899653002701926653003201953653001501985653002002000653001102020653003102031653002002062653002002082653001702102653002502119653000902144653001902153653003602172653003102208653003302239100002802272700002402300700002302324700002102347700002102368700002402389700002102413700002002434700002502454700002002479700001802499700002502517700002302542700002202565700001802587856015202605 2021 eng d a1476-559400aGenetic modifiers regulating DNA replication and double-strand break repair are associated with differences in mammary tumors in mouse models of Li-Fraumeni syndrome.0 aGenetic modifiers regulating DNA replication and doublestrand br c2021 08 a5026-50370 v403 aBreast cancer is the most common tumor among women with inherited variants in the TP53 tumor suppressor, but onset varies widely suggesting interactions with genetic or environmental factors. Rodent models haploinsufficent for Trp53 also develop a wide variety of malignancies associated with Li-Fraumeni syndrome, but BALB/c mice are uniquely susceptible to mammary tumors and is genetically linked to the Suprmam1 locus on chromosome 7. To define mechanisms that interact with deficiencies in p53 to alter susceptibility to mammary tumors, we fine mapped the Suprmam1 locus in females from an N2 backcross of BALB/cMed and C57BL/6J mice. A major modifier was localized within a 10 cM interval on chromosome 7. The effect of the locus on DNA damage responses was examined in the parental strains and mice that are congenic for C57BL/6J alleles on the BALB/cMed background (SM1-Trp53). The mammary epithelium of C57BL/6J-Trp53 females exhibited little radiation-induced apoptosis compared to BALB/cMed-Trp53 and SM1-Trp53 females indicating that the Suprmam1 alleles could not rescue repair of radiation-induced DNA double-strand breaks mostly relying on non-homologous end joining. In contrast, the Suprmam1 alleles in SM1-Trp53 mice were sufficient to confer the C57BL/6J-Trp53 phenotypes in homology-directed repair and replication fork progression. The Suprmam1 alleles in SM1-Trp53 mice appear to act in trans to regulate a panel of DNA repair and replication genes which lie outside the locus.
10aAnimals10aBreast Neoplasms10aChromosome Mapping10aDisease Models, Animal10aDisease Susceptibility10aDNA Breaks, Double-Stranded10aDNA Repair10aDNA Replication10aFemale10aGene Expression Regulation10aGenes, Modifier10aGenetic Linkage10aGenetic Loci10aLi-Fraumeni Syndrome10aMice10aMice, Knockout10aPolymorphism, Single Nucleotide10aRecombinational DNA Repair10aTumor Suppressor Protein p531 aMajhi, Prabin, Dhangada1 aGriner, Nicholas, B1 aMayfield, Jacob, A1 aCompton, Shannon1 aKane, Jeffrey, J1 aBaptiste, Trevor, A1 aDunphy, Karen, A1 aRoberts, Amy, L1 aSchneider, Sallie, S1 aSavage, Evan, M1 aPatel, Divyen1 aBlackburn, Anneke, C1 aMaurus, Kim, Joana1 aWiesmüller, Lisa1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genetic-modifiers-regulating-dna-replication-and-double-strand-break03116nas a2200361 4500008004100000022001400041245010500055210006900160260001200229300001200241490000700253520194200260653001902202653001402221653001302235653001102248653001102259653002302270653002202293653001402315653004002329653003602369100002702405700002102432700002502453700001602478700002002494700002402514700002102538700002402559700002002583856015102603 2021 eng d a1556-834200aHumoral and Cell-Mediated Immune Response in Colostrum from Women Diagnosed Positive for SARS-CoV-2.0 aHumoral and CellMediated Immune Response in Colostrum from Women c2021 12 a987-9940 v163 aTo evaluate the immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in colostrum from women who tested positive for the virus. Between March and September 2020 we obtained bilateral colostrum samples collected on spot cards within 48 hours of delivery from 15 new mothers who had previously tested positive for SARS-CoV-2. Four of 15 women provided liquid colostrum, which was used for validating results obtained from spot cards. Archived bilateral colostrum samples collected from 8 women during 2011-2013 were used as pre-coronavirus disease 2019 (COVID-19) controls. All samples were tested for reactivity to the receptor binding domain (RBD) of the SARS-CoV-2 spike protein using an enzyme-linked immunosorbent assay that measures SARS-CoV-2 RBD-specific IgA, IgG, and IgM and for levels of 10 inflammatory cytokines (interferon-gamma [IFN-γ], tumor necrosis factor-alpha, interleukin [IL]-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13) using a multiplex electrochemiluminescent sandwich assay. Our validation studies indicate that the levels of SARS-CoV-2-specific antibodies and the associated cytokines measured in liquid colostrum are comparable to levels eluted from spot cards. Bilateral colostrum samples from 73%, 73%, and 33% of the 15 COVID-19 mothers exhibited IgA, IgG, and IgM reactivity to RBD, respectively. In addition, symptomatic COVID-19 mothers had statistically significant elevated levels of 4 of the 10 inflammatory markers (IFN-γ, IL-4, IL-6, and IL-12) compared to asymptomatic COVID-19 mothers. A strong humoral immune response is present in the colostrum of women who were infected with SARS-CoV-2 before delivering. The evolution and duration of the antibody response, as well as dynamics of the cytokine response, remain to be determined. Our results also indicate that future large-scale studies can be conducted with milk easily collected on paper spot cards.
10aBreast Feeding10aColostrum10aCOVID-1910aFemale10aHumans10aImmunity, Cellular10aImmunity, Humoral10aPregnancy10aPregnancy Complications, Infectious10aSpike Glycoprotein, Coronavirus1 aNarayanaswamy, Vignesh1 aPentecost, Brian1 aAlfandari, Dominique1 aChin, Emily1 aMinor, Kathleen1 aKastrinakis, Alyssa1 aLieberman, Tanya1 aArcaro, Kathleen, F1 aLeftwich, Heidi uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/humoral-and-cell-mediated-immune-response-colostrum-women-diagnosed02601nas a2200337 4500008004100000022001400041245008000055210006900135260001300204300001100217490000800228520156900236653001201805653001801817653001801835653001101853653001101864653002501875653001901900653002301919653001401942653001901956100001901975700002101994700001802015700002002033700002302053700002702076700001602103856014402119 2021 eng d a1873-223200aAn intrauterine device with potential to control fertility in feral equids.0 aintrauterine device with potential to control fertility in feral c2021 Aug a1067950 v2313 aFertility control of feral equids is difficult. A 4-month pilot study was conducted with a hormone-free intrauterine device (iUPOD). There was evaluation of i) device retention; ii) contraceptive efficacy; iii) fertility following device removal; iv) effects of device on estrous cycle periodicity and; v) abundance of biofilm on devices after removal from the uterus. The iUPODs were inserted trans-cervically in eight mares at random stages of the estrous cycle. Mares were confined in a paddock with a stallion the following day and remained with the stallion for 120 days. Transabdominal detection of the iUPOD, using a non-invasive handheld magnetic detector wand, was performed weekly. Mares were examined using transrectal ultrasonography on days 0 (Time at insertion = day 0), 14, and 30, and subsequently every third week to assess number and size of follicles, corpora lutea, and whether there was intrauterine fluid (IUF) present. The mares and stallion were observed daily for mating behavior. Weekly samples were assayed for progesterone (P) at day 0 and until 3 weeks subsequent to stallion removal. None of the mares became pregnant while fitted with the iUPOD. Two of four mares conceived within 30 days subsequent to iUPOD removal. Three of eight mares fitted with the device had periods greater than 14 days with P concentrations <1 ng/mL, and seven of eight mares had periods greater than 14 days with P concentrations>1 ng/mL. There was a marked abundance of biofilm on devices of two mares at the time of device removal.
10aAnimals10aContraception10aEstrous Cycle10aFemale10aHorses10aIntrauterine Devices10aPilot Projects10aPopulation Control10aPregnancy10aPregnancy Rate1 aGradil, Carlos1 aJoone, Carolynne1 aHaire, Teresa1 aFowler, Bradley1 aZinchuk, Jacquelyn1 aDavies, Christopher, J1 aBall, Barry uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/intrauterine-device-potential-control-fertility-feral-equids02458nas a2200397 4500008004100000022001400041245013100055210006900186260001500255300001400270490000600284520118100290653001201471653002601483653002601509653002501535653001301560653000901573653002001582100002001602700002001622700002801642700001801670700002301688700002601711700001901737700001501756700001901771700001301790700001701803700001701820700002801837700001701865700002401882856015401906 2021 eng d a2051-635500aNanotherapeutics using all-natural materials. Effective treatment of wound biofilm infections using crosslinked nanoemulsions.0 aNanotherapeutics using allnatural materials Effective treatment c2021 06 01 a1776-17820 v83 aBacterial wound infections are a threat to public health. Although antibiotics currently provide front-line treatments for bacterial infections, the development of drug resistance coupled with the defenses provided through biofilm formation render these infections difficult, if not impossible, to cure. Antimicrobials from natural resources provide unique antimicrobial mechanisms and are generally recognized as safe and sustainable. Herein, an all-natural antimicrobial platform is reported. It is active against bacterial biofilms and accelerates healing of wound biofilm infections . This antimicrobial platform uses gelatin stabilized by photocrosslinking using riboflavin (vitamin B) as a photocatalyst, and carvacrol (the primary constituent of oregano oil) as the active antimicrobial. The engineered nanoemulsions demonstrate broad-spectrum antimicrobial activity towards drug-resistant bacterial biofilms and significantly expedite wound healing in an murine wound biofilm model. The antimicrobial activity, wound healing promotion, and biosafety of these nanoemulsions provide a readily translatable and sustainable strategy for managing wound infections.
10aAnimals10aAnti-Bacterial Agents10aAnti-Infective Agents10aBacterial Infections10aBiofilms10aMice10aWound Infection1 aLi, Cheng-Hsuan1 aLandis, Ryan, F1 aMakabenta, Jessa, Marie1 aNabawy, Ahmed1 aTronchet, Tiphaine1 aArchambault, Danielle1 aLiu, Yuanchang1 aHuang, Rui1 aGolan, Morgane1 aCui, Wei1 aMager, Jesse1 aGupta, Akash1 aSchmidt-Malan, Suzannah1 aPatel, Robin1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/nanotherapeutics-using-all-natural-materials-effective-treatment-wound00646nas a2200181 4500008004100000245011600041210006900157300001400226490000800240100002700248700002500275700002300300700002500323700002600348700001600374700002500390856004900415 2021 eng d00aNeutralizing Antibodies and Cytokines in Breast Milk After Coronavirus Disease 2019 (COVID-19) mRNA Vaccination0 aNeutralizing Antibodies and Cytokines in Breast Milk After Coron a181–1910 v1391 aNarayanaswamy, Vignesh1 aPentecost, Brian, T.1 aSchoen, Corina, N.1 aAlfandari, Dominique1 aSchneider, Sallie, S.1 aBaker, Ryan1 aArcaro, Kathleen, F. uhttps://doi.org/10.1097/aog.000000000000466102897nas a2200265 4500008004100000022001400041245010000055210006900155260001500224300001400239490000700253520194900260653002502209653003002234653004502264653001302309653002202322100002602344700002502370700002002395700002302415700002002438700002002458856015302478 2021 eng d a1526-460200aNon-Covalent Carrier Hydrophobicity as a Universal Predictor of Intracellular Protein Activity.0 aNonCovalent Carrier Hydrophobicity as a Universal Predictor of I c2021 07 12 a2850-28630 v223 aOver the past decade, extensive optimization of polymeric cell-penetrating peptide (CPP) mimics (CPPMs) by our group has generated a substantial library of broadly effective carriers which circumvent the need for covalent conjugation often required by CPPs. In this study, design rules learned from CPPM development were applied to reverse-engineer the first library of simple amphiphilic block copolypeptides for non-covalent protein delivery, namely, poly(alanine--arginine), poly(phenylalanine--arginine), and poly(tryptophan--arginine). This new CPP library was screened for enhanced green fluorescent protein and Cre recombinase delivery alongside a library of CPPMs featuring equivalent side-chain configurations. Due to the added hydrophobicity imparted by the polymer backbone as compared to the polypeptide backbone, side-chain functionality was not a universal predictor of carrier performance. Rather, overall carrier hydrophobicity predicted the top performers for both internalization and activity of protein cargoes, regardless of backbone identity. Furthermore, comparison of protein uptake and function revealed carriers which facilitated high gene recombination despite remarkably low Cre internalization, leading us to formalize the concept of intracellular availability (IA) of the delivered cargo. IA, a measure of cargo activity per quantity of cargo internalized, provides valuable insight into the physical relationship between cellular internalization and bioavailability, which can be affected by bottlenecks such as endosomal escape and cargo release. Importantly, carriers with maximal IA existed within a narrow hydrophobicity window, more hydrophilic than those exhibiting maximal cargo uptake. Hydrophobicity may be used as a scaffold-independent predictor of protein uptake, function, and IA, enabling identification of new, effective carriers which would be overlooked by uptake-based screening methods.
10aBiological Transport10aCell-Penetrating Peptides10aHydrophobic and Hydrophilic Interactions10aPolymers10aProtein Transport1 aHango, Christopher, R1 aBacklund, Coralie, M1 aDavis, Hazel, C1 aPosey, Nicholas, D1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/non-covalent-carrier-hydrophobicity-universal-predictor-intracellular02281nas a2200145 4500008004100000022001400041245010600055210006900161260000900230300001100239490000600250520171200256100001301968856015401981 2021 eng d a2296-634X00aOocyte Spontaneous Activation: An Overlooked Cellular Event That Impairs Female Fertility in Mammals.0 aOocyte Spontaneous Activation An Overlooked Cellular Event That c2021 a6480570 v93 aIn mammals, including humans, mature oocytes are ovulated into the oviduct for fertilization. Normally, these oocytes are arrested at metaphase of the second meiosis (MII), and this arrest can be maintained for a certain period, which is essential for fertilization and oocyte manipulations , such as assisted reproduction in clinics and nuclear/spindle transfer in laboratories. However, in some species and under certain circumstances, exit from MII occurs spontaneously without any obvious stimulation or morphological signs, which is so-called oocyte spontaneous activation (OSA). This mini-review summarizes two types of OSA. In the first type (e.g., most rat strains), oocytes can maintain MII arrest , but once removed out, oocytes undergo OSA with sister chromatids separated and eventually scattered in the cytoplasm. Because the stimulation is minimal (oocyte collection itself), this OSA is incomplete and cannot force oocytes into interphase. Notably, once re-activated by sperm or chemicals, those scattered chromatids will form multiple pronuclei (MPN), which may recapitulate certain MPN and aneuploidy cases observed in fertility clinics. The second type of OSA occurs in ovarian oocytes (e.g., certain mouse strains and dromedary camel). Without ovulation or fertilization, these OSA-oocytes can initiate intrafollicular development, but these parthenotes cannot develop to term due to aberrant genomic imprinting. Instead, they either degrade or give rise to ovarian teratomas, which have also been reported in female patients. Last but not the least, genetic models displaying OSA phenotypes and the lessons we can learn from animal OSA for human reproduction are also discussed.
1 aCui, Wei uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/oocyte-spontaneous-activation-overlooked-cellular-event-impairs-female03043nas a2200373 4500008004100000022001400041245009600055210006900151260001200220300001100232490000800243520190100251653001202152653002702164653002002191653002602211653001402237653001102251653000902262653000902271653002302280653001902303653001402322653001602336100002702352700002002379700001602399700001702415700002402432700002302456700001702479700002102496856015202517 2021 eng d a1873-675000aPaternal preconception phthalate exposure alters sperm methylome and embryonic programming.0 aPaternal preconception phthalate exposure alters sperm methylome c2021 10 a1066930 v1553 aPreconception environmental conditions have been demonstrated to shape sperm epigenetics and subsequently offspring health and development. Our previous findings in humans showed that urinary anti-androgenic phthalate metabolites in males were associated with altered sperm methylation and blastocyst-stage embryo development. To corroborate this, we examined the effect of preconception exposure to di(2-ethylhexyl) phthalate (DEHP) on genome-wide DNA methylation and gene expression profiles in mice. Eight-week old C57BL/6J male mice were exposed to either a vehicle control, low, or high dose of DEHP (2.5 and 25 mg/kg/weight, respectively) for 67 days (~2 spermatogenic cycles) and were subsequently mated with unexposed females. Reduced representation bisulfite sequencing (RRBS) of epididymal sperm was performed and gastrulation stage embryos were collected for RRBS and transcriptome analyses in both embryonic and extra-embryonic lineages. Male preconception DEHP exposure resulted in 704 differentially methylated regions (DMRs; q-value < 0.05; ≥10% methylation change) in sperm, 1,716 DMRs in embryonic, and 3,181 DMRs in extra-embryonic tissue. Of these, 29 DMRs overlapped between sperm and F1 tissues, half of which showed concordant methylation changes between F0 and F1 generations. F1 transcriptomes at E7.5 were also altered by male preconception DEHP exposure including developmental gene families such as Hox, Gata, and Sox. Additionally, gene ontology analyses of DMRs and differentially expressed genes showed enrichment of multiple developmental processes including embryonic development, pattern specification and morphogenesis. These data indicate that spermatogenesis in adult may represent a sensitive window in which exposure to DEHP alters the sperm methylome as well as DNA methylation and gene expression in the developing embryo.
10aAnimals10aDiethylhexyl Phthalate10aDNA Methylation10aEmbryonic Development10aEpigenome10aFemale10aMale10aMice10aMice, Inbred C57BL10aPhthalic Acids10aPregnancy10aSpermatozoa1 aOluwayiose, Oladele, A1 aMarcho, Chelsea1 aWu, Haotian1 aHoule, Emily1 aKrawetz, Stephen, A1 aSuvorov, Alexander1 aMager, Jesse1 aPilsner, Richard uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/paternal-preconception-phthalate-exposure-alters-sperm-methylome-and03404nas a2200493 4500008004100000022001400041245016100055210006900216260001200285300001400297490000700311520180500318653000902123653002102132653002502153653002402178653001602202653002002218653002502238653001802263653001102281653001102292653005002303653001502353653002202368653003202390653001602422653002702438653002402465100002302489700001902512700001902531700001802550700001502568700002002583700001802603700001902621700002602640700002302666700002402689700002002713700002402733856015302757 2021 eng d a1538-775500aPrediagnostic White Blood Cell DNA Methylation and Risk of Breast Cancer in the Prostate Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) Cohort.0 aPrediagnostic White Blood Cell DNA Methylation and Risk of Breas c2021 08 a1575-15810 v303 aBACKGROUND: White blood cell (WBC) DNA may contain methylation patterns that are associated with subsequent breast cancer risk. Using a high-throughput array and samples collected, on average, 1.3 years prior to diagnosis, a case-cohort analysis nested in the prospective Sister Study identified 250 individual CpG sites that were differentially methylated between breast cancer cases and noncases. We examined five of the top 40 CpG sites in a case-control study nested in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) Cohort.
METHODS: We investigated the associations between prediagnostic WBC DNA methylation in 297 breast cancer cases and 297 frequency-matched controls. Two WBC DNA specimens from each participant were used: a proximate sample collected 1 to 2.9 years and a distant sample collected 4.2-7.3 years prior to diagnosis in cases or the comparable timepoints in controls. WBC DNA methylation level was measured using targeted bisulfite amplification sequencing. We used logistic regression to obtain ORs and 95% confidence intervals (CI).
RESULTS: A one-unit increase in percent methylation in in proximate WBC DNA was associated with increased breast cancer risk (adjusted OR = 1.29; 95% CI, 1.06-1.57). However, a one-unit increase in percent methylation in in distant WBC DNA was inversely associated with breast cancer risk (adjusted OR = 0.83; 95% CI, 0.69-0.98). None of the other ORs met the threshold for statistical significance.
CONCLUSIONS: There was no convincing pattern between percent methylation in the five CpG sites and breast cancer risk.
IMPACT: The link between prediagnostic WBC DNA methylation marks and breast cancer, if any, is poorly understood.
10aAged10aBreast Neoplasms10aCase-Control Studies10aCell Cycle Proteins10aCpG Islands10aDNA Methylation10aDNA-Binding Proteins10aEndonucleases10aFemale10aHumans10aIntracellular Signaling Peptides and Proteins10aLeukocytes10aMembrane Proteins10aMembrane Transport Proteins10aMiddle Aged10aMitochondrial Proteins10aProspective Studies1 aSturgeon, Susan, R1 aSela, David, A1 aBrowne, Eva, P1 aEinson, Jonah1 aRani, Asha1 aHalabi, Mohamed1 aKania, Thomas1 aKeezer, Andrew1 aBalasubramanian, Raji1 aZiegler, Regina, G1 aSchairer, Catherine1 aKelsey, Karl, T1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/prediagnostic-white-blood-cell-dna-methylation-and-risk-breast-cancer00871nas a2200253 4500008004100000245016000041210006900201300001600270490000700286100002400293700002000317700002000337700001800357700001500375700002000390700001800410700001900428700002600447700002400473700002400497700002100521700002500542856005000567 2021 eng d00aPrediagnostic White Blood Cell DNA Methylation and Risk of Breast Cancer in the Prostate Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) Cohort0 aPrediagnostic White Blood Cell DNA Methylation and Risk of Breas a1575–15810 v301 aSturgeon, Susan, R.1 aSela, David, A.1 aBrowne, Eva, P.1 aEinson, Jonah1 aRani, Asha1 aHalabi, Mohamed1 aKania, Thomas1 aKeezer, Andrew1 aBalasubramanian, Raji1 aZiegler, Regina, G.1 aSchairer, Catherine1 aKelsey, Karl, T.1 aArcaro, Kathleen, F. uhttps://doi.org/10.1158/1055-9965.epi-20-171700654nas a2200169 4500008004100000245015700041210006900198300001100267490000800278100002100286700002300307700002500330700002100355700002600376700002800402856005400430 2021 eng d00aPreliminary study of the contraceptive effect of a self-assembling intrauterine device (iUPODs) in mares maintained in a paddock with a fertile stallion0 aPreliminary study of the contraceptive effect of a selfassemblin a1068810 v2351 aHoopes, Karl, H.1 aGradil, Carlos, M.1 aVanderwall, Dirk, K.1 aMason, Holly, M.1 aSarnecky, Brendan, A.1 aDavies, Christopher, J. uhttps://doi.org/10.1016/j.anireprosci.2021.10688101770nas a2200289 4500008004100000022001400041245014800055210006900203260001500272300001600287490000700303520079200310653001501102653002101117653001501138653001101153653001801164653001801182653002101200100001301221700002001234700001601254700001801270700002401288700002001312856014801332 2021 eng d a1521-377300aProtein-Antibody Conjugates (PACs): A Plug-and-Play Strategy for Covalent Conjugation and Targeted Intracellular Delivery of Pristine Proteins.0 aProteinAntibody Conjugates PACs A PlugandPlay Strategy for Coval c2021 06 01 a12813-128180 v603 aWe report here on protein-antibody conjugates (PACs) that are used for antibody-directed delivery of protein therapeutics to specific cells. PACs have the potential to judiciously combine the merits of two prolific therapeutic approaches-biologics and antibody-drug conjugates. We utilize spherical polymer brushes to construct PACs using the combination of two simple and efficient functionally orthogonal click chemistries. In addition to the synthesis and characterization of these nanoparticles, we demonstrate that PACs are indeed capable of specifically targeting cells based on the presence of target antigen on the cell surface to deliver proteins. The potentially broad adaptability of PACs opens up new opportunities for targeted biologics in therapeutics and sensing.
10aAntibodies10aCell Line, Tumor10aFullerenes10aHumans10aNanoparticles10aParticle Size10aReceptor, erbB-21 aLiu, Bin1 aSingh, Khushboo1 aGong, Shuai1 aCanakci, Mine1 aOsborne, Barbara, A1 aThayumanavan, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/protein-antibody-conjugates-pacs-plug-and-play-strategy-covalent01702nas a2200313 4500008004100000022001400041245005100055210004900106260001200155300001200167490000700179520077800186653001200964653002600976653002601002653001101028653001801039653001401057653002601071653002101097653002401118100002301142700002201165700001901187700002001206700002401226700001701250856012101267 2021 eng d a1474-178400aTargeting Notch in oncology: the path forward.0 aTargeting Notch in oncology the path forward c2021 02 a125-1440 v203 aNotch signalling is involved in many aspects of cancer biology, including angiogenesis, tumour immunity and the maintenance of cancer stem-like cells. In addition, Notch can function as an oncogene and a tumour suppressor in different cancers and in different cell populations within the same tumour. Despite promising preclinical results and early-phase clinical trials, the goal of developing safe, effective, tumour-selective Notch-targeting agents for clinical use remains elusive. However, our continually improving understanding of Notch signalling in specific cancers, individual cancer cases and different cell populations, as well as crosstalk between pathways, is aiding the discovery and development of novel investigational Notch-targeted therapeutics.
10aAnimals10aAntineoplastic Agents10aDrug Delivery Systems10aHumans10aImmunotherapy10aNeoplasms10aNeoplastic Stem Cells10aReceptors, Notch10aSignal Transduction1 aMajumder, Samarpan1 aCrabtree, Judy, S1 aGolde, Todd, E1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/targeting-notch-oncology-path-forward01704nas a2200313 4500008004100000022001400041245005100055210004900106260001200155300001200167490000700179520077800186653001200964653002600976653002601002653001101028653001801039653001401057653002601071653002101097653002401118100002301142700002201165700001901187700002001206700002401226700001701250856012301267 2021 eng d a1474-178400aTargeting Notch in oncology: the path forward.0 aTargeting Notch in oncology the path forward c2021 02 a125-1440 v203 aNotch signalling is involved in many aspects of cancer biology, including angiogenesis, tumour immunity and the maintenance of cancer stem-like cells. In addition, Notch can function as an oncogene and a tumour suppressor in different cancers and in different cell populations within the same tumour. Despite promising preclinical results and early-phase clinical trials, the goal of developing safe, effective, tumour-selective Notch-targeting agents for clinical use remains elusive. However, our continually improving understanding of Notch signalling in specific cancers, individual cancer cases and different cell populations, as well as crosstalk between pathways, is aiding the discovery and development of novel investigational Notch-targeted therapeutics.
10aAnimals10aAntineoplastic Agents10aDrug Delivery Systems10aHumans10aImmunotherapy10aNeoplasms10aNeoplastic Stem Cells10aReceptors, Notch10aSignal Transduction1 aMajumder, Samarpan1 aCrabtree, Judy, S1 aGolde, Todd, E1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/targeting-notch-oncology-path-forward-000543nas a2200157 4500008004100000245010500041210006900146260001800215300001200233490000800245100001500253700002300268700002500291700002100316856004800337 2021 eng d00aTrim-Away mediated knock down uncovers a new function for Lbh during gastrulation of Xenopus laevis.0 aTrimAway mediated knock down uncovers a new function for Lbh dur bElsevier {BV} a74–830 v4701 aWeir, Emma1 aMcLinden, Gretchen1 aAlfandari, Dominique1 aCousin, Hélène uhttps://doi.org/10.1016/j.ydbio.2020.10.01400648nas a2200205 4500008004100000245008100041210006900122260001000191100001800201700002000219700002400239700002400263700001700287700002100304700001300325700002100338700002300359700002200382856003800404 2021 eng d00aTSSK3, a novel target for male contraception, is required for spermiogenesis0 aTSSK3 a novel target for male contraception is required for sper bWiley1 aNayyab, Saman1 aGervasi, M., G.1 aTourzani, Darya, A.1 aCaraballo, Diego, A1 aJha, Kula, N1 aTeves, Maria, E.1 aCui, Wei1 aGeorg, Gunda, I.1 aVisconti, Pablo, E1 aSalicioni, Ana, M uhttps://doi.org/10.1002/mrd.2353902034nas a2200253 4500008004100000022001400041245004700055210004600102260000900148300001100157490000600168520126700174100002001441700002601461700002001487700001901507700001901526700002301545700002101568700002401589700002001613700002201633856012501655 2021 eng d a2296-634X00aWhen Viruses Cross Developmental Pathways.0 aWhen Viruses Cross Developmental Pathways c2021 a6916440 v93 aAberrant regulation of developmental pathways plays a key role in tumorigenesis. Tumor cells differ from normal cells in their sustained proliferation, replicative immortality, resistance to cell death and growth inhibition, angiogenesis, and metastatic behavior. Often they acquire these features as a consequence of dysregulated Hedgehog, Notch, or WNT signaling pathways. Human tumor viruses affect the cancer cell hallmarks by encoding oncogenic proteins, and/or by modifying the microenvironment, as well as by conveying genomic instability to accelerate cancer development. In addition, viral immune evasion mechanisms may compromise developmental pathways to accelerate tumor growth. Viruses achieve this by influencing both coding and non-coding gene regulatory pathways. Elucidating how oncogenic viruses intersect with and modulate developmental pathways is crucial to understanding viral tumorigenesis. Many currently available antiviral therapies target viral lytic cycle replication but with low efficacy and severe side effects. A greater understanding of the cross-signaling between oncogenic viruses and developmental pathways will improve the efficacy of next-generation inhibitors and pave the way to more targeted antiviral therapies.
1 aTrivedi, Pankaj1 aPatel, Sandesh, Kumar1 aBellavia, Diana1 aMessina, Elena1 aPalermo, Rocco1 aCeccarelli, Simona1 aMarchese, Cinzia1 aAnastasiadou, Eleni1 aMinter, Lisa, M1 aFelli, Maria, Pia uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/when-viruses-cross-developmental-pathways02320nas a2200337 4500008004100000022001400041245009700055210006900152260001500221300001200236490000800248520126000256653001201516653002301528653001501551653002501566653002401591653002601615653001101641653004601652653001301698653000901711653001901720653001301739100001601752700001701768700002601785700001701811700001301828856014101841 2021 eng d a1529-726800aZC3H4-a novel CCCH-type zinc finger protein-is essential for early embryogenesis in mice†.0 aZC3H4a novel CCCHtype zinc finger proteinis essential for early c2021 02 11 a325-3350 v1043 aZinc finger domains of the Cys-Cys-Cys-His (CCCH) class are evolutionarily conserved proteins that bind nucleic acids and are involved in various biological processes. Nearly 60 CCCH-type zinc finger proteins have been identified in humans and mice, most have not been functionally characterized. Here, we provide the first in vivo functional characterization of ZC3H4-a novel CCCH-type zinc finger protein. Our results show that although Zc3h4 mutant embryos exhibit normal morphology at E3.5 blastocyst stage, they cannot be recovered at E7.5 early post-gastrulation stage, suggesting implantation failure. Outgrowth assays reveal that mutant blastocysts either fail to hatch from the zona pellucida, or can hatch but do not form a typical inner cell mass colony, the source of embryonic stem cells (ESCs). Although there is no change in levels of reactive oxygen species, Zc3h4 mutants display severe DNA breaks and reduced cell proliferation. Analysis of lineage specification reveals that both epiblast and primitive endoderm lineages are compromised with severe reductions in cell number and/or specification in the mutant blastocysts. In summary, these findings demonstrate the essential role of ZC3H4 during early mammalian embryogenesis.
10aAnimals10aCell Proliferation10aDNA Breaks10aDNA-Binding Proteins10aEmbryo Implantation10aEmbryonic Development10aFemale10aGene Expression Regulation, Developmental10aGenotype10aMice10aMice, Knockout10aMutation1 aSu, Jianmin1 aMiao, Xiaosu1 aArchambault, Danielle1 aMager, Jesse1 aCui, Wei uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/zc3h4-novel-ccch-type-zinc-finger-protein-essential-early00789nas a2200241 4500008004100000245011600041210006900157260002500226490000600251100001700257700001500274700002100289700002000310700002400330700001600354700002300370700001700393700002300410700002300433700001900456700002700475856004500502 2020 eng d00aAcheron/Larp6 Is a Survival Protein That Protects Skeletal Muscle From Programmed Cell Death During Development0 aAcheronLarp6 Is a Survival Protein That Protects Skeletal Muscle bFrontiers Media {SA}0 v81 aSheel, Ankur1 aShao, Rong1 aBrown, Christine1 aJohnson, Joanne1 aHamilton, Alexandra1 aSun, Danhui1 aOppenheimer, Julia1 aSmith, Wendy1 aVisconti, Pablo, E1 aMarkstein, Michele1 aBigelow, Carol1 aSchwartz, Lawrence, M. uhttps://doi.org/10.3389/fcell.2020.0062200608nas a2200193 4500008004100000245006300041210006300104260001000167100002300177700002000200700002700220700002200247700002100269700002600290700002000316700001700336700002300353856003800376 2020 eng d00aCapacitation increases glucose consumption in murine sperm0 aCapacitation increases glucose consumption in murine sperm bWiley1 aHidalgo, David, M.1 aRomarowski, Ana1 aGervasi, Maria, Gracia1 aNavarrete, Felipe1 aBalbach, Melanie1 aSalicioni, Ana, Maria1 aLevin, Lonny, R1 aBuck, Jochen1 aVisconti, Pablo, E uhttps://doi.org/10.1002/mrd.2342100771nas a2200253 4500008004100000245006500041210006500106260002500171490000700196100001800203700002900221700002400250700002000274700001800294700001900312700002000331700001400351700001900365700001700384700002700401700002000428700002400448856004500472 2020 eng d00aCD28 Signaling Drives Notch Ligand Expression on CD4 T Cells0 aCD28 Signaling Drives Notch Ligand Expression on CD4 T Cells bFrontiers Media {SA}0 v111 aMitra, Ankita1 aShanthalingam, Sudarvili1 aSherman, Heather, L1 aSingh, Khushboo1 aCanakci, Mine1 aTorres, Joe, A1 aLawlor, Rebecca1 aRan, Yong1 aGolde, Todd, E1 aMiele, Lucio1 aThayumanavan, Sankaran1 aMinter, Lisa, M1 aOsborne, Barbara, A uhttps://doi.org/10.3389/fimmu.2020.0073500683nas a2200193 4500008004100000245013800041210006900179260001800248300001600266490000700282100001600289700002900305700002400334700001900358700002400377700002000401700002000421856004800441 2020 eng d00aCell-Penetrating Anti-Protein Kinase C Theta Antibodies Act Intracellularly to Generate Stable, Highly Suppressive Regulatory T Cells0 aCellPenetrating AntiProtein Kinase C Theta Antibodies Act Intrac bElsevier {BV} a1987–20060 v281 aOzay, Ilker1 aShanthalingam, Sudarvili1 aSherman, Heather, L1 aTorres, Joe, A1 aOsborne, Barbara, A1 aTew, Gregory, N1 aMinter, Lisa, M uhttps://doi.org/10.1016/j.ymthe.2020.05.02000905nas a2200277 4500008004100000245009900041210006900140260003900209300001300248490000700261100002100268700002300289700002600312700001800338700001300356700002200369700002400391700002200415700002600437700002200463700002400485700001700509700002200526700003000548856004900578 2020 eng d00aContained Mycobacterium tuberculosis infection induces concomitant and heterologous protection0 aContained Mycobacterium tuberculosis infection induces concomita bPublic Library of Science ({PLoS}) ae10086550 v161 aNemeth, Johannes1 aOlson, Gregory, S.1 aRothchild, Alissa, C.1 aJahn, Ana, N.1 aMai, Dat1 aDuffy, Fergal, J.1 aDelahaye, Jared, L.1 aSrivatsan, Sanjay1 aPlumlee, Courtney, R.1 aUrdahl, Kevin, B.1 aGold, Elizabeth, S.1 aAderem, Alan1 aDiercks, Alan, H.1 aSassetti, Christopher, M. uhttps://doi.org/10.1371/journal.ppat.100865500547nas a2200133 4500008004100000245014000041210006900181260003600250100002100286700002000307700002300327700002000350856004300370 2020 eng d00aCRISPR/CAS9-mediated amino acid substitution reveals phosphorylation residues of RSPH6A are not essential for male fertility in mice†0 aCRISPRCAS9mediated amino acid substitution reveals phosphorylati bOxford University Press ({OUP})1 aMiyata, Haruhiko1 aAbbasi, Ferheen1 aVisconti, Pablo, E1 aIkawa, Masahito uhttps://doi.org/10.1093/biolre/ioaa16100583nas a2200169 4500008004100000245007900041210006900120260003500189300001400224490000700238100002100245700002200266700003000288700002300318700002500341856004700366 2020 eng d00aDifferences and Similarities: The Richness of Comparative Sperm Physiology0 aDifferences and Similarities The Richness of Comparative Sperm P bAmerican Physiological Society a196–2080 v351 aDarszon, Alberto1 aNishigaki, Takuya1 aLópez-González, Ignacio1 aVisconti, Pablo, E1 aTreviño, Claudia, L uhttps://doi.org/10.1152/physiol.00033.201900874nas a2200253 4500008004100000245012700041210006900168260002500237490000700262100002600269700002500295700002700320700002200347700002000369700002300389700002100412700002200433700002300455700002000478700002400498700002700522700002600549856004500575 2020 eng d00aDifferentiation of Pathogenic Th17 Cells Is Negatively Regulated by Let-7 MicroRNAs in a Mouse Model of Multiple Sclerosis0 aDifferentiation of Pathogenic Th17 Cells Is Negatively Regulated bFrontiers Media {SA}0 v101 aAngelou, Constance, C1 aWells, Alexandria, C1 aVijayaraghavan, Jyothi1 aDougan, Carey, E.1 aLawlor, Rebecca1 aIverson, Elizabeth1 aLazarevic, Vanja1 aKimura, Motoko, Y1 aPeyton, Shelly, R.1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aPobezinskaya, Elena, L1 aPobezinsky, Leonid, A uhttps://doi.org/10.3389/fimmu.2019.0312503026nas a2200397 4500008004100000022001400041245008600055210006900141260001200210300001100222490000700233520185000240653001202090653001202102653002102114653002902135653002202164653001802186653001702204653001602221653001402237653001402251653001902265653001102284653001202295653000902307653001402316653002502330100002002355700002002375700001802395700001802413700002102431700002302452856015302475 2020 eng d a1532-199100aDivalent cation influx and calcium homeostasis in germinal vesicle mouse oocytes.0 aDivalent cation influx and calcium homeostasis in germinal vesic c2020 05 a1021810 v873 aPrior to maturation, mouse oocytes are arrested at the germinal vesicle (GV) stage during which they experience constitutive calcium (Ca) influx and spontaneous Ca oscillations. The oscillations cease during maturation but Ca influx continues, as the oocytes' internal stores attain maximal content at the culmination of maturation, the metaphase II stage. The identity of the channel(s) that underlie this Ca influx has not been completely determined. GV and matured oocytes are known to express three Ca channels, Ca3.2, TRPV3 and TRPM7, but females null for each of these channels are fertile and their oocytes display minor modifications in Ca homeostasis, suggesting a complex regulation of Ca influx. To define the contribution of these channels at the GV stage, we used different divalent cations, pharmacological inhibitors and genetic models. We found that the three channels are active at this stage. Ca3.2 and TRPM7 channels contributed the majority of Ca influx, as inhibitors and oocytes from homologous knockout (KO) lines showed severely reduced Ca entry. Sr influx was promoted by Ca3.2 channels, as Sr oscillations were negligible in Ca3.2-KO oocytes but robust in control and Trpv3-KO GV oocytes. Mn entry relied on expression of Ca3.2 and TRPM7 channels, but Ni entry depended on the latter. Ca3.2 and TRPV3 channels combined to fill the Ca stores, although Ca3.2 was the most impactful. Studies with pharmacological inhibitors effectively blocked the influx of divalent cations, but displayed off-target effects, and occasionally agonist-like properties. In conclusion, GV oocytes express channels mediating Ca and other divalent cation influx that are pivotal for fertilization and early development. These channels may serve as targets for intervention to improve the success of assisted reproductive technologies.
10aAnimals10aCalcium10aCalcium Channels10aCalcium Channels, T-Type10aCations, Divalent10aCell Membrane10aFluorescence10aHomeostasis10aManganese10aMetaphase10aMice, Knockout10aNickel10aOocytes10aOvum10aStrontium10aTRPM Cation Channels1 aArdestani, Goli1 aMehregan, Aujan1 aFleig, Andrea1 aHorgen, David1 aCarvacho, Ingrid1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/divalent-cation-influx-and-calcium-homeostasis-germinal-vesicle-mouse03271nas a2200397 4500008004100000022001400041245013900055210006900194260001200263300001000275490000800285520198800293653001202281653001802293653002102311653002902332653002102361653001102382653000902393653001302402653002202415653002402437653001902461100002802480700001702508700002002525700002302545700002302568700002002591700001902611700002502630700002502655700002102680700001802701856015402719 2020 eng d a1552-992400aEffects of Benzophenone-3 and Propylparaben on Estrogen Receptor-Dependent R-Loops and DNA Damage in Breast Epithelial Cells and Mice.0 aEffects of Benzophenone3 and Propylparaben on Estrogen ReceptorD c2020 01 a170020 v1283 aBACKGROUND: Endocrine-disrupting chemicals have been shown to have broad effects on development, but their mutagenic actions that can lead to cancer have been less clearly demonstrated. Physiological levels of estrogen have been shown to stimulate DNA damage in breast epithelial cells through mechanisms mediated by estrogen-receptor alpha (). Benzophenone-3 (BP-3) and propylparaben (PP) are xenoestrogens found in the urine of of U.S.
POPULATION:
OBJECTIVES: We investigated the effect of BP-3 and PP on estrogen receptor-dependent transactivation and DNA damage at concentrations relevant to exposures in humans.
METHODS: In human breast epithelial cells, DNA damage following treatment with (), BP-3, and PP was determined by immunostaining with antibodies against and 53BP1. Estrogenic responses were determined using luciferase reporter assays and gene expression. Formation of R-loops was determined with DNA: RNA hybrid-specific S9.6 antibody. Short-term exposure to the chemicals was also studied in ovariectomized mice. Immunostaining of mouse mammary epithelium was performed to quantify R-loops and DNA damage .
RESULTS: Concentrations of and BP-3 or PP increased DNA damage similar to that of treatment in a manner. However, BP-3 and PP had limited transactivation of target genes at and concentrations. BP-3 and PP exposure caused R-loop formation in a normal human breast epithelial cell line when was introduced. R-loops and DNA damage were also detected in mammary epithelial cells of mice treated with BP-3 and PP.
CONCLUSIONS: Acute exposure to xenoestrogens (PP and BP-3) in mice induce DNA damage mediated by formation of R-loops at concentrations 10-fold lower than those required for transactivation. Exposure to these xenoestrogens may cause deleterious estrogenic responses, such as DNA damage, in susceptible individuals. https://doi.org/10.1289/EHP5221.
10aAnimals10aBenzophenones10aCell Line, Tumor10aEnvironmental Pollutants10aEpithelial Cells10aHumans10aMice10aParabens10aR-Loop Structures10aReceptors, Estrogen10aToxicity Tests1 aMajhi, Prabin, Dhangada1 aSharma, Aman1 aRoberts, Amy, L1 aDaniele, Elizabeth1 aMajewski, Aliza, R1 aChuong, Lynn, M1 aBlack, Amye, L1 aVandenberg, Laura, N1 aSchneider, Sallie, S1 aDunphy, Karen, A1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/effects-benzophenone-3-and-propylparaben-estrogen-receptor-dependent-r00828nas a2200241 4500008004100000245014000041210007100181260003800252300001100290490000800301100002800309700001700337700002000354700002300374700002300397700002000420700002000440700002600460700002500486700002100511700001800532856003600550 2020 eng d00aEffects of Benzophenone-3 and Propylparaben on Estrogen Receptor–Dependent R-Loops and DNA Damage in Breast Epithelial Cells and Mice0 aEffects of Benzophenone3 and Propylparaben on Estrogen Receptor– bEnvironmental Health Perspectives a0170020 v1281 aMajhi, Prabin, Dhangada1 aSharma, Aman1 aRoberts, Amy, L1 aDaniele, Elizabeth1 aMajewski, Aliza, R1 aChuong, Lynn, M1 aBlack, Amye, L.1 aVandenberg, Laura, N.1 aSchneider, Sallie, S1 aDunphy, Karen, A1 aJerry, Joseph uhttps://doi.org/10.1289/ehp522100725nas a2200217 4500008004100000245012100041210006900162260003800231300001600269490000700285100001600292700001400308700001800322700002400340700001800364700002000382700002300402700001800425700001900443856004500462 2020 eng d00aEffects of Linoleic Acid-Rich Diet on Plasma Profiles of Eicosanoids and Development of Colitis in Il-10–/– Mice0 aEffects of Linoleic AcidRich Diet on Plasma Profiles of Eicosano bAmerican Chemical Society ({ACS}) a7641–76470 v681 aXie, Minhao1 aYang, Jun1 aZhang, Jianan1 aSherman, Heather, L1 aZhang, Zhenyu1 aMinter, Lisa, M1 aHammock, Bruce, D.1 aPark, Yeonhwa1 aZhang, Guodong uhttps://doi.org/10.1021/acs.jafc.0c0302400873nas a2200253 4500008004100000245020000041210006900241260003900310300001600349490000800365100002100373700001800394700001700412700001600429700001800445700001200463700001500475700001600490700001900506700001500525700001900540700001400559856004600573 2020 eng d00aEffects of Two Types of Calorie Restriction on Methylation Levels of Adiponectin Receptor 1 (AdipoR1) and Leptin Receptor Overlapping Transcript (Leprot) in a MMTV-TGF-a Breast Cancer Mouse Model0 aEffects of Two Types of Calorie Restriction on Methylation Level bCambridge University Press ({CUP}) a1079–10790 v1231 aCicekdal, M., B.1 aKazan, B., T.1 aTuna, B., G.1 aOzorhan, U.1 aEkici, I., D.1 aZhu, F.1 aSuakar, O.1 aKuskucu, A.1 aBayrak, O., F.1 aArcaro, K.1 aCleary, M., P.1 aDogan, S. uhttps://doi.org/10.1017/s000711452000047100841nas a2200241 4500008004100000245014700041210006900188260004700257300001400304490000700318100002100325700002200346700002000368700001500388700002800403700001700431700002000448700002100468700002400489700002300513700001900536856004400555 2020 eng d00aEndoplasmic reticulum transmembrane protein TMTC3 contributes to O-mannosylation of E-cadherin, cellular adherence, and embryonic gastrulation0 aEndoplasmic reticulum transmembrane protein TMTC3 contributes to bAmerican Society for Cell Biology ({ASCB}) a167–1830 v311 aGraham, Jill, B.1 aSunryd, Johan, C.1 aMathavan, Ketan1 aWeir, Emma1 aLarsen, Ida, Signe Bohs1 aHalim, Adnan1 aClausen, Henrik1 aCousin, Hélène1 aAlfandari, Dominque1 aHebert, Daniel, N.1 aMartin, Thomas uhttps://doi.org/10.1091/mbc.e19-07-040800839nas a2200253 4500008004100000245010000041210006900141260002500210300001600235490000700251100002100258700002500279700002100304700002200325700002000347700002300367700002500390700002300415700002100438700002500459700002700484700002400511856005000535 2020 eng d00aEnrichment of CpG island shore region hypermethylation in epigenetic breast field cancerization0 aEnrichment of CpG island shore region hypermethylation in epigen bInforma {UK} Limited a1093–11060 v151 aMuse, Meghan, E.1 aTitus, Alexander, J.1 aSalas, Lucas, A.1 aWilkins, Owen, M.1 aMullen, Chelsey1 aGregory, Kelly, J.1 aSchneider, Sallie, S1 aCrisi, Giovanna, M1 aJawale, Rahul, M1 aOtis, Christopher, N1 aChristensen, Brock, C.1 aArcaro, Kathleen, F uhttps://doi.org/10.1080/15592294.2020.174774802273nas a2200373 4500008004100000022001400041245012600055210006900181260001200250300001000262490000700272520105800279653001501337653001201352653001801364653002301382653002101405653002801426653001101454653002201465653002001487653001401507653000901521653002701530653002801557653002401585653001401609653003801623653002401661100002201685700001801707700002501725856014901750 2020 eng d a1873-170800aExposure to low doses of oxybenzone during perinatal development alters mammary gland morphology in male and female mice.0 aExposure to low doses of oxybenzone during perinatal development c2020 03 a66-770 v923 aOxybenzone (benzophenone-3) is an ultraviolet radiation filter commonly used in personal care products including sunscreens, textiles and inks, and food and beverage containers, among others. Due to its widespread use, human exposures to oxybenzone are widespread. Oxybenzone is considered an endocrine disrupting chemical due to its antiestrogenic and antiandrogenic properties. We evaluated the effects of oral exposures to oxybenzone on the growth and morphology of the mammary gland, body weight and anogenital distance in BALB/c mice exposed to 30, 212 or 3000 μg/kg/day in utero and during lactation. Developmental exposures to oxybenzone reduced the size and growth of mammary gland in males prior to and during puberty. In exposed females, oxybenzone reduced mammary cell proliferation, decreased the number of cells expressing estrogen receptor α, and altered mammary gland morphology in adulthood. These results suggest that even low doses of oxybenzone can disrupt hormone sensitive organs during critical windows of development.
10aAnal Canal10aAnimals10aBenzophenones10aCell Proliferation10aEpithelial Cells10aEstrogen Receptor alpha10aFemale10aGenitalia, Female10aGenitalia, Male10aLactation10aMale10aMammary Glands, Animal10aMaternal-Fetal Exchange10aMice, Inbred BALB C10aPregnancy10aPrenatal Exposure Delayed Effects10aSunscreening Agents1 aMatouskova, Klara1 aJerry, Joseph1 aVandenberg, Laura, N uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/exposure-low-doses-oxybenzone-during-perinatal-development-alters00577nas a2200145 4500008004100000245017600041210006900217260002400286490000700310100002200317700001300339700001700352700001800369856004400387 2020 eng d00aFlow Cytometry Analysis and Fluorescence-activated Cell Sorting of Myeloid Cells from Lung and Bronchoalveolar Lavage Samples from Mycobacterium tuberculosis-infected Mice0 aFlow Cytometry Analysis and Fluorescenceactivated Cell Sorting o bBio-Protocol, {LLC}0 v101 aRothchild, Alissa1 aMai, Dat1 aAderem, Alan1 aDiercks, Alan uhttps://doi.org/10.21769/bioprotoc.363002899nas a2200505 4500008004100000022001400041245008100055210006900136260001200205300001000217490000700227520140700234653001501641653001001656653000901666653001401675653002201689653002101711653002301732653001401755653001101769653004301780653001101823653001601834653001401850653002401864653002601888653001601914100002101930700001901951700002001970700001701990700001302007700001802020700001902038700002402057700002702081700001902108700002502127700002502152700002502177700002302202700001802225856015002243 2020 eng d a1573-703900aInter-Individual Variation in Response to Estrogen in Human Breast Explants.0 aInterIndividual Variation in Response to Estrogen in Human Breas c2020 03 a51-680 v253 aExposure to estrogen is strongly associated with increased breast cancer risk. While all women are exposed to estrogen, only 12% are expected to develop breast cancer during their lifetime. These women may be more sensitive to estrogen, as rodent models have demonstrated variability in estrogen sensitivity. Our objective was to determine individual variation in expression of estrogen receptor (ER) and estrogen-induced responses in the normal human breast. Human breast tissue from female donors undergoing reduction mammoplasty surgery were collected for microarray analysis of ER expression. To examine estrogen-induced responses, breast tissue from 23 female donors were cultured ex- vivo in basal or 10 nM 17β-estradiol (E2) media for 4 days. Expression of ER genes (ESR1 and ESR2) increased significantly with age. E2 induced consistent increases in global gene transcription, but expression of target genes AREG, PGR, and TGFβ2 increased significantly only in explants from nulliparous women. E2-treatment did not induce consistent changes in proliferation or radiation induced apoptosis. Responses to estrogen are highly variable among women and not associated with levels of ER expression, suggesting differences in intracellular signaling among individuals. The differences in sensitivity to E2-stimulated responses may contribute to variation in risk of breast cancer.
10aAdolescent10aAdult10aAged10aApoptosis10aBiomarkers, Tumor10aBreast Neoplasms10aCell Proliferation10aEstrogens10aFemale10aGene Expression Regulation, Neoplastic10aHumans10aMiddle Aged10aPrognosis10aReceptors, Estrogen10aTumor Cells, Cultured10aYoung Adult1 aDunphy, Karen, A1 aBlack, Amye, L1 aRoberts, Amy, L1 aSharma, Aman1 aLi, Zida1 aSuresh, Sneha1 aBrowne, Eva, P1 aArcaro, Kathleen, F1 aSer-Dolansky, Jennifer1 aBigelow, Carol1 aTroester, Melissa, A1 aSchneider, Sallie, S1 aMakari-Judson, Grace1 aCrisi, Giovanna, M1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/inter-individual-variation-response-estrogen-human-breast-explants00906nas a2200289 4500008004100000245008000041210006900121260004600190300001200236490000700248100002100255700002000276700002000296700001700316700001300333700001800346700001900364700002400383700002700407700001900434700002500453700002500478700002500503700002300528700001800551856004700569 2020 eng d00aInter-Individual Variation in Response to Estrogen in Human Breast Explants0 aInterIndividual Variation in Response to Estrogen in Human Breas bSpringer Science and Business Media {LLC} a51–680 v251 aDunphy, Karen, A1 aBlack, Amye, L.1 aRoberts, Amy, L1 aSharma, Aman1 aLi, Zida1 aSuresh, Sneha1 aBrowne, Eva, P1 aArcaro, Kathleen, F1 aSer-Dolansky, Jennifer1 aBigelow, Carol1 aTroester, Melissa, A1 aSchneider, Sallie, S1 aMakari-Judson, Grace1 aCrisi, Giovanna, M1 aJerry, Joseph uhttps://doi.org/10.1007/s10911-020-09446-300473nas a2200145 4500008004100000245008800041210006900129260001000198100001700208700001500225700001900240700001700259700001300276856003800289 2020 eng d00aLoss of POLR1D results in embryonic lethality prior to blastocyst formation in mice0 aLoss of POLR1D results in embryonic lethality prior to blastocys bWiley1 aMiao, Xiaosu1 aSun, Tieqi1 aGolan, Morgane1 aMager, Jesse1 aCui, Wei uhttps://doi.org/10.1002/mrd.2342700498nas a2200157 4500008004100000245008800041210006900129300001600198490000700214100001700221700001500238700001900253700001700272700001300289856003800302 2020 eng d00aLoss of POLR1D results in embryonic lethality prior to blastocyst formation in mice0 aLoss of POLR1D results in embryonic lethality prior to blastocys a1152–11580 v871 aMiao, Xiaosu1 aSun, Tieqi1 aGolan, Morgane1 aMager, Jesse1 aCui, Wei uhttps://doi.org/10.1002/mrd.2342700554nas a2200145 4500008004100000245013700041210006900178260003600247100001700283700001500300700002000315700001700335700001300352856004300365 2020 eng d00aLoss of RBBP4 results in defective inner cell mass, severe apoptosis, hyperacetylated histones and preimplantation lethality in mice0 aLoss of RBBP4 results in defective inner cell mass severe apopto bOxford University Press ({OUP})1 aMiao, Xiaosu1 aSun, Tieqi1 aBarletta, Holly1 aMager, Jesse1 aCui, Wei uhttps://doi.org/10.1093/biolre/ioaa04602406nas a2200385 4500008004100000022001400041245014100055210006900196260001500265300001000280490000800290520121800298653001601516653001201532653001401544653001501558653002401573653001601597653002601613653001301639653001101652653002001663653001301683653000901696653000901705653002301714653001901737653003701756100001701793700001501810700002001825700001701845700001301862856014501875 2020 eng d a1529-726800aLoss of RBBP4 results in defective inner cell mass, severe apoptosis, hyperacetylated histones and preimplantation lethality in mice†.0 aLoss of RBBP4 results in defective inner cell mass severe apopto c2020 06 23 a13-230 v1033 aRetinoblastoma-binding protein 4 (RBBP4) (also known as chromatin-remodeling factor RBAP48) is an evolutionarily conserved protein that has been involved in various biological processes. Although a variety of functions have been attributed to RBBP4 in vitro, mammalian RBBP4 has not been studied in vivo. Here we report that RBBP4 is essential during early mouse embryo development. Although Rbbp4 mutant embryos exhibit normal morphology at E3.5 blastocyst stage, they cannot be recovered at E7.5 early post-gastrulation stage, suggesting an implantation failure. Outgrowth (OG) assays reveal that mutant blastocysts cannot hatch from the zona or can hatch but then arrest without further development. We find that while there is no change in proliferation or levels of reactive oxygen species, both apoptosis and histone acetylation are significantly increased in mutant blastocysts. Analysis of lineage specification reveals that while the trophoblast is properly specified, both epiblast and primitive endoderm lineages are compromised with severe reductions in cell number and/or specification. In summary, these findings demonstrate the essential role of RBBP4 during early mammalian embryogenesis.
10aAcetylation10aAnimals10aApoptosis10aBlastocyst10aEmbryo Implantation10aEmbryo Loss10aEmbryonic Development10aEndoderm10aFemale10aGene Expression10aHistones10aMale10aMice10aMice, Inbred C57BL10aMice, Knockout10aRetinoblastoma-Binding Protein 41 aMiao, Xiaosu1 aSun, Tieqi1 aBarletta, Holly1 aMager, Jesse1 aCui, Wei uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/loss-rbbp4-results-defective-inner-cell-mass-severe-apoptosis00661nas a2200205 4500008004100000245008300041210006900124260001800193300001200211490000800223100002200231700002000253700002100273700002100294700002500315700002200340700002500362700002000387856004800407 2020 eng d00aMcrs1 interacts with Six1 to influence early craniofacial and otic development0 aMcrs1 interacts with Six1 to influence early craniofacial and ot bElsevier {BV} a39–500 v4671 aNeilson, Karen, M1 aKeer, Stephanie1 aBousquet, Nicole1 aMacrorie, Olivia1 aMajumdar, Himani, D.1 aKenyon, Kristy, L1 aAlfandari, Dominique1 aMoody, Sally, A uhttps://doi.org/10.1016/j.ydbio.2020.08.01302385nas a2200409 4500008004100000022001400041245008200055210006900137260001200206300000900218490000800227520113000235653001201365653003101377653002501408653001701433653002201450653002601472653002501498653001101523653004601534653001601580653000901596653000901605653002301614653001901637653001301656653002501669100001301694700001801707700002001725700002001745700001401765700002601779700001701805856015301822 2020 eng d a1741-789900aMCRS1 is essential for epiblast development during early mouse embryogenesis.0 aMCRS1 is essential for epiblast development during early mouse e c2020 01 a1-130 v1593 aMicrospherule protein 1 (MCRS1, also known as MSP58) is an evolutionarily conserved protein that has been implicated in various biological processes. Although a variety of functions have been attributed to MCRS1 in vitro, mammalian MCRS1 has not been studied in vivo. Here we report that MCRS1 is essential during early murine development. Mcrs1 mutant embryos exhibit normal morphology at the blastocyst stage but cannot be recovered at gastrulation, suggesting an implantation failure. Outgrowth (OG) assays reveal that mutant blastocysts do not form a typical inner cell mass (ICM) colony, the source of embryonic stem cells (ESCs). Surprisingly, cell death and histone H4 acetylation analysis reveal that apoptosis and global H4 acetylation are normal in mutant blastocysts. However, analysis of lineage specification reveals that while the trophoblast and primitive endoderm are properly specified, the epiblast lineage is compromised and exhibits a severe reduction in cell number. In summary, our study demonstrates the indispensable role of MCRS1 in epiblast development during early mammalian embryogenesis.
10aAnimals10aBlastocyst Inner Cell Mass10aCell Differentiation10aCell Lineage10aEmbryo, Mammalian10aEmbryonic Development10aEmbryonic Stem Cells10aFemale10aGene Expression Regulation, Developmental10aGerm Layers10aMale10aMice10aMice, Inbred C57BL10aMice, Knockout10aMutation10aRNA-Binding Proteins1 aCui, Wei1 aCheong, Agnes1 aWang, Yongsheng1 aTsuchida, Yuran1 aLiu, Yong1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mcrs1-essential-epiblast-development-during-early-mouse-embryogenesis00527nas a2200157 4500008004100000245005700041210005700098260003600155100002100191700002700212700002700239700002300266700002000289700001700309856004300326 2020 eng d00aMetabolic changes in mouse sperm during capacitation0 aMetabolic changes in mouse sperm during capacitation bOxford University Press ({OUP})1 aBalbach, Melanie1 aGervasi, Maria, Gracia1 aHidalgo, David, Martin1 aVisconti, Pablo, E1 aLevin, Lonny, R1 aBuck, Jochen uhttps://doi.org/10.1093/biolre/ioaa11400625nas a2200181 4500008004100000245010700041210006900148260003000217300001400247490000800261100001900269700002100288700001800309700003100327700002500358700002100383856003900404 2020 eng d00aMMP14 is required for delamination of chick neural crest cells independently of its catalytic activity0 aMMP14 is required for delamination of chick neural crest cells i bThe Company of Biologists adev1839540 v1471 aAndrieu, Cyril1 aMontigny, Audrey1 aBibonne, Anne1 aDespin-Guitard, Evangeline1 aAlfandari, Dominique1 aThéveneau, Eric uhttps://doi.org/10.1242/dev.18395402680nas a2200301 4500008004100000022001400041245010300055210006900158260001200227300001400239490000700253520169500260653001501955653001201970653002101982653002702003653001102030653001102041653001702052653002502069653002702094653000902121100002502130700002402155700002702179700001802206856015402224 2020 eng d a1543-015400aThe Mouse Mammary Gland: a Tool to Inform Adolescents About Environmental Causes of Breast Cancer.0 aMouse Mammary Gland a Tool to Inform Adolescents About Environme c2020 12 a1094-11000 v353 aAdolescence is a vulnerable period of breast development, and environmental chemical exposures that occur during this period can increase the risk of breast cancer in adulthood. Discussing breast health with adolescent girls can be difficult for several reasons. In this project, we worked to not only inform adolescent researchers about environmental risks for breast cancer but to also involve them in research studies. We taught adolescents about the stages of mammary gland development using samples collected from mice, with a specific focus on pre-pubertal and pubertal stages of development. Our analysis shows that adolescent researchers, with relatively modest training, can collect reliable and reproducible data on aspects of mammary gland biology that are known to be disrupted by environmental chemicals, with coefficients of variation < 2.5% for basic mammary gland parameters and 5-7% for more complex measures. Finally, we provided these adolescents with information about environmental risk factors for breast cancer that they could share with their peers and community and action items to potentially modify their individual risk. We hope that researchers working in this field will engage adolescent researchers in projects to evaluate chemicals that influence breast cancer risk. Summer research programs that inform young adolescents about breast cancer risk factors not only benefit these novice researchers individually but also benefit their communities when they are encouraged to talk about the value of basic science studies, discuss vulnerable periods of mammary gland development, and share what they have learned about cancer and the environment.
10aAdolescent10aAnimals10aBreast Neoplasms10aEnvironmental Exposure10aFemale10aHumans10aLaboratories10aLaboratory Personnel10aMammary Glands, Animal10aMice1 aVandenberg, Laura, N1 aKolla, SriDurgaDevi1 aLaPlante, Charlotte, D1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mouse-mammary-gland-tool-inform-adolescents-about-environmental-causes03739nas a2200973 4500008004100000022001400041245008900055210006900144260001500213300001800228490000800246520107400254653001501328653001001343653000901353653002201362653001301384653001101397653001301408653001101421653000901432653001601441653001201457653001501469653003001484653002501514100001501539700001701554700001401571700002501585700001901610700002201629700002201651700002401673700002101697700002001718700002101738700001901759700001701778700002401795700002401819700001301843700001301856700001501869700002201884700001501906700001601921700001701937700001601954700001901970700001601989700001802005700001602023700001402039700001602053700001802069700001502087700001502102700001602117700001702133700002002150700002502170700002202195700002302217700002202240700001302262700002102275700001502296700002302311700001402334700002102348700002102369700001602390700001702406700002602423700002102449700002502470700002202495700001902517700002202536700002002558710004002578856014702618 2020 eng d a1097-417200aMulti-Omics Resolves a Sharp Disease-State Shift between Mild and Moderate COVID-19.0 aMultiOmics Resolves a Sharp DiseaseState Shift between Mild and c2020 12 10 a1479-1495.e200 v1833 aWe present an integrated analysis of the clinical measurements, immune cells, and plasma multi-omics of 139 COVID-19 patients representing all levels of disease severity, from serial blood draws collected during the first week of infection following diagnosis. We identify a major shift between mild and moderate disease, at which point elevated inflammatory signaling is accompanied by the loss of specific classes of metabolites and metabolic processes. Within this stressed plasma environment at moderate disease, multiple unusual immune cell phenotypes emerge and amplify with increasing disease severity. We condensed over 120,000 immune features into a single axis to capture how different immune cell classes coordinate in response to SARS-CoV-2. This immune-response axis independently aligns with the major plasma composition changes, with clinical metrics of blood clotting, and with the sharp transition between mild and moderate disease. This study suggests that moderate disease may provide the most effective setting for therapeutic intervention.
10aAdolescent10aAdult10aAged10aAged, 80 and over10aCOVID-1910aFemale10aGenomics10aHumans10aMale10aMiddle Aged10aRNA-Seq10aSARS-CoV-210aSeverity of Illness Index10aSingle-Cell Analysis1 aSu, Yapeng1 aChen, Daniel1 aYuan, Dan1 aLausted, Christopher1 aChoi, Jongchan1 aDai, Chengzhen, L1 aVoillet, Valentin1 aDuvvuri, Venkata, R1 aScherler, Kelsey1 aTroisch, Pamela1 aBaloni, Priyanka1 aQin, Guangrong1 aSmith, Brett1 aKornilov, Sergey, A1 aRostomily, Clifford1 aXu, Alex1 aLi, Jing1 aDong, Shen1 aRothchild, Alissa1 aZhou, Jing1 aMurray, Kim1 aEdmark, Rick1 aHong, Sunga1 aHeath, John, E1 aEarls, John1 aZhang, Rongyu1 aXie, Jingyi1 aLi, Sarah1 aRoper, Ryan1 aJones, Lesley1 aZhou, Yong1 aRowen, Lee1 aLiu, Rachel1 aMackay, Sean1 aO'Mahony, Shane1 aDale, Christopher, R1 aWallick, Julie, A1 aAlgren, Heather, A1 aZager, Michael, A1 aWei, Wei1 aPrice, Nathan, D1 aHuang, Sui1 aSubramanian, Naeha1 aWang, Kai1 aMagis, Andrew, T1 aHadlock, Jenn, J1 aHood, Leroy1 aAderem, Alan1 aBluestone, Jeffrey, A1 aLanier, Lewis, L1 aGreenberg, Philip, D1 aGottardo, Raphael1 aDavis, Mark, M1 aGoldman, Jason, D1 aHeath, James, R1 aISB-Swedish COVID19 Biobanking Unit uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/multi-omics-resolves-sharp-disease-state-shift-between-mild-and00553nas a2200157 4500008004100000245009100041210006900132260003000201100001800231700002600249700001800275700002300293700002300316700001700339856003900356 2020 eng d00aNuclear encoded mitochondrial ribosomal proteins are required to initiate gastrulation0 aNuclear encoded mitochondrial ribosomal proteins are required to bThe Company of Biologists1 aCheong, Agnes1 aArchambault, Danielle1 aDegani, Rinat1 aIverson, Elizabeth1 aTremblay, Kimberly1 aMager, Jesse uhttps://doi.org/10.1242/dev.18871400639nas a2200181 4500008004100000245010400041210006900145260004600214300001600260490000800276100001300284700001800297700002800315700002400343700002000367700002300387856004700410 2020 eng d00aPolymeric nanoassemblies for enrichment and detection of peptides and proteins in human breast milk0 aPolymeric nanoassemblies for enrichment and detection of peptide bSpringer Science and Business Media {LLC} a1027–10350 v4121 aZhao, Bo1 aGao, Jingjing1 aSerrano, Mahalia, A. C.1 aArcaro, Kathleen, F1 aThayumanavan, S1 aVachet, Richard, W uhttps://doi.org/10.1007/s00216-019-02342-800693nas a2200205 4500008004100000245009300041210006900134260003600203300001400239490000700253100002100260700002200281700001900303700002500322700002500347700002500372700002400397700002700421856003900448 2020 eng d00aPrediagnostic breast milk DNA methylation alterations in women who develop breast cancer0 aPrediagnostic breast milk DNA methylation alterations in women w bOxford University Press ({OUP}) a662–6730 v291 aSalas, Lucas, A.1 aLundgren, Sara, N1 aBrowne, Eva, P1 aPunska, Elizabeth, C1 aAnderton, Douglas, L1 aKaragas, Margaret, R1 aArcaro, Kathleen, F1 aChristensen, Brock, C. uhttps://doi.org/10.1093/hmg/ddz30102787nas a2200337 4500008004100000022001400041245010900055210006900164260001500233300001400248490000700262520154600269653003001815653003501845653003101880653004601911653003001957653002001987653005702007653002702064653002202091653002402113653003002137100001602167700002902183700001902212700002402231700002002255700002002275856015402295 2020 eng d a1525-002400aProtein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulate FOXP3 Stability in Regulatory T Cells.0 aProtein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulat c2020 10 07 a2220-22360 v283 aT cell receptor signaling, together with cytokine-induced signals, can differentially regulate RNA processing to influence T helper versus regulatory T cell fate. Protein kinase C family members have been shown to function in alternative splicing and RNA processing in various cell types. T cell-specific protein kinase C theta, a molecular regulator of T cell receptor downstream signaling, has been shown to phosphorylate splicing factors and affect post-transcriptional control of T cell gene expression. In this study, we explored how using a synthetic cell-penetrating peptide mimic for intracellular anti-protein kinase C theta delivery fine-tunes differentiation of induced regulatory T cells through its differential effects on RNA processing. We identified protein kinase C theta signaling as a critical modulator of two key RNA regulatory factors, heterogeneous nuclear ribonucleoprotein L (hnRNPL) and protein-l-isoaspartate O-methyltransferase-1 (PCMT1), and loss of protein kinase C theta function initiated a "switch" in post-transcriptional organization in induced regulatory T cells. More interestingly, we discovered that protein-l-isoaspartate O- methyltransferase-1 acts as an instability factor in induced regulatory T cells, by methylating the forkhead box P3 (FOXP3) promoter. Targeting protein-l-isoaspartate O-methyltransferase-1 using a cell-penetrating antibody revealed an efficient means of modulating RNA processing to confer a stable regulatory T cell phenotype.
10aCell-Penetrating Peptides10aForkhead Transcription Factors10aGene Expression Regulation10aHeterogeneous-Nuclear Ribonucleoprotein L10aPromoter Regions, Genetic10aProtein Binding10aProtein D-Aspartate-L-Isoaspartate Methyltransferase10aProtein Kinase C-theta10aProtein Stability10aSignal Transduction10aT-Lymphocytes, Regulatory1 aOzay, Ilker1 aShanthalingam, Sudarvili1 aTorres, Joe, A1 aOsborne, Barbara, A1 aTew, Gregory, N1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/protein-kinase-c-theta-modulates-pcmt1-through-hnrnpl-regulate-foxp3-002785nas a2200337 4500008004100000022001400041245010900055210006900164260001500233300001400248490000700262520154600269653003001815653003501845653003101880653004601911653003001957653002001987653005702007653002702064653002202091653002402113653003002137100001602167700002902183700001902212700002402231700002002255700002002275856015202295 2020 eng d a1525-002400aProtein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulate FOXP3 Stability in Regulatory T Cells.0 aProtein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulat c2020 10 07 a2220-22360 v283 aT cell receptor signaling, together with cytokine-induced signals, can differentially regulate RNA processing to influence T helper versus regulatory T cell fate. Protein kinase C family members have been shown to function in alternative splicing and RNA processing in various cell types. T cell-specific protein kinase C theta, a molecular regulator of T cell receptor downstream signaling, has been shown to phosphorylate splicing factors and affect post-transcriptional control of T cell gene expression. In this study, we explored how using a synthetic cell-penetrating peptide mimic for intracellular anti-protein kinase C theta delivery fine-tunes differentiation of induced regulatory T cells through its differential effects on RNA processing. We identified protein kinase C theta signaling as a critical modulator of two key RNA regulatory factors, heterogeneous nuclear ribonucleoprotein L (hnRNPL) and protein-l-isoaspartate O-methyltransferase-1 (PCMT1), and loss of protein kinase C theta function initiated a "switch" in post-transcriptional organization in induced regulatory T cells. More interestingly, we discovered that protein-l-isoaspartate O- methyltransferase-1 acts as an instability factor in induced regulatory T cells, by methylating the forkhead box P3 (FOXP3) promoter. Targeting protein-l-isoaspartate O-methyltransferase-1 using a cell-penetrating antibody revealed an efficient means of modulating RNA processing to confer a stable regulatory T cell phenotype.
10aCell-Penetrating Peptides10aForkhead Transcription Factors10aGene Expression Regulation10aHeterogeneous-Nuclear Ribonucleoprotein L10aPromoter Regions, Genetic10aProtein Binding10aProtein D-Aspartate-L-Isoaspartate Methyltransferase10aProtein Kinase C-theta10aProtein Stability10aSignal Transduction10aT-Lymphocytes, Regulatory1 aOzay, Ilker1 aShanthalingam, Sudarvili1 aTorres, Joe, A1 aOsborne, Barbara, A1 aTew, Gregory, N1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/protein-kinase-c-theta-modulates-pcmt1-through-hnrnpl-regulate-foxp300617nas a2200181 4500008004100000245010800041210006900149260001800218300001600236490000700252100001600259700002900275700001900304700002400323700002000347700002000367856004800387 2020 eng d00aProtein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulate FOXP3 Stability in Regulatory T Cells0 aProtein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulat bElsevier {BV} a2220–22360 v281 aOzay, Ilker1 aShanthalingam, Sudarvili1 aTorres, Joe, A1 aOsborne, Barbara, A1 aTew, Gregory, N1 aMinter, Lisa, M uhttps://doi.org/10.1016/j.ymthe.2020.06.01200631nas a2200169 4500008004100000245015600041210006900197260001800266300001100284490000800295100002600303700001800329700002300347700002600370700001700396856004800413 2020 eng d00aProtein phosphatase 1 regulatory subunit 35 is required for ciliogenesis, notochord morphogenesis, and cell-cycle progression during murine development0 aProtein phosphatase 1 regulatory subunit 35 is required for cili bElsevier {BV} a1–100 v4651 aArchambault, Danielle1 aCheong, Agnes1 aIverson, Elizabeth1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://doi.org/10.1016/j.ydbio.2020.06.01100716nas a2200205 4500008004100000022001400041245007500055210006900130260001200199300001200211490000800223653004900231653001400280653002000294653001100314653000900325653001700334100001300351856014600364 2020 eng d a1556-565300aSOHLHs are essential for fertility regardless of gender or population.0 aSOHLHs are essential for fertility regardless of gender or popul c2020 08 a283-2840 v11410aBasic Helix-Loop-Helix Transcription Factors10aFertility10aGender Identity10aHumans10aMale10aReproduction1 aCui, Wei uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sohlhs-are-essential-fertility-regardless-gender-or-population00633nas a2200169 4500008004100000245012100041210006900162260003600231100002200267700002300289700001900312700002400331700001800355700002400373700002300397856004300420 2020 eng d00aTestis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception†0 aTestisspecific serine kinase protein family in male fertility an bOxford University Press ({OUP})1 aSalicioni, Ana, M1 aGervasi, Maria, G.1 aSosnik, Julian1 aTourzani, Darya, A.1 aNayyab, Saman1 aCaraballo, Diego, A1 aVisconti, Pablo, E uhttps://doi.org/10.1093/biolre/ioaa06402834nas a2200337 4500008004100000022001400041245013900055210006900194260001200263300001200275490000700287520170600294653001802000653001102018653001802029653002502047653002702072653001102099653001102110653002602121653001602147653003002163100002202193700002402215700001902239700002702258700002502285700001802310700002502328856014302353 2020 eng d a1440-171100aThe use of patient-derived breast tissue explants to study macrophage polarization and the effects of environmental chemical exposure.0 ause of patientderived breast tissue explants to study macrophage c2020 11 a883-8960 v983 aEx vivo mammary explant systems are an excellent model to study interactions between epithelium and stromal cell types because they contain physiologically relevant heterotypic interactions in the background of genetically diverse patients. The intact human mammary tissue, termed patient-derived explant (PDE), can be used to investigate cellular responses to a wide variety of external stimuli in situ. For this study, we examined the impact of cytokines or environmental chemicals on macrophage phenotypes. We demonstrate that we can polarize macrophages within human breast tissue PDEs toward M1 or M2 through the addition of interferon-γ (IFNγ) + lipopolysaccharide (LPS) or interleukin (IL)-4 + IL-13, respectively. Elevated expression levels of M(IFNγ + LPS) markers (HLADRA and CXCL10) or M(IL-4 + IL-13) markers (CD209 and CCL18) were observed in cytokine-treated tissues. We also examined the impact of the endocrine-disrupting chemical, benzophenone-3, on PDEs and measured significant, yet varying effects on macrophage polarization. Furthermore, a subset of the PDEs respond to IL-4 + IL-13 through downregulation of E-cadherin and upregulation of vimentin which is reminiscent of epithelial-to-mesenchymal transition (EMT) changes. Finally, we were able to show immortalized nonmalignant breast epithelial cells can exhibit EMT characteristics when exposed to growth factors secreted by M(IL-4 + IL-13) macrophages. Taken together, the PDE model system is an outstanding preclinical model to study early tissue-resident immune responses and effects on epithelial and stromal responses to stimuli found both endogenously in the breast and exogenously as a result of exposures.
10aBenzophenones10aBreast10aCell Polarity10aEndocrine Disruptors10aEnvironmental Exposure10aFemale10aHumans10aMacrophage Activation10aMacrophages10aTissue Culture Techniques1 aGregory, Kelly, J1 aMorin, Stephanie, M1 aKubosiak, Alex1 aSer-Dolansky, Jennifer1 aSchalet, Benjamin, J1 aJerry, Joseph1 aSchneider, Sallie, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/use-patient-derived-breast-tissue-explants-study-macrophage00688nas a2200193 4500008004100000245012000041210006900161260004600230490000700276100002600283700002300309700002100332700002000353700002400373700001700397700001900414700001400433856004700447 2020 eng d00ay-Secretase modulators exhibit selectivity for modulation of APP cleavage but inverse y-secretase modulators do not0 aySecretase modulators exhibit selectivity for modulation of APP bSpringer Science and Business Media {LLC}0 v121 aLessard, Christian, B1 aRodriguez, Edgardo1 aLadd, Thomas, B.1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio1 aGolde, Todd, E1 aRan, Yong uhttps://doi.org/10.1186/s13195-020-00622-500519nas a2200145 4500008004100000245009500041210006900136260003600205100001600241700001700257700002600274700001700300700001300317856004300330 2020 eng d00aZC3H4, a novel CCCH-type zinc finger protein, is essential for early embryogenesis in mice0 aZC3H4 a novel CCCHtype zinc finger protein is essential for earl bOxford University Press ({OUP})1 aSu, Jianmin1 aMiao, Xiaosu1 aArchambault, Danielle1 aMager, Jesse1 aCui, Wei uhttps://doi.org/10.1093/biolre/ioaa21500714nas a2200217 4500008004100000245009300041210006900134260002500203490000700228100002400235700002100259700002500280700002100305700002100326700002200347700002400369700002000393700001700413700002100430856004500451 2019 eng d00aAdenosine A2A Receptor Stimulation Inhibits TCR-Induced Notch1 Activation in CD8+T-Cells0 aAdenosine A2A Receptor Stimulation Inhibits TCRInduced Notch1 Ac bFrontiers Media {SA}0 v101 aSorrentino, Claudia1 aHossain, Fokhrul1 aRodriguez, Paulo, C.1 aSierra, Rosa, A.1 aPannuti, Antonio1 aHatfield, Stephen1 aOsborne, Barbara, A1 aMinter, Lisa, M1 aMiele, Lucio1 aMorello, Silvana uhttps://doi.org/10.3389/fimmu.2019.0016200876nas a2200265 4500008004100000245012900041210006900170260001800239300001600257490000700273100001900280700002000299700001700319700002300336700002200359700002900381700002000410700002100430700002400451700002400475700002000499700002300519700002000542856004800562 2019 eng d00aAlpha-1 Antitrypsin-Expressing Mesenchymal Stromal Cells Confer a Long-Term Survival Benefit in a Mouse Model of Lethal GvHD0 aAlpha1 AntitrypsinExpressing Mesenchymal Stromal Cells Confer a bElsevier {BV} a1436–14510 v271 aGeiger, Sabine1 aOzay, Emrah, I.1 aGeumann, Ulf1 aHereth, Marina, K.1 aMagnusson, Terese1 aShanthalingam, Sudarvili1 aHirsch, Daniela1 aKälin, Stefanie1 aGünther, Christine1 aOsborne, Barbara, A1 aTew, Gregory, N1 aHermann, Felix, G.1 aMinter, Lisa, M uhttps://doi.org/10.1016/j.ymthe.2019.05.00700736nas a2200205 4500008004100000245012400041210006900165260006500234300001300299490000600312100002600318700002300344700002100367700001900388700001300407700002400420700002200444700001700466856004700483 2019 eng d00aAlveolar macrophages generate a noncanonical NRF2-driven transcriptional response to Mycobacterium tuberculosis in vivo0 aAlveolar macrophages generate a noncanonical NRF2driven transcri bAmerican Association for the Advancement of Science ({AAAS}) aeaaw66930 v41 aRothchild, Alissa, C.1 aOlson, Gregory, S.1 aNemeth, Johannes1 aAmon, Lynn, M.1 aMai, Dat1 aGold, Elizabeth, S.1 aDiercks, Alan, H.1 aAderem, Alan uhttps://doi.org/10.1126/sciimmunol.aaw669300727nas a2200217 4500008004100000245010600041210006900147260002500216490000700241100002400248700002100272700002500293700002100318700002100339700002200360700002400382700002000406700001700426700002100443856004500464 2019 eng d00aCorrigendum: Adenosine A2A Receptor Stimulation Inhibits TCR-Induced Notch1 Activation in CD8+T-Cells0 aCorrigendum Adenosine A2A Receptor Stimulation Inhibits TCRInduc bFrontiers Media {SA}0 v101 aSorrentino, Claudia1 aHossain, Fokhrul1 aRodriguez, Paulo, C.1 aSierra, Rosa, A.1 aPannuti, Antonio1 aHatfield, Stephen1 aOsborne, Barbara, A1 aMinter, Lisa, M1 aMiele, Lucio1 aMorello, Silvana uhttps://doi.org/10.3389/fimmu.2019.0093502550nas a2200433 4500008004100000022001400041245012300055210007100178260001200249300001100261490000700272520110800279653001201387653002701399653001101426653002601437653004401463653001101507653003501518653004201553653002701595653000901622653002101631100001601652700002701668700002901695700002901724700002401753700002401777700002301801700001901824700002401843700002001867700002101887700002301908700001801931700002001949856014701969 2019 eng d a1876-775300aCymerus™ iPSC-MSCs significantly prolong survival in a pre-clinical, humanized mouse model of Graft-vs-host disease.0 aCymerus™ iPSCMSCs significantly prolong survival in a preclinica c2019 03 a1014010 v353 aThe immune-mediated tissue destruction of graft-vs-host disease (GvHD) remains a major barrier to greater use of hematopoietic stem cell transplantation (HSCT). Mesenchymal stem cells (MSCs) have intrinsic immunosuppressive qualities and are being actively investigated as a therapeutic strategy for treating GvHD. We characterized Cymerus™ MSCs, which are derived from adult, induced pluripotent stem cells (iPSCs), and show they display surface markers and tri-lineage differentiation consistent with MSCs isolated from bone marrow (BM). Administering iPSC-MSCs altered phosphorylation and cellular localization of the T cell-specific kinase, Protein Kinase C theta (PKCθ), attenuated disease severity, and prolonged survival in a humanized mouse model of GvHD. Finally, we evaluated a constellation of pro-inflammatory molecules on circulating PBMCs that correlated closely with disease progression and which may serve as biomarkers to monitor therapeutic response. Altogether, our data suggest Cymerus iPSC-MSCs offer the potential for an off-the-shelf, cell-based therapy to treat GvHD.
10aAnimals10aDisease Models, Animal10aFemale10aGraft vs Host Disease10aHematopoietic Stem Cell Transplantation10aHumans10aInduced Pluripotent Stem Cells10aMesenchymal Stem Cell Transplantation10aMesenchymal Stem Cells10aMice10aMice, Inbred NOD1 aOzay, Ilker1 aVijayaraghavan, Jyothi1 aGonzalez-Perez, Gabriela1 aShanthalingam, Sudarvili1 aSherman, Heather, L1 aGarrigan, Daniel, T1 aChandiran, Karthik1 aTorres, Joe, A1 aOsborne, Barbara, A1 aTew, Gregory, N1 aSlukvin, Igor, I1 aMacdonald, Ross, A1 aKelly, Kilian1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cymerustm-ipsc-mscs-significantly-prolong-survival-pre-clinical00562nas a2200169 4500008004100000245007100041210006700112260003900179300001300218490000700231100002200238700002600260700002600286700001700312700001400329856004900343 2019 eng d00aThe elongation factor Elof1 is required for mammalian gastrulation0 aelongation factor Elof1 is required for mammalian gastrulation bPublic Library of Science ({PLoS}) ae02194100 v141 aTellier, Adam, P.1 aArchambault, Danielle1 aTremblay, Kimberly, D1 aMager, Jesse1 aWu, Qiang uhttps://doi.org/10.1371/journal.pone.021941000830nas a2200241 4500008004100000245011600041210006900157260004600226490000700272100002200279700002700301700002400328700001700352700001700369700001800386700002800404700002300432700002300455700002500478700001500503700002400518856004600542 2019 eng d00aEnvironmental exposures during windows of susceptibility for breast cancer: a framework for prevention research0 aEnvironmental exposures during windows of susceptibility for bre bSpringer Science and Business Media {LLC}0 v211 aTerry, Mary, Beth1 aMichels, and, Karin B.1 aBrody, Julia, Green1 aByrne, Celia1 aChen, Shiuan1 aJerry, Joseph1 aMalecki, Kristen, M. C.1 aMartin, Mary, Beth1 aMiller, Rachel, L.1 aNeuhausen, Susan, L.1 aSilk, Kami1 aTrentham-Dietz, Amy uhttps://doi.org/10.1186/s13058-019-1168-204080nas a2200445 4500008004100000022001400041245011700055210006900172260001500241300000700256490000700263520267800270653001202948653002102960653002702981653002703008653001103035653001103046653002203057653001403079653001403093653001203107653001303119653001703132653001703149100002203166700002203188700002403210700001703234700001703251700001803268700002603286700002303312700002203335700002403357700001503381700002403396710006303420856015103483 2019 eng d a1465-542X00aEnvironmental exposures during windows of susceptibility for breast cancer: a framework for prevention research.0 aEnvironmental exposures during windows of susceptibility for bre c2019 08 20 a960 v213 aBACKGROUND: The long time from exposure to potentially harmful chemicals until breast cancer occurrence poses challenges for designing etiologic studies and for implementing successful prevention programs. Growing evidence from animal and human studies indicates that distinct time periods of heightened susceptibility to endocrine disruptors exist throughout the life course. The influence of environmental chemicals on breast cancer risk may be greater during several windows of susceptibility (WOS) in a woman's life, including prenatal development, puberty, pregnancy, and the menopausal transition. These time windows are considered as specific periods of susceptibility for breast cancer because significant structural and functional changes occur in the mammary gland, as well as alterations in the mammary micro-environment and hormone signaling that may influence risk. Breast cancer research focused on these breast cancer WOS will accelerate understanding of disease etiology and prevention.
MAIN TEXT: Despite the plausible heightened mechanistic influences of environmental chemicals on breast cancer risk during time periods of change in the mammary gland's structure and function, most human studies of environmental chemicals are not focused on specific WOS. This article reviews studies conducted over the past few decades that have specifically addressed the effect of environmental chemicals and metals on breast cancer risk during at least one of these WOS. In addition to summarizing the broader evidence-base specific to WOS, we include discussion of the NIH-funded Breast Cancer and the Environment Research Program (BCERP) which included population-based and basic science research focused on specific WOS to evaluate associations between breast cancer risk and particular classes of endocrine-disrupting chemicals-including polycyclic aromatic hydrocarbons, perfluorinated compounds, polybrominated diphenyl ethers, and phenols-and metals. We outline ways in which ongoing transdisciplinary BCERP projects incorporate animal research and human epidemiologic studies in close partnership with community organizations and communication scientists to identify research priorities and effectively translate evidence-based findings to the public and policy makers.
CONCLUSIONS: An integrative model of breast cancer research is needed to determine the impact and mechanisms of action of endocrine disruptors at different WOS. By focusing on environmental chemical exposure during specific WOS, scientists and their community partners may identify when prevention efforts are likely to be most effective.
10aAnimals10aBreast Neoplasms10aDisease Susceptibility10aEnvironmental Exposure10aFemale10aHumans10aMaternal Exposure10aMenopause10aPregnancy10aPuberty10aResearch10aRisk Factors10aTime Factors1 aTerry, Mary, Beth1 aMichels, Karin, B1 aBrody, Julia, Green1 aByrne, Celia1 aChen, Shiuan1 aJerry, Joseph1 aMalecki, Kristen, M C1 aMartin, Mary, Beth1 aMiller, Rachel, L1 aNeuhausen, Susan, L1 aSilk, Kami1 aTrentham-Dietz, Amy1 aBreast Cancer and the Environment Research Program (BCERP) uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/environmental-exposures-during-windows-susceptibility-breast-cancer00751nas a2200241 4500008004100000245013100041210006900172260002500241300001400266490000700280100001700287700001700304700001400321700001600335700001300351700001500364700001300379700001700392700001800409700001700427700001500444856005000459 2019 eng d00aEssential roles of HDAC1 and 2 in lineage development and genome-wide DNA methylation during mouse preimplantation development0 aEssential roles of HDAC1 and 2 in lineage development and genome bInforma {UK} Limited a369–3850 v151 aZhao, Panpan1 aWang, Huanan1 aWang, Han1 aDang, Yanna1 aLuo, Lei1 aLi, Shuang1 aShi, Yan1 aWang, Lefeng1 aWang, Shaohua1 aMager, Jesse1 aZhang, Kun uhttps://doi.org/10.1080/15592294.2019.166937500492nas a2200121 4500008004100000245012500041210006900166260001800235100002200253700001800275700002600293856005100319 2019 eng d00aExposure to low doses of oxybenzone during perinatal development alters mammary gland morphology in male and female mice0 aExposure to low doses of oxybenzone during perinatal development bElsevier {BV}1 aMatouskova, Klara1 aJerry, Joseph1 aVandenberg, Laura, N. uhttps://doi.org/10.1016/j.reprotox.2019.08.00204302nas a2200601 4500008004100000022001400041245008600055210006900141260001500210300000700225490000700232520249400239653001002733653001202743653001502755653002202770653002102792653002702813653002402840653001102864653003002875653003102905653002902936653001102965653001602976653005002992653002603042653002203068653000903090653001903099653001603118653002403134653001803158100002203176700002003198700002003218700002303238700001903261700002303280700002303303700002103326700002503347700002003372700001203392700002203404700001603426700002403442700001703466700002203483700002503505700001803530856015203548 2019 eng d a1465-542X00aGene expression signature of atypical breast hyperplasia and regulation by SFRP1.0 aGene expression signature of atypical breast hyperplasia and reg c2019 06 27 a760 v213 aBACKGROUND: Atypical breast hyperplasias (AH) have a 10-year risk of progression to invasive cancer estimated at 4-7%, with the overall risk of developing breast cancer increased by ~ 4-fold. AH lesions are estrogen receptor alpha positive (ERα+) and represent risk indicators and/or precursor lesions to low grade ERα+ tumors. Therefore, molecular profiles of AH lesions offer insights into the earliest changes in the breast epithelium, rendering it susceptible to oncogenic transformation.
METHODS: In this study, women were selected who were diagnosed with ductal or lobular AH, but no breast cancer prior to or within the 2-year follow-up. Paired AH and histologically normal benign (HNB) tissues from patients were microdissected. RNA was isolated, amplified linearly, labeled, and hybridized to whole transcriptome microarrays to determine gene expression profiles. Genes that were differentially expressed between AH and HNB were identified using a paired analysis. Gene expression signatures distinguishing AH and HNB were defined using AGNES and PAM methods. Regulation of gene networks was investigated using breast epithelial cell lines, explant cultures of normal breast tissue and mouse tissues.
RESULTS: A 99-gene signature discriminated the histologically normal and AH tissues in 81% of the cases. Network analysis identified coordinated alterations in signaling through ERα, epidermal growth factor receptors, and androgen receptor which were associated with the development of both lobular and ductal AH. Decreased expression of SFRP1 was also consistently lower in AH. Knockdown of SFRP1 in 76N-Tert cells resulted altered expression of 13 genes similarly to that observed in AH. An SFRP1-regulated network was also observed in tissues from mice lacking Sfrp1. Re-expression of SFRP1 in MCF7 cells provided further support for the SFRP1-regulated network. Treatment of breast explant cultures with rSFRP1 dampened estrogen-induced progesterone receptor levels.
CONCLUSIONS: The alterations in gene expression were observed in both ductal and lobular AH suggesting shared underlying mechanisms predisposing to AH. Loss of SFRP1 expression is a significant regulator of AH transcriptional profiles driving previously unidentified changes affecting responses to estrogen and possibly other pathways. The gene signature and pathways provide insights into alterations contributing to AH breast lesions.
10aAdult10aAnimals10aBiomarkers10aBiomarkers, Tumor10aBreast Neoplasms10aDisease Models, Animal10aDisease Progression10aFemale10aGene Expression Profiling10aGene Expression Regulation10aGene Regulatory Networks10aHumans10aHyperplasia10aIntercellular Signaling Peptides and Proteins10aMammary Glands, Human10aMembrane Proteins10aMice10aMice, Knockout10aMiddle Aged10aSignal Transduction10aTranscriptome1 aGregory, Kelly, J1 aRoberts, Amy, L1 aConlon, Erin, M1 aMayfield, Jacob, A1 aHagen, Mary, J1 aCrisi, Giovanna, M1 aBentley, Brooke, A1 aKane, Jeffrey, J1 aMakari-Judson, Grace1 aMason, Holly, S1 aYu, Jun1 aZhu, Lihua, Julie1 aSimin, Karl1 aJohnson, Jacob, P S1 aKhan, Ashraf1 aSchneider, Ben, R1 aSchneider, Sallie, S1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/gene-expression-signature-atypical-breast-hyperplasia-and-regulation00987nas a2200313 4500008004100000245008500041210006900126260004600195490000700241100002300248700002000271700002100291700002300312700001900335700002300354700002300377700002200400700002500422700002100447700001200468700002200480700001600502700002600518700001700544700002300561700002500584700001800609856004600627 2019 eng d00aGene expression signature of atypical breast hyperplasia and regulation by SFRP10 aGene expression signature of atypical breast hyperplasia and reg bSpringer Science and Business Media {LLC}0 v211 aGregory, Kelly, J.1 aRoberts, Amy, L1 aConlon, Erin, M.1 aMayfield, Jacob, A1 aHagen, Mary, J1 aCrisi, Giovanna, M1 aBentley, Brooke, A1 aKane, Jeffrey, J.1 aMakari-Judson, Grace1 aMason, Holly, S.1 aYu, Jun1 aZhu, Lihua, Julie1 aSimin, Karl1 aJohnson, Jacob, P. S.1 aKhan, Ashraf1 aSchneider, Ben, R.1 aSchneider, Sallie, S1 aJerry, Joseph uhttps://doi.org/10.1186/s13058-019-1157-500746nas a2200205 4500008004100000245012500041210006900166260007000235300001800305490000800323100002600331700002300357700002100380700002000401700002400421700001700445700001900462700001400481856004500495 2019 eng d00aIndividual and combined presenilin 1 and 2 knockouts reveal that both have highly overlapping functions in HEK293T cells0 aIndividual and combined presenilin 1 and 2 knockouts reveal that bAmerican Society for Biochemistry {&} Molecular Biology ({ASBMB}) a11276–112850 v2941 aLessard, Christian, B1 aRodriguez, Edgardo1 aLadd, Thomas, B.1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio1 aGolde, Todd, E1 aRan, Yong uhttps://doi.org/10.1074/jbc.ra119.00804102810nas a2200373 4500008004100000022001400041245012600055210006900181260001500250300001600265490000800281520159100289653002201880653004101902653001201943653002301955653003001978653001702008653001102025653001502036653000902051653001702060653001702077653002602094100002602120700002302146700002002169700002002189700002402209700001702233700001902250700001402269856015302283 2019 eng d a1083-351X00aIndividual and combined presenilin 1 and 2 knockouts reveal that both have highly overlapping functions in HEK293T cells.0 aIndividual and combined presenilin 1 and 2 knockouts reveal that c2019 07 19 a11276-112850 v2943 aPresenilins 1 and 2 (PS1 and 2) are the catalytic subunits of γ-secretase, a multiprotein protease that cleaves amyloid protein precursor and other type I transmembrane proteins. Previous studies with mouse models or cells have indicated differences in PS1 and PS2 functions. We have recently reported that clinical γ-secretase inhibitors (GSIs), initially developed to manage Alzheimer's disease and now being considered for other therapeutic interventions, are both pharmacologically and functionally distinct. Here, using CRISPR/Cas9-based gene editing, we established human HEK 293T cell lines in which endogenous PS1, PS2, or both have been knocked out. Using these knockout lines to examine differences in PS1- and PS2-mediated cleavage events, we confirmed that PS2 generates more intracellular β-amyloid than does PS1. Moreover, we observed subtle differences in PS1- and PS2-mediated cleavages of select substrates. In exploring the question of whether differences in activity among clinical GSIs could be attributed to differential inhibition of PS1 or PS2, we noted that select GSIs inhibit PS1 and PS2 activities on specific substrates with slightly different potencies. We also found that endoproteolysis of select PS1 FAD-linked variants in human cells is more efficient than what has been previously reported for mouse cell lines. Overall, these results obtained with HEK293T cells suggest that selective PS1 or PS2 inhibition by a given GSI does not explain the previously observed differences in functional and pharmacological properties among various GSIs.
10aAlzheimer Disease10aAmyloid Precursor Protein Secretases10aAnimals10aCRISPR-Cas Systems10aGene Knockdown Techniques10aHEK293 Cells10aHumans10aHydrolysis10aMice10aPresenilin-110aPresenilin-210aSubstrate Specificity1 aLessard, Christian, B1 aRodriguez, Edgardo1 aLadd, Thomas, B1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio1 aGolde, Todd, E1 aRan, Yong uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/individual-and-combined-presenilin-1-and-2-knockouts-reveal-both-have00640nas a2200157 4500008004100000245013200041210006900173260004600242490000700288100002900295700003400324700001900358700002300377700003600400856004600436 2019 eng d00aManipulation of bicarbonate concentration in sperm capacitation media improves in vitro fertilization output in porcine species0 aManipulation of bicarbonate concentration in sperm capacitation bSpringer Science and Business Media {LLC}0 v101 aSoriano-Úbeda, Cristina1 aRomero-Aguirregomezcorta, Jon1 aMatás, Carmen1 aVisconti, Pablo, E1 aGarcía-Vázquez, Francisco, A. uhttps://doi.org/10.1186/s40104-019-0324-y00534nas a2200133 4500008004100000245010200041210006900143260004600212100002600258700002400284700002700308700001800335856004700353 2019 eng d00aThe Mouse Mammary Gland: a Tool to Inform Adolescents About Environmental Causes of Breast Cancer0 aMouse Mammary Gland a Tool to Inform Adolescents About Environme bSpringer Science and Business Media {LLC}1 aVandenberg, Laura, N.1 aKolla, SriDurgaDevi1 aLaPlante, Charlotte, D1 aJerry, Joseph uhttps://doi.org/10.1007/s13187-019-01563-w00491nas a2200133 4500008004100000245009300041210006900134260003600203100001800239700001800257700002600275700001700301856003900318 2019 eng d00aA null allele of Dnaaf2 displays embryonic lethality and mimics human ciliary dyskinesia0 anull allele of Dnaaf2 displays embryonic lethality and mimics hu bOxford University Press ({OUP})1 aCheong, Agnes1 aDegani, Rinat1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://doi.org/10.1093/hmg/ddz10600751nas a2200241 4500008004100000245009900041210006900140260001100209490000700220100002400227700001800251700002600269700002800295700001900323700001300342700001300355700002000368700002200388700001700410700001900427700002100446856004200467 2019 eng d00aPath-seq identifies an essential mycolate remodeling program for mycobacterial host adaptation0 aPathseq identifies an essential mycolate remodeling program for b{EMBO}0 v151 aPeterson, Eliza, JR1 aBailo, Rebeca1 aRothchild, Alissa, C.1 aArrieta-Ortiz, Mario, L1 aKaur, Amardeep1 aPan, Min1 aMai, Dat1 aAbidi, Abrar, A1 aCooper, Charlotte1 aAderem, Alan1 aBhatt, Apoorva1 aBaliga, Nitin, S uhttps://doi.org/10.15252/msb.2018858400525nas a2200157 4500008004100000245008200041210007100123260003800194300001600232490000700248100001700255700001300272700001700285700002000302856004500322 2019 eng d00aPostfunctionalization of Noncationic RNA–Polymer Complexes for RNA Delivery0 aPostfunctionalization of Noncationic RNA–Polymer Complexes for R bAmerican Chemical Society ({ACS}) a6982–69910 v581 aJiang, Ziwen1 aCui, Wei1 aMager, Jesse1 aThayumanavan, S uhttps://doi.org/10.1021/acs.iecr.9b0066600593nas a2200169 4500008004100000245008800041210006900129260003800198300001600236490000700252100002600259700002200285700001700307700002000324700002500344856005400369 2019 eng d00aProtein Transduction Domain Mimics Facilitate Rapid Antigen Delivery into Monocytes0 aProtein Transduction Domain Mimics Facilitate Rapid Antigen Deli bAmerican Chemical Society ({ACS}) a2462–24690 v161 aBacklund, Coralie, M.1 aParhamifar, Ladan1 aMinter, Lisa1 aTew, Gregory, N1 aAndresen, Thomas, L. uhttps://doi.org/10.1021/acs.molpharmaceut.9b0007000503nas a2200145 4500008004100000245009300041210006900134260001800203300001100221490000700232100002200239700002800261700002100289856004700310 2019 eng d00aSelf-Assembling Intrauterine Device (Upod) Modulation of the Reproductive Cycle in Mares0 aSelfAssembling Intrauterine Device Upod Modulation of the Reprod bElsevier {BV} a1026900 v831 aGradil, Carlos, M1 aUricchio, Cassandra, K.1 aSchwarz, Allison uhttps://doi.org/10.1016/j.jevs.2019.02.00900686nas a2200193 4500008004100000245012200041210006900163260003800232300001800270490000700288100002000295700002300315700002500338700002500363700001700388700002400405700002000429856004300449 2019 eng d00aSymbiotic Self-Assembly Strategy toward Lipid-Encased Cross-Linked Polymer Nanoparticles for Efficient Gene Silencing0 aSymbiotic SelfAssembly Strategy toward LipidEncased CrossLinked bAmerican Chemical Society ({ACS}) a24971–249830 v111 aDutta, Kingshuk1 aBochicchio, Davide1 aRibbe, Alexander, E.1 aAlfandari, Dominique1 aMager, Jesse1 aPavan, Giovanni, M.1 aThayumanavan, S uhttps://doi.org/10.1021/acsami.9b0473101242nas a2200397 4500008004100000245014300041210006900184260005400253300001600307490000700323100001800330700001400348700002200362700001600384700001400400700001500414700001700429700002000446700001300466700002700479700001800506700002400524700002700548700001700575700001800592700002400610700002500634700002000659700001800679700001500697700001700712700002300729700002300752700001900775856005000794 2019 eng d00aTargeted Metabolomics Identifies the Cytochrome P450 Monooxygenase Eicosanoid Pathway as a Novel Therapeutic Target of Colon Tumorigenesis0 aTargeted Metabolomics Identifies the Cytochrome P450 Monooxygena bAmerican Association for Cancer Research ({AACR}) a1822–18300 v791 aWang, Weicang1 aYang, Jun1 aEdin, Matthew, L.1 aWang, Yuxin1 aLuo, Ying1 aWan, Debin1 aYang, Haixia1 aSong, Chun-Qing1 aXue, Wen1 aSanidad, Katherine, Z.1 aSong, Mingyue1 aBisbee, Heather, A.1 aBradbury, Jennifer, A.1 aNan, Guanjun1 aZhang, Jianan1 aShih, Pei-an, Betty1 aLee, Kin, Sing Steph1 aMinter, Lisa, M1 aKim, Daeyoung1 aXiao, Hang1 aLiu, Jun-Yan1 aHammock, Bruce, D.1 aZeldin, Darryl, C.1 aZhang, Guodong uhttps://doi.org/10.1158/0008-5472.can-18-322100728nas a2200205 4500008004100000245014500041210006900186260002300255300000800278490000700286100003600293700002300329700002300352700002600375700002500401700002300426700002500449700001200474856003600486 2019 eng d00aTissue plasminogen activator (tPA) of paternal origin is necessary for the success of in vitro but not of in vivo fertilization in the mouse0 aTissue plasminogen activator tPA of paternal origin is necessary b{CSIRO} Publishing a4330 v311 aGarcía-Vázquez, Francisco, A.1 aSoriano-Úbeda, C.1 aLaguna-Barraza, R.1 aIzquierdo-Rico, José1 aNavarrete, Felipe, A1 aVisconti, Pablo, E1 aGutiérrez-Adán, A.1 aCoy, P. uhttps://doi.org/10.1071/rd1817500938nas a2200289 4500008004100000245009000041210006900131260002500200490000600225100002500231700001700256700002600273700002400299700002600323700002000349700003600369700002000405700001700425700002100442700002400463700001700487700002300504700002600527700002700553700002300580856004500603 2019 eng d00aTransient Sperm Starvation Improves the Outcome of Assisted Reproductive Technologies0 aTransient Sperm Starvation Improves the Outcome of Assisted Repr bFrontiers Media {SA}0 v71 aNavarrete, Felipe, A1 aAguila, Luis1 aMartin-Hidalgo, David1 aTourzani, Darya, A.1 aLuque, Guillermina, M1 aArdestani, Goli1 aGarcía-Vázquez, Francisco, A.1 aLevin, Lonny, R1 aBuck, Jochen1 aDarszon, Alberto1 aBuffone, Mariano, G1 aMager, Jesse1 aFissore, Rafael, A1 aSalicioni, Ana, Maria1 aGervasi, Maria, Gracia1 aVisconti, Pablo, E uhttps://doi.org/10.3389/fcell.2019.0026200622nas a2200193 4500008004100000245008800041210006900129260003600198300001200234490000800246100001600254700001800270700001800288700002400306700002000330700001700350700001900367856004200386 2019 eng d00aTriclocarban Exposure Exaggerates Spontaneous Colonic Inflammation in Il-10-/- Mice0 aTriclocarban Exposure Exaggerates Spontaneous Colonic Inflammati bOxford University Press ({OUP}) a92–990 v1741 aXie, Minhao1 aZhang, Hongna1 aWang, Weicang1 aSherman, Heather, L1 aMinter, Lisa, M1 aCai, Zongwei1 aZhang, Guodong uhttps://doi.org/10.1093/toxsci/kfz24800614nas a2200169 4500008004100000245010900041210006900150260004300219100001700262700001300279700001900292700002200311700002700333700001700360700002000377856004700397 2018 eng d00aBait-and-Switch Supramolecular Strategy To Generate Noncationic RNA–Polymer Complexes for RNA Delivery0 aBaitandSwitch Supramolecular Strategy To Generate Noncationic RN bAmerican Chemical Society ({ACS})cdec1 aJiang, Ziwen1 aCui, Wei1 aPrasad, Priyaa1 aTouve, Mollie, A.1 aGianneschi, Nathan, C.1 aMager, Jesse1 aThayumanavan, S uhttps://doi.org/10.1021/acs.biomac.8b0132100542nas a2200157 4500008004100000245008000041210006900121260003000190490000600220100002400226700001800250700002500268700002300293700002300316856004500339 2018 eng d00aChanges in Protein O-GlcNAcylation During Mouse Epididymal Sperm Maturation0 aChanges in Protein OGlcNAcylation During Mouse Epididymal Sperm bFrontiers Media {SA}cjun0 v61 aTourzani, Darya, A.1 aPaudel, Bidur1 aMiranda, Patricia, V1 aVisconti, Pablo, E1 aGervasi, Maria, G. uhttps://doi.org/10.3389/fcell.2018.0006001127nas a2200361 4500008004100000245011800041210006900159260006500228300001300293490000700306100001700313700001800330700002800348700001200376700002700388700001800415700001400433700002200447700002100469700001700490700002200507700001600529700001600545700001800561700002500579700001500604700002000619700001900639700001700658700002200675700001900697856004900716 2018 eng d00aA common antimicrobial additive increases colonic inflammation and colitis-associated colon tumorigenesis in mice0 acommon antimicrobial additive increases colonic inflammation and bAmerican Association for the Advancement of Science ({AAAS}) aeaan41160 v101 aYang, Haixia1 aWang, Weicang1 aRomano, Kymberleigh, A.1 aGu, Min1 aSanidad, Katherine, Z.1 aKim, Daeyoung1 aYang, Jun1 aSchmidt, Birgitta1 aPanigrahy, Dipak1 aPei, Ruisong1 aMartin, Derek, A.1 aOzay, Ilker1 aWang, Yuxin1 aSong, Mingyue1 aBolling, Bradley, W.1 aXiao, Hang1 aMinter, Lisa, M1 aYang, Guang-Yu1 aLiu, Zhenhua1 aRey, Federico, E.1 aZhang, Guodong uhttps://doi.org/10.1126/scitranslmed.aan411602599nas a2200289 4500008004100000022001400041245012400055210006900179260001300248300001400261490000700275520160500282653001201887653003501899653000801934653002801942653002001970653002701990653001702017653001502034100002202049700002002071700002202091700001902113700002602132856015102158 2018 eng d a1526-499800aComparative CYP-omic analysis between the DDT-susceptible and -resistant Drosophila melanogaster strains 91-C and 91-R.0 aComparative CYPomic analysis between the DDTsusceptible and resi c2018 Nov a2530-25430 v743 aBACKGROUND: Cytochrome P450 monooxygenases (P450s) are involved in the biosynthesis of endogenous intracellular compounds and the metabolism of xenobiotics, including chemical insecticides. We investigated the structural and expression level variance across all P450 genes with respect to the evolution of insecticide resistance under multigenerational dichlorodiphenyltrichloroethane (DDT) selection.
RESULTS: RNA-sequencing (RNA-seq) and reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) indicated that the transcript levels of seven P450 genes were significantly up-regulated and three P450 genes were down-regulated in the DDT-resistant strain 91-R, as compared to the control strain 91-C. The overexpression of Cyp6g1 was associated with the presence of an Accord and an HMS-Beagle element insertion in the 5' upstream region in conjunction with copy number variation in the 91-R strain, but not in the 91-C strain. A total of 122 (50.2%) fixed nonsynonymous (amino acid-changing) mutations were found between 91-C and 91-R, and 20 (8.2%) resulted in amino acid changes within functional domains. Three P450 proteins were truncated as a result of premature stop codons and fixed between strains.
CONCLUSION: Our results demonstrate that a combination of changes in P450 protein-coding regions and transcript levels are possibly associated with DDT resistance, and thereby suggest that selection for variant function may occur within this gene family in response to chronic DDT exposure. © 2018 Society of Chemical Industry.
10aAnimals10aCytochrome P-450 Enzyme System10aDDT10aDrosophila melanogaster10aInsect Proteins10aInsecticide Resistance10aInsecticides10aProteomics1 aSeong, Keon, Mook1 aCoates, Brad, S1 aBerenbaum, May, R1 aClark, John, M1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/comparative-cyp-omic-analysis-between-ddt-susceptible-and-resistant00586nas a2200157 4500008004100000245014200041210006900183260001500252300001600267490000700283100002300290700002700313700002400340700002100364856004300385 2018 eng d00aComparative two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of human milk to identify dysregulated proteins in breast cancer0 aComparative twodimensional polyacrylamide gel electrophoresis 2D bWileycjun a1723–17340 v391 aAslebagh, Roshanak1 aChannaveerappa, Devika1 aArcaro, Kathleen, F1 aDarie, Costel, C uhttps://doi.org/10.1002/elps.20180002500394nas a2200133 4500008004100000245003400041210003400075260003400109300001900143490000900162100002100171700002500192856004300217 2018 eng d00aCranial Neural Crest Explants0 aCranial Neural Crest Explants bCold Spring Harbor Laboratory apdb.prot0973940 v20181 aCousin, Hélène1 aAlfandari, Dominique uhttps://doi.org/10.1101/pdb.prot09739400368nas a2200121 4500008004100000245003700041210003700078260003900115300001900154490000900173100002100182856004300203 2018 eng d00aCranial Neural Crest Transplants0 aCranial Neural Crest Transplants bCold Spring Harbor Laboratorycjan apdb.prot0974020 v20181 aCousin, Hélène uhttps://doi.org/10.1101/pdb.prot09740200454nas a2200133 4500008004100000245009500041210006900136260001000205300001100215490000700226100002500233700002400258856003800282 2018 eng d00aCut loose and run: The complex role of ADAM proteases during neural crest cell development0 aCut loose and run The complex role of ADAM proteases during neur bWiley ae230950 v561 aAlfandari, Dominique1 aTaneyhill, Lisa, A. uhttps://doi.org/10.1002/dvg.2309500517nas a2200145 4500008004100000245007900041210006900120260003900189100002000228700001600248700002300264700002300287700002600310856003500336 2018 eng d00aDevelopment of a genetically encoded sensor for endogenous CaMKII activity0 aDevelopment of a genetically encoded sensor for endogenous CaMKI bCold Spring Harbor Laboratorycjun1 aArdestani, Goli1 aWest, Megan1 aMaresca, Thomas, J1 aFissore, Rafael, A1 aStratton, Margaret, M uhttps://doi.org/10.1101/35939800768nas a2200205 4500008004100000245016400041210006900205260002300274100002100297700001900318700002500337700002100362700002000383700002000403700002500423700002000448700002300468700002400491856004700515 2018 eng d00aDietary Intervention to Increase Fruit and Vegetable Consumption in Breastfeeding Women: A Pilot Randomized Trial Measuring Inflammatory Markers in Breast Milk0 aDietary Intervention to Increase Fruit and Vegetable Consumption bElsevier {BV}csep1 aEssa, Angela, R.1 aBrowne, Eva, P1 aPunska, Elizabeth, C1 aPerkins, Katelyn1 aBoudreau, Emily1 aWiggins, Hilary1 aAnderton, Douglas, L1 aSibeko, Lindiwe1 aSturgeon, Susan, R1 aArcaro, Kathleen, F uhttps://doi.org/10.1016/j.jand.2018.06.01500565nas a2200157 4500008004100000245010000041210006900141260001500210100002500225700002100250700002500271700002500296700002400321700002400345856003800369 2018 eng d00aDNA methylation in breast cancers: Differences based on estrogen receptor status and recurrence0 aDNA methylation in breast cancers Differences based on estrogen bWileycsep1 aWilliams, Kristin, E1 aJawale, Rahul, M1 aSchneider, Sallie, S1 aOtis, Christopher, N1 aPentecost, Brian, T1 aArcaro, Kathleen, F uhttps://doi.org/10.1002/jcb.2743100854nas a2200253 4500008004100000245017800041210006900219260003600288300001400324490000800338100002700346700001700373700001800390700001600408700001400424700001200438700001800450700001800468700001800486700002000504700001500524700001900539856004200558 2018 eng d00aEffects of Consumer Antimicrobials Benzalkonium Chloride, Benzethonium Chloride, and Chloroxylenol on Colonic Inflammation and Colitis-Associated Colon Tumorigenesis in Mice0 aEffects of Consumer Antimicrobials Benzalkonium Chloride Benzeth bOxford University Press ({OUP}) a490–4990 v1631 aSanidad, Katherine, Z.1 aYang, Haixia1 aWang, Weicang1 aOzay, Ilker1 aYang, Jun1 aGu, Min1 aKarner, Emmet1 aZhang, Jianan1 aKim, Daeyoung1 aMinter, Lisa, M1 aXiao, Hang1 aZhang, Guodong uhttps://doi.org/10.1093/toxsci/kfy04502338nas a2200385 4500008004100000022001400041245011600055210006900171260001300240300001200253490000800265520116100273653001201434653001901446653003701465653001201502653001601514653001701530653002301547653002101570653000901591653000901600653002301609653001501632100001501647700001701662700001301679700001801692700001601710700001801726700002101744700001901765700001801784856015001802 2018 eng d a1873-635100aExposure to permethrin promotes high fat diet-induced weight gain and insulin resistance in male C57BL/6J mice.0 aExposure to permethrin promotes high fat dietinduced weight gain c2018 Jan a405-4160 v1113 aPermethrin is a pyrethroid pesticide that was previously reported to promote fat accumulation and insulin resistance in vitro. A recent study in female mice also found that permethrin could promote high fat-induced insulin resistance. The effects of permethrin on glucose and lipid metabolisms in male mice, however, remain unknown. The purpose of this study was to investigate the effects and interactions of permethrin exposure (50, 500, and 5000 μg/kg body weight/day) and dietary fat (low fat, 4% w/w; high fat, 20% w/w) on development of obesity and insulin resistance in male C57BL/6J mice. Our results showed that permethrin treatment significantly increased body weight, fat mass, and insulin resistance with high fat diet, but not with low fat diet, without influencing energy intake. Permethrin treatment also significantly increased serum levels of insulin, glucose, leptin, triglycerides and cholesterol. Further results showed that permethrin inhibited AMP-activated protein kinase in white adipose tissue. These results suggest that permethrin interacts with dietary fat to alter lipid and glucose metabolisms in male C57BL/6J mice.
10aAnimals10aDiet, High-Fat10aDose-Response Relationship, Drug10aGlucose10aHomeostasis10aInsecticides10aInsulin Resistance10aLipid Metabolism10aMale10aMice10aMice, Inbred C57BL10aPermethrin1 aXiao, Xiao1 aSun, Quancai1 aKim, Yoo1 aYang, Szu-Hao1 aQi, Weipeng1 aKim, Daeyoung1 aYoon, Kyong, Sup1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/exposure-permethrin-promotes-high-fat-diet-induced-weight-gain-and03055nas a2200409 4500008004100000022001400041245007400055210006900129260001200198300001000210490000700220520179600227653001202023653002102035653001402056653001402070653001102084653003802095653002202133653001102155653002602166653003002192653000902222653002802231653001702259100001802276700002002294700002302314700002202337700002102359700002502380700002502405700002802430700001702458700002402475856014602499 2018 eng d a1432-177700aGenetic variation in sensitivity to estrogens and breast cancer risk.0 aGenetic variation in sensitivity to estrogens and breast cancer c2018 02 a24-370 v293 aBreast cancer risk is intimately intertwined with exposure to estrogens. While more than 160 breast cancer risk loci have been identified in humans, genetic interactions with estrogen exposure remain to be established. Strains of rodents exhibit striking differences in their responses to endogenous ovarian estrogens (primarily 17β-estradiol). Similar genetic variation has been observed for synthetic estrogen agonists (ethinyl estradiol) and environmental chemicals that mimic the actions of estrogens (xenoestrogens). This review of literature highlights the extent of variation in responses to estrogens among strains of rodents and compiles the genetic loci underlying pathogenic effects of excessive estrogen signaling. Genetic linkage studies have identified a total of the 35 quantitative trait loci (QTL) affecting responses to 17β-estradiol or diethylstilbestrol in five different tissues. However, the QTL appear to act in a tissue-specific manner with 9 QTL affecting the incidence or latency of mammary tumors induced by 17β-estradiol or diethylstilbestrol. Mammary gland development during puberty is also exquisitely sensitive to the actions of endogenous estrogens. Analysis of mammary ductal growth and branching in 43 strains of inbred mice identified 20 QTL. Regions in the human genome orthologous to the mammary development QTL harbor loci associated with breast cancer risk or mammographic density. The data demonstrate extensive genetic variation in regulation of estrogen signaling in rodent mammary tissues that alters susceptibility to tumors. Genetic variants in these pathways may identify a subset of women who are especially sensitive to either endogenous estrogens or environmental xenoestrogens and render them at increased risk of breast cancer.
10aAnimals10aBreast Neoplasms10aEstradiol10aEstrogens10aFemale10aGenetic Predisposition to Disease10aGenetic Variation10aHumans10aMammary Glands, Human10aMammary Neoplasms, Animal10aMice10aQuantitative Trait Loci10aRisk Factors1 aJerry, Joseph1 aShull, James, D1 aHadsell, Darryl, L1 aRijnkels, Monique1 aDunphy, Karen, A1 aSchneider, Sallie, S1 aVandenberg, Laura, N1 aMajhi, Prabin, Dhangada1 aByrne, Celia1 aTrentham-Dietz, Amy uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genetic-variation-sensitivity-estrogens-and-breast-cancer-risk00729nas a2200229 4500008004100000245007300041210006900114260002500183300001200208490000700220100001800227700002100245700002400266700002200290700002100312700002500333700002600358700002800384700001700412700002400429856004600453 2018 eng d00aGenetic variation in sensitivity to estrogens and breast cancer risk0 aGenetic variation in sensitivity to estrogens and breast cancer bSpringer Naturecfeb a24–370 v291 aJerry, Joseph1 aShull, James, D.1 aHadsell, Darryl, L.1 aRijnkels, Monique1 aDunphy, Karen, A1 aSchneider, Sallie, S1 aVandenberg, Laura, N.1 aMajhi, Prabin, Dhangada1 aByrne, Celia1 aTrentham-Dietz, Amy uhttps://doi.org/10.1007/s00335-018-9741-z00467nas a2200133 4500008004100000245007800041210006900119260002900188300001600217490000700233100002600240700002500266856004200291 2018 eng d00aLet's fight cancer: let-7 is a tool to enhance antitumor immune responses0 aLets fight cancer let7 is a tool to enhance antitumor immune res bFuture Medicine Ltdcmay a1141–11450 v141 aPobezinsky, Leonid, A1 aWells, Alexandria, C uhttps://doi.org/10.2217/fon-2018-003703072nas a2200445 4500008004100000022001400041245013600055210006900191260001200260300001000272490000700282520172200289653001602011653001202027653001502039653002702054653002202081653002602103653001102129653001702140653003502157653001302192653002102205653001202226653003202238653001202270653001402282653001302296653001502309100001602324700001302340700001802353700001502371700001502386700001502401700002002416700001802436700001802454856015402472 2018 eng d a2152-499800aMC1568 Enhances Histone Acetylation During Oocyte Meiosis and Improves Development of Somatic Cell Nuclear Transfer Embryos in Pig.0 aMC1568 Enhances Histone Acetylation During Oocyte Meiosis and Im c2018 02 a55-650 v203 aAn increasing number of studies have revealed that histone deacetylase (HDAC) mediated histone deacetylation is important for mammalian oocyte development. However, nonselective HDAC inhibitors (HDACi) were applied in most studies; the precise functions of specific HDAC classes during meiosis are poorly defined. In this study, the class IIa-specific HDACi MC1568 was used to reveal a crucial role of class IIa HDACs in the regulation of histone deacetylation during porcine oocyte meiosis. Besides, the functions of HDACs and histone acetyltransferases in regulating the balance of histone acetylation/deacetylation were also confirmed during oocyte maturation. After the validation of nontoxicity of MC1568 in maturation rate, spindle morphology, and chromosome alignment, effects of MC1568 on developmental competence of porcine somatic cell nuclear transfer (SCNT) embryos were evaluated, and data indicated that treatment with 10 μM MC1568 for 12 hours following electrical activation significantly enhanced the blastocyst rate and cell numbers. Moreover, results showed that optimal MC1568 treatment increased the H4K12 acetylation level in SCNT one cells and two cells. In addition, MC1568 treatment stimulated expression of the development-related genes OCT4, CDX2, SOX2, and NANOG in SCNT blastocysts. Collectively, our investigation uncovered a critical role of class IIa HDACs in the regulation of histone deacetylation during oocyte meiosis. Furthermore, for the first time, we showed that MC1568 can improve the in vitro development of porcine SCNT embryos. These findings provide an alternative HDACi for improving animal cloning efficiency and may shed more light on nuclear reprogramming.
10aAcetylation10aAnimals10aBlastocyst10aCellular Reprogramming10aCloning, Organism10aEmbryonic Development10aFemale10aHistone Code10aHistone Deacetylase Inhibitors10aHistones10aHydroxamic Acids10aMeiosis10aNuclear Transfer Techniques10aOocytes10aPregnancy10aPyrroles10aSus scrofa1 aWang, Huili1 aCui, Wei1 aMeng, Chunhua1 aZhang, Jun1 aLi, Yinxia1 aQian, Yong1 aXing, Guangdong1 aZhao, Dongmin1 aCao, Shaoxian uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mc1568-enhances-histone-acetylation-during-oocyte-meiosis-and-improves00592nas a2200181 4500008004100000245007800041210006900119260002400188100001300212700002000225700002000245700001800265700001900283700002600302700002500328700001700353856004000370 2018 eng d00aMed20 is essential for early embryogenesis and regulates Nanog expression0 aMed20 is essential for early embryogenesis and regulates Nanog e bBioscientificacdec1 aCui, Wei1 aMarcho, Chelsea1 aWang, Yongsheng1 aDegani, Rinat1 aGolan, Morgane1 aTremblay, Kimberly, D1 aRivera-Pérez, Jaime1 aMager, Jesse uhttps://doi.org/10.1530/rep-18-050800627nas a2200193 4500008004100000245007800041210006900119260002500188490000600213100002100219700002300240700002000263700002500283700001900308700002000327700002400347700001700371856004500388 2018 eng d00aNotch Signaling in Myeloid Cells as a Regulator of Tumor Immune Responses0 aNotch Signaling in Myeloid Cells as a Regulator of Tumor Immune bFrontiers Media {SA}0 v91 aHossain, Fokhrul1 aMajumder, Samarpan1 aUcar, Deniz, A.1 aRodriguez, Paulo, C.1 aGolde, Todd, E1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio uhttps://doi.org/10.3389/fimmu.2018.0128801002nas a2200301 4500008004100000245015500041210006900196260002500265490000600290100002100296700002400317700002000341700001400361700002500375700002500400700001900425700002200444700002100466700002000487700001900507700002900526700002100555700002000576700001900596700002400615700001700639856004400656 2018 eng d00aNotch Signaling Regulates Mitochondrial Metabolism and NF-kB Activity in Triple-Negative Breast Cancer Cells via IKKa-Dependent Non-canonical Pathways0 aNotch Signaling Regulates Mitochondrial Metabolism and NFkB Acti bFrontiers Media {SA}0 v81 aHossain, Fokhrul1 aSorrentino, Claudia1 aUcar, Deniz, A.1 aPeng, Yin1 aMatossian, Margarite1 aWyczechowska, Dorota1 aCrabtree, Judy1 aZabaleta, Jovanny1 aMorello, Silvana1 aDel Valle, Luis1 aBurow, Matthew1 aCollins-Burow, Bridgette1 aPannuti, Antonio1 aMinter, Lisa, M1 aGolde, Todd, E1 aOsborne, Barbara, A1 aMiele, Lucio uhttps://doi.org/10.3389/fonc.2018.0057502567nas a2200409 4500008004100000022001400041245010300055210006900158260001200227300001200239490000700251520126900258653001201527653003101539653002501570653002101595653000901616653002301625653001401648653001501662653001801677653002101695653002401716653001401740653002601754653002701780100002301807700002001830700002101850700003001871700002301901700001901924700001701943700002401960700002001984856015302004 2018 eng d a1872-914200aNotch1 primes CD4 T cells for T helper type I differentiation through its early effects on miR-29.0 aNotch1 primes CD4 T cells for T helper type I differentiation th c2018 07 a191-1980 v993 aThe transmembrane receptor, Notch1 plays an important role during the differentiation of CD4 T cells into T helper (Th) subsets in the presence of appropriate cytokines, including differentiation into Th1 cells. MicroRNAs have also been shown to be important regulators of immune responses, including negatively regulating cytokine production by Th1 cells. The miR-29 family of microRNAs can act to inhibit tbx21 and ifng transcription, two important pro-inflammatory genes that are abundantly expressed in Th1 cells. Here we show that Notch1 may prime CD4 T cells to be responsive to Th1-polarizing cues through its early repressive effects on the miR-29 family of microRNAs. Using a combination of cell lines and primary cells, we demonstrate that Notch1 can repress miR-29a, miR-29b, and miR-29c transcription through a mechanism that is independent of NF-κB. We further show that this repression is mediated by canonical Notch signaling and requires active Mastermind like (MAML) 1, but this process is superseded by positive regulation of miR-29 in response to IFNγ at later stages of CD4 T cell activation and differentiation. Collectively, our data suggest an additional mechanism by which Notch1 signaling may fine-tune Th1 cell differentiation.
10aAnimals10aCD4-Positive T-Lymphocytes10aCell Differentiation10aInterferon-gamma10aMice10aMice, Inbred C57BL10aMicroRNAs10aNF-kappa B10aNIH 3T3 Cells10aReceptor, Notch110aSignal Transduction10aTh1 Cells10aTranscription Factors10aTranscription, Genetic1 aChandiran, Karthik1 aLawlor, Rebecca1 aPannuti, Antonio1 aPerez, Gabriela, Gonzalez1 aSrinivasan, Janani1 aGolde, Todd, E1 aMiele, Lucio1 aOsborne, Barbara, A1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch1-primes-cd4-t-cells-t-helper-type-i-differentiation-through-its00715nas a2200217 4500008004100000245010200041210006900143260001800212300001400230490000700244100002300251700002000274700002100294700003000315700002300345700001900368700001700387700002400404700002000428856004900448 2018 eng d00aNotch1 primes CD4 T cells for T helper type I differentiation through its early effects on miR-290 aNotch1 primes CD4 T cells for T helper type I differentiation th bElsevier {BV} a191–1980 v991 aChandiran, Karthik1 aLawlor, Rebecca1 aPannuti, Antonio1 aPerez, Gabriela, Gonzalez1 aSrinivasan, Janani1 aGolde, Todd, E1 aMiele, Lucio1 aOsborne, Barbara, A1 aMinter, Lisa, M uhttps://doi.org/10.1016/j.molimm.2018.05.00200790nas a2200229 4500008004100000245011100041210006900152260001500221100002600236700002600262700002600288700002200314700002500336700002000361700002600381700003200407700001900439700001700458700002300475700002400498856003800522 2018 eng d00aOnly a subpopulation of mouse sperm displays a rapid increase in intracellular calcium during capacitation0 aOnly a subpopulation of mouse sperm displays a rapid increase in bWileycjun1 aLuque, Guillermina, M1 aDalotto-Moreno, Tomas1 aMartin-Hidalgo, David1 aRitagliati, Carla1 aMolina, Lis, C. Puga1 aRomarowski, Ana1 aBalestrini, Paula, A.1 aSchiavi-Ehrenhaus, Liza, J.1 aGilio, Nicolas1 aKrapf, Dario1 aVisconti, Pablo, E1 aBuffone, Mariano, G uhttps://doi.org/10.1002/jcp.2688300576nas a2200169 4500008004100000245009400041210006900135260003100204300001400235490000600249100002700255700001700282700002100299700001800320700002600338856004200364 2018 eng d00aOxybenzone Alters Mammary Gland Morphology in Mice Exposed During Pregnancy and Lactation0 aOxybenzone Alters Mammary Gland Morphology in Mice Exposed Durin bThe Endocrine Societycmay a903–9210 v21 aLaPlante, Charlotte, D1 aBansal, Ruby1 aDunphy, Karen, A1 aJerry, Joseph1 aVandenberg, Laura, N. uhttps://doi.org/10.1210/js.2018-0002400574nas a2200169 4500008004100000245011200041210006900153260001500222300001400237490000700251100002000258700002100278700002400299700001700323700002600340856003800366 2018 eng d00aPatterning of the hepato-pancreatobiliary boundary by BMP reveals heterogeneity within the murine liver bud0 aPatterning of the hepatopancreatobiliary boundary by BMP reveals bWileycmay a274–2880 v681 aPalaria, Amrita1 aAngelo, Jesse, R1 aGuertin, Taylor, M.1 aMager, Jesse1 aTremblay, Kimberly, D uhttps://doi.org/10.1002/hep.2976900837nas a2200229 4500008004100000245014400041210006900185260003800254100001900292700002200311700002800333700002200361700001900383700002500402700002000427700001900447700002500466700002500491700002400516700002100540856004600561 2018 eng d00aPro-inflammatory cytokines and growth factors in human milk: an exploratory analysis of racial differences to inform breast cancer etiology0 aProinflammatory cytokines and growth factors in human milk an ex bSpringer Nature America, Inccaug1 aMurphy, Jeanne1 aPfeiffer, Ruth, M1 aLynn, Brittny, C. Davis1 aCaballero, Ana, I1 aBrowne, Eva, P1 aPunska, Elizabeth, C1 aYang, Hannah, P1 aFalk, Roni, T.1 aAnderton, Douglas, L1 aGierach, Gretchen, L1 aArcaro, Kathleen, F1 aSherman, Mark, E uhttps://doi.org/10.1007/s10549-018-4907-700516nas a2200157 4500008004100000245007400041210006900115260001500184300001400199490000700213100002300220700002700243700002400270700002100294856004300315 2018 eng d00aProteomics analysis of human breast milk to assess breast cancer risk0 aProteomics analysis of human breast milk to assess breast cancer bWileycfeb a653–6650 v391 aAslebagh, Roshanak1 aChannaveerappa, Devika1 aArcaro, Kathleen, F1 aDarie, Costel, C uhttps://doi.org/10.1002/elps.20170012302636nas a2200325 4500008004100000022001400041245011700055210006900172260001200241300001200253490000700265520160800272653001201880653002801892653001901920653001101939653002801950653002701978653001702005653001502022653002102037100001402058700001702072700001102089700001202100700001502112700001502127700001602142856015202158 2018 eng d a1365-258300aRNA interference validation of detoxification genes involved in ivermectin tolerance in Drosophila melanogaster.0 aRNA interference validation of detoxification genes involved in c2018 10 a651-6600 v273 aPreviously, we observed increased transcription levels of specific cytochrome P450 monooxygenase (P450) and adenosine triphosphate binding cassette (ABC) transporter genes in human body lice, Pediculus humanus humanus, following exposure to ivermectin using the non-invasive induction assay, which resulted in tolerance. To confirm the roles of these genes in induction and tolerance, the robust genetic model insect Drosophila melanogaster was chosen. Orthologous genes corresponding to the body louse P450 (Cyp9f2, Cyp6g2 and Cyp9h1) and ABC transporter (Mrp1, GC1824 as an ABCB type and CG3327 as an ABCG type) genes were selected for in vivo bioassay. Following a brief treatment with a sublethal dose of ivermectin, the mortality response was significantly slower, indicating the presence of tolerance. Concurrently, the transcription levels of Cyp9f2 and Mrp1 at 3 h and those of Cyp6g2, Cyp9h1, Mrp1, CG1824 and CG3327 at 6 h post-treatment were upregulated, indicating gene induction. In behavioural bioassay using GAL4/UAS-RNA interference transgenic fly lines, increased susceptibility to ivermectin was observed following heat shock in the Cyp9f2 , Cyp6g2 , Cyp9h1 , Mrp1 or CG3327-knockdown flies. Considering that these five genes are orthologous to those which had the largest over-expression level following ivermectin-induced tolerance in the body louse, the current results suggest that they are also associated with ivermectin detoxification in D. melanogaster and that body lice and D. melanogaster are likely to share, in part, similar mechanisms of tolerance to ivermectin.
10aAnimals10aDrosophila melanogaster10aDrug Tolerance10aFemale10aInactivation, Metabolic10aInsecticide Resistance10aInsecticides10aIvermectin10aRNA Interference1 aKim, J, H1 aMoreau, J, A1 aAli, Y1 aRazo, P1 aHong, K, B1 aYoon, K, S1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/rna-interference-validation-detoxification-genes-involved-ivermectin00562nas a2200157 4500008004100000245009000041210006900131260003800200300001600238490000700254100002400261700002700285700002000312700002000332856005200352 2018 eng d00aThe Role of Cargo Binding Strength in Polymer-Mediated Intracellular Protein Delivery0 aRole of Cargo Binding Strength in PolymerMediated Intracellular bAmerican Chemical Society ({ACS}) a2679–26900 v291 aPosey, Nicholas, D.1 aHango, Christopher, R.1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1021/acs.bioconjchem.8b0036300719nas a2200193 4500008004100000245017400041210006900215260002500284490000600309100002300315700002400338700002000362700002000382700001700402700002000419700002100439700002000460856004500480 2018 eng d00aRotenone Treatment Reveals a Role for Electron Transport Complex I in the Subcellular Localization of Key Transcriptional Regulators During T Helper Cell Differentiation0 aRotenone Treatment Reveals a Role for Electron Transport Complex bFrontiers Media {SA}0 v91 aOzay, Emrah, Ilker1 aSherman, Heather, L1 aMello, Victoria1 aTrombley, Grace1 aLerman, Adam1 aTew, Gregory, N1 aYadava, Nagendra1 aMinter, Lisa, M uhttps://doi.org/10.3389/fimmu.2018.0128400675nas a2200193 4500008004100000245012000041210006900161260003500230300001400265490000800279100002600287700002400313700002400337700001700361700002100378700001700399700002600416856003900442 2018 eng d00aSingle-cell murine genetic fate mapping reveals bipotential hepatoblasts and novel multi-organ endoderm progenitors0 aSinglecell murine genetic fate mapping reveals bipotential hepat bThe Company of Biologistscsep adev1686580 v1451 aSebae, Gabriel, K. El1 aMalatos, Joseph, M.1 aCone, Mary-Kate, E.1 aRhee, Siyeon1 aAngelo, Jesse, R1 aMager, Jesse1 aTremblay, Kimberly, D uhttps://doi.org/10.1242/dev.16865800689nas a2200193 4500008004100000245011200041210006900153260004100222100001800263700002700281700001900308700002100327700002000348700001700368700001900385700002600404700002300430856004200453 2018 eng d00aSperm capacitation is associated with phosphorylation of the testis-specific radial spoke protein Rsph6a†0 aSperm capacitation is associated with phosphorylation of the tes bOxford University Press ({OUP})csep1 aPaudel, Bidur1 aGervasi, Maria, Gracia1 aPorambo, James1 aCaraballo, Diego1 aTourzani, Darya1 aMager, Jesse1 aPlatt, Mark, D1 aSalicioni, Ana, Maria1 aVisconti, Pablo, E uhttps://doi.org/10.1093/biolre/ioy20200693nas a2200193 4500008004100000245010900041210006900150260004100219100001800260700002700278700001900305700002400324700002400348700001700372700001900389700002600408700002300434856004200457 2018 eng d00aSperm capacitation is associated with phosphorylation of the testis-specific radial spoke protein Rsph6a0 aSperm capacitation is associated with phosphorylation of the tes bOxford University Press ({OUP})csep1 aPaudel, Bidur1 aGervasi, Maria, Gracia1 aPorambo, James1 aCaraballo, Diego, A1 aTourzani, Darya, A.1 aMager, Jesse1 aPlatt, Mark, D1 aSalicioni, Ana, Maria1 aVisconti, Pablo, E uhttps://doi.org/10.1093/biolre/ioy20201872nas a2200181 4500008004100000022001400041245011300055210007000168260001600238520116300254100002301417700002001440700002101460700002001481700001801501700002101519856015001540 2018 eng d a2769-667700aSterilization of Silastic Capsules Containing 17β-Estradiol for Effective Hormone Delivery in Mus musculus.0 aSterilization of Silastic Capsules Containing 17βEstradiol for E c2018 Oct 123 aSilastic capsules are frequently used to study the physiologic effects of estrogen exposure in animal models. The Officeof Laboratory Animal Welfare requires the sterilization of nonpharmaceutical-grade compounds before use. We compared 2commonly used terminal sterilization methods-ionizing radiation (IR) and ethylene oxide (EO)-for their utility in sterilizingsilastic capsules containing 0.05 or 0.1 mg 17β-estradiol (E2). E2-specific ELISA demonstrated that serum estrogen levelsdid not differ between mice implanted with 0.05-mg E2 capsules that were sterilized with IR or EO and those implanted withnonsterilized capsules. Likewise, mammary gland morphology and progesterone receptor expression and proliferation inmammary epithelium were similar among mice treated with E2 capsules, regardless of sterilization method, and pregnant day15 mice. In addition, IR-sterilized 0.1-mg E2 pellets provided high serum E2. We conclude that neither ionizing radiation norethylene oxide degraded E2 or the cellulose matrix, suggesting that these methods of sterilization are appropriate to provideeffective sterile hormone capsules for animal research.
1 aMajewski, Aliza, R1 aChuong, Lynn, M1 aNeill, Hannah, M1 aRoberts, Amy, L1 aJerry, Joseph1 aDunphy, Karen, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sterilization-silastic-capsules-containing-17v-estradiol-effective00611nas a2200157 4500008004100000245011200041210007000153260005500223100002300278700002000301700002100321700002000342700001800362700002100380856005200401 2018 eng d00aSterilization of Silastic Capsules Containing 17β-Estradiol for Effective Hormone Delivery in Mus musculus0 aSterilization of Silastic Capsules Containing 17βEstradiol for E bAmerican Association for Laboratory Animal Science1 aMajewski, Aliza, R1 aChuong, Lynn, M1 aNeill, Hannah, M1 aRoberts, Amy, L1 aJerry, Joseph1 aDunphy, Karen, A uhttps://doi.org/10.30802/aalas-jaalas-18-00003000979nas a2200289 4500008004100000245012100041210006900162260003000231300001400261490000800275100002000283700003100303700003200334700002400366700001500390700002600405700003200431700003200463700003300495700001700528700002300545700001700568700002000585700002100605700002400626856003900650 2018 eng d00aSuper-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis0 aSuperresolution imaging of live sperm reveals dynamic changes of bThe Company of Biologists ajcs2189580 v1311 aRomarowski, Ana1 aFélix, Ángel, G. Velasco1 aRodri'guez, Paulina, Torres1 aGervasi, Mari'a, G.1 aXu, Xinran1 aLuque, Guillermina, M1 aContreras-Jiménez, Gastón1 aSánchez-Cárdenas, Claudia1 aRami'rez-Gómez, Héctor, V.1 aKrapf, Diego1 aVisconti, Pablo, E1 aKrapf, Dario1 aGuerrero, Adán1 aDarszon, Alberto1 aBuffone, Mariano, G uhttps://doi.org/10.1242/jcs.21895802600nas a2200325 4500008004100000022001400041245012500055210006900180260000900249300001300258490000700271520155300278653001401831653001501845653003301860653002101893653001101914653002601925653003101951100001601982700001801998700001602016700001502032700001402047700001602061700001602077700002002093700001302113856014802126 2018 eng d a1932-620300aTauroursodeoxycholic acid (TUDCA) alleviates endoplasmic reticulum stress of nuclear donor cells under serum starvation.0 aTauroursodeoxycholic acid TUDCA alleviates endoplasmic reticulum c2018 ae01967850 v133 aSerum starvation is a routine protocol for synchronizing nuclear donor cells to G0/G1 phase during somatic cell nuclear transfer (SCNT). However, abrupt serum deprivation can cause serious stress to the cells cultured in vitro, which might result in endoplasmic reticulum (ER) stress, chromosome damage, and finally reduce the success rate of SCNT. In the present study, the effects of tauroursodeoxycholic acid (TUDCA), an effective ER stress-relieving drug, on the nuclear donor cells under serum deprivation condition as well as following SCNT procedures were first assessed in the bovine. The results showed that TUDCA significantly reduced ER stress and cell apoptosis in those nuclear donor cells. Moreover, it significantly decreased the expression of Hdac1 and Dnmt1, and increased the level of H3K9 acetylation in nuclear donor cells compared with control group. SCNT reconstructed embryos cloned from TUDCA-treated donor cells showed significantly higher fusion, cleavage, blastocyst formation rate, total cell number in day 7 blastocysts, and lower apoptotic index than that from control group. In addition, the expression of Hdac1, Dnmt1 and Bax was significantly lower in blastocysts derived from TUDCA-treated donor cells than that from control group. In conclusion, TUDCA significantly reduced the ER stress of nuclear donor cells under serum starvation condition, and significantly improved the developmental competence of following SCNT reconstructed embryos when these TUDCA-treated cells were used as the nuclear donors.
10aApoptosis10aCell Cycle10aEndoplasmic Reticulum Stress10aFood Deprivation10aHumans10aStress, Physiological10aTaurochenodeoxycholic Acid1 aZhang, Ying1 aQu, Pengxiang1 aMa, Xiaonan1 aQiao, Fang1 aMa, Yefei1 aQing, Suzhu1 aZhang, Yong1 aWang, Yongsheng1 aCui, Wei uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/tauroursodeoxycholic-acid-tudca-alleviates-endoplasmic-reticulum00481nas a2200121 4500008004100000245011600041210006900157260004300226300001200269100001300281700001700294856004800311 2018 eng d00aTranscriptional Regulation and Genes Involved in First Lineage Specification During Preimplantation Development0 aTranscriptional Regulation and Genes Involved in First Lineage S bSpringer International Publishingcnov a31–461 aCui, Wei1 aMager, Jesse uhttps://doi.org/10.1007/978-3-319-63187-5_400609nas a2200181 4500008004100000245007400041210006900115260002500184490000700209100002600216700001900242700002500261700002100286700002500307700002500332700002400357856004600381 2018 eng d00aTROP2 methylation and expression in tamoxifen-resistant breast cancer0 aTROP2 methylation and expression in tamoxifenresistant breast ca bSpringer Naturecjul0 v181 aZimmers, Stephanie, M1 aBrowne, Eva, P1 aWilliams, Kristin, E1 aJawale, Rahul, M1 aOtis, Christopher, N1 aSchneider, Sallie, S1 aArcaro, Kathleen, F uhttps://doi.org/10.1186/s12935-018-0589-900700nas a2200217 4500008004100000245009500041210006900136260003000205300001400235490000800249100001600257700001900273700002000292700002800312700002100340700002000361700002300381700002500404700001400429856003900443 2018 eng d00aXenopusADAM19 regulates Wnt signaling and neural crest specification by stabilizing ADAM130 aXenopusADAM19 regulates Wnt signaling and neural crest specifica bThe Company of Biologists adev1581540 v1451 aLi, Jiejing1 aPerfetto, Mark1 aNeuner, Russell1 aBahudhanapati, Harinath1 aChristian, Laura1 aMathavan, Ketan1 aBridges, Lance, C.1 aAlfandari, Dominique1 aWei, Shuo uhttps://doi.org/10.1242/dev.15815402428nas a2200349 4500008004100000022001400041245013400055210006900189260001300258300001400271490000700285520131400292653001201606653001401618653000801632653003801640653002001678653001701698653000901715653002301724653001401747653001701761653003301778100001701811700002101828700001301849700002101862700002601883700001801909700002201927856012901949 2017 eng d a1097-001000a4,4'-Dichlorodiphenyltrichloroethane (DDT) and 4,4'-dichlorodiphenyldichloroethylene (DDE) inhibit myogenesis in C2C12 myoblasts.0 a44Dichlorodiphenyltrichloroethane DDT and 44dichlorodiphenyldich c2017 Dec a5176-51850 v973 aBACKGROUND: Most countries have banned the use of 4,4'-dichlorodiphenyltrichloroethane (DDT). However, owing to its extremely high lipophilic characteristics, DDT and its metabolite 4,4'-dichlorodiphenyldichloroethylene (DDE) are ubiquitous in the environment and in many types of food. The positive correlation between exposure to insecticides, including DDT and DDE, and weight gain, resulting in impaired energy metabolism in offspring following perinatal DDT and DDE exposure, was previously reported. Therefore the influence of DDT and DDE on myogenesis using C2C12 myoblasts was investigated in this study.
RESULTS: DDT and DDE decreased myotube formation dose- and time-dependently. Among myogenic regulatory factors, DDT and DDE mainly decreased MyoD1 and Myf5 expression. DDT and DDE treatment also altered Myostatin expression, phosphorylation of protein kinase B, p70 ribosomal protein S6 kinase, forkhead box O protein 3 and mammalian target of rapamycin, resulting in attenuation of myotube formation.
CONCLUSION: These results may have significant implications for understanding the effects of developmental exposure of DDT and DDE on myogenesis and development of obesity and type 2 diabetes later in life. © 2017 Society of Chemical Industry.
10aAnimals10aCell Line10aDDT10aDichlorodiphenyl Dichloroethylene10aGene Expression10aInsecticides10aMice10aMuscle Development10aMyoblasts10aMyoD Protein10aMyogenic Regulatory Factor 51 aKim, Jonggun1 aPark, Min, Young1 aKim, Yoo1 aYoon, Kyong, Sup1 aClark, John, Marshall1 aPark, Yeonhwa1 aWhang, Kwang-Youn uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/44-dichlorodiphenyltrichloroethane-ddt-and-4400597nas a2200157 4500008004100000245009200041210007100133260007000204300001800274490000800292100002600300700002300326700002500349700002100374856004400395 2017 eng d00aActive site–adjacent phosphorylation at Tyr-397 by c-Abl kinase inactivates caspase-90 aActive site–adjacent phosphorylation at Tyr397 by cAbl kinase in bAmerican Society for Biochemistry {&} Molecular Biology ({ASBMB}) a21352–213650 v2921 aSerrano, Banyuhay, P.1 aSzydlo, Hannah, S.1 aAlfandari, Dominique1 aHardy, Jeanne, A uhttps://doi.org/10.1074/jbc.m117.81197600588nas a2200181 4500008004100000245011400041210006900155260001500224300001600239490000700255100001400262700001600276700002500292700001400317700001700331700001900348856003900367 2017 eng d00aBlood Glucose and Insulin Concentrations after Octreotide Administration in Horses With Insulin Dysregulation0 aBlood Glucose and Insulin Concentrations after Octreotide Admini bWileycmay a1188–11920 v311 aFrank, N.1 aHermida, P.1 aSanchez-Londoño, A.1 aSingh, R.1 aGradil, C.M.1 aUricchio, C.K. uhttps://doi.org/10.1111/jvim.1471800475nas a2200121 4500008004100000245013300041210006900174260002300243300001200266490000800278100002100286856004600307 2017 eng d00aCadherins function during the collective cell migration of Xenopus Cranial Neural Crest cells: revisiting the role of E-cadherin0 aCadherins function during the collective cell migration of Xenop bElsevier {BV}cdec a79–880 v1481 aCousin, Hélène uhttps://doi.org/10.1016/j.mod.2017.04.00602034nas a2200421 4500008004100000022001400041245006600055210006200121260001500183300001400198490000800212520076800220653001800988653002601006653002701032653002101059653000901080653001101089653001501100653002501115653002401140653002401164653001901188653001401207100001801221700002701239700001601266700001801282700002101300700001801321700002101339700001801360700002101378700002301399700002201422700002401444856014401468 2017 eng d a1520-512600aCancer Cell Discrimination Using Host-Guest "Doubled" Arrays.0 aCancer Cell Discrimination Using HostGuest Doubled Arrays c2017 06 14 a8008-80120 v1393 aWe report a nanosensor that uses cell lysates to rapidly profile the tumorigenicity of cancer cells. This sensing platform uses host-guest interactions between cucurbit[7]uril and the cationic headgroup of a gold nanoparticle to non-covalently modify the binding of three fluorescent proteins of a multi-channel sensor in situ. This approach doubles the number of output channels to six, providing single-well identification of cell lysates with 100% accuracy. Significantly, this classification could be extended beyond the training set, determining the invasiveness of novel cell lines. The unique fingerprint of these cell lysates required minimal sample quantity (200 ng, ∼1000 cells), making the methodology compatible with microbiopsy technology.
10aBinding Sites10aBiosensing Techniques10aBridged-Ring Compounds10aCell Line, Tumor10aGold10aHumans10aImidazoles10aLuminescent Proteins10aMetal Nanoparticles10aMolecular Structure10aNanotechnology10aNeoplasms1 aLe, Ngoc, D B1 aTonga, Gulen, Yesilbag1 aMout, Rubul1 aKim, Sung-Tae1 aWille, Marcos, E1 aRana, Subinoy1 aDunphy, Karen, A1 aJerry, Joseph1 aYazdani, Mahdieh1 aRamanathan, Rajesh1 aRotello, Caren, M1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cancer-cell-discrimination-using-host-guest-doubled-arrays-000795nas a2200253 4500008004100000245007100041210006500112260004800177300001600225490000800241100002000249700002700269700001600296700001800312700002200330700001800352700002100370700001800391700002100409700002300430700002300453700002400476856004100500 2017 eng d00aCancer Cell Discrimination Using Host–Guest “Doubled” Arrays0 aCancer Cell Discrimination Using Host–Guest Doubled Arrays bAmerican Chemical Society ({ACS})cjun/2017 a8008–80120 v1391 aLe, Ngoc, D. B.1 aTonga, Gulen, Yesilbag1 aMout, Rubul1 aKim, Sung-Tae1 aWille, Marcos, E.1 aRana, Subinoy1 aDunphy, Karen, A1 aJerry, Joseph1 aYazdani, Mahdieh1 aRamanathan, Rajesh1 aRotello, Caren, M.1 aRotello, Vincent, M uhttps://doi.org/10.1021/jacs.7b0365703063nas a2200349 4500008004100000022001400041245015500055210006900210260001500279300001400294490000600308520188200314653001202196653000802208653002802216653002402244653001102268653003002279653003102309653001902340653002702359653001702386653000902403653002402412653002602436100002202462700002002484700001602504700001902520700002602539856014802565 2017 eng d a1759-665300aChanges in Neuronal Signaling and Cell Stress Response Pathways are Associated with a Multigenic Response of Drosophila melanogaster to DDT Selection.0 aChanges in Neuronal Signaling and Cell Stress Response Pathways c2017 12 01 a3356-33720 v93 aThe adaptation of insect populations to insecticidal control is a continual threat to human health and sustainable agricultural practices, but many complex genomic mechanisms involved in this adaption remain poorly understood. This study applied a systems approach to investigate the interconnections between structural and functional variance in response to dichlorodiphenyltrichloroethane (DDT) within the Drosophila melanogaster strain 91-R. Directional selection in 6 selective sweeps coincided with constitutive gene expression differences in DDT resistant flies, including the most highly upregulated transcript, Unc-115 b, which plays a central role in axon guidance, and the most highly downregulated transcript, the angiopoietin-like CG31832, which is involved in directing vascular branching and dendrite outgrowth but likely may be under trans-regulatory control. Direct functions and protein-protein interactions mediated by differentially expressed transcripts control changes in cell migration, signal transduction, and gene regulatory cascades that impact the nervous system. Although changes to cellular stress response pathways involve 8 different cytochrome P450s, stress response, and apoptosis is controlled by a multifacetted regulatory mechanism. These data demonstrate that DDT selection in 91-R may have resulted in genome-wide adaptations that impacts genetic and signal transduction pathways that converge to modify stress response, cell survival, and neurological functions. This study implicates the involvement of a multigenic mechanism in the adaptation to a chemical insecticide, which impact interconnected regulatory cascades. We propose that DDT selection within 91-R might act systemically, wherein pathway interactions function to reinforce the epistatic effects of individual adaptive changes on an additive or nonadditive basis.
10aAnimals10aDDT10aDrosophila melanogaster10aDrosophila Proteins10aFemale10aGene Expression Profiling10aGene Expression Regulation10aGenome, Insect10aInsecticide Resistance10aInsecticides10aMale10aSignal Transduction10aStress, Physiological1 aSeong, Keon, Mook1 aCoates, Brad, S1 aSun, Weilin1 aClark, John, M1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/changes-neuronal-signaling-and-cell-stress-response-pathways-are02387nas a2200337 4500008004100000022001400041245012600055210006900181260001200250300001200262490000700274520130300281653001201584653002401596653002701620653003101647653001101678653001901689653001401708653002801722653001701750100001401767700001801781700001501799700001501814700001801829700002201847700001401869700001601883856015001899 2017 eng d a1365-258300aComparison of the proliferation and excretion of Bartonella quintana between body and head lice following oral challenge.0 aComparison of the proliferation and excretion of Bartonella quin c2017 06 a266-2760 v263 aHuman body and head lice are highly related haematophagous ectoparasites but only the body louse has been shown to transmit Bartonella quintana, the causative agent of trench fever. The mechanisms by which body lice became a vector for B. quintana, however, are poorly understood. Following oral challenge, green fluorescent protein-expressing B. quintana proliferated over 9 days postchallenge with the number of bacteria being significantly higher in whole body vs. head lice. The numbers of B. quintana detected in faeces from infected lice, however, were approximately the same in both lice. Nevertheless, the viability of B. quintana was significantly higher in body louse faeces. Comparison of immune responses in alimentary tract tissues revealed that basal transcription levels of peptidoglycan recognition protein and defensins were lower in body lice and the transcription of defensin 1 was up-regulated by oral challenge with wild-type B. quintana in head but not in body lice. In addition, the level of cytotoxic reactive oxygen species generated by epithelial cells was significantly lower in body lice. Although speculative at this time, the reduced immune response is consistent with the higher vector competence seen in body vs. head lice in terms of B. quintana infection.
10aAnimals10aBartonella quintana10aGastrointestinal Tract10aGreen Fluorescent Proteins10aHumans10aInsect Vectors10aPediculus10aReactive Oxygen Species10aTrench Fever1 aKim, J, H1 aPrevite, D, J1 aYoon, K, S1 aMurenzi, E1 aKoehler, J, E1 aPittendrigh, B, R1 aLee, S, H1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/comparison-proliferation-and-excretion-bartonella-quintana-between00658nas a2200205 4500008004100000245008500041210006900126260002600195300001400221490000800235100001700243700002000260700002400280700002200304700002600326700002000352700002000372700002000392856004000412 2017 eng d00aDefective sperm head decondensation undermines the success of ICSI in the bovine0 aDefective sperm head decondensation undermines the success of IC b{BioScientifica}cjul a207–2180 v1541 aAguila, Luis1 aFelmer, Ricardo1 aArias, Maria, Elena1 aNavarrete, Felipe1 aMartin-Hidalgo, David1 aLee, Hoi, Chang1 aVisconti, Pablo1 aFissore, Rafael uhttps://doi.org/10.1530/rep-17-027002734nas a2200385 4500008004100000022001400041245009700055210006900152260001300221300001200234490000800246520155000254653001701804653002701821653001501848653001701863653003401880653001201914653002701926653003601953653001701989653001502006653000902021653001302030653002002043653001502063653001802078100001602096700002202112700001502134700001702149700001902166700001802185856014502203 2017 eng d a1873-635100aDeltamethrin increases the fat accumulation in 3T3-L1 adipocytes and Caenorhabditis elegans.0 aDeltamethrin increases the fat accumulation in 3T3L1 adipocytes c2017 Mar a149-1560 v1013 aResearch has shown that permethrin, a Type-I pyrethroid, increases triglyceride (fat) accumulation in adipocytes. Little is known, however, about any similar effect of deltamethrin, a Type-II pyrethroid, which produces a distinct syndrome of poisoning in mammals compared with permethrin. This study was therefore aimed to explore the role of deltamethrin on fat accumulation in 3T3-L1 adipocytes and Caenorhabditis elegans. Deltamethrin (10 μM) significantly increased the fat accumulation in 3T3-L1 adipocytes and wild type C. elegans compared to respective controls. Deltamethrin decreased the ratio of phosphorylated AMP-activated kinase (pAMPKα) over AMPKα and phosphorylated acetyl-CoA carboxylase (ACC) over ACC, while it increased expression of CCAAT/enhancer-binding protein (C/EBPα) and peroxisome proliferator-activated receptor-γ (PPARγ) in 3T3-L1 adipocytes. Similarly, deltamethrin potentiated fat accumulation in C. elegans without affecting growth or pharyngeal pumping rate. Moreover, deltamethrin significantly reduced the total progeny number and locomotive activities in C. elegans in a dose-dependent manner. Deltamethrin increased fat accumulation via aak-2 (an ortholog of AMPKα) and nhr-49 (a homolog of peroxisome proliferator-activated receptor-α and also downstream target of aak-2) mediated mechanisms. The current work is the first report of the effects of deltamethrin on increased fat storage by 3T3- L1 adipocytes and C. elegans via aak-2 (AMPKα ortholog)-mediated mechanism.
10a3T3-L1 Cells10aAcetyl-CoA Carboxylase10aAdipocytes10aAdipogenesis10aAMP-Activated Protein Kinases10aAnimals10aCaenorhabditis elegans10aCCAAT-Enhancer-Binding Proteins10aInsecticides10aLocomotion10aMice10aNitriles10aPhosphorylation10aPyrethrins10aTriglycerides1 aShen, Peiyi1 aHsieh, Tsung-Hsiu1 aYue, Yiren1 aSun, Quancai1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/deltamethrin-increases-fat-accumulation-3t3-l1-adipocytes-and00661nas a2200169 4500008004100000245015900041210006900200260004500269490000600314100002200320700002600342700002000368700001900388700001900407700002500426856004000451 2017 eng d00aDual control of pcdh8l/PCNS expression and function in Xenopus laevis neural crest cells by adam13/33 via the transcription factors tfap2alpha and arid3a.0 aDual control of pcdh8lPCNS expression and function in Xenopus la b{eLife} Sciences Organisation, Ltd.caug0 v61 aKhedgikar, Vikram1 aAbbruzzese, Genevieve1 aMathavan, Ketan1 aSzydlo, Hannah1 aCousin, Helene1 aAlfandari, Dominique uhttps://doi.org/10.7554/elife.2689800624nas a2200169 4500008004100000245013100041210006900172260003900241300001300280490000700293100002000300700002200320700002100342700002500363700001700388856004900405 2017 eng d00aThe ectodomain of cadherin-11 binds to erbB2 and stimulates Akt phosphorylation to promote cranial neural crest cell migration0 aectodomain of cadherin11 binds to erbB2 and stimulates Akt phosp bPublic Library of Science ({PLoS}) ae01889630 v121 aMathavan, Ketan1 aKhedgikar, Vikram1 aBartolo, Vanessa1 aAlfandari, Dominique1 aAhmad, Aamir uhttps://doi.org/10.1371/journal.pone.018896303182nas a2200313 4500008004100000022001400041245010900055210006900164260001600233300001400249490000700263520220200270653001502472653002302487653001802510653001002528653001002538653002302548653001502571653001202586653001802598100001702616700002102633700001402654700002302668700001902691700001302710856014502723 2017 eng d a1520-511800aEffectiveness of Commercial and Homemade Washing Agents in Removing Pesticide Residues on and in Apples.0 aEffectiveness of Commercial and Homemade Washing Agents in Remov c2017 Nov 08 a9744-97520 v653 aRemoval of pesticide residues from fresh produce is important to reduce pesticide exposure to humans. This study investigated the effectiveness of commercial and homemade washing agents in the removal of surface and internalized pesticide residues from apples. Surface-enhanced Raman scattering (SERS) mapping and liquid chromatography tandem mass spectrometry (LC-MS/MS) methods were used to determine the effectiveness of different washing agents in removing pesticide residues. Surface pesticide residues were most effectively removed by sodium bicarbonate (baking soda, NaHCO) solution when compared to either tap water or Clorox bleach. Using a 10 mg/mL NaHCO washing solution, it took 12 and 15 min to completely remove thiabendazole or phosmet surface residues, respectively, following a 24 h exposure to these pesticides, which were applied at a concentration of 125 ng/cm. LC-MS/MS results showed, however, that 20% of applied thiabendazole and 4.4% of applied phosmet had penetrated into the apples following the 24 h exposure. Thiabendazole, a systemic pesticide, penetrated 4-fold deeper into the apple peel than did phosmet, a non-systemic pesticide, which led to more thiabendazole residues inside the apples, which could not be washed away using the NaHCO washing solution. This study gives us the information that the standard postharvest washing method using Clorox bleach solution for 2 min is not an effective means to completely remove pesticide residues on the surface of apples. The NaHCO method is more effective in removing surface pesticide residues on apples. In the presence of NaHCO, thiabendazole and phosmet can degrade, which assists the physical removal force of washing. However, the NaHCO method was not completely effective in removing residues that have penetrated into the apple peel. The overall effectiveness of the method to remove all pesticide residues diminished as pesticides penetrated deeper into the fruit. In practical application, washing apples with NaHCO solution can reduce pesticides mostly from the surface. Peeling is more effective to remove the penetrated pesticides; however, bioactive compounds in the peels will become lost too.
10aDetergents10aFood Contamination10aFood Handling10aFruit10aMalus10aPesticide Residues10aPesticides10aPhosmet10aThiabendazole1 aYang, Tianxi1 aDoherty, Jeffery1 aZhao, Bin1 aKinchla, Amanda, J1 aClark, John, M1 aHe, Lili uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/effectiveness-commercial-and-homemade-washing-agents-removing00745nas a2200253 4500008004100000245008300041210006900124260004400193300001300237490000700250100001800257700001600275700001500291700001600306700001700322700001500339700001200354700001300366700001600379700001300395700002000408700001400428856004900442 2017 eng d00aEffects of embryo-derived exosomes on the development of bovine cloned embryos0 aEffects of embryoderived exosomes on the development of bovine c bPublic Library of Science ({PLoS})cmar ae01745350 v121 aQu, Pengxiang1 aQing, Suzhu1 aLiu, Ruiqi1 aQin, Hongyu1 aWang, Weiwei1 aQiao, Fang1 aGe, Hui1 aLiu, Jun1 aZhang, Yong1 aCui, Wei1 aWang, Yongsheng1 aShen, Wei uhttps://doi.org/10.1371/journal.pone.017453503808nas a2200337 4500008004100000022001400041245019000055210006900245260001300314300001200327490000700339520260600346653001202952653003302964653003302997653001103030653001703041653001503058653001203073653002503085653001503110653003403125653003403159653001903193100001903212700002303231700002503254700002103279700001903300856015103319 2017 eng d a1872-971100aEvaluation of microtransplantation of rat brain neurolemma into Xenopus laevis oocytes as a technique to study the effect of neurotoxicants on endogenous voltage-sensitive ion channels.0 aEvaluation of microtransplantation of rat brain neurolemma into c2017 May a260-2730 v603 aMicrotransplantation of mammalian brain neurolemma into the plasma membrane of Xenopus oocytes is used to study ion channels in their native form as they appear in the central nervous system. Use of microtransplanted neurolemma is advantageous for various reasons: tissue can be obtained from various sources and at different developmental stages; ion channels and receptors are present in their native configuration in their proper lipid environment along with appropriate auxiliary subunits; allowing the evaluation of numerous channelpathies caused by neurotoxicants in an ex vivo state. Here we show that Xenopus oocytes injected with post-natal day 90 (PND90) rat brain neurolemma fragments successfully express functional ion channels. Using a high throughput two electrode voltage clamp (TEVC) electrophysiological system, currents that were sensitive to tetrodotoxin, ω-conotoxin MVIIC, and tetraethylammonium were detected, indicating the presence of multiple voltage-sensitive ion channels (voltage-sensitive sodium (VSSC), calcium and potassium channels, respectively). The protein expression pattern for nine different VSSC isoforms (Na1.1-Na1.9) was determined in neurolemma using automated western blotting, with the predominant isoforms expressed being Na1.2 and Na1.6. VSSC were also successfully detected in the plasma membrane of Xenopus oocytes microtransplanted with neurolemma. Using this approach, a "proof-of-principle" experiment was conducted where a well-established structure-activity relationship between the neurotoxicant, 1,1,1-trichloro-2,2-di(4-chlorophenyl)ethane (DDT) and its non-neurotoxic metabolite, 1,1-bis-(4-chlorophenyl)-2,2-dichloroethene (DDE) was examined. A differential sensitivity of DDT and DDE on neurolemma-injected oocytes was determined where DDT elicited a concentration-dependent increase in TTX-sensitive inward sodium current upon pulse-depolarization whereas DDE resulted in no significant effect. Additionally, DDT resulted in a slowing of sodium channel inactivation kinetics whereas DDE was without effect. These results are consistent with the findings obtained using heterologous expression of single isoforms of rat brain VSSCs in Xenopus oocytes and with many other electrophysiological approaches, validating the use of the microtransplantation procedure as a toxicologically-relevant ex vivo assay. Once fully characterized, it is likely that this approach could be expanded to study the role of environmental toxicants and contaminants on various target tissues (e.g. neural, reproductive, developmental) from many species.
10aAnimals10aBrain Tissue Transplantation10aDrug Evaluation, Preclinical10aFemale10aIon Channels10aNeurilemma10aOocytes10aRats, Sprague-Dawley10aToxicology10aTransplantation, Heterologous10aVoltage-Gated Sodium Channels10aXenopus laevis1 aMurenzi, Edwin1 aToltin, Abigail, C1 aSymington, Steven, B1 aMorgan, Molly, M1 aClark, John, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evaluation-microtransplantation-rat-brain-neurolemma-xenopus-laevis02973nas a2200445 4500008004100000022001400041245016200055210006900217260001600286300001400302490000700316520159400323653001501917653001701932653001401949653003401963653001201997653001902009653001102028653001102039653001502050653001702065653000902082653000902091653002302100653001902123653002002142653001202162653003402174653002402208100001702232700001602249700001502265700001802280700001802298700002102316700001902337700001802356856015302374 2017 eng d a1520-511800aImidacloprid Promotes High Fat Diet-Induced Adiposity in Female C57BL/6J Mice and Enhances Adipogenesis in 3T3-L1 Adipocytes via the AMPKα-Mediated Pathway.0 aImidacloprid Promotes High Fat DietInduced Adiposity in Female C c2017 Aug 09 a6572-65810 v653 aImidacloprid, a neonicotinoid insecticide, was previously reported to enhance adipogenesis and resulted in insulin resistance in cell culture models. It was also reported to promote high fat diet-induced obesity and insulin resistance in male C57BL/6J mice. Thus, the goal of the present study was to determine the effects of imidacloprid and dietary fat interaction on the development of adiposity and insulin resistance in female C57BL/6J mice. Mice were fed with a low (4% w/w) or high fat (20% w/w) diet containing imidacloprid (0.06, 0.6, or 6 mg/kg bw/day) for 12 weeks. Mice fed with imidacloprid (0.6 mg/kg bw/day) significantly enhanced high fat diet-induced weight gain and adiposity. Treatment with imidacloprid significantly increased serum insulin levels with high fat diet without effects on other markers of glucose homeostasis. AMPKα activation was significantly inhibited by 0.6 and 6 mg imidacloprid/kg bw/day in white adipose tissue. Moreover, AMPKα activation with 5-aminoimidazole-4-carboxamide ribonucleotide abolished the effects of imidacloprid (10 μM) on enhanced adipogenesis in 3T3-L1 adipocytes. N-Acetyl cysteine also partially reversed the effects of imidacloprid on reduced phosphorylation of protein kinase B (AKT) in C2C12 myotubes. These results indicate that imidacloprid may potentiate high fat diet-induced adiposity in female C57BL/6J mice and enhance adipogenesis in 3T3-L1 adipocytes via the AMPKα-mediated pathway. Imidacloprid might also influence glucose homeostasis partially by inducing cellular oxidative stress in C2C12 myotubes.
10aAdipocytes10aAdipogenesis10aAdiposity10aAMP-Activated Protein Kinases10aAnimals10aDiet, High-Fat10aFemale10aHumans10aImidazoles10aInsecticides10aMale10aMice10aMice, Inbred C57BL10aNeonicotinoids10aNitro Compounds10aObesity10aProto-Oncogene Proteins c-akt10aSignal Transduction1 aSun, Quancai1 aQi, Weipeng1 aXiao, Xiao1 aYang, Szu-Hao1 aKim, Daeyoung1 aYoon, Kyong, Sup1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/imidacloprid-promotes-high-fat-diet-induced-adiposity-female-c57bl-6j00816nas a2200241 4500008004100000245012200041210006900163260003000232300001600262490000800278100002500286700002700311700002100338700002500359700001800384700002500402700002300427700002300450700002100473700002100494700002100515856003800536 2017 eng d00aIndividual-specific variation in the respiratory activities of HMECs and their bioenergetic response to IGF1 and TNFa0 aIndividualspecific variation in the respiratory activities of HM bWiley-Blackwellcmay/2017 a2750–27650 v2321 aSchneider, Sallie, S1 aHenchey, Elizabeth, M.1 aSultana, Nazneen1 aMorin, Stephanie, M.1 aJerry, Joseph1 aMakari-Judson, Grace1 aCrisi, Giovanna, M1 aArenas, Richard, B1 aJohnson, Melissa1 aMason, Holly, S.1 aYadava, Nagendra uhttps://doi.org/10.1002/jcp.2593203272nas a2200493 4500008004100000022001400041245012400055210006900179260001300248300001400261490000800275520173200283653001002015653000902025653002102034653002102055653002202076653002102098653001102119653001102130653003302141653002602174653001702200653001602217653002402233653001402257653001702271653001702288653002602305653003202331653001602363100002502379700002602404700002102430700002402451700001802475700002502493700002302518700002302541700002102564700002002585700002102605856015202626 2017 eng d a1097-465200aIndividual-specific variation in the respiratory activities of HMECs and their bioenergetic response to IGF1 and TNFα.0 aIndividualspecific variation in the respiratory activities of HM c2017 Oct a2750-27650 v2323 aMetabolic reprograming is a hallmark of cancer cells. However, the roles of pre-existing differences in normal cells metabolism toward cancer risk is not known. In order to assess pre-existing variations in normal cell metabolism, we have quantified the inter-individual variation in oxidative metabolism of normal primary human mammary epithelial cells (HMECs). We then assessed their response to selected cytokines such as insulin growth factor 1 (IGF1) and tumor necrosis factor alpha (TNFα), which are associated with breast cancer risk. Specifically, we compared the oxidative metabolism of HMECs obtained from women with breast cancer and without cancer. Our data show considerable inter-individual variation in respiratory activities of HMECs from different women. A bioenergetic parameter called pyruvate-stimulated respiration (PySR) was identified as a key distinguishing feature of HMECs from women with breast cancer and without cancer. Samples showing PySR over 20% of basal respiration rate were considered PySR and the rest as PySR . By this criterion, HMECs from tumor-affected breasts (AB) and non-tumor affected breasts (NAB) of cancer patients were mostly PySR (88% and 89%, respectively), while HMECs from non-cancer patients were mostly PySR (57%). This suggests that PySR phenotypes are individual-specific and are not caused by field effects due to the presence of tumor. The effects of IGF1 and TNFα treatments on HMECs revealed that both suppressed respiration and extracellular acidification. In addition, IGF1 altered PySR phenotypes. These results reveal individual-specific differences in pyruvate metabolism of normal breast epithelial cells and its association with breast cancer risk.
10aAdult10aAged10aBreast Neoplasms10aCell Respiration10aEnergy Metabolism10aEpithelial Cells10aFemale10aHumans10aInsulin-Like Growth Factor I10aMammary Glands, Human10aMetabolomics10aMiddle Aged10aOxidation-Reduction10aPhenotype10aPyruvic Acid10aTime Factors10aTumor Cells, Cultured10aTumor Necrosis Factor-alpha10aYoung Adult1 aSchneider, Sallie, S1 aHenchey, Elizabeth, M1 aSultana, Nazneen1 aMorin, Stephanie, M1 aJerry, Joseph1 aMakari-Judson, Grace1 aCrisi, Giovanna, M1 aArenas, Richard, B1 aJohnson, Melissa1 aMason, Holly, S1 aYadava, Nagendra uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/individual-specific-variation-respiratory-activities-hmecs-and-their02845nas a2200277 4500008004100000022001400041245014300055210006900198260001600267300001400283490000700297520188200304653000902186653002402195653002102219653002302240653001702263653002902280653001802309100001702327700001402344700002302358700001902381700001302400856015402413 2017 eng d a1520-511800aInvestigation of Pesticide Penetration and Persistence on Harvested and Live Basil Leaves Using Surface-Enhanced Raman Scattering Mapping.0 aInvestigation of Pesticide Penetration and Persistence on Harves c2017 May 03 a3541-35500 v653 aUnderstanding pesticide behavior in plants is important for effectively applying pesticides and in reducing pesticide exposures from ingestion. This study aimed to investigate the penetration and persistence of pesticides applied on harvested and live basil leaves. Surface-enhanced Raman scattering (SERS) mapping was applied for in situ and real-time tracking of pesticides over time using gold nanoparticles as probes. The results showed that, after surface exposure of 30 min to 48 h, pesticides (10 mg/L) penetrated more rapidly and deeply into the live leaves than the harvested leaves. The systemic pesticide thiabendazole and the nonsystemic pesticide ferbam can penetrate into the live leaves with depths of 225 and 130 μm, respectively, and the harvested leaves with depths of 180 and 18 μm, respectively, after 48 h of exposure. The effects of leaf integrity and age on thiabendazole penetration were also evaluated on live basil leaves after 24 h of exposure. Thiabendazole (10 mg/L) when applied onto intact leaves penetrated deeper (170 μm) than when applied onto damaged leaves (80 μm) prepared with 20 scrapes on the top surface of the leaves. Older leaves with a wet mass of 0.204 ± 0.019 g per leaf (45 days after leaf out) allowed more rapid and deeper penetration of pesticides (depth of 165 μm) than younger leaves with a wet mass of 0.053 ± 0.007 g per leaf (15 days after leaf out, depth of 95 μm). The degradation of thiabendazole on live leaves was detected after 1 week, whereas the apparent degradation of ferbam was detected after 2 weeks. In addition, the removal of pesticides from basil was more efficient when compared with other fresh produce possibly due to the specific gland structure of basil leaves. The information obtained here provides a better understanding of the behavior and biological fate of pesticides on plants.
10aGold10aMetal Nanoparticles10aOcimum basilicum10aPesticide Residues10aPlant Leaves10aSpectrum Analysis, Raman10aThiabendazole1 aYang, Tianxi1 aZhao, Bin1 aKinchla, Amanda, J1 aClark, John, M1 aHe, Lili uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/investigation-pesticide-penetration-and-persistence-harvested-and-live00740nas a2200217 4500008004100000245008400041210006900125260004500194490000600239100002500245700002200270700002600292700002000318700002100338700002300359700002500382700002200407700002700429700002600456856004000482 2017 eng d00aModulation of let-7 miRNAs controls the differentiation of effector CD8 T cells0 aModulation of let7 miRNAs controls the differentiation of effect b{eLife} Sciences Organisation, Ltd.cjul0 v61 aWells, Alexandria, C1 aDaniels, Keith, A1 aAngelou, Constance, C1 aFagerberg, Eric1 aBurnside, Amy, S1 aMarkstein, Michele1 aAlfandari, Dominique1 aWelsh, Raymond, M1 aPobezinskaya, Elena, L1 aPobezinsky, Leonid, A uhttps://doi.org/10.7554/elife.2639800464nas a2200133 4500008004100000245009700041210006900138260002500207300001400232490000600246100002000252700001900272856003900291 2017 eng d00aMolecular changes and signaling events occurring in spermatozoa during epididymal maturation0 aMolecular changes and signaling events occurring in spermatozoa bWiley-Blackwellcmar a204–2180 v51 aGervasi, M., G.1 aVisconti, P, E uhttps://doi.org/10.1111/andr.1232000715nas a2200169 4500008004100000245020300041210006900244260003000313300001000343100002600353700002400379700002200403700002400425700002300449700002300472856005000495 2017 eng d00aMolecular Characterization and Comparison of Phospholipase C zeta (PLCZ1) Gene Between Swamp (Bubalus carabanensis) and Riverine (Bubalus bubalis) Buffaloes: Its Implications and Future Perspectives0 aMolecular Characterization and Comparison of Phospholipase C zet bInforma {UK} Limitedcaug a1–91 aAtabay, Eufrocina, P.1 aTadeo, Roseline, D.1 aAtabay, Edwin, C.1 aVenturina, Emma, V.1 aFissore, Rafael, A1 aMingala, Claro, N. uhttps://doi.org/10.1080/10495398.2017.135068900641nas a2200193 4500008004100000245009100041210006900132260002300201300001400224490000800238100002200246700002700268700001900295700002100314700001900335700002500354700002000379856004800399 2017 eng d00aPa2G4 is a novel Six1 co-factor that is required for neural crest and otic development0 aPa2G4 is a novel Six1 cofactor that is required for neural crest bElsevier {BV}cjan a171–1820 v4211 aNeilson, Karen, M1 aAbbruzzesse, Genevieve1 aKenyon, Kristy1 aBartolo, Vanessa1 aKrohn, Patrick1 aAlfandari, Dominique1 aMoody, Sally, A uhttps://doi.org/10.1016/j.ydbio.2016.11.02100871nas a2200217 4500008004100000245022800041210006900269260003000338300001600368490000600384100002300390700002500413700002000438700002400458700002400482700002200506700002800528700001700556700003000573856005000603 2017 eng d00aPassive therapy with humanized anti-staphylococcal enterotoxin B antibodies attenuates systemic inflammatory response and protects from lethal pneumonia caused by staphylococcal enterotoxin B-producing Staphylococcus aureus0 aPassive therapy with humanized antistaphylococcal enterotoxin B bInforma {UK} Limitedcjan a1148–11590 v81 aKarau, Melissa, J.1 aTilahun, Mulualem, E1 aKrogman, Ashton1 aOsborne, Barbara, A1 aGoldsby, Richard, A1 aDavid, Chella, S.1 aMandrekar, Jayawant, N.1 aPatel, Robin1 aRajagopalan, Govindarajan uhttps://doi.org/10.1080/21505594.2016.126789402579nas a2200361 4500008004100000022001400041245017000055210006900225260001300294300001200307490000800319520143200327653001201759653001601771653001701787653001801804653001101822653001201833653001601845653001701861653002301878653000901901653002301910653001901933653001501952100001501967700001301982700001801995700002102013700001902034700001802053856014602071 2017 eng d a1873-635100aPermethrin alters glucose metabolism in conjunction with high fat diet by potentiating insulin resistance and decreases voluntary activities in female C57BL/6J mice.0 aPermethrin alters glucose metabolism in conjunction with high fa c2017 Oct a161-1700 v1083 aPermethrin, a type 1 pyrethroid insecticide, was previously reported to promote adipogenesis in 3T3-L1 adipocytes and insulin resistance in C2C12 muscle cells; however, the effects of permethrin exposure on glucose and lipid metabolisms in vivo remain unknown. The purpose of this study was to investigate the effects of permethrin exposure on glucose and lipid homeostasis as well as voluntary movement in female mice in response to dietary fat. We tested three doses of permethrin (50, 500, & 5000 μg/kg body weight/day) in low fat diet-fed (4% w/w of diet) and high fat diet-fed (20% w/w of diet) female C57BL/6 J mice for twelve weeks. Our results demonstrated that permethrin treatment potentiated high fat diet-induced insulin resistance as indicated by insulin tolerance tests, glucose tolerance tests, and homeostasis model assessment - insulin resistance (HOMA-IR) without altering weight or fat mass. Permethrin treatment significantly decreased voluntary movement and elevated blood glucose and insulin levels. Western blot results further showed that permethrin impaired insulin signaling via the Akt signaling pathway in the gastrocnemius muscle. Taken together, these results suggest that oral administration of permethrin potentiated high fat diet-induced insulin resistance, possibly increasing the risk of type 2 diabetes without altering weight gain in female C57BL/6 J mice.
10aAnimals10aBody Weight10aDietary Fats10aEnergy Intake10aFemale10aGlucose10aHomeostasis10aInsecticides10aInsulin Resistance10aMice10aMice, Inbred C57BL10aMotor Activity10aPermethrin1 aXiao, Xiao1 aKim, Yoo1 aKim, Daeyoung1 aYoon, Kyong, Sup1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/permethrin-alters-glucose-metabolism-conjunction-high-fat-diet02410nas a2200361 4500008004100000022001400041245019300055210006900248260001300317300001100330490000800341520115700349653003401506653001201540653003101552653001701583653001201600653000901612653003901621653002801660653001501688653002001703653003401723653002401757100001701781700001301798700001601811700001301827700001901840700001801859700001801877856015301895 2017 eng d a1873-635100aPermethrin decreased insulin-stimulated AKT phosphorylation dependent on extracellular signal-regulated kinase-1 (ERK), but not AMP-activated protein kinase α (AMPKα), in C2C12 myotubes.0 aPermethrin decreased insulinstimulated AKT phosphorylation depen c2017 Nov a95-1010 v1093 aPreviously 10 μM permethrin (38.7% cis and 59.4% trans isomers), a pyrethroid insecticide widely used in agriculture and household products for pest control, was reported to reduce insulin-stimulated glucose uptake and phosphorylation of protein kinase B (p-AKT) in C2C12 mouse myotubes. The underlying mechanisms on how permethrin decreases insulin-stimulated AKT phosphorylation, however, are unknown. Thus, the goal of this study was to determine the possible mechanism(s) through which permethrin reduced insulin-stimulated AKT phosphorylation in C2C12 myotubes. Permethrin treatment, at 10 μM, decreased insulin-stimulated membrane glucose transporter type 4 (GLUT4) and AKT phosphorylation, and increased insulin receptor substrate 1 (IRS1) Ser307 phosphorylation in the presence of insulin. The inactivation of AKT by permethrin was independent of AMPKα. ERK inactivation by U0126, however, restored insulin-stimulated AKT phosphorylation, which was decreased by permethrin treatment. These results suggest that permethrin decreased insulin-stimulated AKT phosphorylation via ERK activation, but not by AMPKα inactivation.
10aAMP-Activated Protein Kinases10aAnimals10aGlucose Transporter Type 410aInsecticides10aInsulin10aMice10aMitogen-Activated Protein Kinase 310aMuscle Fibers, Skeletal10aPermethrin10aPhosphorylation10aProto-Oncogene Proteins c-akt10aSignal Transduction1 aSun, Quancai1 aPeng, Ye1 aQi, Weipeng1 aKim, Yoo1 aClark, John, M1 aKim, Daeyoung1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/permethrin-decreased-insulin-stimulated-akt-phosphorylation-dependent02190nas a2200313 4500008004100000022001400041245014100055210006900196260001300265300001200278490000800290520116700298653001701465653001501482653001701497653001201514653001201526653002501538653002601563653003301589653001701622653000901639653001501648100001501663700001601678700001901694700001801713856014501731 2017 eng d a1873-635100aPermethrin potentiates adipogenesis via intracellular calcium and endoplasmic reticulum stress-mediated mechanisms in 3T3-L1 adipocytes.0 aPermethrin potentiates adipogenesis via intracellular calcium an c2017 Nov a123-1290 v1093 aPermethrin, a pyrethroid insecticide, was previously reported to promote adipogenesis in vitro and weight gain in vivo. The mechanism by which permethrin promotes adipogenesis/obesity, however, has not been fully explored. Intracellular calcium and endoplasmic reticulum (ER) stress have been reported to be linked with adipogenesis and obesity. Because pyrethroid insecticides have been determined to influence intracellular calcium and ER stress in vitro, the purpose of this current study was to investigate whether permethrin potentiates adipogenesis via a change in intracellular calcium, leading to endoplasmic reticulum (ER) stress in 3T3-L1 adipocytes. 3T3-L1 cells were exposed to four different concentrations of permethrin (0.01, 0.1, 1 & 10 μM) for 6 days during differentiation. Treatment of permethrin increased intracellular calcium level in a concentration-dependent manner. Similarly, permethrin treatment increased protein levels of ER stress markers in a concentration-dependent manner. These data suggest that intracellular calcium and ER stress may be involved in permethrin-induced adipogenesis of 3T3-L1 cells.
10a3T3-L1 Cells10aAdipocytes10aAdipogenesis10aAnimals10aCalcium10aCell Differentiation10aEndoplasmic Reticulum10aEndoplasmic Reticulum Stress10aInsecticides10aMice10aPermethrin1 aXiao, Xiao1 aQi, Weipeng1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/permethrin-potentiates-adipogenesis-intracellular-calcium-and00630nas a2200169 4500008004100000245015100041210006900192260002500261100001900286700002000305700002100325700001800346700002200364700001600386700002000402856003800422 2017 eng d00aPharmacokinetics of intravenous lithium chloride and assessment of agreement between two methods of lithium concentration measurement in the horse0 aPharmacokinetics of intravenous lithium chloride and assessment bWiley-Blackwellcdec1 aMartin, L., M.1 aBukoski, A., D.1 aWhelchel, D., D.1 aEvans, T., J.1 aWiedmeyer, C., E.1 aBlack, S, J1 aJohnson, P., J. uhttps://doi.org/10.1111/evj.1277801682nas a2200241 4500008004100000022001400041245009900055210006900154260001300223300001200236490000800248520087200256653001201128653003001140653002701170653001101197653001701208653001201225100001501237700001901252700001801271856015101289 2017 eng d a1873-635100aPotential contribution of insecticide exposure and development of obesity and type 2 diabetes.0 aPotential contribution of insecticide exposure and development o c2017 Jul a456-4740 v1053 aThe introduction of insecticides has greatly improved agricultural productivity and human nutrition; however, the wide use of insecticides has also sparked growing concern over their health impacts. Increased rate of cancers, neurodegenerative disorders, reproductive dysfunction, birth defects, respiratory diseases, cardiovascular diseases and aging have been linked with insecticide exposure. Meanwhile, a growing body of evidence is suggesting that exposure to insecticides can also potentiate the risk of obesity and type 2 diabetes. This review summarizes the relationship between insecticide exposure and development of obesity and type 2 diabetes using epidemiological and rodent animal studies, including potential mechanisms. The evidence as a whole suggests that exposure to insecticides is linked to increased risk of obesity and type 2 diabetes.
10aAnimals10aDiabetes Mellitus, Type 210aEnvironmental Exposure10aHumans10aInsecticides10aObesity1 aXiao, Xiao1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/potential-contribution-insecticide-exposure-and-development-obesity00550nas a2200157 4500008004100000245011300041210006900154260002000223300001100243490000900254100002500263700001600288700002400304700002300328856004100351 2017 eng d00aRelationships between Global DNA Methylation in Circulating White Blood Cells and Breast Cancer Risk Factors0 aRelationships between Global DNA Methylation in Circulating Whit bHindawi Limited a1–250 v20171 aChopra-Tandon, Nayha1 aWu, Haotian1 aArcaro, Kathleen, F1 aSturgeon, Susan, R uhttps://doi.org/10.1155/2017/270586000836nas a2200241 4500008004100000245012700041210006900168260003800237490000600275100002600281700002000307700001800327700002600345700001900371700003200390700003000422700001900452700001800471700001600489700002200505700002500527856004200552 2017 eng d00aRole of Granulocyte-Macrophage Colony-Stimulating Factor Production by T Cells during Mycobacterium tuberculosis Infection0 aRole of GranulocyteMacrophage ColonyStimulating Factor Productio bAmerican Society for Microbiology0 v81 aRothchild, Alissa, C.1 aStowell, Britni1 aGoyal, Girija1 aNunes-Alves, Cláudio1 aYang, Qianting1 aPapavinasasundaram, Kadamba1 aSassetti, Christopher, M.1 aDranoff, Glenn1 aChen, Xinchun1 aLee, Jinhee1 aBehar, Samuel, M.1 aPirofski, Liise-anne uhttps://doi.org/10.1128/mbio.01514-1700568nas a2200169 4500008004100000245009800041210006900139260001000208300001600218490000700234100002200241700002600263700002600289700002000315700002000335856004300355 2017 eng d00aROMP- and RAFT-Based Guanidinium-Containing Polymers as Scaffolds for Protein Mimic Synthesis0 aROMP and RAFTBased GuanidiniumContaining Polymers as Scaffolds f bWiley a6858–68630 v231 aSarapas, Joel, M.1 aBacklund, Coralie, M.1 adeRonde, Brittany, M.1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1002/chem.20170042300583nas a2200181 4500008004100000245006400041210006300105260002800168300001400196490000800210100002400218700002600242700001700268700002600285700002000311700002000331856005000351 2017 eng d00aSequence segregation improves non-covalent protein delivery0 aSequence segregation improves noncovalent protein delivery bElsevier {BV}cmay/2017 a131–1360 v2541 aSgolastra, Federica1 aBacklund, Coralie, M.1 aOzay, E., I.1 adeRonde, Brittany, M.1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1016/j.jconrel.2017.03.38700627nas a2200193 4500008004100000245011100041210006900152260002500221490000600246100001800252700001400270700001800284700001600302700001500318700001600333700001700349700002000366856004700386 2017 eng d00aSperm-borne miR-449b influences cleavage, epigenetic reprogramming and apoptosis of SCNT embryos in bovine0 aSpermborne miR449b influences cleavage epigenetic reprogramming bSpringer Naturecoct0 v71 aWang, Mengyun1 aGao, Yang1 aQu, Pengxiang1 aQing, Suzhu1 aQiao, Fang1 aZhang, Yong1 aMager, Jesse1 aWang, Yongsheng uhttps://doi.org/10.1038/s41598-017-13899-800741nas a2200229 4500008004100000245010200041210006900143260002500212300001600237490000700253100001900260700002300279700002100302700002500323700002400348700002100372700002000393700001900413700001700432700002500449856003700474 2017 eng d00aTools to minimize interlaboratory variability in vitellogenin gene expression monitoring programs0 aTools to minimize interlaboratory variability in vitellogenin ge bWiley-Blackwellcaug a3102–31070 v361 aJastrow, Aaron1 aGordon, Denise, A.1 aAuger, Kasie, M.1 aPunska, Elizabeth, C1 aArcaro, Kathleen, F1 aKeteles, Kristen1 aWinkelman, Dana1 aLattier, David1 aBiales, Adam1 aLazorchak, James, M. uhttps://doi.org/10.1002/etc.388503924nas a2200481 4500008004100000022001400041245011200055210006900167260001600236490000600252520226900258653001202527653004202539653003602581653003002617653003002647653002602677653002702703653003302730653001902763653001302782653001302795653000902808653002302817653002102840653001902861653002102880653003602901653003502937653003302972653005003005653002403055653003303079653004103112100002003153700002603173700001603199700001803215700001803233700001803251700002003269856015303289 2017 eng d a2047-998000aUse of p53-Silenced Endothelial Progenitor Cells to Treat Ischemia in Diabetic Peripheral Vascular Disease.0 aUse of p53Silenced Endothelial Progenitor Cells to Treat Ischemi c2017 Apr 010 v63 aBACKGROUND: Peripheral vascular disease is a major diabetes mellitus-related complication. In this study, we noted that expressions of proapoptotic p53 gene and its downstream cascade gene such as p21 are upregulated in hyperglycemia. Therefore, we investigated whether p53- and p21-silenced endothelial progenitor cells (EPCs) were able to survive in hyperglycemic milieu, and whether transplantation of either p53 knockout (KO) or p21KO or p53- and p21-silenced EPCs could improve collateral vessel formation and blood flow in diabetic vaso-occlusive peripheral vascular disease mouse models.
METHODS AND RESULTS: We transplanted p53 and p21KO mouse EPCs (mEPCs) into streptozotocin-induced diabetic (type 1 diabetes mellitus model) C57BL/6J and db/db (B6.BKS(D)-Leprdb/J) (type 2 model) post-femoral artery occlusion. Similarly, Ad-p53-silenced and Ad-p21-silenced human EPCs (CD34+) cells were transplanted into streptozotocin-induced diabetic NOD.CB17-Prkdcscid/J mice. We measured blood flow at 3, 7, and 10 days and hindlimb muscles were obtained postsacrifice for mRNA estimation and CD31 staining. Enhanced blood flow was noted with delivery of p53 and p21KO mEPCs in streptozotocin-induced diabetic C57BL/6J mice. Similar results were obtained when human Ad-p53shEPCs(CD34+) and Ad-p21shEPCs(CD34+) were transplanted into streptozotocin-induced nonobese diabetic severe combined immunodeficiency mice. Gene expression analysis of p53 and p21KO EPCs transplanted hindlimb muscles showed increased expression of endothelial markers such as endothelial nitric oxide synthase, vascular endothelial growth factor A, and platelet endothelial cell adhesion molecule 1. Similarly, quantitative reverse transcriptase polymerase chain reaction of human Ad-p53shEPCs (CD34+)- and Ad-p21shEPCs (CD34+)-transplanted hindlimb muscles also showed increased expression of endothelial markers such as vascular endothelial growth factor A, noted primarily in the p53-silenced EPCs group. However, such beneficial effect was not noted in the db/db type 2 diabetic mouse models.
CONCLUSIONS: Transient silencing of p53 using adenoviral vector in EPCs may have a therapeutic role in diabetic peripheral vascular disease.
10aAnimals10aCyclin-Dependent Kinase Inhibitor p2110aDiabetes Mellitus, Experimental10aDiabetes Mellitus, Type 110aDiabetes Mellitus, Type 210aDiabetic Angiopathies10aDisease Models, Animal10aEndothelial Progenitor Cells10aGene Silencing10aHindlimb10aIschemia10aMice10aMice, Inbred C57BL10aMice, Inbred NOD10aMice, Knockout10aMuscle, Skeletal10aNeovascularization, Physiologic10aNitric Oxide Synthase Type III10aPeripheral Vascular Diseases10aPlatelet Endothelial Cell Adhesion Molecule-110aRegional Blood Flow10aTumor Suppressor Protein p5310aVascular Endothelial Growth Factor A1 aKundu, Nabanita1 aDomingues, Cleyton, C1 aChou, Cyril1 aAhmadi, Neeki1 aHouston, Sara1 aJerry, Joseph1 aSen, Sabyasachi uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/use-p53-silenced-endothelial-progenitor-cells-treat-ischemia-diabetic00670nas a2200193 4500008004100000245011100041210006900152260005600221300001200277490000600289100002000295700002700315700001600342700001800358700001800376700001800394700002000412856004400432 2017 eng d00aUse of p53-Silenced Endothelial Progenitor Cells to Treat Ischemia in Diabetic Peripheral Vascular Disease0 aUse of p53Silenced Endothelial Progenitor Cells to Treat Ischemi bOvid Technologies (Wolters Kluwer Health)capr/2017 ae0051460 v61 aKundu, Nabanita1 aDomingues, Cleyton, C.1 aChou, Cyril1 aAhmadi, Neeki1 aHouston, Sara1 aJerry, Joseph1 aSen, Sabyasachi uhttps://doi.org/10.1161/jaha.116.00514600828nas a2200241 4500008004100000245014000041210006900181260002500250490000700275100002300282700002100305700002400326700001700350700001600367700001700383700002500400700002000425700002300445700002400468700002600492700002200518856004600540 2017 eng d00aWhite blood cell DNA methylation and risk of breast cancer in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO)0 aWhite blood cell DNA methylation and risk of breast cancer in th bSpringer Naturecaug0 v191 aSturgeon, Susan, R1 aPilsner, Richard1 aArcaro, Kathleen, F1 aIkuma, Kaoru1 aWu, Haotian1 aKim, Soon-Mi1 aChopra-Tandon, Nayha1 aKarpf, Adam, R.1 aZiegler, Regina, G1 aSchairer, Catherine1 aBalasubramanian, Raji1 aReckhow, David, A uhttps://doi.org/10.1186/s13058-017-0886-600521nas a2200157 4500008004100000245007900041210006900120260003500189300001400224490000700238100002500245700001800270700001500288700001700303856004300320 2017 eng d00aYY1 Is Required for Posttranscriptional Stability of SOX2 and OCT4Proteins0 aYY1 Is Required for Posttranscriptional Stability of SOX2 and OC bMary Ann Liebert Inccaug/2017 a263–2690 v191 aWallingford, Mary, C1 aHiller, Jacob1 aZhang, Kun1 aMager, Jesse uhttps://doi.org/10.1089/cell.2017.000200726nas a2200229 4500008004100000245010300041210007000144260001600214300001400230490000600244100001400250700002100264700002100285700002600306700002200332700001800354700002000372700002400392700001700416700001900433856004400452 2017 eng d00aγ-Secretase inhibitors in cancer clinical trials are pharmacologically and functionally distinct.0 aγSecretase inhibitors in cancer clinical trials are pharmacologi b{EMBO}cmay a950–9660 v91 aRan, Yong1 aHossain, Fokhrul1 aPannuti, Antonio1 aLessard, Christian, B1 aLadd, Gabriela, Z1 aJung, Joo, In1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aMiele, Lucio1 aGolde, Todd, E uhttps://doi.org/10.15252/emmm.20160726500554nas a2200157 4500008004100000245010300041210006900144260002300213300001400236490000800250100002600258700002100284700001800305700002500323856004800348 2016 eng d00aADAM13 cleavage of cadherin-11 promotes CNC migration independently of the homophilic binding site0 aADAM13 cleavage of cadherin11 promotes CNC migration independent bElsevier {BV}cjul a383–3900 v4151 aAbbruzzese, Genevieve1 aBecker, Sarah, F1 aKashef, Jubin1 aAlfandari, Dominique uhttps://doi.org/10.1016/j.ydbio.2015.07.01802031nas a2200265 4500008004100000022001400041245011500055210006900170260001500239300001200254490000700266520115200273653000901425653002401434653001501458653002901473653000801502100001501510700001801525700001901543700001701562700001901579700001301598856015401611 2016 eng d a1520-585100aAlteration of the Nonsystemic Behavior of the Pesticide Ferbam on Tea Leaves by Engineered Gold Nanoparticles.0 aAlteration of the Nonsystemic Behavior of the Pesticide Ferbam o c2016 06 21 a6216-230 v503 aA model system consisting of a nonsystemic pesticide (ferbam), engineered gold nanoparticles (AuNPs) and a plant tissue (tea leaves) was investigated using surface enhanced Raman spectroscopy (SERS). Ferbam has no ability by itself to penetrate into tea leaves. When AuNPs were placed with ferbam onto the surface of tea leaves, however, the SERS signal of the ferbam-AuNPs complex was observed inside of the tea leaves. Within 1 h, the ferbam-AuNPs complex rapidly penetrated into the leaf to a depth of approximately 190 μm, about (1)/3 to (1)/2 of the leaf's thickness. The rate of penetration was dependent on the size of AuNPs, with 30 nm AuNPs-ferbam penetrating more rapidly when compared with complexes made with the 50 and 69 nm AuNPs. These results clearly demonstrated an alteration of the nonsystemic behavior of ferbam in the combined presence with AuNPs. This finding might lead to the development of some new pesticide formulations. Conversely, new toxicity issues may arise as the behaviors and fate of pesticides are altered significantly upon interaction with engineered NPs in the pesticide formulation or environment.
10aGold10aMetal Nanoparticles10aPesticides10aSpectrum Analysis, Raman10aTea1 aHou, Ruyan1 aZhang, Zhiyun1 aPang, Shintaro1 aYang, Tianxi1 aClark, John, M1 aHe, Lili uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/alteration-nonsystemic-behavior-pesticide-ferbam-tea-leaves-engineered00767nas a2200241 4500008004100000245008400041210007100125260002500196300001000221490000600231100002200237700002200259700002200281700002100303700002300324700002000347700002600367700002500393700002600418700002300444700001800467856004000485 2016 eng d00aCadherin-11 localizes to focal adhesions and promotes cell–substrate adhesion0 aCadherin11 localizes to focal adhesions and promotes cell–substr bSpringer Naturecmar a109090 v71 aLanghe, Rahul, P.1 aGudzenko, Tetyana1 aBachmann, Michael1 aBecker, Sarah, F1 aGonnermann, Carina1 aWinter, Claudia1 aAbbruzzese, Genevieve1 aAlfandari, Dominique1 aKratzer, Marie-Claire1 aFranz, Clemens, M.1 aKashef, Jubin uhttps://doi.org/10.1038/ncomms1090900893nas a2200289 4500008004100000245013900041210006900180260002300249300001600272490000800288100001600296700001800312700001500330700001300345700001600358700001700374700001800391700002500409700001800434700002100452700002100473700001700494700001600511700001300527700001400540856004900554 2016 eng d00aCellular Prion Protein Mediates Pancreatic Cancer Cell Survival and Invasion through Association with and Enhanced Signaling of Notch10 aCellular Prion Protein Mediates Pancreatic Cancer Cell Survival bElsevier {BV}cnov a2945–29560 v1861 aWang, Yiwei1 aYu, Shuiliang1 aHuang, Dan1 aCui, Min1 aHu, Huankai1 aZhang, Lihua1 aWang, Weihuan1 aParameswaran, Neetha1 aJackson, Mark1 aOsborne, Barbara1 aBedogni, Barbara1 aLi, Chaoyang1 aSy, Man-Sun1 aXin, Wei1 aZhou, Lan uhttps://doi.org/10.1016/j.ajpath.2016.07.01001376nas a2200121 4500008004100000245006200041210005900103260001500162520088900177100002701066700002301093856013801116 2016 ENG d00aChang's meaning of capacitation: A molecular perspective.0 aChangs meaning of capacitation A molecular perspective c2016 Jun 33 aDr. Min Chue Chang's contributions to the field of reproductive biology set the stage for the development of the contraceptive pill and in vitro fertilization. Throughout his publications, Dr. Chang was also able to transmit his view of the fertilization process in ways that organized research for newer generations of reproductive biologists. Particularly relevant for the achievement of in vitro fertilization in mammals was the discovery that the sperm required a period of residence in the female tract to become fertilization-competent; Dr. Chang and Dr. Austin, in Australia, independently reported this process, known as sperm capacitation. This review discusses Dr. Chang's views on capacitation, and puts them in the context of recent advances in the understanding of the molecular basis of this process. This article is protected by copyright. All rights reserved.
1 aGervasi, Maria, Gracia1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/changs-meaning-of-capacitation-a-molecular-perspective02367nas a2200181 4500008004100000245007500041210006900116260001300185300001100198490000700209520171000216100002001926700002401946700002401970700001501994700002502009856015102034 2016 eng d00aDepression, Antidepressant Use, and Postmenopausal Breast Cancer Risk.0 aDepression Antidepressant Use and Postmenopausal Breast Cancer R c2016 Jan a158-640 v253 aBACKGROUND: Whether depression and antidepressant (AD) use might influence breast cancer risk is unclear, and these exposures have not been evaluated together in a single, prospective cohort study of breast cancer risk. METHODS: Among 71,439 postmenopausal women in the Women's Health Initiative Observational Study (WHI-OS), we estimated multivariable-adjusted HRs for the independent and joint effects of depressive symptoms and AD use on breast cancer risk using Cox proportional hazards regression. RESULTS: When analyzed separately, neither depressive symptoms nor AD use at baseline were associated with a significantly increased risk of total breast cancer (HR = 0.96, 95% CI, 0.85-1.08; HR = 1.04, 95% CI, 0.92-1.20, respectively) or invasive breast cancer (HR = 0.98, 95% CI, 0.86-1.12; HR = 1.00, 95% CI, 0.86-1.16, respectively). Current AD use was associated with a borderline-significant increase of in situ breast cancer (HR = 1.30, 95% CI, 0.99-1.75) after adjustment for depressive symptoms; however, this relationship was attenuated after adjustment for mammographic screening (HR = 1.08, 95% CI, 0.76-1.51). No significant variation in total breast cancer risk was observed when the separate and joint effects of depressive symptoms and AD use were explored (P for interaction = 0.14). CONCLUSION: We found no evidence that either depression or AD use influences breast cancer risk. An elevated risk of in situ disease among AD users could not be ruled out, though is likely due to increased screening in this subgroup. IMPACT: Given the high prevalence of these exposures, these results may provide reassurance to the millions of women who are depressed and/or use ADs each year.
1 aBrown, Susan, B1 aHankinson, Susan, E1 aArcaro, Kathleen, F1 aQian, Jing1 aReeves, Katherine, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/depression-antidepressant-use-and-postmenopausal-breast-cancer-risk03272nas a2200241 4500008004100000245007200041210006900113260000900182300000600191490000700197520244600204100002402650700001902674700002302693700002502716700002002741700002502761700002402786700002102810700002502831700002402856856015002880 2016 eng d00aELF5 and DOK7 regulation in anti-estrogen treated cells and tumors.0 aELF5 and DOK7 regulation in antiestrogen treated cells and tumor c2016 a80 v163 aBACKGROUND: Most women with primary breast cancers that express estrogen receptor alpha (ER or ESR1) are treated with endocrine therapies including the anti-estrogen tamoxifen, but resistance to these anti-endocrine therapies often develops. This study characterizes the expression of hormone receptors, and the mRNA and DNA methylation levels of docking protein 7 (DOK7), and E74-like factor 5 (ELF5), in 21 novel tamoxifen-resistant cell lines and extends the findings to primary and recurrent human breast tumors. METHODS: Twenty-one tamoxifen-selected cell lines were developed through cloning by limiting dilution of an MCF-7 cell culture treated with 1 μM tamoxifen for 6 months. The parent (MCF-7) and tamoxifen-selected cell lines were characterized for protein expression of ER, progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) using immunohistochemistry (IHC). The mRNA levels of ER, DOK7, and ELF5 were assessed using quantitative RT-PCR. Promoter methylation levels of DOK7 and ELF5 were determined by pyrosequencing of bisulfite-modified DNA. The relationship between hormone receptor status and promoter methylation of DOK7 and ELF5 was further examined using available methylation array data (Illumina HM450) from a set of paired primary and second breast tumors from 24 women. RESULTS: All 21 of the novel tamoxifen-selected cell lines are ER-positive, and HER2-negative, and 18 of the cell lines are PR-negative while the MCF-7 cells were scored as ER-positive, modestly PR-positive and HER2 negative. Expression of DOK7 and ELF5 is significantly up-regulated in half of the tamoxifen-selected cell lines as compared to the parental MCF-7. In contrast, the previously established ER-negative TMX2-28 cell line has decreased expression of both DOK7 and ELF5 and increased DNA methylation in the transcriptional start site region of these genes. ELF5 methylation was lower in second versus primary tumors in women who received anti-estrogen treatment, in PR-negative versus PR-positive tumors, and in the subset of PR-positive first tumors from the group of women who had second PR-negative tumors as compared to those who had second PR-positive tumors. CONCLUSIONS: The distinct ELF5 methylation of PR-positive primary tumors from women who had a PR-negative recurrence indicates the possibility of stratification of women for tailored treatment in the early stages of disease.
1 aFitzgerald, Lily, M1 aBrowne, Eva, P1 aChristie, Kevin, D1 aPunska, Elizabeth, C1 aSimmons, Leo, O1 aWilliams, Kristin, E1 aPentecost, Brian, T1 aJawale, Rahul, M1 aOtis, Christopher, N1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/elf5-and-dok7-regulation-in-anti-estrogen-treated-cells-and-tumors00768nas a2200217 4500008004100000245013100041210006900172260002500241300001600266490000800282100002500290700002400315700003100339700002000370700002900390700002300419700002100442700002500463700002400488856003800512 2016 eng d00aEssential Role of CFTR in PKA-Dependent Phosphorylation, Alkalinization, and Hyperpolarization During Human Sperm Capacitation0 aEssential Role of CFTR in PKADependent Phosphorylation Alkaliniz bWiley-Blackwellcoct a1404–14140 v2321 aMolina, Lis, C. Puga1 aPinto, Nicolás, A.1 aRodriguez, Paulina, Torres1 aRomarowski, Ana1 aSanchez, Alberto, Vicens1 aVisconti, Pablo, E1 aDarszon, Alberto1 aTreviño, Claudia, L1 aBuffone, Mariano, G uhttps://doi.org/10.1002/jcp.2563402031nas a2200217 4500008004100000022001400041245013400055210006900189260001300258300001600271490000700287520124600294100001701540700001401557700001501571700001801586700002301604700001901627700001301646856015401659 2016 eng d a1750-384100aEvaluation of the Penetration of Multiple Classes of Pesticides in Fresh Produce Using Surface-Enhanced Raman Scattering Mapping.0 aEvaluation of the Penetration of Multiple Classes of Pesticides c2016 Nov aT2891-T29010 v813 aUnderstanding pesticide penetration is important for effectively applying pesticides and in reducing pesticide exposures from food. This study aims to evaluate multiclass systemic and nonsystemic pesticide penetration in 3 representative fresh produce (apples, grapes, and spinach leaves). Surface-enhanced Raman scattering mapping was applied for in situ and real-time tracking of pesticide penetration over time. The results show that 100 mg/L of systemic pesticides, thiabendazole and acetamiprid, penetrated more rapidly and deeply with maximum depth around 220 μm after 48-h exposure into the tested fresh produce than 100 mg/L of nonsystemic pesticides, ferbam and phosmet, with maximum depth about 80 μm. The fact that 2 nonsystemic pesticides were also able to penetrate over time into all 3 fresh produce tested may raise additional food safety concerns. Comparatively, grapes were generally more resistant for pesticide penetration with all pesticides penetration depth below 80 μm compared to apples and spinach leaves. The information obtained here could provide technical support and guidance for accurate, effective, and safe application of pesticides and for the reduction of pesticide exposure from fresh produce.
1 aYang, Tianxi1 aZhao, Bin1 aHou, Ruyan1 aZhang, Zhiyun1 aKinchla, Amanda, J1 aClark, John, M1 aHe, Lili uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evaluation-penetration-multiple-classes-pesticides-fresh-produce-using02510nas a2200469 4500008004100000022001400041245008500055210007000140260001300210300001100223490000700234520102700241653001701268653002701285653001501312653001701327653003101344653003401375653001201409653004101421653002501462653002001487653003101507653001901538653001701557653000901574653001501583653001401598653004001612653005201652653002001704653001901724653002401743653001801767100001701785700001601802700002001818700002101838700001901859700001801878856014401896 2016 eng d a1873-635100aFipronil promotes adipogenesis via AMPKα-mediated pathway in 3T3-L1 adipocytes.0 aFipronil promotes adipogenesis via AMPKαmediated pathway in 3T3L c2016 Jun a217-230 v923 aEmerging evidence suggests that organochlorine, organophosphorus and neonicotinoid insecticide exposure may be linked to the development of obesity and type 2 diabetes. However, there is no knowledge of the potential influence of fipronil, which belongs to the phenylpyrazole chemical family, on obesity. Thus, the goal of this study was to determine the role of fipronil in adipogenesis using 3T3-L1 adipocytes. Fipronil treatment, at 10 μM, increased fat accumulation in 3T3-L1 adipocytes as well as promoted key regulators of adipocyte differentiation (CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor gamma-γ), and key regulators of lipogenesis (acetyl-CoA carboxylase and fatty acid synthase). The activation of AMPKα with 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) abolished effects of fipronil on increased adipogenesis. These results suggest that fipronil alters adipogenesis and results in increased lipid accumulation through a AMPKα-mediated pathway.
10a3T3-L1 Cells10aAcetyl-CoA Carboxylase10aAdipocytes10aAdipogenesis10aAminoimidazole Carboxamide10aAMP-Activated Protein Kinases10aAnimals10aCCAAT-Enhancer-Binding Protein-alpha10aCell Differentiation10aCells, Cultured10aGene Expression Regulation10aImmunoblotting10aInsecticides10aMice10aPPAR gamma10aPyrazoles10aReal-Time Polymerase Chain Reaction10aReverse Transcriptase Polymerase Chain Reaction10aRibonucleotides10aRNA, Messenger10aSignal Transduction10aTriglycerides1 aSun, Quancai1 aQi, Weipeng1 aYang, Jeremy, J1 aYoon, Kyong, Sup1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/fipronil-promotes-adipogenesis-ampka-mediated-pathway-3t3-l100826nas a2200241 4500008004100000245010700041210006900148260005700217300001800274490000800292100002700300700001300327700002300340700002100363700001900384700002400403700002200427700002300449700002400472700002000496700002400516856004400540 2016 eng d00aHIF-KDM3A-MMP12 regulatory circuit ensures trophoblast plasticity and placental adaptations to hypoxia0 aHIFKDM3AMMP12 regulatory circuit ensures trophoblast plasticity bProceedings of the National Academy of Sciencescnov aE7212–E72210 v1131 aChakraborty, Damayanti1 aCui, Wei1 aRosario, Gracy, X.1 aScott, Regan, L.1 aDhakal, Pramod1 aRenaud, Stephen, J.1 aTachibana, Makoto1 aRumi, M., A. Karim1 aMason, Clifford, W.1 aKrieg, Adam, J.1 aSoares, Michael, J. uhttps://doi.org/10.1073/pnas.161262611302349nas a2200445 4500008004100000022001400041245010400055210006900159260001600228300001400244490000700258520104900265653002601314653001401340653003401354653001201388653001801400653001901418653001101437653001501448653001701463653001201480653002301492653001001515653000901525653000901534653002301543653001901566653002001585653001201605653001601617100001701633700001501650700001301665700001801678700002101696700001901717700001801736856014901754 2016 eng d a1520-511800aImidacloprid Promotes High Fat Diet-Induced Adiposity and Insulin Resistance in Male C57BL/6J Mice.0 aImidacloprid Promotes High Fat DietInduced Adiposity and Insulin c2016 Dec 14 a9293-93060 v643 aImidacloprid, a neonicotinoid insecticide widely used in agriculture worldwide, has been reported to promote adipogenesis and cause insulin resistance in vitro. The purpose of the current study was to determine the effects of imidacloprid and its interaction with dietary fat in the development of adiposity and insulin resistance using male C57BL/6J mice. Imidacloprid (0.06, 0.6, or 6 mg/kg bw/day) was mixed in a low-fat (4% w/w) or high-fat (20% w/w) diet and given to mice ad libitum for 12 weeks. Imidacloprid significantly promoted high fat diet-induced body weight gain and adiposity. In addition, imidacloprid treatment with the high fat diet resulted in impaired glucose metabolism. Consistently, there were significant effects of imidacloprid on genes regulating lipid and glucose metabolisms, including the AMP-activated protein kinase-α (AMPKα) pathway in white adipose tissue and liver. These results suggest that imidacloprid may potentiate high fat diet-induced adiposity and insulin resistance in male C57BL/6J mice.
10aAdipose Tissue, White10aAdiposity10aAMP-Activated Protein Kinases10aAnimals10aBlood Glucose10aDiet, High-Fat10aHumans10aImidazoles10aInsecticides10aInsulin10aInsulin Resistance10aLiver10aMale10aMice10aMice, Inbred C57BL10aNeonicotinoids10aNitro Compounds10aObesity10aWeight Gain1 aSun, Quancai1 aXiao, Xiao1 aKim, Yoo1 aKim, Daeyoung1 aYoon, Kyoon, Sup1 aClark, John, M1 aPark, Yeonhwa uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/imidacloprid-promotes-high-fat-diet-induced-adiposity-and-insulin01690nas a2200229 4500008004100000022001400041245010400055210006900159260000900228300001200237490000600249520088600255100002201141700001901163700001701182700001701199700002301216700002001239700002401259700002401283856015301307 2016 eng d a2451-929400aImmunomodulatory effects of coated gold nanoparticles in LPS-stimulated and murine model systems.0 aImmunomodulatory effects of coated gold nanoparticles in LPSstim c2016 a320-3270 v13 aThe ability of nanoparticle surface functionalities to regulate immune responses during an immunological challenge (i. e. inflammation) would open new doors for their use in non-prophylactic therapeutics. We report here the use of functionalized 2 nm core gold nanoparticles to control the immunological responses of and systems presented with an inflammatory challenge. The results showed that NPs bearing a hydrophobic zwitterionic functionality boost inflammatory outcomes while hydrophilic zwitterionic NPs generate minimal immunological responses. Surprisingly, tetra(ethylene glycol) headgroups generate a significant anti-inflammatory response both and . These results demonstrate the ability of simple surface ligands to provide immunomodulatory properties, making them promising leads for the therapeutic usage of nanomaterials in diseases involving inflammation.
1 aMoyano, Daniel, F1 aLiu, Yuanchang1 aAyaz, Furkan1 aHou, Singyuk1 aPuangploy, Premsak1 aDuncan, Bradley1 aOsborne, Barbara, A1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/immunomodulatory-effects-coated-gold-nanoparticles-lps-stimulated-and00635nas a2200193 4500008004100000245008100041210006900122260003900191300001600230490000600246100002600252700002400278700001700302700002000319700002400339700001900363700002000382856003900402 2016 eng d00aIncreased hydrophobic block length of PTDMs promotes protein internalization0 aIncreased hydrophobic block length of PTDMs promotes protein int bRoyal Society of Chemistry ({RSC}) a7514–75210 v71 aBacklund, Coralie, M.1 aSgolastra, Federica1 aOtter, Ronja1 aMinter, Lisa, M1 aTakeuchi, Toshihide1 aFutaki, Shiroh1 aTew, Gregory, N uhttps://doi.org/10.1039/c6py01615d00793nas a2200241 4500008004100000245011000041210006900151260002300220300001600243490000700259100001700266700002900283700001900312700002700331700002000358700002400378700002400402700002100426700002400447700002000471700002000491856004000511 2016 eng d00aIntracellular Delivery of Anti-pPKC0 (Thr538) via Protein Transduction Domain Mimics for Immunomodulation0 aIntracellular Delivery of AntipPKC0 Thr538 via Protein Transduct bElsevier {BV}cdec a2118–21300 v241 aOzay, E., I.1 aGonzalez-Perez, Gabriela1 aTorres, Joe, A1 aVijayaraghavan, Jyothi1 aLawlor, Rebecca1 aSherman, Heather, L1 aGarrigan, Daniel, T1 aBurnside, Amy, S1 aOsborne, Barbara, A1 aTew, Gregory, N1 aMinter, Lisa, M uhttps://doi.org/10.1038/mt.2016.17703165nas a2200469 4500008004100000022001400041245011200055210007000167260001300237300001400250490000700264520165700271653001201928653003801940653002501978653002302003653003002026653002602056653001102082653002102093653001502114653002802129653002602157653000902183653002002192653002102212653002702233653002402260653001402284100001602298700002902314700001902343700002702362700002002389700002402409700002402433700002102457700002402478700002002502700002002522856015302542 2016 eng d a1525-002400aIntracellular Delivery of Anti-pPKCθ (Thr538) via Protein Transduction Domain Mimics for Immunomodulation.0 aIntracellular Delivery of AntipPKCθ Thr538 via Protein Transduct c2016 Dec a2118-21300 v243 aTargeting cellular proteins with antibodies, to better understand cellular signaling pathways in the context of disease modulation, is a fast-growing area of investigation. Humanized antibodies are increasingly gaining attention for their therapeutic potential, but the collection of cellular targets is limited to those secreted from cells or expressed on the cell surface. This approach leaves a wealth of intracellular proteins unexplored as putative targets for antibody binding. Protein kinase Cθ (PKCθ) is essential to T cell activation, proliferation, and differentiation, and its phosphorylation at specific residues is required for its activity. Here we report on the design, synthesis, and characterization of a protein transduction domain mimic capable of efficiently delivering an antibody against phosphorylated PKCθ (Thr538) into human peripheral mononuclear blood cells and altering expression of downstream indicators of T cell activation and differentiation. We used a humanized, lymphocyte transfer model of graft-versus-host disease, to evaluate the durability of protein transduction domain mimic:Anti-pPKCθ modulation, when delivered into human peripheral mononuclear blood cells ex vivo. We demonstrate that protein transduction domain mimic:Antibody complexes can be readily introduced with high efficacy into hard-to-transfect human peripheral mononuclear blood cells, eliciting a biological response sufficient to alter disease progression. Thus, protein transduction domain mimic:Antibody delivery may represent an efficient ex vivo approach to manipulating cellular responses by targeting intracellular proteins.
10aAnimals10aAntibodies, Monoclonal, Humanized10aCell Differentiation10aCell Proliferation10aCell-Penetrating Peptides10aGraft vs Host Disease10aHumans10aImmunomodulation10aIsoenzymes10aLeukocytes, Mononuclear10aLymphocyte Activation10aMice10aPhosphorylation10aProtein Kinase C10aProtein Kinase C-theta10aSignal Transduction10aTh1 Cells1 aOzay, Ilker1 aGonzalez-Perez, Gabriela1 aTorres, Joe, A1 aVijayaraghavan, Jyothi1 aLawlor, Rebecca1 aSherman, Heather, L1 aGarrigan, Daniel, T1 aBurnside, Amy, S1 aOsborne, Barbara, A1 aTew, Gregory, N1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/intracellular-delivery-anti-ppkcth-thr538-protein-transduction-domain00514nas a2200169 4500008004100000245005700041210005400098260002000152300001000172490000900182100002100191700002300212700002700235700002100262700002000283856004100303 2016 eng d00aMale Infertility: Genetics, Mechanism, and Therapies0 aMale Infertility Genetics Mechanism and Therapies bHindawi Limited a1–10 v20161 aCoutton, Charles1 aFissore, Rafael, A1 aPalermo, Gianpiero, D.1 aStouffs, Katrien1 aTouré, Aminata uhttps://doi.org/10.1155/2016/737236202976nas a2200445 4500008004100000022001400041245008400055210006900139260001300208300001100221490000700232520162800239653002801867653001201895653002301907653002201930653001101952653002601963653001101989653001402000653001702014653001502031653002202046653001502068653000902083653001402092653002002106653002202126653001802148653004702166100001602213700002602229700001902255700002302274700002102297700002202318700001902340700002102359856015002380 2016 eng d a1525-147000aManagement of Head Louse Infestations in the United States-A Literature Review.0 aManagement of Head Louse Infestations in the United StatesA Lite c2016 Sep a466-720 v333 aUNLABELLED: Head lice are a source of scalp irritation, social disruption, and loss of school time. Health care providers need authoritative information to help avoid the costs and risks of ineffective treatment. A review was completed to provide relevant information on infestation treatments available in the United States. Three major biomedical databases were searched from 1985, when current products were first available, to 2014, focusing on U.S.
REPORTS: A total of 579 references remained after duplicates were removed. A search of the U.S. Food and Drug Administration website and labels of approved products were reviewed. A marked decline in the effectiveness of permethrin and synergized pyrethrins was found, probably because of resistance arising from widespread and indiscriminate use, and the emergence of knockdown resistance mutations. The potential toxicity of lindane in the setting of readily available, safer, and more effective alternatives, should limit its use. Prescription products shown to be safe and effective with a single application, without nit combing, are topical ivermectin, malathion, and spinosad, whereas benzyl alcohol requires two applications. Home remedies such as mayonnaise, and essential oils, have not been demonstrated to be safe or effective, and may carry potential for severe adverse events. The high risk of failure of over-the-counter treatments in eliminating head louse infestations drives a need for health care provider recognition of the limitations of current treatments and for judicious use of treatments that remain effective.
10aAdministration, Topical10aAnimals10aDatabases, Factual10aDrug Combinations10aFemale10aHexachlorocyclohexane10aHumans10aIncidence10aInsecticides10aIvermectin10aLice Infestations10aMacrolides10aMale10aPediculus10aRisk Assessment10aTreatment Outcome10aUnited States10aUnited States Food and Drug Administration1 aKoch, Ellen1 aClark, John, Marshall1 aCohen, Bernard1 aMeinking, Terri, L1 aRyan, William, G1 aStevenson, Audrey1 aYetman, Robert1 aYoon, Kyong, Sup uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/management-head-louse-infestations-united-states-literature-review00556nas a2200157 4500008004100000245009200041210006900133260003800202300001600240490000700256100002200263700002600285700002000311700002000331856004700351 2016 eng d00aMapping Optimal Charge Density and Length of ROMP-Based PTDMs for siRNA Internalization0 aMapping Optimal Charge Density and Length of ROMPBased PTDMs for bAmerican Chemical Society ({ACS}) a3205–32120 v171 aCaffrey, Leah, M.1 adeRonde, Brittany, M.1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1021/acs.biomac.6b0090000910nas a2200265 4500008004100000245013800041210007100179260005200250300001800302490000800320100002600328700002300354700002100377700002600398700001700424700001300441700002000454700002100474700002100495700002400516700002100540700002200561700001700583856004400600 2016 eng d00aMiR-155–regulated molecular network orchestrates cell fate in the innate and adaptive immune response to Mycobacterium tuberculosis0 aMiR155–regulated molecular network orchestrates cell fate in the bProceedings of the National Academy of Sciences aE6172–E61810 v1131 aRothchild, Alissa, C.1 aSissons, James, R.1 aShafiani, Shahin1 aPlaisier, Christopher1 aMin, Deborah1 aMai, Dat1 aGilchrist, Mark1 aPeschon, Jacques1 aLarson, Ryan, P.1 aBergthaler, Andreas1 aBaliga, Nitin, S1 aUrdahl, Kevin, B.1 aAderem, Alan uhttps://doi.org/10.1073/pnas.160825511300654nas a2200181 4500008004100000245012500041210006900166260003800235300001600273490000700289100002600296700002400322700001700346700002200363700002000385700002000405856004700425 2016 eng d00aOptimal Hydrophobicity in Ring-Opening Metathesis Polymerization-Based Protein Mimics Required for siRNA Internalization0 aOptimal Hydrophobicity in RingOpening Metathesis PolymerizationB bAmerican Chemical Society ({ACS}) a1969–19770 v171 adeRonde, Brittany, M.1 aPosey, Nicholas, D.1 aOtter, Ronja1 aCaffrey, Leah, M.1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1021/acs.biomac.6b0013800659nas a2200181 4500008004100000245012500041210006900166260004300235300001600278490000700294100002600301700002400327700001700351700002200368700002000390700002000410856004700430 2016 eng d00aOptimal Hydrophobicity in Ring-Opening Metathesis Polymerization-Based Protein Mimics Required for siRNA Internalization0 aOptimal Hydrophobicity in RingOpening Metathesis PolymerizationB bAmerican Chemical Society ({ACS})cjun a1969–19770 v171 adeRonde, Brittany, M.1 aPosey, Nicholas, D.1 aOtter, Ronja1 aCaffrey, Leah, M.1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1021/acs.biomac.6b0013802078nas a2200253 4500008004100000245011600041210006900157260001300226300001000239490000800249520118700257100001901444700002101463700001901484700002201503700002501525700002201550700002001572700001701592700001701609700002501626700002401651856014901675 2016 eng d00aPotential of breastmilk analysis to inform early events in breast carcinogenesis: rationale and considerations.0 aPotential of breastmilk analysis to inform early events in breas c2016 May a13-220 v1573 aThis review summarizes methods related to the study of human breastmilk in etiologic and biomarkers research. Despite the importance of reproductive factors in breast carcinogenesis, factors that act early in life are difficult to study because young women rarely require breast imaging or biopsy, and analysis of critical circulating factors (e.g., hormones) is often complicated by the requirement to accurately account for menstrual cycle date. Accordingly, novel approaches are needed to understand how events such as pregnancy, breastfeeding, weaning, and post-weaning breast remodeling influence breast cancer risk. Analysis of breastmilk offers opportunities to understand mechanisms related to carcinogenesis in the breast, and to identify risk markers that may inform efforts to identify high-risk women early in the carcinogenic process. In addition, analysis of breastmilk could have value in early detection or diagnosis of breast cancer. In this article, we describe the potential for using breastmilk to characterize the microenvironment of the lactating breast with the goal of advancing research on risk assessment, prevention, and detection of breast cancer.
1 aMurphy, Jeanne1 aSherman, Mark, E1 aBrowne, Eva, P1 aCaballero, Ana, I1 aPunska, Elizabeth, C1 aPfeiffer, Ruth, M1 aYang, Hannah, P1 aLee, Maxwell1 aYang, Howard1 aGierach, Gretchen, L1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/potential-of-breastmilk-analysis-to-inform-early-events-in-breast00436nas a2200133 4500008004100000245007000041210006900111260002500180300001400205490000700219100001600226700002200242856003800264 2016 eng d00aProspects for vaccination against pathogenic African trypanosomes0 aProspects for vaccination against pathogenic African trypanosome bWiley-Blackwellcdec a735–7430 v381 aBlack, S, J1 aMansfield, J., M. uhttps://doi.org/10.1111/pim.1238700709nas a2200205 4500008004100000245010900041210006900150260002300219300001200242490000800254100002500262700002600287700002200313700002800335700002200363700002300385700002200408700002500430856004800455 2016 eng d00aRab3A, a possible marker of cortical granules, participates in cortical granule exocytosis in mouse eggs0 aRab3A a possible marker of cortical granules participates in cor bElsevier {BV}csep a42–510 v3471 aBello, Oscar, Daniel1 aCappa, Andrea, Isabel1 ade Paola, Matilde1 aZanetti, Maria, Natalia1 aFukuda, Mitsunori1 aFissore, Rafael, A1 aMayorga, Luis, S.1 aMichaut, Marcela, A. uhttps://doi.org/10.1016/j.yexcr.2016.07.00502261nas a2200289 4500008004100000022001400041245012500055210006900180260001500249300001200264490000700276520130700283653000901590653002401599653001501623653001701638653002901655653002201684100001701706700001801723700001401741700001501755700002001770700001901790700001301809856014901822 2016 eng d a1520-688200aReal-Time and in Situ Monitoring of Pesticide Penetration in Edible Leaves by Surface-Enhanced Raman Scattering Mapping.0 aRealTime and in Situ Monitoring of Pesticide Penetration in Edib c2016 05 17 a5243-500 v883 aUnderstanding of the penetration behaviors of pesticides in fresh produce is of great significance for effectively applying pesticides and minimizing pesticide residues in food. There is lack, however, of an effective method that can measure pesticide penetration. Herein, we developed a novel method for real-time and in situ monitoring of pesticide penetration behaviors in spinach leaves based on surface-enhanced Raman scattering (SERS) mapping. Taking advantage of penetrative gold nanoparticles (AuNPs) as probes to enhance the internalized pesticide signals in situ, we have successfully obtained the internal signals from thiabendazole, a systemic pesticide, following its penetration into spinach leaves after removing surface pesticide residues. Comparatively, ferbam, a nonsystemic pesticide, did not show internal signals after removing surface pesticide residues, demonstrating its nonsystemic behavior. In both cases, if the surface pesticides were not removed, copenetration of both AuNPs and pesticides was observed. These results demonstrate a successful application of SERS as an effective method for measuring pesticides penetration in fresh produce in situ. The information obtained could provide useful guidance for effective and safe applications of pesticides on plants.
10aGold10aMetal Nanoparticles10aPesticides10aPlant Leaves10aSpectrum Analysis, Raman10aSpinacia oleracea1 aYang, Tianxi1 aZhang, Zhiyun1 aZhao, Bin1 aHou, Ruyan1 aKinchla, Amanda1 aClark, John, M1 aHe, Lili uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/real-time-and-situ-monitoring-pesticide-penetration-edible-leaves00677nas a2200205 4500008004100000245007200041210006900113260005700182300001600239490000800255100001800263700001300281700001900294700002300313700002300336700002000359700002400379700002400403856004400427 2016 eng d00aRethinking progesterone regulation of female reproductive cyclicity0 aRethinking progesterone regulation of female reproductive cyclic bProceedings of the National Academy of Sciencescmar a4212–42170 v1131 aKubota, Kaiyu1 aCui, Wei1 aDhakal, Pramod1 aWolfe, Michael, W.1 aRumi, M., A. Karim1 aVivian, Jay, L.1 aRoby, Katherine, F.1 aSoares, Michael, J. uhttps://doi.org/10.1073/pnas.160182511301853nas a2200193 4500008004100000245006500041210006400106260000900170300001100179490000800190520118900198100001901387700002801406700001801434700002401452700002301476700001701499856014301516 2016 eng d00aSperm Capacitation and Acrosome Reaction in Mammalian Sperm.0 aSperm Capacitation and Acrosome Reaction in Mammalian Sperm c2016 a93-1060 v2203 aPhysiological changes that endow mammalian sperm with fertilizing capacity are known as sperm capacitation. As part of capacitation, sperm develop an asymmetrical flagellar beating known as hyperactivation and acquire the ability to undergo the acrosome reaction. Together, these processes promote fertilizing competence in sperm. At the molecular level, capacitation involves a series of signal transduction events which include activation of cAMP-dependent phosphorylation pathways, removal of cholesterol, hyperpolarization of the sperm plasma membrane, and changes in ion permeability. In recent years, new technologies have aided in the study of sperm signaling molecules with better resolution, at both spatial and temporal levels, unraveling how different cascades integrate and cooperate to render a fertilizing sperm. Despite this new information, the molecular mechanisms connecting capacitation with acrosomal exocytosis and hyperactivation are not well understood. This review brings together results obtained in mammalian species in the field of sperm capacitation with special focus on those pathways involved in the preparation to undergo the acrosomal reaction.
1 aStival, Cintia1 aMolina, Lis, Del C Puga1 aPaudel, Bidur1 aBuffone, Mariano, G1 aVisconti, Pablo, E1 aKrapf, Dario uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sperm-capacitation-and-acrosome-reaction-in-mammalian-sperm02430nas a2200349 4500008004100000022001400041245010500055210006900160260001600229300001000245490000600255520125900261653002501520653002401545653001601569653001201585653003401597653005001631653000801681653002801689653002401717653002701741653001301768653002401781653002101805653002701826100002201853700001601875700001901891700002601910856014401936 2016 eng d a2045-232200aSplice form variant and amino acid changes in MDR49 confers DDT resistance in transgenic Drosophila.0 aSplice form variant and amino acid changes in MDR49 confers DDT c2016 Mar 22 a233550 v63 aThe ATP-binding cassette (ABC) transporters represent a superfamily of proteins that have important physiological roles in both prokaryotes and eukaryotes. In insects, ABC transporters have previously been implicated in insecticide resistance. The 91-R strain of Drosophila melanogaster has been intensely selected with DDT over six decades. A recent selective sweeps analysis of 91-R implicated the potential role of MDR49, an ABC transporter, in DDT resistance, however, to date the details of how MDR49 may play a role in resistance have not been elucidated. In this study, we investigated the impact of structural changes and an alternative splicing event in MDR49 on DDT-resistance in 91-R, as compared to the DDT susceptible strain 91-C. We observed three amino acid differences in MDR49 when 91-R was compared with 91-C, and only one isoform (MDR49B) was implicated in DDT resistance. A transgenic Drosophila strain containing the 91-R-MDR49B isoform had a significantly higher LD50 value as compared to the 91-C-MDR49B isoform at the early time points (6 h to 12 h) during DDT exposure. Our data support the hypothesis that the MDR49B isoform, with three amino acid mutations, plays a role in the early aspects of DDT resistance in 91-R.
10aAlternative Splicing10aAmino Acid Sequence10aAmino Acids10aAnimals10aAnimals, Genetically Modified10aATP Binding Cassette Transporter, Subfamily B10aDDT10aDrosophila melanogaster10aDrosophila Proteins10aInsecticide Resistance10aMutation10aOpen Reading Frames10aProtein Isoforms10aSequence Analysis, RNA1 aSeong, Keon, Mook1 aSun, Weilin1 aClark, John, M1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/splice-form-variant-and-amino-acid-changes-mdr49-confers-ddt00597nas a2200181 4500008004100000245010800041210006900149260002500218490000600243100001300249700001900262700002000281700001800301700001500319700002600334700001700360856003800377 2016 eng d00aTowards Functional Annotation of the Preimplantation Transcriptome: An RNAi Screen in Mammalian Embryos0 aTowards Functional Annotation of the Preimplantation Transcripto bSpringer Naturecnov0 v61 aCui, Wei1 aDai, Xiangpeng1 aMarcho, Chelsea1 aHan, Zhengbin1 aZhang, Kun1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://doi.org/10.1038/srep3739600808nas a2200253 4500008004100000245009900041210006900140260002500209490000600234100002500240700001900265700002000284700002000304700001700324700003100341700002100372700001700393700001700410700002300427700002200450700002100472700002300493856003800516 2016 eng d00aTransient exposure to calcium ionophore enables in vitro fertilization in sterile mouse models0 aTransient exposure to calcium ionophore enables in vitro fertili bSpringer Naturecsep0 v61 aNavarrete, Felipe, A1 aAlvau, Antonio1 aLee, Hoi, Chang1 aLevin, Lonny, R1 aBuck, Jochen1 aDe Leon, Patricia, Martin-1 aSanti, Celia, M.1 aKrapf, Dario1 aMager, Jesse1 aFissore, Rafael, A1 aSalicioni, Ana, M1 aDarszon, Alberto1 aVisconti, Pablo, E uhttps://doi.org/10.1038/srep3358900603nas a2200169 4500008004100000245012200041210006900163260002500232490000600257100002100263700002000284700002000304700002200324700002300346700002600369856003800395 2016 eng d00aTRPM7-like channels are functionally expressed in oocytes and modulate post-fertilization embryo development in mouse0 aTRPM7like channels are functionally expressed in oocytes and mod bSpringer Naturecsep0 v61 aCarvacho, Ingrid1 aArdestani, Goli1 aLee, Hoi, Chang1 aMcGarvey, Kaitlyn1 aFissore, Rafael, A1 aLykke-Hartmann, Karin uhttps://doi.org/10.1038/srep3423600549nas a2200169 4500008004100000245007000041210006800111260002300179300001200202490000700214100002000221700002100241700002600262700002300288700002100311856004700332 2016 eng d00aTRPV3 channels mediate Ca2+ influx induced by 2-APB in mouse eggs0 aTRPV3 channels mediate Ca2 influx induced by 2APB in mouse eggs bElsevier {BV}cjan a21–310 v591 aLee, Hoi, Chang1 aYoon, Sook-Young1 aLykke-Hartmann, Karin1 aFissore, Rafael, A1 aCarvacho, Ingrid uhttps://doi.org/10.1016/j.ceca.2015.12.00100693nas a2200217 4500008004100000245006700041210006600108260004400174300001300218490000700231100002100238700001900259700002300278700001900301700001900320700002500339700001800364700002100382700002300403856004900426 2016 eng d00aTrypanosoma brucei Co-opts NK Cells to Kill Splenic B2 B Cells0 aTrypanosoma brucei Coopts NK Cells to Kill Splenic B2 B Cells bPublic Library of Science ({PLoS})cjul ae10057330 v121 aFrenkel, Deborah1 aZhang, Fengqiu1 aGuirnalda, Patrick1 aHaynes, Carole1 aBockstal, Viki1 aRadwanska, Magdalena1 aMagez, Stefan1 aBlack, Samuel, J1 aUzonna, Jude, Ezeh uhttps://doi.org/10.1371/journal.ppat.100573302437nas a2200265 4500008004100000245012900041210006900170260001600239520142800255100001901683700003201702700002701734700002501761700001501786700003201801700003501833700002201868700001701890700002101907700001701928700002601945700002301971700002301994856015402017 2016 ENG d00aThe tyrosine kinase FER is responsible for the capacitation-associated increase in tyrosine phosphorylation in murine sperm.0 atyrosine kinase FER is responsible for the capacitationassociate c2016 May 253 aSperm capacitation is required for fertilization. At the molecular level, this process is associated with a fast activation of protein kinase A. Downstream of this event, capacitating conditions lead to an increase in tyrosine phosphorylation. The identity of the tyrosine kinase(s) mediating this process has not been conclusively demonstrated. Recent experiments using stallion and human sperm have suggested a role for PYK2 based on the use of small molecule inhibitors directed against this kinase. However, critical loss-of-function experiments have not been reported. Here, we used both pharmacological inhibitors and genetically modified mice models to investigate the identity of the tyrosine kinase(s) mediating the increase in tyrosine phosphorylation in mouse sperm. Similar to stallion and human, PF431396 blocks the capacitation-associated increase in tyrosine phosphorylation. Yet, sperm from Pyk2(-/-) mice displayed normal increase in tyrosine phosphorylation, implying that PYK2 is not responsible for this phosphorylation process. Here we show that PF431396 can also inhibit FER, a tyrosine kinase known to be present in sperm. Sperm from mice targeted with a kinase inactivating mutation in Fer failed to undergo capacitation-associated increases in tyrosine phosphorylation. While these mice are fertile, their sperm displayed a reduced ability to fertilize metaphase-II arrested eggs in vitro.
1 aAlvau, Antonio1 aBattistone, Maria, Agustina1 aGervasi, Maria, Gracia1 aNavarrete, Felipe, A1 aXu, Xinran1 aSánchez-Cárdenas, Claudia1 aDe la Vega-Beltran, Jose, Luis1 aDa Ros, Vanina, G1 aGreer, Peter1 aDarszon, Alberto1 aKrapf, Diego1 aSalicioni, Ana, Maria1 aCuasnicu, Patricia1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-tyrosine-kinase-fer-is-responsible-for-the-capacitation-associated01881nas a2200145 4500008004100000245010400041210006900145260001600214520126200230100002601492700002101518700001801539700002501557856015301582 2015 ENG d00aADAM13 cleavage of cadherin-11 promotes CNC migration independently of the homophilic binding site.0 aADAM13 cleavage of cadherin11 promotes CNC migration independent c2015 Jul 213 aThe cranial neural crest (CNC) is a highly motile population of cells that is responsible for forming the face and jaw in all vertebrates and perturbing their migration can lead to craniofacial birth defects. Cell motility requires a dynamic modification of cell-cell and cell-matrix adhesion. In the CNC, cleavage of the cell adhesion molecule cadherin-11 by ADAM13 is essential for cell migration. This cleavage generates a shed extracellular fragment of cadherin-11 (EC1-3) that possesses pro-migratory activity via an unknown mechanism. Cadherin-11 plays an important role in modulating contact inhibition of locomotion (CIL) in the CNC to regulate directional cell migration. Here, we show that while the integral cadherin-11 requires the homophilic binding site to promote CNC migration in vivo, the EC1-3 fragment does not. In addition, we show that increased ADAM13 activity or expression of the EC1-3 fragment increases CNC invasiveness in vitro and blocks the repulsive CIL response in colliding cells. This activity requires the presence of an intact homophilic binding site on the EC1-3 suggesting that the cleavage fragment may function as a competitive inhibitor of cadherin-11 adhesion in CIL but not to promote cell migration in vivo.
1 aAbbruzzese, Genevieve1 aBecker, Sarah, F1 aKashef, Jubin1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/adam13-cleavage-of-cadherin-11-promotes-cnc-migration-independently-002588nas a2200325 4500008004100000022001400041245004400055210004300099260001300142300001200155490000700167520165200174653002501826653001201851653001801863653001801881653001101899653001401910653001701924100002701941700003201968700001302000700001902013700001902032700002402051700002602075700002002101700002202121856011902143 2015 eng d a1537-171900aAlternative Splice in Alternative Lice.0 aAlternative Splice in Alternative Lice c2015 Oct a2749-590 v323 aGenomic and transcriptomics analyses have revealed human head and body lice to be almost genetically identical; although con-specific, they nevertheless occupy distinct ecological niches and have differing feeding patterns. Most importantly, while head lice are not known to be vector competent, body lice can transmit three serious bacterial diseases; epidemictyphus, trench fever, and relapsing fever. In order to gain insights into the molecular bases for these differences, we analyzed alternative splicing (AS) using next-generation sequencing data for one strain of head lice and one strain of body lice. We identified a total of 3,598 AS events which were head or body lice specific. Exon skipping AS events were overrepresented among both head and body lice, whereas intron retention events were underrepresented in both. However, both the enrichment of exon skipping and the underrepresentation of intron retention are significantly stronger in body lice compared with head lice. Genes containing body louse-specific AS events were found to be significantly enriched for functions associated with development of the nervous system, salivary gland, trachea, and ovarian follicle cells, as well as regulation of transcription. In contrast, no functional categories were overrepresented among genes with head louse-specific AS events. Together, our results constitute the first evidence for transcript pool differences in head and body lice, providing insights into molecular adaptations that enabled human lice to adapt to clothing, and representing a powerful illustration of the pivotal role AS can play in functional adaptation.
10aAlternative Splicing10aAnimals10aGene Ontology10aGenes, Insect10aHumans10aPediculus10aPhthiraptera1 aTovar-Corona, Jaime, M1 aCastillo-Morales, Atahualpa1 aChen, Lu1 aOlds, Brett, P1 aClark, John, M1 aReynolds, Stuart, E1 aPittendrigh, Barry, R1 aFeil, Edward, J1 aUrrutia, Araxi, O uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/alternative-splice-alternative-lice02858nas a2200361 4500008004100000245009700041210006900138260000900207300001100216490000700227520168400234653001601918653002101934653001601955653001401971653002001985653004302005653002802048653001102076653003002087653002402117100001802141700001502159700001502174700002502189700002502214700001802239700002402257700002002281700001702301700002402318856015402342 2015 eng d00aAnalysis of DNA methylation and gene expression in radiation-resistant head and neck tumors.0 aAnalysis of DNA methylation and gene expression in radiationresi c2015 a545-610 v103 aResistance to radiation therapy constitutes a significant challenge in the treatment of head and neck squamous cell cancer (HNSCC). Alteration in DNA methylation is thought to play a role in this resistance. Here, we analyzed DNA methylation changes in a matched model of radiation resistance for HNSCC using the Illumina HumanMethylation450 BeadChip. Our results show that compared to radiation-sensitive cells (SCC-61), radiation-resistant cells (rSCC-61) had a significant increase in DNA methylation. After combining these results with microarray gene expression data, we identified 84 differentially methylated and expressed genes between these 2 cell lines. Ingenuity Pathway Analysis revealed ILK signaling, glucocorticoid receptor signaling, fatty acid α-oxidation, and cell cycle regulation as top canonical pathways associated with radiation resistance. Validation studies focused on CCND2, a protein involved in cell cycle regulation, which was identified as hypermethylated in the promoter region and downregulated in rSCC-61 relative to SCC-61 cells. Treatment of rSCC-61 and SCC-61 with the DNA hypomethylating agent 5-aza-2'deoxycitidine increased CCND2 levels only in rSCC-61 cells, while treatment with the control reagent cytosine arabinoside did not influence the expression of this gene. Further analysis of HNSCC data from The Cancer Genome Atlas found increased methylation in radiation-resistant tumors, consistent with the cell culture data. Our findings point to global DNA methylation status as a biomarker of radiation resistance in HNSCC, and suggest a need for targeted manipulation of DNA methylation to increase radiation response in HNSCC.
10aAzacitidine10aCell Line, Tumor10aCpG Islands10aCyclin D210aDNA Methylation10aGene Expression Regulation, Neoplastic10aHead and Neck Neoplasms10aHumans10aPromoter Regions, Genetic10aRadiation Tolerance1 aChen, Xiaofei1 aLiu, Liang1 aMims, Jade1 aPunska, Elizabeth, C1 aWilliams, Kristin, E1 aZhao, Weiling1 aArcaro, Kathleen, F1 aTsang, Allen, W1 aZhou, Xiaobo1 aFurdui, Cristina, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/analysis-of-dna-methylation-and-gene-expression-in-radiation-resistant02584nas a2200241 4500008004100000245009100041210007000132260001300202300001100215490000700226520172800233100002001961700002901981700002702010700001902037700001902056700002502075700002102100700002502121700002102146700002402167856015102191 2015 eng d00aAssociation of TGF-β2 levels in breast milk with severity of breast biopsy diagnosis.0 aAssociation of TGFβ2 levels in breast milk with severity of brea c2015 Mar a345-540 v263 aPURPOSE: TGF-β plays a dual role in breast carcinogenesis, acting at early stages as tumor-suppressors and later as tumor-promoters. TGF-β isoforms are expressed in breast tissues and secreted in milk, suggesting that analysis of levels in milk might be informative for breast cancer risk. Accordingly, we assessed TGF-β2 levels in milk from women who had undergone a breast biopsy and related the concentrations to diagnosis. METHODS: Milk donated by women who had undergone or were scheduled for a breast biopsy was shipped on ice for processing and testing. Breast cancer risk factors were obtained through a self-administered questionnaire, and biopsy diagnoses were extracted from pathology reports. TGF-β2 levels in milk, assessed as absolute levels and in relation to total protein, were analyzed in bilateral samples donated by 182 women. Linear regression was used to estimate relationships of log-transformed TGF-β2 levels and TGF-β2/ total protein ratios to biopsy category. RESULTS: Milk TGF-β2 levels from biopsied and non-biopsied breasts within women were highly correlated (r (2) = 0.77). Higher mean TGF-β2 milk levels (based on average of bilateral samples) were marginally associated with more severe breast pathological diagnosis, after adjusting for duration of nursing current child (adjusted p trend = 0.07). CONCLUSIONS: Our exploratory analysis suggests a borderline significant association between higher mean TGF-β2 levels in breast milk and more severe pathologic diagnoses. Further analysis of TGF-β signaling in milk may increase understanding of postpartum remodeling and advance efforts to analyze milk as a means of assessing risk of breast pathology.
1 aYang, Hannah, P1 aSchneider, Sallie, Smith1 aChisholm, Christina, M1 aBrowne, Eva, P1 aMahmood, Sidra1 aGierach, Gretchen, L1 aLenington, Sarah1 aAnderton, Douglas, L1 aSherman, Mark, E1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/association-of-tgf-v2-levels-in-breast-milk-with-severity-of-breast02596nas a2200205 4500008004100000245007700041210006900118260001600187520182300203100002502026700003602051700001902087700002302106700001702129700003202146700002202178700002102200700002302221856014602244 2015 ENG d00aBiphasic Role of Calcium in Mouse Sperm Capacitation Signaling Pathways.0 aBiphasic Role of Calcium in Mouse Sperm Capacitation Signaling P c2015 Jan 173 aMammalian sperm acquire fertilizing ability in the female tract in a process known as capacitation. At the molecular level, capacitation is associated with up-regulation of a cAMP-dependent pathway, changes in intracellular pH, intracellular Ca(2+) and an increase in tyrosine phosphorylation. How these signaling systems interact during capacitation is not well understood. Results presented in this study indicate that Ca(2+) ions have a biphasic role in the regulation of cAMP-dependent signaling. Media without added Ca(2+) salts (nominal zero Ca(2+) ) still contain micromolar concentrations of this ion. Sperm incubated in this medium did not undergo PKA activation or the increase in tyrosine phosphorylation suggesting that these phosphorylation pathways require Ca(2+) . However, chelation of the extracellular Ca(2+) traces by EGTA induced both cAMP-dependent phosphorylation and the increase in tyrosine phosphorylation. The EGTA effect in nominal zero Ca(2+) media was mimicked by two calmodulin antagonists, W7 and calmidazolium, and by the calcineurin inhibitor cyclosporine A. These results suggest that Ca(2+) ions regulate sperm cAMP and tyrosine phosphorylation pathways in a biphasic manner and that some of its effects are mediated by calmodulin. Interestingly, contrary to wild type mouse sperm, sperm from CatSper1 KO mice underwent PKA activation and an increase in tyrosine phosphorylation upon incubation in nominal zero Ca(2+) media. Therefore, sperm lacking Catsper Ca(2+) channels behave as wild-type sperm incubated in the presence of EGTA. This latter result suggests that Catsper transports the Ca(2+) involved in the regulation of cAMP-dependent and tyrosine phosphorylation pathways required for sperm capacitation. This article is protected by copyright. All rights reserved.
1 aNavarrete, Felipe, A1 aGarcía-Vázquez, Francisco, A.1 aAlvau, Antonio1 aEscoffier, Jessica1 aKrapf, Dario1 aSánchez-Cárdenas, Claudia1 aSalicioni, Ana, M1 aDarszon, Alberto1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/biphasic-role-of-calcium-in-mouse-sperm-capacitation-signaling00618nas a2200205 4500008004100000245008900041210006900130260002500199300001400224490000800238100002200246700001400268700001900282700001700301700001200318700001200330700001700342700001400359856003900373 2015 eng d00acis-regulatory control of Mesp1 expression by YY1 and SP1 during mouse embryogenesis0 acisregulatory control of Mesp1 expression by YY1 and SP1 during bWiley-Blackwellcnov a379–3870 v2451 aBeketaev, Ilimbek1 aZhang, Yi1 aWeng, Kuo-Chan1 aRhee, Siyeon1 aYu, Wei1 aLiu, Yu1 aMager, Jesse1 aWang, Jun uhttps://doi.org/10.1002/dvdy.2434902089nas a2200193 4500008004100000245009000041210006900131260001600200520140200216100002201618700001401640700001901654700001701673700001201690700001201702700001701714700001401731856015001745 2015 ENG d00acis-regulatory control of Mesp1 expression by YY1 and SP1 during mouse embryogenesis.0 acisregulatory control of Mesp1 expression by YY1 and SP1 during c2015 Sep 183 aBACKGROUND: Mesp1 is critical for early cardiomyocyte differentiation and heart development. We previously observed down-regulation of Mesp1 expression in YY1-ablated mouse embryonic hearts. However, how Mesp1 expression is mediated by YY1 is not well understood. RESULTS: We excised YY1 in the murine embryos using Sox2-cre and found that Mesp1 was down-regulated in the embryonic day (E) 7.5 mutant embryos. Also, YY1 activated the 6 kb Mesp1 regulatory element fused to a luciferase reporter. We identified two putative YY1 binding sites in the proximal promoter region of Mesp1 gene, and found that mutation of these sites significantly reduced YY1-induced activation of the Mesp1 promoter. We also uncovered one cognitive site for SP1, one of the earliest binding partners of YY1 identified. Mutation of this SP1 site repressed SP1-induced activation of the Mesp1 promoter. Moreover, YY1 and SP1 synergistically activated the Mesp1 promoter. Consistently, while Lacz expression driven by the wild-type 6 kb regulatory element of Mesp1 gene was robust in E7.5 mouse embryos, the mutation of these binding sites in the context of this 6 kb sequence substantially reduced the LacZ expression during embryogenesis. CONCLUSIONS: YY1 and SP1 independently and cooperatively govern the Mesp1 expression during embryogenesis. Developmental Dynamics, 2015. © 2015 Wiley Periodicals, Inc.
1 aBeketaev, Ilimbek1 aZhang, Yi1 aWeng, Kuo-Chan1 aRhee, Siyeon1 aYu, Wei1 aLiu, Yu1 aMager, Jesse1 aWang, Jun uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cis-regulatory-control-of-mesp1-expression-by-yy1-and-sp1-during-000552nas a2200145 4500008004100000245012200041210006900163260004000232300001600272490000800288100001900296700002300315700001900338856004900357 2015 eng d00aCXCR4 expression on pathogenic T cells facilitates their bone marrow infiltration in a mouse model of aplastic anemia0 aCXCR4 expression on pathogenic T cells facilitates their bone ma bAmerican Society of Hematologycfeb a2087–20940 v1251 aKuksin, Arieta1 aGonzalez-Perez, G.1 aMinter, L., M. uhttps://doi.org/10.1182/blood-2014-08-59479600565nas a2200157 4500008004100000245009900041210006900140260004300209300001600252490000700268100002600275700001900301700002000320700002000340856004700360 2015 eng d00aDevelopment of Guanidinium-Rich Protein Mimics for Efficient siRNA Delivery into Human T Cells0 aDevelopment of GuanidiniumRich Protein Mimics for Efficient siRN bAmerican Chemical Society ({ACS})coct a3172–31790 v161 adeRonde, Brittany, M.1 aTorres, Joe, A1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://doi.org/10.1021/acs.biomac.5b0079500616nas a2200169 4500008004100000245011700041210006900158260004300227300001400270490000700284100003100291700001900322700002200341700002400363700001800387856004100405 2015 eng d00aDirect Image-Based Enumeration of Clostridium phytofermentans Cells on Insoluble Plant Biomass Growth Substrates0 aDirect ImageBased Enumeration of Clostridium phytofermentans Cel bAmerican Society for Microbiologycdec a972–9780 v821 aAlvelo-Maurosa, Jesús, G.1 aLee, Scott, J.1 aHazen, Samuel, P.1 aLeschine, Susan, B.1 aKelly, R., M. uhttps://doi.org/10.1128/aem.03119-1500538nas a2200157 4500008004100000245004500041210004300086260001500129300000600144490000800150100002200158700002500180700003200205700002100237856012200258 2015 eng d00aEditorial: Cell adhesion in development.0 aEditorial Cell adhesion in development c2015 May 1 a10 v4011 aBajanca, Fernanda1 aAlfandari, Dominique1 aThorsteinsdóttir, Sólveig1 aThéveneau, Eric uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/editorial-cell-adhesion-in-development00445nas a2200145 4500008004100000245005900041210005900100260002600159300001600185490000800201100002000209700001300229700001700242856004000259 2015 eng d00aEpigenetic dynamics during preimplantation development0 aEpigenetic dynamics during preimplantation development b{BioScientifica}cjun aR109–R1200 v1501 aMarcho, Chelsea1 aCui, Wei1 aMager, Jesse uhttps://doi.org/10.1530/rep-15-018002055nas a2200205 4500008004100000245015800041210006900199260001600268300001100284490000800295520128400303100001801587700001701605700001601622700001601638700001301654700001701667700001401684856015101698 2015 eng d00aExpression patterns of long noncoding RNAs from Dlk1-Dio3 imprinted region and the potential mechanisms of Gtl2 activation during blastocyst development.0 aExpression patterns of long noncoding RNAs from Dlk1Dio3 imprint c2015 Jul 31 a167-730 v4633 aThe function of long noncoding RNAs (lncRNAs) in cell differentiation and development have begun to be revealed in recent years. However, the expression pattern and mechanisms regulating lncRNAs are largely unknown during mammalian preimplantation development. LncRNAs expressed from Dlk1-Dio3 imprinted region have been linked to pluripotency of induced pluripotent cells (iPSCs). In this study we show that these lncRNAs (Gtl2, Rian and Mirg) are first expressed at the morula stage and gradually restricted to the inner cell mass (ICM) as the embryo differentiates into the blastocyst. Analysis of DNA methylation at IG-DMR and Gtl2-DMR showed no change during preimplantation while the presence of the activating histone modification H3K4me3 increased significantly from 8-cell to blastocyst stage, which may explain the expression activation. Additionally, knockdown of transcription factors (Oct4, Sox2 and Nanog) in blastocyst reduced the expression of Gtl2, indicating pluripotency factors regulate transcription of these lncRNAs. This study provides the spatiotemporal expression and dynamic changes of lncRNAs from Dlk1-Dio3 imprinted region in mouse preimplantation stage embryos and offers insight into the potential mechanisms responsible for Gtl2 activation.
1 aHan, Zhengbin1 aYu, Changwei1 aTian, Yijun1 aZeng, Tiebo1 aCui, Wei1 aMager, Jesse1 aWu, Qiong uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/expression-patterns-of-long-noncoding-rnas-from-dlk1-dio3-imprinted01167nas a2200361 4500008004100000245012500041210006900166260004400235300001300279490000700292100001600299700002500315700002100340700002400361700002000385700002400405700002200429700001600451700002400467700002100491700002500512700001700537700001800554700002000572700002300592700001600615700002300631700002300654700002400677700002600701700002900727856004900756 2015 eng d00aGenome and Transcriptome of Clostridium phytofermentans, Catalyst for the Direct Conversion of Plant Feedstocks to Fuels0 aGenome and Transcriptome of Clostridium phytofermentans Catalyst bPublic Library of Science ({PLoS})cjun ae01182850 v101 aPetit, Elsa1 aCoppi, Maddalena, V.1 aHayes, James, C.1 aTolonen, Andrew, C.1 aWarnick, Thomas1 aLatouf, William, G.1 aAmisano, Danielle1 aBiddle, Amy1 aMukherjee, Supratim1 aIvanova, Natalia1 aLykidis, Athanassios1 aLand, Miriam1 aHauser, Loren1 aKyrpides, Nikos1 aHenrissat, Bernard1 aLau, Joanne1 aSchnell, Danny, J.1 aChurch, George, M.1 aLeschine, Susan, B.1 aBlanchard, Jeffrey, L1 avan Berkel, Willem, J.H. uhttps://doi.org/10.1371/journal.pone.011828502885nas a2200433 4500008004100000022001400041245017200055210006900227260000900296300001300305490000700318520145000325653002401775653001201799653003401811653002301845653003501868653002801903653002401931653002101955653001101976653002001987653001202007653000902019653002802028653002902056653001402085653002402099100001602123700001902139700002202158700002102180700001802201700001902219700001902238700001702257700002602274856015102300 2015 eng d a1932-620300aA glycine insertion in the estrogen-related receptor (ERR) is associated with enhanced expression of three cytochrome P450 genes in transgenic Drosophila melanogaster.0 aglycine insertion in the estrogenrelated receptor ERR is associa c2015 ae01187790 v103 aInsecticide-resistant Drosophila melanogaster strains represent a resource for the discovery of the underlying molecular mechanisms of cytochrome P450 constitutive over-expression, even if some of these P450s are not directly involved in the resistance phenotype. For example, in select 4,4'-dichlorodiphenyltrichloroethane (DDT) resistant strains the glucocorticoid receptor-like (GR-like) potential transcription factor binding motifs (TFBMs) have previously been shown to be associated with constitutively differentially-expressed cytochrome P450s, Cyp12d1, Cyp6g2 and Cyp9c1. However, insects are not known to have glucocorticoids. The only ortholog to the mammalian glucocorticoid receptor (GR) in D. melanogaster is an estrogen-related receptor (ERR) gene, which has two predicted alternative splice isoforms (ERRa and ERRb). Sequencing of ERRa and ERRb in select DDT susceptible and resistant D. melanogaster strains has revealed a glycine (G) codon insertion which was only observed in the ligand binding domain of ERR from the resistant strains tested (ERR-G). Transgenic flies, expressing the ERRa-G allele, constitutively over-expressed Cyp12d1, Cyp6g2 and Cyp9c1. Only Cyp12d1 and Cyp6g2 were over-expressed in the ERRb-G transgenic flies. Phylogenetic studies show that the G-insertion appeared to be located in a less conserved domain in ERR and this insertion is found in multiple species across the Sophophora subgenera.
10aAmino Acid Sequence10aAnimals10aAnimals, Genetically Modified10aConserved Sequence10aCytochrome P-450 Enzyme System10aDrosophila melanogaster10aDrosophila Proteins10aEnzyme Induction10aFemale10aGene Expression10aGlycine10aMale10aMolecular Sequence Data10aMutagenesis, Insertional10aPhylogeny10aReceptors, Estrogen1 aSun, Weilin1 aValero, Carmen1 aSeong, Keon, Mook1 aSteele, Laura, D1 aHuang, I-Ting1 aLee, Chien-Hui1 aClark, John, M1 aQiu, Xinghui1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/glycine-insertion-estrogen-related-receptor-err-associated-enhanced00944nas a2200277 4500008004100000245010200041210006900143260003900212300001300251490000700264100002600271700002300297700002700320700002600347700002600373700002500399700002500424700002800449700002300477700002000500700002100520700002900541700002200570700002500592856004900617 2015 eng d00aHuman and Murine Clonal CD8+ T Cell Expansions Arise during Tuberculosis Because of TCR Selection0 aHuman and Murine Clonal CD8 T Cell Expansions Arise during Tuber bPublic Library of Science ({PLoS}) ae10048490 v111 aNunes-Alves, Cláudio1 aBooty, Matthew, G.1 aCarpenter, Stephen, M.1 aRothchild, Alissa, C.1 aMartin, Constance, J.1 aDesjardins, Danielle1 aSteblenko, Katherine1 aKløverpris, Henrik, N.1 aMadansein, Rajhmun1 aRamsuran, Duran1 aLeslie, Alasdair1 aCorreia-Neves, Margarida1 aBehar, Samuel, M.1 aLewinsohn, David, M. uhttps://doi.org/10.1371/journal.ppat.100484900603nas a2200157 4500008004100000245014500041210006900186260005100255300001600306490000800322100001800330700001900348700001800367700001500385856004500400 2015 eng d00aImpact of Notch1 Deletion in Macrophages on Proinflammatory Cytokine Production and the Outcome of Experimental Autoimmune Encephalomyelitis0 aImpact of Notch1 Deletion in Macrophages on Proinflammatory Cyto bThe American Association of Immunologistscoct a5337–53460 v1951 aWongchana, W.1 aLawlor, R., G.1 aOsborne, B, A1 aPalaga, T. uhttps://doi.org/10.4049/jimmunol.140177001961nas a2200253 4500008004100000245014300041210006900184260001300253300001100266490000700277520103300284100002601317700002301343700002201366700001601388700001901404700002301423700002201446700002201468700002201490700002301512700001901535856015301554 2015 eng d00aLet-7 microRNAs target the lineage-specific transcription factor PLZF to regulate terminal NKT cell differentiation and effector function.0 aLet7 microRNAs target the lineagespecific transcription factor P c2015 May a517-240 v163 aLethal-7 (let-7) microRNAs (miRNAs) are the most abundant miRNAs in the genome, but their role in developing thymocytes is unclear. We found that let-7 miRNAs targeted Zbtb16 mRNA, which encodes the lineage-specific transcription factor PLZF, to post-transcriptionally regulate PLZF expression and thereby the effector functions of natural killer T cells (NKT cells). Dynamic upregulation of let-7 miRNAs during the development of NKT thymocytes downregulated PLZF expression and directed their terminal differentiation into interferon-γ (IFN-γ)-producing NKT1 cells. Without upregulation of let-7 miRNAs, NKT thymocytes maintained high PLZF expression and terminally differentiated into interleukin 4 (IL-4)-producing NKT2 cells or IL-17-producing NKT17 cells. Upregulation of let-7 miRNAs in developing NKT thymocytes was signaled by IL-15, vitamin D and retinoic acid. Such targeting of a lineage-specific transcription factor by miRNA represents a previously unknown level of developmental regulation in the thymus.
1 aPobezinsky, Leonid, A1 aEtzensperger, Ruth1 aJeurling, Susanna1 aAlag, Amala1 aKadakia, Tejas1 aMcCaughtry, Tom, M1 aKimura, Motoko, Y1 aSharrow, Susan, O1 aGuinter, Terry, I1 aFeigenbaum, Lionel1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/let-7-micrornas-target-the-lineage-specific-transcription-factor-plzf00457nas a2200121 4500008004100000245009500041210006900136260003000205490000600235100003000241700002000271856004400291 2015 eng d00aThe Link between Autoimmunity and Lymphoma: Does NOTCH Signaling Play a Contributing Role?0 aLink between Autoimmunity and Lymphoma Does NOTCH Signaling Play bFrontiers Media {SA}cfeb0 v51 aKuksin, Christina, Arieta1 aMinter, Lisa, M uhttps://doi.org/10.3389/fonc.2015.0005102645nas a2200493 4500008004100000245009700041210006900138260001500207300001000222490000800232520115800240653001201398653001201410653001401422653003101436653002501467653002301492653001801515653002001533653002101553653002401574653002501598653001101623653003001634653001901664653001301683653001601696653000901712653000901721653001601730653001301746653002801759653002701787653001501814653002601829100002601855700002101881700001501902700002001917700001801937700001701955700002901972856015002001 2015 eng d00aMga is essential for the survival of pluripotent cells during peri-implantation development.0 aMga is essential for the survival of pluripotent cells during pe c2015 Jan 1 a31-400 v1423 aThe maintenance and control of pluripotency is of great interest in stem cell biology. The dual specificity T-box/basic-helix-loop-helix-zipper transcription factor Mga is expressed in the pluripotent cells of the inner cell mass (ICM) and epiblast of the peri-implantation mouse embryo, but its function has not been investigated previously. Here, we use a loss-of-function allele and RNA knockdown to demonstrate that Mga depletion leads to the death of proliferating pluripotent ICM cells in vivo and in vitro, and the death of embryonic stem cells (ESCs) in vitro. Additionally, quiescent pluripotent cells lacking Mga are lost during embryonic diapause. Expression of Odc1, the rate-limiting enzyme in the conversion of ornithine into putrescine in the synthesis of polyamines, is reduced in Mga mutant cells, and the survival of mutant ICM cells as well as ESCs is rescued in culture by the addition of exogenous putrescine. These results suggest a mechanism whereby Mga influences pluripotent cell survival through regulation of the polyamine pool in pluripotent cells of the embryo, whether they are in a proliferative or quiescent state.
10aAlleles10aAnimals10aApoptosis10aBlastocyst Inner Cell Mass10aCell Differentiation10aCell Proliferation10aCell Survival10aCells, Cultured10aCrosses, Genetic10aEmbryo Implantation10aEmbryonic Stem Cells10aFemale10aGene Knockdown Techniques10aGene Targeting10aGenotype10aGerm Layers10aMale10aMice10aMutagenesis10aMutation10aOrnithine Decarboxylase10aPluripotent Stem Cells10aPolyamines10aTranscription Factors1 aWashkowitz, Andrew, J1 aSchall, Caroline1 aZhang, Kun1 aWurst, Wolfgang1 aFloss, Thomas1 aMager, Jesse1 aPapaioannou, Virginia, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mga-is-essential-for-the-survival-of-pluripotent-cells-during-peri03110nas a2200433 4500008004100000245016900041210006900210260001600279300001100295490000800306520167400314653002501988653001202013653002102025653001802046653001402064653001102078653004602089653002702135653000902162653002602171653004402197653001402241653000902255653002502264653002002289653002502309653001902334653002402353653001802377100002002395700001602415700001802431700001602449700001902465700002402484700001902508856014902527 2015 eng d00aMicroarray analysis of neonatal rat anteroventral periventricular transcriptomes identifies the proapoptotic Cugbp2 gene as sex-specific and regulated by estradiol.0 aMicroarray analysis of neonatal rat anteroventral periventricula c2015 Sep 10 a312-220 v3033 aSexually dimorphic neural structures regulate numerous gender-specific functions including luteinizing hormone (LH) release patterns. The female cyclic surge pattern of release is controlled by the anteroventral periventricular nucleus (AVPV), a preoptic area (POA) region that is significantly smaller in males. The prevailing hypothesis used to explain these differences in structure and function is that a "default" feminine AVPV is defeminized by exposure to estradiol (E2), a metabolite of testosterone (T) produced by the perinatal testes. E2 exposure then culminates in apoptosis in the male AVPV, but the upstream pathways are poorly understood. To address this issue, we compared AVPV transcriptomes of postnatal day 2 (PND2) males and females with those of females treated with E2 or vehicle. Only six of 89 sex-specific genes were also regulated by E2 in the PND2 AVPV and E2 regulated over 280 genes not found to be sex-specific. Of targets that changed similarly in males and E2-treated females, the gene encoding CUG triplet repeat, RNA-binding protein 2 (Cugbp2), a proapoptotic protein, showed the highest fold-changes. Quantitative polymerase chain reaction (QPCR) studies confirmed higher mRNA levels in PND2 male and E2-treated female AVPVs wherein E2 induces apoptosis. POA mapping studies detected Cugbp2 mRNA in the AVPV and in the sexually dimorphic nucleus of the POA (SDN-POA); however, sex differences and E2 effects occurred only in the AVPV. Combined with evidence that Cugbp2 regulates splicing and translation of mRNAs linked to sexual differentiation, we propose that this gene mediates E2-dependent effects on AVPV defeminization.
10aAnalysis of Variance10aAnimals10aAnimals, Newborn10aCELF Proteins10aEstradiol10aFemale10aGene Expression Regulation, Developmental10aHypothalamus, Anterior10aMale10aNerve Tissue Proteins10aOligonucleotide Array Sequence Analysis10aPregnancy10aRats10aRats, Sprague-Dawley10aReceptors, GABA10aReceptors, Glutamate10aRNA, Messenger10aSex Differentiation10aTranscriptome1 aSans, Del, Pino1 aKrishnan, S1 aAggison, L, K1 aAdams, H, L1 aShrikant, M, M1 aLópez-Giráldez, F1 aPetersen, S, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/microarray-analysis-of-neonatal-rat-anteroventral-periventricular01880nas a2200169 4500008004100000245006400041210006300105260001500168520128200183100001301465700002001478700001801498700002001516700001501536700001701551856014201568 2015 ENG d00aNop2 is required for mammalian preimplantation development.0 aNop2 is required for mammalian preimplantation development c2015 Dec 33 aNucleolar protein 2 (NOP2) is evolutionarily conserved from yeast to human, and has been found to play an important role in accelerating cell proliferation, cell-cycle progression, and tumor aggressiveness. The expression pattern and function of Nop2 during early mammalian embryo development, however, has not been investigated. We identified Nop2 as an essential gene for development to the blastocyst stage while performing an RNA interference (RNAi)-based screen in mouse preimplantation embryos. Nop2 is expressed throughout preimplantation development, with highest mRNA and protein accumulation at the 8-cell and morula stages, respectively. RNAi-mediated knockdown of Nop2 results in embryos that arrest as morula. NOP2-deficient embryos exhibit reduced blastomere numbers, greatly increased apoptosis, and impaired cell-lineage specification. Furthermore, knockdown of Nop2 results in global reduction of all RNA species, including rRNA, small nuclear RNA, small nucleolar RNA, and mRNA. Taken together, our results demonstrate that Nop2 is an essential gene for blastocyst formation, and is required for RNA processing and/or stability in vivo during preimplantation embryo development in the mouse. This article is protected by copyright. All rights reserved.
1 aCui, Wei1 aPizzollo, Jason1 aHan, Zhengbin1 aMarcho, Chelsea1 aZhang, Kun1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/nop2-is-required-for-mammalian-preimplantation-development02180nas a2200313 4500008004100000022001400041245007400055210006900129260001300198300001000211490000700221520122000228653001201448653003501460653001101495653002201506653001901528653001201547653001401559653002301573653001001596653001901606100002201625700001501647700001901662700001901681700002001700856014601720 2015 eng d a1879-024000aOdorant receptor-based discovery of natural repellents of human lice.0 aOdorant receptorbased discovery of natural repellents of human l c2015 Nov a103-90 v663 aThe body louse, Pediculus humanus humanus, is an obligate blood-feeding ectoparasite and an important insect vector that mediates the transmission of diseases to humans. The analysis of the body louse genome revealed a drastic reduction of the chemosensory gene repertoires when compared to other insects, suggesting specific olfactory adaptations to host specialization and permanent parasitic lifestyle. Here, we present for the first time functional evidence for the role of odorant receptors (ORs) in this insect, with the objective to gain insight into the chemical ecology of this vector. We identified seven putative full-length ORs, in addition to the odorant receptor co-receptor (Orco), and expressed four of them in the Xenopus laevis oocytes system. When screened with a panel of ecologically-relevant odorants, PhumOR2 responded to a narrow set of compounds. At the behavior level, both head and body lice were repelled by the physiologically-active chemicals. This study presents the first evidence of the OR pathway being functional in lice and identifies PhumOR2 as a sensitive receptor of natural repellents that could be used to develop novel efficient molecules to control these insects.
10aAnimals10aElectrophysiological Phenomena10aFemale10aInsect Repellents10aInsect Vectors10aOocytes10aPediculus10aReceptors, Odorant10aSmell10aXenopus laevis1 aPelletier, Julien1 aXu, Pingxi1 aYoon, Kyong, S1 aClark, John, M1 aLeal, Walter, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/odorant-receptor-based-discovery-natural-repellents-human-lice00657nas a2200181 4500008004100000245016200041210006900203260003000272100002200302700001300324700001400337700001400351700001600365700001700381700001700398700001700415856004300432 2015 eng d00aOptimized Protocols for In Vitro Maturation of Rat Oocytes Dramatically Improve Their Developmental Competence to a Level Similar to That of Ovulated Oocytes0 aOptimized Protocols for In Vitro Maturation of Rat Oocytes Drama bMary Ann Liebert Inccdec1 aJiao, Guang-Zhong1 aCui, Wei1 aYang, Rui1 aLin, Juan1 aGong, Shuai1 aLian, Hua-Yu1 aSun, Ming-Ju1 aTan, Jing-He uhttps://doi.org/10.1089/cell.2015.005503157nas a2200469 4500008004100000245010600041210006900147260001600216300001100232490000800243520166400251653002201915653001101937653001201948653001401960653002101974653004101995653001502036653003102051653001602082653000902098653000902107653002402116653002302140653002102163653002902184653002002213653002402233653002302257653001602280100002002296700002502316700002602341700002802367700002602395700002502421700002602446700002302472700001702495700002402512856015102536 2015 eng d00aPKA-dependent phosphorylation of LIMK1 and Cofilin is essential for mouse sperm acrosomal exocytosis.0 aPKAdependent phosphorylation of LIMK1 and Cofilin is essential f c2015 Sep 15 a237-490 v4053 aMammalian sperm must acquire their fertilizing ability after a series of biochemical modifications in the female reproductive tract collectively called capacitation to undergo acrosomal exocytosis, a process that is essential for fertilization. Actin dynamics play a central role in controlling the process of exocytosis in somatic cells as well as in sperm from several mammalian species. In somatic cells, small GTPases of the Rho family are widely known as master regulators of actin dynamics. However, the role of these proteins in sperm has not been studied in detail. In the present work we characterized the participation of small GTPases of the Rho family in the signaling pathway that leads to actin polymerization during mouse sperm capacitation. We observed that most of the proteins of this signaling cascade and their effector proteins are expressed in mouse sperm. The activation of the signaling pathways of cAMP/PKA, RhoA/C and Rac1 is essential for LIMK1 activation by phosphorylation on Threonine 508. Serine 3 of Cofilin is phosphorylated by LIMK1 during capacitation in a transiently manner. Inhibition of LIMK1 by specific inhibitors (BMS-3) resulted in lower levels of actin polymerization during capacitation and a dramatic decrease in the percentage of sperm that undergo acrosomal exocytosis. Thus, we demonstrated for the first time that the master regulators of actin dynamics in somatic cells are present and active in mouse sperm. Combining the results of our present study with other results from the literature, we have proposed a working model regarding how LIMK1 and Cofilin control acrosomal exocytosis in mouse sperm.
10aAcrosome Reaction10aActins10aAnimals10aCofilin 110aCrosses, Genetic10aCyclic AMP-Dependent Protein Kinases10aExocytosis10aGene Expression Regulation10aLim Kinases10aMale10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aMice, Transgenic10aMicroscopy, Fluorescence10aPhosphorylation10aSignal Transduction10aSperm Capacitation10aSpermatozoa1 aRomarowski, Ana1 aBattistone, Maria, A1 aLa Spina, Florenza, A1 aMolina, Lis, Del C Puga1 aLuque, Guillermina, M1 aVitale, Alejandra, M1 aCuasnicu, Patricia, S1 aVisconti, Pablo, E1 aKrapf, Dario1 aBuffone, Mariano, G uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pka-dependent-phosphorylation-of-limk1-and-cofilin-is-essential-for01708nas a2200205 4500008004100000245012100041210006900162260001500231300001100246490000700257520095100264100001901215700002801234700001301262700001801275700002101293700001701314700002001331856015101351 2015 eng d00aPolyamide Nanogels from Generally Recognized as Safe Components and Their Toxicity in Mouse Preimplantation Embryos.0 aPolyamide Nanogels from Generally Recognized as Safe Components c2015 Nov 9 a3491-80 v163 aSafe delivery systems that can not only encapsulate hydrophobic drug molecules, but also release them in response to specific triggers are important in several therapeutic and biomedical applications. In this paper, we have designed a nanogel based on molecules that are generally recognized as safe (GRAS). We have shown that the resultant polymeric nanogels exhibit responsive molecular release and also show high in vitro cellular viability on HEK 293T, HeLa, MCF 7, and A549 cell lines. The toxicity of these nanogels was further evaluated with a highly sensitive assay using mouse preimplantation embryo development, where blastocysts were formed after 4 days of in vitro culture, and live pups were born when morulae/early blastocysts were transferred to the uteri of surrogate recipients. Our results indicate that these nanogels are nontoxic during mammalian development and do not alter normal growth or early embryo success rate.
1 aPrasad, Priyaa1 aMolla, Mijanur, Rahaman1 aCui, Wei1 aCanakci, Mine1 aOsborne, Barbara1 aMager, Jesse1 aThayumanavan, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/polyamide-nanogels-from-generally-recognized-as-safe-components-and01712nas a2200217 4500008004100000245009400041210006900135260001600204300001200220490000700232520096100239100001401200700001601214700002001230700001701250700001801267700002001285700001601305700002401321856014901345 2015 eng d00aProstate tumorigenesis induced by PTEN deletion involves estrogen receptor β repression.0 aProstate tumorigenesis induced by PTEN deletion involves estroge c2015 Mar 31 a1982-910 v103 aThe role of ERβ in prostate cancer is unclear, although loss of ERβ is associated with aggressive disease. Given that mice deficient in ERβ do not develop prostate cancer, we hypothesized that ERβ loss occurs as a consequence of tumorigenesis caused by other oncogenic mechanisms and that its loss is necessary for tumorigenesis. In support of this hypothesis, we found that ERβ is targeted for repression in prostate cancer caused by PTEN deletion and that loss of ERβ is important for tumor formation. ERβ transcription is repressed by BMI-1, which is induced by PTEN deletion and important for prostate tumorigenesis. This finding provides a mechanism for how ERβ expression is regulated in prostate cancer. Repression of ERβ contributes to tumorigenesis because it enables HIF-1/VEGF signaling that sustains BMI-1 expression. These data reveal a positive feedback loop that is activated in response to PTEN loss and sustains BMI-1.
1 aMak, Paul1 aLi, Jiarong1 aSamanta, Sanjoy1 aChang, Cheng1 aJerry, Joseph1 aDavis, Roger, J1 aLeav, Irwin1 aMercurio, Arthur, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/prostate-tumorigenesis-induced-by-pten-deletion-involves-estrogen02111nas a2200361 4500008004100000022001400041245009400055210006900149260001600218300001200234490000700246520096300253653001201216653002101228653003601249653002701285653004301312653001101355653000901366653000901375653002401384653002601408653002401434100001401458700001601472700002001488700001701508700001801525700002001543700001601563700002401579856014601603 2015 eng d a2211-124700aProstate tumorigenesis induced by PTEN deletion involves estrogen receptor β repression.0 aProstate tumorigenesis induced by PTEN deletion involves estroge c2015 Mar 31 a1982-910 v103 aThe role of ERβ in prostate cancer is unclear, although loss of ERβ is associated with aggressive disease. Given that mice deficient in ERβ do not develop prostate cancer, we hypothesized that ERβ loss occurs as a consequence of tumorigenesis caused by other oncogenic mechanisms and that its loss is necessary for tumorigenesis. In support of this hypothesis, we found that ERβ is targeted for repression in prostate cancer caused by PTEN deletion and that loss of ERβ is important for tumor formation. ERβ transcription is repressed by BMI-1, which is induced by PTEN deletion and important for prostate tumorigenesis. This finding provides a mechanism for how ERβ expression is regulated in prostate cancer. Repression of ERβ contributes to tumorigenesis because it enables HIF-1/VEGF signaling that sustains BMI-1 expression. These data reveal a positive feedback loop that is activated in response to PTEN loss and sustains BMI-1.
10aAnimals10aCell Line, Tumor10aCell Transformation, Neoplastic10aEstrogen Receptor beta10aGene Expression Regulation, Neoplastic10aHumans10aMale10aMice10aProstatic Neoplasms10aPTEN Phosphohydrolase10aSignal Transduction1 aMak, Paul1 aLi, Jiarong1 aSamanta, Sanjoy1 aChang, Cheng1 aJerry, Joseph1 aDavis, Roger, J1 aLeav, Irwin1 aMercurio, Arthur, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/prostate-tumorigenesis-induced-pten-deletion-involves-estrogen02794nas a2200289 4500008004100000022001400041245018600055210006900241260000900310300001300319490000700332520176400339653001202103653000802115653002802123653001102151653002702162100002102189700001702210700001902227700001602246700002202262700002102284700001902305700002602324856015402350 2015 eng d a1932-620300aSelective sweep analysis in the genomes of the 91-R and 91-C Drosophila melanogaster strains reveals few of the 'usual suspects' in dichlorodiphenyltrichloroethane (DDT) resistance.0 aSelective sweep analysis in the genomes of the 91R and 91C Droso c2015 ae01230660 v103 aAdaptation of insect phenotypes for survival after exposure to xenobiotics can result from selection at multiple loci with additive genetic effects. To the authors' knowledge, no selective sweep analysis has been performed to identify such loci in highly dichlorodiphenyltrichloroethane (DDT) resistant insects. Here we compared a highly DDT resistant phenotype in the Drosophila melanogaster (Drosophila) 91-R strain to the DDT susceptible 91-C strain, both of common origin. Whole genome re-sequencing data from pools of individuals was generated separately for 91-R and 91-C, and mapped to the reference Drosophila genome assembly (v. 5.72). Thirteen major and three minor effect chromosome intervals with reduced nucleotide diversity (π) were identified only in the 91-R population. Estimates of Tajima's D (D) showed corresponding evidence of directional selection in these same genome regions of 91-R, however, no similar reductions in π or D estimates were detected in 91-C. An overabundance of non-synonymous proteins coding to synonymous changes were identified in putative open reading frames associated with 91-R. Except for NinaC and Cyp4g1, none of the identified genes were the 'usual suspects' previously observed to be associated with DDT resistance. Additionally, up-regulated ATP-binding cassette transporters have been previously associated with DDT resistance; however, here we identified a structurally altered MDR49 candidate resistance gene. The remaining fourteen genes have not previously been shown to be associated with DDT resistance. These results suggest hitherto unknown mechanisms of DDT resistance, most of which have been overlooked in previous transcriptional studies, with some genes having orthologs in mammals.
10aAnimals10aDDT10aDrosophila melanogaster10aGenome10aInsecticide Resistance1 aSteele, Laura, D1 aCoates, Brad1 aValero, Carmen1 aSun, Weilin1 aSeong, Keon, Mook1 aMuir, William, M1 aClark, John, M1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/selective-sweep-analysis-genomes-91-r-and-91-c-drosophila-melanogaster02645nas a2200169 4500008004100000245007700041210006900118260000900187300000700196490000600203520203800209100002002247700002302267700002602290700001702316856014202333 2015 eng d00aTissue-specific regulation of Igf2r/Airn imprinting during gastrulation.0 aTissuespecific regulation of Igf2rAirn imprinting during gastrul c2015 a100 v83 aBACKGROUND: Appropriate epigenetic regulation of gene expression during lineage allocation and tissue differentiation is required for normal development. One example is genomic imprinting, which is defined as parent-of-origin mono-allelic gene expression. Imprinting is established largely due to epigenetic differences arriving in the zygote from sperm and egg haploid genomes. In the mouse, there are approximately 150 known imprinted genes, many of which occur in imprinted gene clusters that are regulated together. One imprinted cluster includes the maternally expressed Igf2r, Slc22a2, and Slc22a3 genes and the paternally expressed long non-coding RNA (lncRNA) Airn. Although it is known that Igf2r and Airn are reciprocally imprinted, the timing of imprinted expression and accompanying epigenetic changes have not been well characterized in vivo. RESULTS: Here we show lineage- and temporal-specific regulation of DNA methylation and histone modifications at the Igf2r/Airn locus correlating with differential establishment of imprinted expression during gastrulation. Our results show that Igf2r is expressed from both alleles in the E6.5 epiblast. After gastrulation commences, the locus becomes imprinted in the embryonic lineage with the lncRNA Airn expressed from the paternal allele and Igf2r restricted to maternal allele expression. We document differentially enriched allele-specific histone modifications in extraembryonic and embryonic tissues. We also document for the first time allele-specific spreading of DNA methylation during gastrulation concurrent with establishment of imprinted expression of Igf2r. Importantly, we show that imprinted expression does not change in the extraembryonic lineage even though maternal DMR2 methylation spreading does occur, suggesting distinct mechanisms at play in embryonic and extraembryonic lineages. CONCLUSIONS: These results indicate that similar to preimplantation, gastrulation represents a window of dynamic lineage-specific epigenetic regulation in vivo.
1 aMarcho, Chelsea1 aBevilacqua, Ariana1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/tissue-specific-regulation-of-igf2r-airn-imprinting-during02315nas a2200229 4500008004100000245009400041210006900135260001500204300001100219490000800230520150000238100002001738700002401758700002201782700002501804700001501829700002401844700002101868700002301889700002501912856014801937 2015 eng d00aUrinary melatonin concentration and the risk of breast cancer in Nurses' Health Study II.0 aUrinary melatonin concentration and the risk of breast cancer in c2015 Feb 1 a155-620 v1813 aExperimental and epidemiologic data support a protective role for melatonin in breast cancer etiology, yet studies in premenopausal women are scarce. In a case-control study nested within the Nurses' Health Study II cohort, we measured the concentration of melatonin's major urinary metabolite, 6-sulfatoxymelatonin (aMT6s), in urine samples collected between 1996 and 1999 among 600 breast cancer cases and 786 matched controls. Cases were predominantly premenopausal women who were diagnosed with incident breast cancer after urine collection and before June 1, 2007. Using multivariable conditional logistic regression, we computed odds ratios and 95% confidence intervals. Melatonin levels were not significantly associated with total breast cancer risk (for the fourth (top) quartile (Q4) of aMT6s vs. the first (bottom) quartile (Q1), odds ratio (OR) = 0.91, 95% confidence interval (CI): 0.64, 1.28; Ptrend = 0.38) or risk of invasive or in situ breast cancer. Findings did not vary by body mass index, smoking status, menopausal status, or time between urine collection and diagnosis (all Pinteraction values ≥ 0.12). For example, the odds ratio for total breast cancer among women with ≤5 years between urine collection and diagnosis was 0.74 (Q4 vs. Q1; 95% CI: 0.45, 1.20; Ptrend = 0.09), and it was 1.20 (Q4 vs. Q1; 95% CI: 0.72, 1.98; Ptrend = 0.70) for women with >5 years. Our data do not support an overall association between urinary melatonin levels and breast cancer risk.1 aBrown, Susan, B1 aHankinson, Susan, E1 aEliassen, Heather1 aReeves, Katherine, W1 aQian, Jing1 aArcaro, Kathleen, F1 aWegrzyn, Lani, R1 aWillett, Walter, C1 aSchernhammer, Eva, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/urinary-melatonin-concentration-and-the-risk-of-breast-cancer-in00570nas a2200169 4500008004100000245008800041210006900129260002300198300001200221490000800233100002000241700002200261700002200283700002500305700002300330856004700353 2015 eng d00aUsing Xenopus to discover new genes involved in branchiootorenal spectrum disorders0 aUsing Xenopus to discover new genes involved in branchiootorenal bElsevier {BV}cdec a16–240 v1781 aMoody, Sally, A1 aNeilson, Karen, M1 aKenyon, Kristy, L1 aAlfandari, Dominique1 aPignoni, Francesca uhttps://doi.org/10.1016/j.cbpc.2015.06.00701963nas a2200157 4500008004100000245008900041210006900130260001600199520132800215100002001543700002201563700002201585700002501607700002301632856015001655 2015 ENG d00aUsing Xenopus to discover new genes involved in Branchiootorenal spectrum disorders.0 aUsing Xenopus to discover new genes involved in Branchiootorenal c2015 Jun 243 aCongenital hearing loss is an important clinical problem because, without early intervention, affected children do not properly acquire language and consequently have difficulties developing social skills. Although most newborns in the US are screened for hearing deficits, even earlier diagnosis can be made with prenatal genetic screening. Genetic screening that identifies the relevant mutated gene can also warn about potential congenital defects in organs not related to hearing. We will discuss efforts to identify new candidate genes that underlie the Branchiootorenal spectrum disorders in which affected children have hearing deficits and are also at risk for kidney defects. Mutations in two genes, SIX1 and EYA1, have been identified in about half of the patients tested. To uncover new candidate genes, we have used the aquatic animal model, Xenopus laevis, to identify genes that are part of the developmental genetic pathway of Six1 during otic and kidney development. We have already identified a large number of potential Six1 transcriptional targets and candidate co-factor proteins that are expressed at the right time and in the correct tissues to interact with Six1 during development. We discuss the advantages of using this system for gene discovery in a human congenital hearing loss syndrome.
1 aMoody, Sally, A1 aNeilson, Karen, M1 aKenyon, Kristy, L1 aAlfandari, Dominique1 aPignoni, Francesca uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/using-xenopus-to-discover-new-genes-involved-in-branchiootorenal-002091nas a2200301 4500008004100000022001400041245010200055210006900157260001300226300001100239490000800250520111400258653001201372653002101384653001901405653001101424653002101435653002701456653001701483653001401500653001501514100002001529700002101549700001901570700002001589700002601609856015401635 2015 eng d a1095-993900aUtilization of the human louse genome to study insecticide resistance and innate immune response.0 aUtilization of the human louse genome to study insecticide resis c2015 May a125-320 v1203 aSince sequencing the human body louse genome, substantial advances have occurred in the utilization of the information gathered from louse genomes and transcriptomes. Comparatively, the body louse genome contains far fewer genes involved in environmental response, such as xenobiotic detoxification and innate immune response. Additionally, the body louse maintains a primary bacterial endosymbiont, Candidatus Riesia pediculicola, and a number of bacterial pathogens that it vectors, which have genomes that are also reduced in size. Thus, human louse genomes offer unique information and tools for use in advancing our understanding of coevolution among vectors, endosymbionts and pathogens. In this review, we summarize the current literature on the extent of pediculicide resistance, the availability of new pediculicides and information establishing this organism as an efficient model to study how xenobiotic metabolism, which is involved in insecticide resistance, is induced and how insects modify their innate immune response upon bacterial challenge resulting in enhanced vector competence.
10aAnimals10aBiological Assay10aGenome, Insect10aHumans10aImmunity, Innate10aInsecticide Resistance10aInsecticides10aPediculus10aPyrethrins1 aClark, Marshall1 aYoon, Kyong, Sup1 aKim, Ju, Hyeon1 aLee, Si, Hyeock1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/utilization-human-louse-genome-study-insecticide-resistance-and-innate01723nas a2200205 4500008004100000245007500041210006900116260001600185300001200201490000800213520097800221100002601199700002401225700002401249700002101273700001901294700002601313700002501339856015301364 2015 eng d00aThe Wnt receptor Frizzled-4 modulates ADAM13 metalloprotease activity.0 aWnt receptor Frizzled4 modulates ADAM13 metalloprotease activity c2015 Mar 15 a1139-490 v1283 aCranial neural crest (CNC) cells are a transient population of stem cells that originate at the border of the neural plate and the epidermis, and migrate ventrally to contribute to most of the facial structures including bones, cartilage, muscles and ganglia. ADAM13 is a cell surface metalloprotease that is essential for CNC cell migration. Here, we show in Xenopus laevis embryos that the Wnt receptor Fz4 binds to the cysteine-rich domain of ADAM13 and negatively regulates its proteolytic activity in vivo. Gain of Fz4 function inhibits CNC cell migration and can be rescued by gain of ADAM13 function. Loss of Fz4 function also inhibits CNC cell migration and induces a reduction of mature ADAM13, together with an increase in the ADAM13 cytoplasmic fragment that is known to translocate into the nucleus to regulate gene expression. We propose that Fz4 associates with ADAM13 during its transport to the plasma membrane to regulate its proteolytic activity.
1 aAbbruzzese, Genevieve1 aGorny, Anne-Kathrin1 aKaufmann, Lilian, T1 aCousin, Hélène1 aKleino, Iivari1 aSteinbeisser, Herbert1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-wnt-receptor-frizzled-4-modulates-adam13-metalloprotease-activity02959nas a2200229 4500008004100000245010200041210006900143260001500212300000800227490000700235520214600242100002202388700001802410700001902428700002502447700002402472700002102496700002202517700002402539700002002563856014602583 2014 ENG d00aActivation of epidermal growth factor receptor is required for Chlamydia trachomatis development.0 aActivation of epidermal growth factor receptor is required for C c2014 Dec 4 a2770 v143 aBackground Chlamydia trachomatis (C. trachomatis) is a clinically significant human pathogen and one of the leading causative agents of sexually transmitted diseases. As obligate intracellular bacteria, C. trachomatis has evolved strategies to redirect the host¿s signaling and resources for its own survival and propagation. Despite the clinical notoriety of Chlamydia infections, the molecular interactions between C. trachomatis and its host cell proteins remain elusive.ResultsIn this study, we focused on the involvement of the host cell epidermal growth factor receptor (EGFR) in C. trachomatis attachment and development. A combination of molecular approaches, pharmacological agents and cell lines were used to demonstrate distinct functional requirements of EGFR in C. trachomatis infection. We show that C. trachomatis increases the phosphorylation of EGFR and of its downstream effectors PLC¿1, Akt and STAT5. While both EGFR and platelet-derived growth factor receptor-ß (PDGFRß) are partially involved in bacterial attachment to the host cell surface, it is only the knockdown of EGFR and not PDGFRß that affects the formation of C. trachomatis inclusions in the host cells. Inhibition of EGFR results in small immature inclusions, and prevents C. trachomatis-induced intracellular calcium mobilization and the assembly of the characteristic F-actin ring at the inclusion periphery. By using complementary approaches, we demonstrate that the coordinated regulation of both calcium mobilization and F-actin assembly by EGFR are necessary for maturation of chlamydial inclusion within the host cells. A particularly important finding of this study is the co-localization of EGFR with the F-actin at the periphery of C. trachomatis inclusion where it may function to nucleate the assembly of signaling protein complexes for cytoskeletal remodeling required for C. trachomatis development.ConclusionCumulatively, the data reported here connect the function of EGFR to C. trachomatis attachment and development in the host cells, and this could lead to new venues for targeting C. trachomatis infections and associated diseases.1 aPatel, Achchhe, L1 aChen, Xiaofei1 aWood, Scott, T1 aStuart, Elizabeth, S1 aArcaro, Kathleen, F1 aMolina, Doris, P1 aPetrovic, Snezana1 aFurdui, Cristina, M1 aTsang, Allen, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/activation-of-epidermal-growth-factor-receptor-is-required-for02089nas a2200205 4500008004100000245012300041210006900164260001300233300001100246490000800257520130300265100002601568700001901594700002401613700002501637700002101662700002201683700002401705856015401729 2014 eng d00aDetermination of free Bisphenol A (BPA) concentrations in breast milk of U.S. women using a sensitive LC/MS/MS method.0 aDetermination of free Bisphenol A BPA concentrations in breast m c2014 Jun a237-430 v1043 aBisphenol A (BPA) is a synthetic, endocrine-disrupting compound. Free BPA has been detected in human samples indicating that humans are internally exposed to estrogenically active BPA. The purpose of this study was to develop a sensitive method to detect free BPA in human breast milk. BPA was isolated from the milk of 21 nursing mothers in the U.S. by solid-phase extraction. It was then derivatized to improve sensitivity and subsequently analyzed by ultra high performance liquid chromatography-tandem mass spectrometry. Free BPA was detected in 62% of the milk samples (≤ 0.22-10.8 ng mL(-1), median 0.68 ng mL(-1), mean 3.13 ng mL(-1)). No statistical difference in BPA concentrations was observed between women with a low or high Body Mass Index (BMI) (<30 (n=11) and>30 (n=10), respectively). However, there was a significant association between BPA concentration and race. Caucasian women had significantly higher levels of free BPA in their breast milk than non-Caucasian women (mean=4.44 (n=14) and 0.52 (n=7), respectively; p<0.05). The difference between races could be attributed to variations in exposure, lifestyle or metabolism and suggests that certain populations should take extra precautions to limit BPA exposure, particularly during pregnancy and lactation.
1 aZimmers, Stephanie, M1 aBrowne, Eva, P1 aO'Keefe, Patrick, W1 aAnderton, Douglas, L1 aKramer, Lawrence1 aReckhow, David, A1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/determination-of-free-bisphenol-a-bpa-concentrations-in-breast-milk-of00930nas a2200157 4500008004100000245003000041210002900071260000900100300001000109490000900119520047100128100002800599700001900627700001800646856010800664 2014 eng d00aDissecting DR3 signaling.0 aDissecting DR3 signaling c2014 a15-220 v11553 aReceptor signaling can be evaluated in multiple ways, including analysis of phosphorylation of downstream molecules and analysis of proteins that are recruited to the receptor upon ligand binding. Majority of studies on the mechanism of DR3 signaling were performed using overexpression systems that can often lead to artifacts. In this chapter we describe how to analyze DR3 downstream events with most attention being paid to endogenous immunoprecipitation.
1 aPobezinskaya, Yelena, L1 aLiu, Zhenggang1 aChoksi, Swati uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/dissecting-dr3-signaling02408nas a2200397 4500008004100000245011500041210006900156260001300225300001200238490000700250520111500257653001501372653001001387653002101397653002001418653001801438653001101456653002801467653001101495653001501506653004201521653001601563653001601579653004001595653003001635653001401665653001601679100002301695700002401718700002401742700002601766700001701792700001801809700002901827856015401856 2014 eng d00aDNA methylation in paired breast epithelial and white blood cells from women undergoing reduction mammoplasty.0 aDNA methylation in paired breast epithelial and white blood cell c2014 Jun a2985-900 v343 aBACKGROUND: The extent to which white blood cell (WBC) DNA methylation provides information on the status of breast epithelial cell DNA is unknown. PATIENTS AND METHODS: We examined the correlation between methylation in Ras-association domain family-1 gene (RASSF1), a tumor-suppressor gene, and methylation in repetitive elements in paired sets of DNA from WBC and breast epithelial cells collected from 32 women undergoing reduction mammoplasty. RESULTS: We observed no evidence of correlation in methylation levels for ALU, long interspersed nuclear element-1 (LINE1) or juxtacentromeric satellite-2 (SAT2) (r=0.02 for LINE1, p=0.98; r=0.28 for ALU, p=0.12; r=0.26 for SAT2, p=0.17) for matched sets of DNA from WBC and breast epithelial cells. Variability in these markers across individuals and in the same tissue was low. Five women had an average methylation level above 5% for RASSF1 in breast epithelial cell DNA; however, average methylation levels in WBC DNA for these women were all below 1%. CONCLUSION: Methylation patterns in WBC DNA did not reflect methylation patterns in the breast.
10aAdolescent10aAdult10aBreast Neoplasms10aDNA Methylation10aDNA, Neoplasm10aFemale10aGenes, Tumor Suppressor10aHumans10aLeukocytes10aLong Interspersed Nucleotide Elements10aMammaplasty10aMiddle Aged10aNeoplasms, Glandular and Epithelial10aPolymerase Chain Reaction10aPrognosis10aYoung Adult1 aSturgeon, Susan, R1 aArcaro, Kathleen, F1 aJohnson, Melissa, A1 aBalasubramanian, Raji1 aZorn, Martha1 aJerry, Joseph1 aSchneider, Sallie, Smith uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/dna-methylation-in-paired-breast-epithelial-and-white-blood-cells-from01574nas a2200217 4500008004100000245010400041210006900145260000900214300000900223490000600232520081900238100001601057700002001073700001901093700001701112700001901129700002401148700002101172700001701193856014601210 2014 ENG d00aEphrinB2 affects apical constriction in Xenopus embryos and is regulated by ADAM10 and flotillin-1.0 aEphrinB2 affects apical constriction in Xenopus embryos and is r c2014 a35160 v53 aThe Eph/ephrin signalling pathways have a critical function in cell adhesion and repulsion, and thus play key roles in various morphogenetic events during development. Here we show that a decrease in ephrinB2 protein causes neural tube closure defects during Xenopus laevis embryogenesis. Such a decrease in ephrinB2 protein levels is observed on the loss of flotillin-1 scaffold protein, a newly identified ephrinB2-binding partner. This dramatic decline in ephrinB2 protein levels on the absence of flotillin-1 expression is specific, and is partly the result of an increased susceptibility to cleavage by the metalloprotease ADAM10. These findings indicate that flotillin-1 regulates ephrinB2 protein levels through ADAM10, and is required for appropriate neural tube morphogenesis in the Xenopus embryo.
1 aJi, Yon, Ju1 aHwang, Yoo-Seok1 aMood, Kathleen1 aCho, Hee-Jun1 aLee, Hyun-Shik1 aWinterbottom, Emily1 aCousin, Hélène1 aDaar, Ira, O uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/ephrinb2-affects-apical-constriction-in-xenopus-embryos-and-is02731nas a2200169 4500008004100000245017000041210006900211260001500280520200700295100002102302700001302323700002002336700001902356700001702375700002002392856014902412 2014 ENG d00aEvidence for the involvement of proline-rich tyrosine kinase 2 in tyrosine phosphorylation downstream of protein kinase A activation during human sperm capacitation.0 aEvidence for the involvement of prolinerich tyrosine kinase 2 in c2014 Sep 13 aSperm capacitation involves an increase in intracellular Ca(2+) concentration as well as in protein kinase A (PKA)-dependent protein tyrosine (Tyr) phosphorylation. Interestingly, in humans, a decrease in extracellular Ca(2+) concentration ([Ca(2+)]e) during capacitation induces an increase in Tyr phosphorylation indicating the complexity of Ca(2+) signaling during this process. In view of this, in the present study we further investigated the Ca(2+)-mediated signaling pathways implicated in Tyr phosphorylation during human sperm capacitation. Results revealed that sperm incubation in a medium without added Ca(2+) (Ө Ca(2+)) increased Tyr phosphorylation but did not modify PKA-mediated phosphorylation. Moreover, inhibition of either PKA or Src family kinase signaling cascades in Ө Ca(2+) downregulated both PKA substrate and Tyr phosphorylations indicating that the [Ca(2+)]e effects on Tyr phosphorylation depend on PKA targets. Inhibition of calmodulin or Ser/Thr protein phosphatase 2B (PP2B) also increased Tyr phosphorylation without affecting PKA-mediated phosphorylation, supporting the potential role of these Ca(2+) downstream effectors in the increase in Tyr phosphorylation observed in Ө Ca(2+). Experiments aimed to identify the kinase responsible for these observations revealed the presence of Proline-rich Tyrosine Kinase 2 (PYK2), a Focal Adhesion Kinase (FAK) family member, in human sperm, and the use of PF431396, a FAK inhibitor, supported the involvement of PYK2 in Tyr phosphorylation downstream of PKA activation. Results also showed that PYK2 was activated in Ө Ca(2+) as well as during capacitation and that PF431396 affected capacitated sperm motility, acrosome reaction, and ability to penetrate both mouse cumulus matrix and zona-free hamster eggs. Together, our observations support PYK2 as an intermediary component of Ca(2+) signaling between PKA-mediated and Tyr phosphorylation that is required for achieving functional human sperm capacitation.
1 aBattistone, M, A1 aAlvau, A1 aSalicioni, A, M1 aVisconti, P, E1 aDa Ros, V, G1 aCuasnicú, P, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evidence-for-the-involvement-of-proline-rich-tyrosine-kinase-2-in02833nas a2200445 4500008004100000022001400041245019100055210006900246260000900315300001100324490000600335520135600341653001201697653002801709653001201737653002301749653002401772653000801796653002801804653002401832653002001856653001101876653001901887653003401906653004201940653000901982653001301991653002402004653003602028100002102064700002102085700002202106700001902128700002402147700001602171700001902187700001702206700002602223856013802249 2014 eng d a1932-620300aGenome-wide sequencing and an open reading frame analysis of dichlorodiphenyltrichloroethane (DDT) susceptible (91-C) and resistant (91-R) Drosophila melanogaster laboratory populations.0 aGenomewide sequencing and an open reading frame analysis of dich c2014 ae985840 v93 aThe Drosophila melanogaster 91-R and 91-C strains are of common origin, however, 91-R has been intensely selected for dichlorodiphenyltrichloroethane (DDT) resistance over six decades while 91-C has been maintained as the non-selected control strain. These fly strains represent a unique genetic resource to understand the accumulation and fixation of mutations under laboratory conditions over decades of pesticide selection. Considerable research has been done to investigate the differential expression of genes associated with the highly DDT resistant strain 91-R, however, with the advent of whole genome sequencing we can now begin to develop an in depth understanding of the genomic changes associated with this intense decades-long xenobiotic selection pressure. Here we present the first whole genome sequencing analysis of the 91-R and 91-C fly strains to identify genome-wide structural changes within the open reading frames. Between-strain changes in allele frequencies revealed a higher percent of new alleles going to fixation for the 91-R strain, as compared to 91-C (P<0.0001). These results suggest that resistance to DDT in the 91-R laboratory strain could potentially be due primarily to new mutations, as well as being polygenic rather than the result of a few major mutations, two hypotheses that remain to be tested.
10aAlleles10aAmino Acid Substitution10aAnimals10aChromosome Mapping10aChromosomes, Insect10aDDT10aDrosophila melanogaster10aDrosophila Proteins10aDrug Resistance10aFemale10aGenome, Insect10aGenome-Wide Association Study10aHigh-Throughput Nucleotide Sequencing10aMale10aMutation10aOpen Reading Frames10aPolymorphism, Single Nucleotide1 aSteele, Laura, D1 aMuir, William, M1 aSeong, Keon, Mook1 aValero, Carmen1 aRangesa, Madhumitha1 aSun, Weilin1 aClark, John, M1 aCoates, Brad1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genome-wide-sequencing-and-open-reading-frame-analysis02003nas a2200145 4500008004100000245009900041210006900140260001500209520138600224100002601610700002101636700002601657700002501683856014901708 2014 ENG d00aGSK3 and Polo-like kinase regulate ADAM13 function during cranial neural crest cell migration.0 aGSK3 and Pololike kinase regulate ADAM13 function during cranial c2014 Oct 83 aADAMs are cell surface metalloproteases that control multiple biological processes by cleaving signaling and adhesion molecules. ADAM13 controls Cranial Neural Crest (CNC) cell migration both by cleaving Cadherin-11 to release a promigratory extracellular fragment and by controlling multiple genes' expression via its cytoplasmic domain. The latter activity is regulated by γ-secretase cleavage and the translocation of the cytoplasmic domain into the nucleus. One of the genes regulated by ADAM13, the protease Calpain8, is essential for CNC migration. While the nuclear function of ADAM13 is evolutionarily conserved, it is unclear if the transcriptional regulation is also performed by other ADAMs and how this process may be regulated. We show that ADAM13 function to promote CNC migration is regulated by two phosphorylation events involving GSK3 and Polo-like kinase (Plk). We further show that inhibition of either kinase blocks CNC migration and that the respective phosphomimetic forms of ADAM13 can rescue these inhibitions. However, these phosphorylations are not required for ADAM13 proteolysis of its substrates, γ-secretase cleavage or nuclear translocation of its cytoplasmic domain. Significantly, migration of the CNC can be restored in the absence of Plk phosphorylation by expression of Calpain-8a, pointing to an impaired nuclear activity of ADAM13.
1 aAbbruzzese, Genevieve1 aCousin, Hélène1 aSalicioni, Ana, Maria1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/gsk3-and-polo-like-kinase-regulate-adam13-function-during-cranial03420nas a2200421 4500008004100000022001400041245011700055210006900172260001300241300001000254490000600264520215400270653001502424653001002439653001002449653001202459653001002471653001102481653002102492653001102513653001102524653002802535653001902563653000902582653001302591100002502604700002102629700002302650700002502673700002002698700002402718700001902742700001902761700002002780700002402800700002102824856015302845 2014 eng d a1935-273500aHeterogeneous feeding patterns of the dengue vector, Aedes aegypti, on individual human hosts in rural Thailand.0 aHeterogeneous feeding patterns of the dengue vector Aedes aegypt c2014 Aug ae30480 v83 aBACKGROUND: Mosquito biting frequency and how bites are distributed among different people can have significant epidemiologic effects. An improved understanding of mosquito vector-human interactions would refine knowledge of the entomological processes supporting pathogen transmission and could reveal targets for minimizing risk and breaking pathogen transmission cycles.
METHODOLOGY AND PRINCIPAL FINDINGS: We used human DNA blood meal profiling of the dengue virus (DENV) vector, Aedes aegypti, to quantify its contact with human hosts and to infer epidemiologic implications of its blood feeding behavior. We determined the number of different people bitten, biting frequency by host age, size, mosquito age, and the number of times each person was bitten. Of 3,677 engorged mosquitoes collected and 1,186 complete DNA profiles, only 420 meals matched people from the study area, indicating that Ae. aegypti feed on people moving transiently through communities to conduct daily business. 10-13% of engorged mosquitoes fed on more than one person. No biting rate differences were detected between high- and low-dengue transmission seasons. We estimate that 43-46% of engorged mosquitoes bit more than one person within each gonotrophic cycle. Most multiple meals were from residents of the mosquito collection house or neighbors. People ≤ 25 years old were bitten less often than older people. Some hosts were fed on frequently, with three hosts bitten nine times. Interaction networks for mosquitoes and humans revealed biologically significant blood feeding hotspots, including community marketplaces.
CONCLUSION AND SIGNIFICANCE: High multiple-feeding rates and feeding on community visitors are likely important features in the efficient transmission and rapid spread of DENV. These results help explain why reducing vector populations alone is difficult for dengue prevention and support the argument for additional studies of mosquito feeding behavior, which when integrated with a greater understanding of human behavior will refine estimates of risk and strategies for dengue control.
10aAdolescent10aAdult10aAedes10aAnimals10aChild10aDengue10aFeeding Behavior10aFemale10aHumans10aInsect Bites and Stings10aInsect Vectors10aMale10aThailand1 aHarrington, Laura, C1 aFleisher, Andrew1 aRuiz-Moreno, Diego1 aVermeylen, Francoise1 aWa, Chrystal, V1 aPoulson, Rebecca, L1 aEdman, John, D1 aClark, John, M1 aJones, James, W1 aKitthawee, Sangvorn1 aScott, Thomas, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/heterogeneous-feeding-patterns-dengue-vector-aedes-aegypti-individual02113nas a2200169 4500008004100000245010000041210006900141260001300210490000600223520144300229100002501672700002501697700002001722700002401742700002401766856015301790 2014 ENG d00aHigh-density array analysis of DNA methylation in tamoxifen-resistant breast cancer cell lines.0 aHighdensity array analysis of DNA methylation in tamoxifenresist c2014 Feb0 v93 aRoughly two-thirds of all breast cancers are ERα-positive and can be treated with the antiestrogen, Tamoxifen, however resistance occurs in 33% of women who take the drug for more than 5 y. Aberrant DNA methylation, an epigenetic mechanism that alters gene expression in cancer, is thought to play a role in this resistance. To develop an understanding of Tamoxifen-resistance and identify novel pathways and targets of aberrant methylation, DNA from MCF-7 breast cancer cells and Tamoxifen-resistant derivatives, TMX2-11 and TMX2-28, were analyzed using the Illumina HumanMethylation450 BeadChip. Normalizing against MCF-7 values, ERα-positive TMX2-11 had 4,000 hypermethylated sites and ERα-negative TMX2-28 had over 33,000. Analysis of CpG sites altered in both TMX2-11 and TMX2-28 revealed that the Tamoxifen-resistant cell lines share 3000 hypermethylated and 200 hypomethylated CpGs. ZNF350 and MAGED1, two genes hypermethylated in both cell lines, were examined in greater detail. Treatment with 5-aza-2'deoxycitidine caused a significant reduction in promoter methylation of both ZNF350 and MAGED1 and a corresponding increase in expression in TMX2-28. A similar relationship between methylation and expression was not detected in TMX2-11. Our findings are indicative of the variable responses to methylation-targeted breast cancer therapy and highlight the need for biomarkers that accurately predict treatment outcome.
1 aWilliams, Kristin, E1 aAnderton, Douglas, L1 aLee, Maxwell, P1 aPentecost, Brian, T1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/high-density-array-analysis-of-dna-methylation-in-tamoxifen-resistant03051nas a2200229 4500008004100000245006200041210005900103260000900162300000700171490000600178520231100184100001802495700001902513700001702532700002402549700002502573700001802598700002202616700002202638700002302660856013802683 2014 eng d00aHuman Mammary Tumor Virus (HMTV) sequences in human milk.0 aHuman Mammary Tumor Virus HMTV sequences in human milk c2014 a200 v93 aBACKGROUND: Retroviral sequences 90-95% homologous to the mouse mammary tumor virus (MMTV) were present in 38% of the breast cancers studied from American women and were not detectable in non-tumor breast tissue from the same patient. The entire proviral structure was described and viral particles were isolated from primary cultures of human breast cancer. This virus was designated as human mammary tumor virus (HMTV). Hormone response elements present in the HMTV Long-Terminal-Repeat (LTR) suggest a mechanism for association of HMTV with hormonally responding tissues. In fact, the incidence of HMTV sequences is higher in gestational breast cancers, which are associated with hormonal changes. Milk epithelial cells are also under hormonal regulation and therefore are excellent specimens for HMTV sequence detection. METHODS: The HMTV sequence was studied in milk samples from lactating women recruited with increased risk of breast cancer because they had undergone breast biopsies (Biopsy-Group) and lactating women without breast biopsies (Reference-Group). RESULTS: HMTV-env sequences were detected by PCR in milk of 7.61% of 92 women of the Reference-Group and in 20.55% of 73 women of the Biopsy-Group (p: 0.015). The sequences were 94-98% homologous to MMTV. HMTV-env and HMTV-env/LTR junction sequences were detected in high-speed pellet RNA, implying the presence of HMTV viral particles. PCR assays to detect the murine mitochondrial cytochrome oxidase gene and intracisternal-A-type particle sequences were performed to rule out mouse mitochondrial or genomic DNA contamination. Eight women of the 73 Biopsy-Group participants had breast cancer and the milk of only one of these eight women had HMTV-env sequences. In the remaining 65 women of the Biopsy-Group, under enough clinical suspicion to lead to biopsy, HMTV was detected in 14, nearly three times the number of milks as compared to the Reference-Group (21.54% versus 7.61%; p: 0.016). CONCLUSION: The significance of HMTV in milk from the Reference-Group, the greater frequency in the milk of women who had undergone a breast biopsy and its possible infectivity for infants are important questions under study. The similarity of HMTV to MMTV is striking and suggests one possible avenue for viral transmission in humans.
1 aNartey, Teiko1 aMoran, Heberth1 aMarin, Tania1 aArcaro, Kathleen, F1 aAnderton, Douglas, L1 aEtkind, Polly1 aHolland, James, F1 aMelana, Stella, M1 aPogo, Beatriz, G-T uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/human-mammary-tumor-virus-hmtv-sequences-in-human-milk02115nas a2200205 4500008004100000245009200041210006900133260001500202300001000217490000700227520136800234100002501602700002001627700002001647700002201667700002601689700002301715700001701738856015401755 2014 ENG d00aIdentification of a novel isoform of the leukemia-associated MLLT1 (ENL/LTG19) protein.0 aIdentification of a novel isoform of the leukemiaassociated MLLT c2014 Dec 3 a11-150 v173 aGenome wide transcriptional profiles offer abundant information regarding mRNA levels in specific tissues, organs or developmental stages. Although these data sets do not offer spatial or cell type-specific information, they can be extremely useful for gene discovery when analyzed by the appropriate techniques. Previously, we proposed and validated the use of combinatorial dataset analysis techniques to identify novel genes required during pre-implantation development. Now we build upon this work to identify genes that have dynamic expression during gametogenesis. Here we present detailed analysis of the expression pattern of leukemia-associated, myeloid/lymphoid or mixed-lineage leukemia; translocated to 1 (Mllt1) gene. We document a novel splice isoform of Mllt1 and confirm that both Mllt1 mRNA isoforms are translated. We provide data supporting that MLLT1 protein isoforms display distinct stage-specific expression during spermiogenesis and adult tissues. Finally, we evaluated genes neighboring the Mllt1 locus, and show dynamic stage specific expression patterns of other genes Catsperd, Prr22, Rfx2 and Slc25a41. We document testes expressed alternative isoforms of Prr22 and Rfx2. These results indicate that transcriptome data mining, combined with specific expression analysis provides a wealth of novel gene expression information.
1 aWallingford, Mary, C1 aFilkins, Rachel1 aAdams, Danielle1 aWalentuk, Melanie1 aSalicioni, Ana, Maria1 aVisconti, Pablo, E1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-a-novel-isoform-of-the-leukemia-associated-mllt1-enl02330nas a2200169 4500008004100000245005500041210005400096260001500150520172500165100002701890700002201917700002501939700001901964700002201983700002202005856013302027 2014 ENG d00aImpaired Sperm Maturation in Rnase9 Knockout Mice.0 aImpaired Sperm Maturation in Rnase9 Knockout Mice c2014 Apr 93 aRibonuclease, RNase A family, 9 (RNASE9) is a ribonuclease A superfamily member that is expressed only in the epididymis. It is a small, secreted polypeptide, it lacks ribonuclease activity, and its function(s) is unknown. However, epididymis-specific expression suggests a role in sperm maturation. We generated Rnase9(-/-) mice to study RNASE9 function in vivo. We confirm that RNASE9 expression is restricted to the epididymis. Within the epididymis, RNASE9 is first detected in mid caput, persists through the distal caput and corpus, and wanes in the cauda. Rnase9(-/-) mice are born at the expected Mendelian ratio, have normal post-natal growth and development, and have no outwardly apparent phenotype. Spermatogenesis is normal and Rnase9 null sperm are morphologically normal. Rnase9(-/-) males have normal fertility in unrestricted mating trials and fertilization rates in in vitro fertilization assays are indistinguishable from wild type mice. Visual observations coupled with analyses of sperm velocities shortly after swim out from the corpus shows that motility of Rnase9 null sperm is significantly impaired. However, no differences between wild type and Rnase9 null sperm are detected by computer-assisted sperm analysis 10-90 min after sperm isolation from the corpus or cauda. Assessment of capacitation-dependent signaling pathways in Rnase9 null sperm showed that, while levels of tyrosine phosphorylation of sperm proteins were normal, there was decreased phosphorylation of protein kinase A substrates upon capacitation compared to wild type mice. In conclusion, RNASE9 is dispensable for fertility, but the absence of RNASE9 during epididymal transit results in impaired sperm maturation.
1 aWestmuckett, Andrew, D1 aNguyen, Edward, B1 aHerlea-Pana, Oana, M1 aAlvau, Antonio1 aSalicioni, Ana, M1 aMoore, Kevin, Lee uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/impaired-sperm-maturation-in-rnase9-knockout-mice02005nas a2200169 4500008004100000245010000041210006900141260001600210300001100226490000700237520135200244100002401596700002001620700002401640700002001664856015101684 2014 eng d00aImportance of sequence specific hydrophobicity in synthetic protein transduction domain mimics.0 aImportance of sequence specific hydrophobicity in synthetic prot c2014 Mar 10 a812-200 v153 aA new series of synthetic protein transduction domain mimics (PTDMs) was designed to analyze the importance of guanidine and phenyl group segregation along the backbone on their membrane interaction and cellular internalization abilities. ROMP was utilized to synthesize three polymers: nonsegregated homopolymers, intermediately segregated gradient copolymers, and strongly segregated block copolymers. In order to understand the role of functional group segregation on activity, it was important to design monomers that enabled these three different polymer topologies, or constitutional macromolecular isomers, to be prepared with identical chemical compositions. The structure-activity relationships were evaluated by both a biophysical assay, using dye-loaded vesicles, and by in vitro cellular uptake studies of fluorescently labeled chains. The results showed that functional group segregation impacts activity. In general, the nonsegregated homopolymer was the most active in both assays but also showed larger, ill-defined aggregates compared to either the gradient or block copolymers. It was also the most cytotoxic of the three isomers. As a result, the gradient copolymer with intermediate segregation optimizes activity and solubility with low cytotoxicity. This study gives new design guidelines for the development of PTDMs.
1 aSgolastra, Federica1 aMinter, Lisa, M1 aOsborne, Barbara, A1 aTew, Gregory, N uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/importance-of-sequence-specific-hydrophobicity-in-synthetic-protein00600nas a2200181 4500008004100000245006200041210006200103260004600165300001400211490000700225100002600232700002300258700002700281700002200308700002600330700002200356856004000378 2014 eng d00aIn search of a new paradigm for protective immunity to TB0 aIn search of a new paradigm for protective immunity to TB bSpringer Science and Business Media {LLC} a289–2990 v121 aNunes-Alves, Cláudio1 aBooty, Matthew, G.1 aCarpenter, Stephen, M.1 aJayaraman, Pushpa1 aRothchild, Alissa, C.1 aBehar, Samuel, M. uhttps://doi.org/10.1038/nrmicro323000580nas a2200169 4500008004100000245007100041210006900112260003900181300001300220490000700233100002600240700002200266700002600288700002200314700002500336856004900361 2014 eng d00aiNKT Cell Production of GM-CSF Controls Mycobacterium tuberculosis0 aiNKT Cell Production of GMCSF Controls Mycobacterium tuberculosi bPublic Library of Science ({PLoS}) ae10038050 v101 aRothchild, Alissa, C.1 aJayaraman, Pushpa1 aNunes-Alves, Cláudio1 aBehar, Samuel, M.1 aLewinsohn, David, M. uhttps://doi.org/10.1371/journal.ppat.100380502799nas a2200361 4500008004100000022001400041245011000055210006900165260001300234300001000247490000700257520165700264653001201921653001101933653001901944653002601963653002701989653001702016653001302033653001402046653001502060653002002075653001802095100002102113700002402134700002502158700002002183700001902203700002602222700002002248700002002268856014902288 2014 eng d a0022-258500aKnockdown resistance allele frequencies in North American head louse (Anoplura: Pediculidae) populations.0 aKnockdown resistance allele frequencies in North American head l c2014 Mar a450-70 v513 aThe study examines the extent and frequency of a knockdown-type resistance allele (kdr type) in North American populations of human head lice. Lice were collected from 32 locations in Canada and the United States. DNA was extracted from individual lice and used to determine their zygosity using the serial invasive signal amplification technique to detect the kdr-type T917I (TI) mutation, which is most responsible for nerve insensitivity that results in the kdr phenotype and permethrin resistance. Previously sampled sites were resampled to determine if the frequency of the TI mutation was changing. The TI frequency was also reevaluated using a quantitative sequencing method on pooled DNA samples from selected sites to validate this population genotyping method. Genotyping substantiated that TI occurs at high levels in North American lice (88.4%). Overall, the TI frequency in U.S. lice was 84.4% from 1999 to 2009, increased to 99.6% from 2007 to 2009, and was 97.1% in Canadian lice in 2008. Genotyping results using the serial invasive signal amplification reaction (99.54%) and quantitative sequencing (99.45%) techniques were highly correlated. Thus, the frequencies of TI in North American head louse populations were found to be uniformly high, which may be due to the high selection pressure from the intensive and widespread use of the pyrethrins- or pyrethroid-based pediculicides over many years, and is likely a main cause of increased pediculosis and failure of pyrethrins- or permethrin-based products in Canada and the United States. Alternative approaches to treatment of head lice infestations are critically needed.
10aAnimals10aCanada10aGene Frequency10aGenotyping Techniques10aInsecticide Resistance10aInsecticides10aMutation10aPediculus10aPermethrin10aSodium Channels10aUnited States1 aYoon, Kyong, Sup1 aPrevite, Domenic, J1 aHodgdon, Hilliary, E1 aPoole, Bryan, C1 aKwon, Deok, Ho1 aEl-Ghar, Gamal, E Abo1 aLee, Si, Hyeock1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/knockdown-resistance-allele-frequencies-north-american-head-louse00708nas a2200193 4500008004100000245010600041210006900147260001200216490003300228653001700261100002100278700002400299700002600323700002600349700002400375700002200399700001400421856007900435 2014 eng d00aMeasuring cytotoxicity by bioluminescence imaging outperforms the standard chromium-51 release assay.0 aMeasuring cytotoxicity by bioluminescence imaging outperforms th c02/20140 v10.1371/journal.pone.008935710acytotoxicity1 aKambayashi, Taku1 aBehrens, Edward, M.1 aBachmann, Michael, H.1 aSalicioni, Ana, Maria1 aBaldwin, Cynthia, L1 aKarimi, Mobin, A.1 aLee, Eric uhttp://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.008935702317nas a2200217 4500008004100000245010200041210006900143260000900212300000700221490000600228520154600234100002001780700002301800700002301823700001901846700001701865700001901882700002001901700002401921856015401945 2014 eng d00aNon-Canonical Notch Signaling Drives Activation and Differentiation of Peripheral CD4(+) T Cells.0 aNonCanonical Notch Signaling Drives Activation and Differentiati c2014 a540 v53 aCleavage of the Notch receptor via a γ-secretase, results in the release of the active intra-cellular domain of Notch that migrates to the nucleus and interacts with RBP-Jκ, resulting in the activation of downstream target genes. This canonical Notch signaling pathway has been documented to influence T cell development and function. However, the mechanistic details underlying this process remain obscure. In addition to RBP-Jκ, the intra-cellular domain of Notch also interacts with other proteins in the cytoplasm and nucleus, giving rise to the possibility of an alternate, RBP-Jκ independent Notch pathway. However, the contribution of such RBP-Jκ independent, "non-canonical" Notch signaling in regulating peripheral T cell responses is unknown. In this report, we specifically demonstrate the requirement of Notch1 for regulating signal strength and signaling events distal to the T cell receptor in peripheral CD4(+) T cells. By using mice with a conditional deletion in Notch1 or RBP-Jκ, we show that Notch1 regulates activation and proliferation of CD4(+) T cells independently of RBP-Jκ. Furthermore, differentiation to TH1 and iTreg lineages although Notch dependent, is RBP-Jκ independent. Our striking observations demonstrate that many of the cell-intrinsic functions of Notch occur independently of RBP-Jκ. Such non-canonical regulation of these processes likely occurs through NF-κ B. This reveals a previously unknown, novel role of non-canonical Notch signaling in regulating peripheral T cell responses.
1 aDongre, Anushka1 aSurampudi, Lalitha1 aLawlor, Rebecca, G1 aFauq, Abdul, H1 aMiele, Lucio1 aGolde, Todd, E1 aMinter, Lisa, M1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/non-canonical-notch-signaling-drives-activation-and-differentiation-of02007nas a2200169 4500008004100000245015200041210006900193260001300262300001200275490000700287520130700294100002001601700002101621700002601642700002101668856014801689 2014 eng d00aA nonsynonymous mutation in the transcriptional regulator lbh is associated with cichlid craniofacial adaptation and neural crest cell development.0 anonsynonymous mutation in the transcriptional regulator lbh is a c2014 Dec a3113-240 v313 aSince the time of Darwin, biologists have sought to understand the origins and maintenance of life's diversity of form. However, the nature of the exact DNA mutations and molecular mechanisms that result in morphological differences between species remains unclear. Here, we characterize a nonsynonymous mutation in a transcriptional coactivator, limb bud and heart homolog (lbh), which is associated with adaptive variation in the lower jaw of cichlid fishes. Using both zebrafish and Xenopus, we demonstrate that lbh mediates migration of cranial neural crest cells, the cellular source of the craniofacial skeleton. A single amino acid change that is alternatively fixed in cichlids with differing facial morphologies results in discrete shifts in migration patterns of this multipotent cell type that are consistent with both embryological and adult craniofacial phenotypes. Among animals, this polymorphism in lbh represents a rare example of a coding change that is associated with continuous morphological variation. This work offers novel insights into the development and evolution of the craniofacial skeleton, underscores the evolutionary potential of neural crest cells, and extends our understanding of the genetic nature of mutations that underlie divergence in complex phenotypes.
1 aPowder, Kara, E1 aCousin, Hélène1 aMcLinden, Gretchen, P1 aAlbertson, Craig uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/a-nonsynonymous-mutation-in-the-transcriptional-regulator-lbh-is02597nas a2200265 4500008004100000245011200041210007000153260000900223300000800232490000600240520167600246100001901922700002501941700001801966700002301984700003002007700002502037700001902062700001902081700001702100700001802117700002402135700002002159856015202179 2014 eng d00aNOTCH1 Can Initiate NF-κB Activation via Cytosolic Interactions with Components of the T Cell Signalosome.0 aNOTCH1 Can Initiate NFκB Activation via Cytosolic Interactions w c2014 a2490 v53 aT cell stimulation requires the input and integration of external signals. Signaling through the T cell receptor (TCR) is known to induce formation of the membrane-tethered CBM complex, comprising CARMA1, BCL10, and MALT1, which is required for TCR-mediated NF-κB activation. TCR signaling has been shown to activate NOTCH proteins, transmembrane receptors also implicated in NF-κB activation. However, the link between TCR-mediated NOTCH signaling and early events leading to induction of NF-κB activity remains unclear. In this report, we demonstrate a novel cytosolic function for NOTCH1 and show that it is essential to CBM complex formation. Using a model of skin allograft rejection, we show in vivo that NOTCH1 acts in the same functional pathway as PKCθ, a T cell-specific kinase important for CBM assembly and classical NF-κB activation. We further demonstrate in vitro NOTCH1 associates physically with PKCθ and CARMA1 in the cytosol. Unexpectedly, when NOTCH1 expression was abrogated using RNAi approaches, interactions between CARMA1, BCL10, and MALT1 were lost. This failure in CBM assembly reduced inhibitor of kappa B alpha phosphorylation and diminished NF-κB-DNA binding. Finally, using a luciferase gene reporter assay, we show the intracellular domain of NOTCH1 can initiate robust NF-κB activity in stimulated T cells, even when NOTCH1 is excluded from the nucleus through modifications that restrict it to the cytoplasm or hold it tethered to the membrane. Collectively, these observations provide evidence that NOTCH1 may facilitate early events during T cell activation by nucleating the CBM complex and initiating NF-κB signaling.
1 aShin, Hyun, Mu1 aTilahun, Mulualem, E1 aCho, Ok, Hyun1 aChandiran, Karthik1 aKuksin, Christina, Arieta1 aKeerthivasan, Shilpa1 aFauq, Abdul, H1 aGolde, Todd, E1 aMiele, Lucio1 aThome, Margot1 aOsborne, Barbara, A1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch1-can-initiate-nf-kb-activation-via-cytosolic-interactions-with02824nas a2200457 4500008004100000245009600041210006900137260000900206300000800215490000700223520145900230653001501689653001001704653000901714653002101723653002501744653002101769653001101790653003001801653001101831653001601842653001501858653001101873653001401884653002401898653001701922653001801939653001601957100002101973700001801994700002102012700002102033700003002054700001802084700002102102700002902123700001802152700002202170700002502192856014902217 2014 eng d00aParity-related molecular signatures and breast cancer subtypes by estrogen receptor status.0 aParityrelated molecular signatures and breast cancer subtypes by c2014 aR740 v163 aINTRODUCTION: Relationships of parity with breast cancer risk are complex. Parity is associated with decreased risk of postmenopausal hormone receptor-positive breast tumors, but may increase risk for basal-like breast cancers and early-onset tumors. Characterizing parity-related gene expression patterns in normal breast and breast tumor tissues may improve understanding of the biological mechanisms underlying this complex pattern of risk. METHODS: We developed a parity signature by analyzing microRNA microarray data from 130 reduction mammoplasty (RM) patients (54 nulliparous and 76 parous). This parity signature, together with published parity signatures, was evaluated in gene expression data from 150 paired tumors and adjacent benign breast tissues from the Polish Breast Cancer Study, both overall and by tumor estrogen receptor (ER) status. RESULTS: We identified 251 genes significantly upregulated by parity status in RM patients (parous versus nulliparous; false discovery rate = 0.008), including genes in immune, inflammation and wound response pathways. This parity signature was significantly enriched in normal and tumor tissues of parous breast cancer patients, specifically in ER-positive tumors. CONCLUSIONS: Our data corroborate epidemiologic data, suggesting that the etiology and pathogenesis of breast cancers vary by ER status, which may have implications for developing prevention strategies for these tumors.
10aAdolescent10aAdult10aAged10aBreast Neoplasms10aCase-Control Studies10aCluster Analysis10aFemale10aGene Expression Profiling10aHumans10aMiddle Aged10aOdds Ratio10aParity10aPregnancy10aReceptors, Estrogen10aRisk Factors10aTranscriptome10aYoung Adult1 aRotunno, Melissa1 aSun, Xuezheng1 aFigueroa, Jonine1 aSherman, Mark, E1 aGarcia-Closas, Montserrat1 aMeltzer, Paul1 aWilliams, Tyisha1 aSchneider, Sallie, Smith1 aJerry, Joseph1 aYang, Xiaohong, R1 aTroester, Melissa, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/parity-related-molecular-signatures-and-breast-cancer-subtypes-by02912nas a2200469 4500008004100000022001400041245009600055210006900151260001600220300000800236490000700244520151700251653001501768653001001783653000901793653002101802653002501823653002101848653001101869653003001880653001101910653001601921653001501937653001101952653001401963653002401977653001702001653001802018653001602036100002102052700001802073700002102091700002102112700003002133700001802163700002102181700002902202700001802231700002202249700002502271856014602296 2014 eng d a1465-542X00aParity-related molecular signatures and breast cancer subtypes by estrogen receptor status.0 aParityrelated molecular signatures and breast cancer subtypes by c2014 Jul 08 aR740 v163 aINTRODUCTION: Relationships of parity with breast cancer risk are complex. Parity is associated with decreased risk of postmenopausal hormone receptor-positive breast tumors, but may increase risk for basal-like breast cancers and early-onset tumors. Characterizing parity-related gene expression patterns in normal breast and breast tumor tissues may improve understanding of the biological mechanisms underlying this complex pattern of risk.
METHODS: We developed a parity signature by analyzing microRNA microarray data from 130 reduction mammoplasty (RM) patients (54 nulliparous and 76 parous). This parity signature, together with published parity signatures, was evaluated in gene expression data from 150 paired tumors and adjacent benign breast tissues from the Polish Breast Cancer Study, both overall and by tumor estrogen receptor (ER) status.
RESULTS: We identified 251 genes significantly upregulated by parity status in RM patients (parous versus nulliparous; false discovery rate = 0.008), including genes in immune, inflammation and wound response pathways. This parity signature was significantly enriched in normal and tumor tissues of parous breast cancer patients, specifically in ER-positive tumors.
CONCLUSIONS: Our data corroborate epidemiologic data, suggesting that the etiology and pathogenesis of breast cancers vary by ER status, which may have implications for developing prevention strategies for these tumors.
10aAdolescent10aAdult10aAged10aBreast Neoplasms10aCase-Control Studies10aCluster Analysis10aFemale10aGene Expression Profiling10aHumans10aMiddle Aged10aOdds Ratio10aParity10aPregnancy10aReceptors, Estrogen10aRisk Factors10aTranscriptome10aYoung Adult1 aRotunno, Melissa1 aSun, Xuezheng1 aFigueroa, Jonine1 aSherman, Mark, E1 aGarcia-Closas, Montserrat1 aMeltzer, Paul1 aWilliams, Tyisha1 aSchneider, Sallie, Smith1 aJerry, Joseph1 aYang, Xiaohong, R1 aTroester, Melissa, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/parity-related-molecular-signatures-and-breast-cancer-subtypes02728nas a2200205 4500008004100000245011600041210006900157260001600226520193900242100001902181700002002200700002502220700001502245700001902260700002402279700002502303700002402328700001902352856015102371 2014 ENG d00aPromoter Methylation in Epithelial-Enriched and Epithelial-Depleted Cell Populations Isolated from Breast Milk.0 aPromoter Methylation in EpithelialEnriched and EpithelialDeplete c2014 Aug 273 aBACKGROUND: Breast cancer is the most frequently diagnosed cancer among Turkish women and both the incidence and associated mortality appear to be increasing. Of particular concern is the percentage of young women diagnosed with breast cancer; roughly 20% of all breast cancer diagnoses in Turkey are in women younger than 40 years. Increased DNA methylation in the promoter region of tumor suppressor genes is a promising molecular biomarker, and human milk provides exfoliated breast epithelial cells appropriate for DNA methylation analyses. Comparisons between DNA methylation patterns in epithelial (epithelial-enriched) and nonepithelial (epithelial-depleted) cell fractions from breast milk have not been reported previously. OBJECTIVE: In the present study, we examined promoter methylation of 3 tumor suppressor genes in epithelial-enriched and epithelial-depleted cell fractions isolated from breast milk of 43 Turkish women. METHODS: Percentage methylation in the promoter region of Rass association domain family 1 (RASSF1), secreted frizzle related protein 1 (SFRP1), and glutathione-S-transferase class pi 1 was determined by pyrosequencing of the epithelial-enriched and epithelial-depleted cell fractions. RESULTS: Pyrosequencing identified a few subjects with significantly increased methylation in 1 or more genes. There was little correlation between the 2 cell fractions within individuals; only 1 woman had increased methylation for 1 gene (SFRP1) in both her enriched and depleted cell fractions. Methylation was positively associated with age for SFRP1 (epithelial-depleted fraction) and with body mass index for RASSF1 (epithelial-enriched cell fraction), respectively. CONCLUSION: Overall, results show that the methylation signals vary between different cell types in breast milk and suggest that breast milk can be used to assess DNA methylation patterns associated with increased breast cancer risk.
1 aBrowne, Eva, P1 aDinc, Signem, E1 aPunska, Elizabeth, C1 aAgus, Sami1 aVitrinel, Ayca1 aErdag, Gulay, Ciler1 aAnderton, Douglas, L1 aArcaro, Kathleen, F1 aYilmaz, Bayram uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/promoter-methylation-in-epithelial-enriched-and-epithelial-depleted00617nas a2200181 4500008004100000245010700041210006900148260002500217300001400242490000700256100002000263700001900283700001700302700002000319700002300339700002700362856004600389 2014 eng d00aProtein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters0 aProtein phospholipase C Zeta1 expression in patients with failed bSpringer Naturecapr a749–7560 v311 aLee, Hoi, Chang1 aArny, Margaret1 aGrow, Daniel1 aDumesic, Daniel1 aFissore, Rafael, A1 aJellerette-Nolan, Teru uhttps://doi.org/10.1007/s10815-014-0229-902428nas a2200433 4500008004100000245008700041210006900128260001500197300000900212490000800221520110600229653001201335653002001347653002401367653002201391653001101413653001601424653001301440653004001453653001101493653001601504653000901520653001901529653002401548653003001572653003001602100001801632700002501650700002001675700002001695700001701715700001501732700002101747700002401768700002201792700001701814700001701831856014601848 2014 eng d00aRLIM is dispensable for X-chromosome inactivation in the mouse embryonic epiblast.0 aRLIM is dispensable for Xchromosome inactivation in the mouse em c2014 Jul 3 a86-90 v5113 aIn female mice, two forms of X-chromosome inactivation (XCI) ensure the selective silencing of female sex chromosomes during mouse embryogenesis. Beginning at the four-cell stage, imprinted XCI (iXCI) exclusively silences the paternal X chromosome. Later, around implantation, epiblast cells of the inner cell mass that give rise to the embryo reactivate the paternal X chromosome and undergo a random form of XCI (rXCI). Xist, a long non-coding RNA crucial for both forms of XCI, is activated by the ubiquitin ligase RLIM (also known as Rnf12). Although RLIM is required for triggering iXCI in mice, its importance for rXCI has been controversial. Here we show that RLIM levels are downregulated in embryonic cells undergoing rXCI. Using mouse genetics we demonstrate that female cells lacking RLIM from pre-implantation stages onwards show hallmarks of XCI, including Xist clouds and H3K27me3 foci, and have full embryogenic potential. These results provide evidence that RLIM is dispensable for rXCI, indicating that in mice an RLIM-independent mechanism activates Xist in the embryo proper.
10aAnimals10aDown-Regulation10aEmbryo Implantation10aEmbryo, Mammalian10aFemale10aGerm Layers10aHistones10aIn Situ Hybridization, Fluorescence10aLysine10aMethylation10aMice10aMice, Knockout10aRNA, Long Noncoding10aUbiquitin-Protein Ligases10aX Chromosome Inactivation1 aShin, JongDae1 aWallingford, Mary, C1 aGallant, Judith1 aMarcho, Chelsea1 aJiao, Baowei1 aByron, Meg1 aBossenz, Michael1 aLawrence, Jeanne, B1 aJones, Stephen, N1 aMager, Jesse1 aBach, Ingolf uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/rlim-is-dispensable-for-x-chromosome-inactivation-in-the-mouse00480nas a2200133 4500008004100000245011100041210006900152260002500221300001600246490000800262100001500270700002300285856003800308 2014 eng d00aRole of Caspase-3 Cleaved IP3R1 on Ca2+ Homeostasis and Developmental Competence of Mouse Oocytes and Eggs0 aRole of Caspase3 Cleaved IP3R1 on Ca2 Homeostasis and Developmen bWiley-Blackwellcjul a1842–18540 v2291 aZhang, Nan1 aFissore, Rafael, A uhttps://doi.org/10.1002/jcp.2463800691nas a2200193 4500008004100000245011700041210006900158260004400227300001100271490000600282100002300288700002900311700001800340700002400358700002100382700002300403700002200426856004900448 2014 eng d00aThe Role of Lipolysis Stimulated Lipoprotein Receptor in Breast Cancer and Directing Breast Cancer Cell Behavior0 aRole of Lipolysis Stimulated Lipoprotein Receptor in Breast Canc bPublic Library of Science ({PLoS})cmar ae917470 v91 aReaves, Denise, K.1 aFagan-Solis, Katerina, D1 aDunphy, Karen1 aOliver, Shannon, D.1 aScott, David, W.1 aFleming, Jodie, M.1 aFanning, Alan, S. uhttps://doi.org/10.1371/journal.pone.009174700691nas a2200217 4500008004100000245009600041210006900137260004400206300001100250490000600261100002100267700001300288700001500301700001500316700002000331700001500351700002200366700001700388700001900405856004900424 2014 eng d00aRole of Na+/Ca2+ Exchanger (NCX) in Modulating Postovulatory Aging of Mouse and Rat Oocytes0 aRole of NaCa2 Exchanger NCX in Modulating Postovulatory Aging of bPublic Library of Science ({PLoS})capr ae934460 v91 aZhang, Chuan-Xin1 aCui, Wei1 aZhang, Min1 aZhang, Jie1 aWang, Tian-Yang1 aZhu, Jiang1 aJiao, Guang-Zhong1 aTan, Jing-He1 aSun, Qing-Yuan uhttps://doi.org/10.1371/journal.pone.009344603471nas a2200541 4500008004100000245013200041210006900173260001300242300001100255490000800266520171900274653001001993653001602003653000902019653002202028653001502050653002302065653001402088653003002102653004202132653002802174653001102202653004302213653001102256653001602267653001602283653002102299653002602320653002002346653002102366653003902387653001102426653002402437653001702461653003702478653003002515653001802545100002902563700002402592700002202616700002302638700002302661700002502684700002502709700002902734700002402763856014202787 2014 eng d00aSKP2 overexpression is associated with increased serine 10 phosphorylation of p27 (pSer10p27) in triple-negative breast cancer.0 aSKP2 overexpression is associated with increased serine 10 phosp c2014 Sep a1160-90 v2293 aS-phase kinase-associated protein 2 (SKP2) is an important cell cycle regulator, targeting the cyclin-dependent kinase (CDK) inhibitor p27 for degradation, and is frequently overexpressed in breast cancer. p27 regulates G1 /S transition by abrogating the activity of cyclin/CDK complexes. p27 can undergo phosphorylation at serine 10 (pSer10p27). This phosphorylation event is associated with increased cell proliferation and poor prognosis in patients with glioma. The relationship between SKP2 and pSer10p27 in breast cancer has not been previously investigated. Immunohistochemistry (IHC) of SKP2, p27, pSer10p27, and other genes involved in this pathway, was analyzed in 188 breast tumors and 50 benign reduction mammoplasty samples. IHC showed SKP2 to be more highly expressed in estrogen receptor α (ERα)-negative breast cancers and demonstrated that triple-negative tumors were more likely to have high expression of SKP2 than were non-triple negative, ERα-negative tumors. A significant positive relationship was discovered for SKP2 and pSer10p27. High levels of SKP2 and pSer10p27 were observed significantly more often in ERα-negative and triple-negative than in ERα-positive breast cancers. Use of the triple-negative TMX2-28 breast cancer cell line to address the role of SKP2 in cell cycle progression confirmed that SKP2 contributes to a more rapid cell cycle progression and may regulates pSer10p27 levels. Together, the results indicate that presence of high SKP2 plus high pSer10p27 levels in triple-negative breast cancers is associated with aggressive growth, and highlight the validity of using SKP2 inhibitors as a therapeutic approach for treating this subset of breast cancers.
10aAdult10aAge Factors10aAged10aAged, 80 and over10aCell Cycle10aCell Proliferation10aCyclin D110aCyclin-Dependent Kinase 210aCyclin-Dependent Kinase Inhibitor p2710aEstrogen Receptor alpha10aFemale10aGene Expression Regulation, Neoplastic10aHumans10aMCF-7 Cells10aMiddle Aged10aNeoplasm Grading10aNeoplasm Invasiveness10aPhosphorylation10aRNA Interference10aS-Phase Kinase-Associated Proteins10aSerine10aSignal Transduction10aTransfection10aTriple Negative Breast Neoplasms10aTumor Markers, Biological10aUp-Regulation1 aFagan-Solis, Katerina, D1 aPentecost, Brian, T1 aGozgit, Joseph, M1 aBentley, Brooke, A1 aMarconi, Sharon, M1 aOtis, Christopher, N1 aAnderton, Douglas, L1 aSchneider, Sallie, Smith1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/skp2-overexpression-is-associated-with-increased-serine-1002339nas a2200169 4500008004100000245014900041210006900190260001600259520161200275100002001887700002001907700002401927700002201951700002401973700001901997856015302016 2014 ENG d00aSpecific Recognition of Mycobacterial Protein and Peptide Antigens by γδ T Cell Subsets following Infection with Virulent Mycobacterium bovis.0 aSpecific Recognition of Mycobacterial Protein and Peptide Antige c2014 Feb 143 aPromoting effective immunity to Mycobacterium bovis infection is a challenge that is of interest to the fields of human and animal medicine alike. We report that γδ T cells from virulent M. bovis-infected cattle respond specifically and directly to complex, protein, and nonprotein mycobacterial Ags. Importantly, to our knowledge, we demonstrate for the first time that bovine γδ T cells specifically recognize peptide Ags derived from the mycobacterial protein complex ESAT6:CFP10 and that this recognition requires direct contact with APCs and signaling through the T cell Ag receptor but is independent of MHC class I or II. Furthermore, we show that M. bovis infection in cattle induces robust IL-17A protein responses. Interestingly, in contrast to results from mice, bovine CD4 T cells, and not γδ T cells, are the predominant source of this critical proinflammatory mediator. Bovine γδ T cells are divided into subsets based upon their expression of Workshop Cluster 1 (WC1), and we demonstrate that the M. bovis-specific γδ T cell response is composed of a heterogeneous mix of WC1-expressing populations, with the serologically defined WC1.1(+) and WC1.2(+) subsets responding in vitro to mycobacterial Ags and accumulating in the lesions of M. bovis-infected animals. The results described in this article enhance our understanding of γδ T cell biology and, because virulent M. bovis infection of cattle represents an excellent model of tuberculosis in humans, contribute to our overall understanding of the role of γδ T cells in the mycobacterial-specific immune response.
1 aMcGill, Jodi, L1 aSacco, Randy, E1 aBaldwin, Cynthia, L1 aTelfer, Janice, C1 aPalmer, Mitchell, V1 aWaters, Ray, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/specific-recognition-of-mycobacterial-protein-and-peptide-antigens-by01756nas a2200193 4500008004100000245013600041210006900177260000900246300000900255490000600264520102000270100001401290700002601304700002201330700001901352700002601371700001901397856014601416 2014 eng d00aT cell receptor stimulation impairs IL-7 receptor signaling by inducing expression of the microRNA miR-17 to target Janus kinase 1.0 aT cell receptor stimulation impairs IL7 receptor signaling by in c2014 ara830 v73 aT cell receptor (TCR)-mediated inhibition of interleukin-7 (IL-7) signaling is important for lineage fate determination in the thymus and for T cell survival in the periphery because uninterrupted IL-7 signaling results in T cell death. The initial event in IL-7 signaling is the transactivation of Janus kinases 1 and 3 (Jak1 and Jak3), which are associated with the cytosolic tails of the IL-7 receptor α chain (IL-7Rα) and the γc subunit, the two cell surface proteins that constitute IL-7R. We found that Jak1 is a highly unstable protein with a half-life of only 1.5 hours, so that continuous Jak1 protein synthesis is required to maintain Jak1 protein in sufficient abundance to support IL-7 signaling. However, we also found that Jak1 protein synthesis was acutely reduced by TCR-responsive microRNAs in the miR-17 family, which targeted Jak1 mRNA (messenger RNA) to inhibit its translation. Thus, this study identifies a molecular mechanism by which TCR engagement acutely disrupts IL-7 signaling.
1 aKatz, Gil1 aPobezinsky, Leonid, A1 aJeurling, Susanna1 aShinzawa, Miho1 aVan Laethem, Francois1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/t-cell-receptor-stimulation-impairs-il-7-receptor-signaling-by00707nas a2200181 4500008004100000245013700041210006900178260007500247300001800322490000800340100002200348700001900370700002100389700002500410700002300435700002300458856004400481 2014 eng d00aTMTC1 and TMTC2 Are Novel Endoplasmic Reticulum Tetratricopeptide Repeat-containing Adapter Proteins Involved in Calcium Homeostasis0 aTMTC1 and TMTC2 Are Novel Endoplasmic Reticulum Tetratricopeptid bAmerican Society for Biochemistry {&} Molecular Biology ({ASBMB})capr a16085–160990 v2891 aSunryd, Johan, C.1 aCheon, Banyoon1 aGraham, Jill, B.1 aGiorda, Kristina, M.1 aFissore, Rafael, A1 aHebert, Daniel, N. uhttps://doi.org/10.1074/jbc.m114.55407101511nas a2200193 4500008004100000245010000041210006900141260000900210300001200219490000800231520078600239100002501025700002301050700002901073700002301102700002101125700002401146856014701170 2014 eng d00aUsing breast milk to assess breast cancer risk: the role of mass spectrometry-based proteomics.0 aUsing breast milk to assess breast cancer risk the role of mass c2014 a399-4080 v8063 aAlthough mammography and treatment advances have led to declines in breast cancer mortality in the United States, breast cancer remains a major cause of morbidity and mortality. Breast cancer in young women is associated with increased mortality and current methods of detecting breast cancers in this group of women have known limitations. Tools for accurately assessing personal breast cancer risk in young women are needed to identify those women who would benefit the most from earlier intervention. Proteomic analysis of breast milk could identify biomarkers of breast cancer risk and provide a tool for identifying women at increased risk. A preliminary analysis of milk from four women provides a proof of concept for using breast milk to assess breast cancer risk.
1 aSchneider, Sallie, S1 aAslebagh, Roshanak1 aWetie, Armand, G Ngounou1 aSturgeon, Susan, R1 aDarie, Costel, C1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/using-breast-milk-to-assess-breast-cancer-risk-the-role-of-mass01716nas a2200169 4500008004100000245008800041210006900129260001600198300001100214490000600225520107800231100002501309700001701334700001801351700002301369856015401392 2014 eng d00aWeight gain following breast cancer diagnosis: Implication and proposed mechanisms.0 aWeight gain following breast cancer diagnosis Implication and pr c2014 Aug 10 a272-820 v53 aWeight gain occurs in the majority of women following breast cancer treatment. An overview of studies describing weight gain amongst women treated with early to modern chemotherapy regimens is included. Populations at higher risk include women who are younger, closer to ideal body weight and who have been treated with chemotherapy. Weight gain ranges between 1 to 5 kg, and may be associated with change in body composition with gain in fat mass and loss in lean body mass. Women are unlikely to return to pre-diagnosis weight. Possible mechanisms including inactivity and metabolic changes are explored. Potential interventions are reviewed including exercise, dietary changes and pharmacologic agents. Although breast cancer prognosis does not appear to be significantly impacted, weight gain has negative consequences on quality of life and overall health. Future studies should explore change in body composition, metabolism and insulin resistance. Avoiding weight gain in breast cancer survivors following initial diagnosis and treatment should be encouraged.
1 aMakari-Judson, Grace1 aBraun, Barry1 aJerry, Joseph1 aMertens, Wilson, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/weight-gain-following-breast-cancer-diagnosis-implication-and-proposed01737nas a2200181 4500008004100000022001400041245008800055210006900143260001600212300001100228490000600239520108000245100002501325700001701350700001801367700002301385856014701408 2014 eng d a2218-433300aWeight gain following breast cancer diagnosis: Implication and proposed mechanisms.0 aWeight gain following breast cancer diagnosis Implication and pr c2014 Aug 10 a272-820 v53 aWeight gain occurs in the majority of women following breast cancer treatment. An overview of studies describing weight gain amongst women treated with early to modern chemotherapy regimens is included. Populations at higher risk include women who are younger, closer to ideal body weight and who have been treated with chemotherapy. Weight gain ranges between 1 to 5 kg, and may be associated with change in body composition with gain in fat mass and loss in lean body mass. Women are unlikely to return to pre-diagnosis weight. Possible mechanisms including inactivity and metabolic changes are explored. Potential interventions are reviewed including exercise, dietary changes and pharmacologic agents. Although breast cancer prognosis does not appear to be significantly impacted, weight gain has negative consequences on quality of life and overall health. Future studies should explore change in body composition, metabolism and insulin resistance. Avoiding weight gain in breast cancer survivors following initial diagnosis and treatment should be encouraged.
1 aMakari-Judson, Grace1 aBraun, Barry1 aJerry, Joseph1 aMertens, Wilson, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/weight-gain-following-breast-cancer-diagnosis-implication-and-000447nas a2200133 4500008004100000245007300041210006900114260002300183300001200206490000700218100001800225700002300243856004700266 2013 eng d00aCa2 homeostas+is and regulation of ER Ca2+ in mammalian oocytes/eggs0 aCa2 homeostasis and regulation of ER Ca2 in mammalian oocytesegg bElsevier {BV}cjan a63–670 v531 aWakai, Takuya1 aFissore, Rafael, A uhttps://doi.org/10.1016/j.ceca.2012.11.01000586nas a2200157 4500008004100000245011800041210006900159260003100228300001400259100002400273700001600297700002600313700001700339700002400356856004800380 2013 eng d00aCHAPTER 7. Technologies to Study Plant Biomass Fermentation Using the Model Bacterium Clostridium Phytofermentans0 aCHAPTER 7 Technologies to Study Plant Biomass Fermentation Using bRoyal Society of Chemistry a114–1391 aTolonen, Andrew, C.1 aPetit, Elsa1 aBlanchard, Jeffrey, L1 aWarnick, Tom1 aLeschine, Susan, B. uhttps://doi.org/10.1039/9781849734738-0011402364nas a2200253 4500008004100000245008100041210006900122260001600191520146600207100002001673700001701693700003001710700003201740700002201772700002001794700002301814700002201837700002001859700001701879700001701896700002101913700002301934856015301957 2013 ENG d00aCompartmentalization of Distinct cAMP Signaling Pathways in Mammalian Sperm.0 aCompartmentalization of Distinct cAMP Signaling Pathways in Mamm c2013 Oct 153 aFertilization competence is acquired in the female tract in a process known as capacitation. Capacitation is needed for the activation of motility (e.g. hyperactivation) and to prepare the sperm for an exocytotic process known as acrosome reaction. While the HCO(3)--dependent soluble adenylyl cyclase Adcy10 plays a role in motility, less is known about the source of cAMP in the sperm head. Transmembrane adenylyl cyclases (tmACs) are another possible source of cAMP. These enzymes are regulated by stimulatory heterotrimeric Gs proteins; however, the presence of Gs or tmACs in mammalian sperm has been controversial. In this manuscript, we used Western blotting and cholera toxin-dependent ADP ribosylation to show Gs presence in the sperm head. Also, we showed that forskolin, a tmAC specific activator, induces cAMP accumulation in sperm from both WT and Adcy10 null mice. This increase is blocked by the tmAC inhibitor SQ-22536 but not by the Adcy10 inhibitor KH7. While Gs immunoreactivity and tmAC activity are detected in the sperm head, PKA is only found in the tail, where Adcy10 was previously shown to reside. Consistent with an acrosomal localization, Gs reactivity is lost in acrosome reacted sperm, and forskolin is able to increase intracellular Ca(2+) and induce the acrosome reaction. Altogether, these data suggest that cAMP pathways are compartmentalized in sperm, with Gs and tmAC in the head and Adcy10 and PKA in the flagellum.
1 aWertheimer, Eva1 aKrapf, Dario1 aVega-Beltran, José, Luis1 aSánchez-Cárdenas, Claudia1 aNavarrete, Felipe1 aHaddad, Douglas1 aEscoffier, Jessica1 aSalicioni, Ana, M1 aLevin, Lonny, R1 aBuck, Jochen1 aMager, Jesse1 aDarszon, Alberto1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/compartmentalization-of-distinct-camp-signaling-pathways-in-mammalian00665nas a2200205 4500008004100000245011300041210006900154260004600223300001400269490000700283100001200290700001400302700001700316700001600333700001400349700001600363700001500379700001500394856005000409 2013 eng d00aControl of Spontaneous Activation of Rat Oocytes by Regulating Plasma Membrane Na+/Ca2+ Exchanger Activities0 aControl of Spontaneous Activation of Rat Oocytes by Regulating P bOxford University Press ({OUP})cmay/2013 a160–1600 v881 aCui, W.1 aZhang, J.1 aZhang, C.-X.1 aJiao, G.-Z.1 aZhang, M.1 aWang, T.-Y.1 aLuo, M.-J.1 aTan, J.-H. uhttps://doi.org/10.1095/biolreprod.113.10826601972nas a2200157 4500008004100000245014100041210006900182260001500251300001000266490000800276520132800284100001501612700002501627700001701652856014501669 2013 eng d00aCTR9/PAF1c regulates molecular lineage identity, histone H3K36 trimethylation and genomic imprinting during preimplantation development.0 aCTR9PAF1c regulates molecular lineage identity histone H3K36 tri c2013 Nov 1 a15-270 v3833 aGenome-wide epigenetic reprogramming is required for successful preimplantation development. Inappropriate or deficient chromatin regulation can result in defective lineage specification and loss of genomic imprinting, compromising normal development. Here we report that two members of the RNA polymerase II associated factor, homolog (Saccharomyces cerevisiae) complex (PAF1 complex) components, Ctr9 and Rtf1, are required during mammalian preimplantation development. We demonstrate that Ctr9-deficient embryos fail to correctly specify lineages at the blastocyst stage. Expression of some lineage specific factors is markedly reduced in Ctr9 knockdown embryos, including Eomes, Elf5 and Sox2, while others are inappropriately expressed (Oct4, Nanog, Gata6, Fgf4 and Sox17). We also show that several imprinted genes (Mest, Peg3, Snrpn and Meg3) are aberrantly expressed although allele specific DNA methylation is not altered. We document a loss of histone H3 lysine 36 trimethylation (H3K36me3) in Ctr9-deficient embryos and confirm that knockdown of either Setd2 or Rtf1 results in similar phenotypes. These findings show that the PAF1 complex is required for mammalian development, likely through regulation of H3K36me3, and indicate functional conservation of the PAF1 complex from yeast to mammals in vivo.
1 aZhang, Kun1 aHaversat, Jocelyn, M1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/ctr9-paf1c-regulates-molecular-lineage-identity-histone-h3k3603303nas a2200181 4500008004100000245015800041210006900199260001600268520256200284100002102846700001702867700002002884700002002904700001302924700001902937700002002956856014502976 2013 ENG d00aFunctional human sperm capacitation requires both bicarbonate-dependent PKA activation and down-regulation of Ser/Thr phosphatases by Src family kinases.0 aFunctional human sperm capacitation requires both bicarbonatedep c2013 May 193 aIn all mammalian species studied so far, sperm capacitation correlates with an increase in protein tyrosine (Tyr) phosphorylation mediated by a bicarbonate-dependent cAMP/protein kinase A (PKA) pathway. Recent studies in mice revealed, however, that a Src family kinase (SFK)-induced inactivation of serine/threonine (Ser/Thr) phosphatases is also involved in the signaling pathways leading to Tyr phosphorylation. In view of these observations and with the aim of getting a better understanding of the signaling pathways involved in human sperm capacitation, in the present work we investigated the involvement of both the cAMP/PKA and SFK/phosphatase pathways in relation to the capacitation state of the cells. For this purpose, different signaling events and sperm functional parameters were analyzed as a function of capacitation time. Results revealed a very early bicarbonate-dependent activation of PKA indicated by the rapid (1 min) increase in both phospho-PKA substrates and cAMP levels (P < 0.05). However, a complete pattern of Tyr phosphorylation was detected only after 6-h incubation at which time sperm exhibited the ability to undergo the acrosome reaction (AR) and to penetrate zona-free hamster oocytes. Sperm capacitated in the presence of the SFK inhibitor SKI606 showed a decrease in both PKA substrate and Tyr phosphorylation levels, which was overcome by exposure of sperm to the Ser/Thr phosphatase inhibitor okadaic acid (OA). However, OA was unable to induce phosphorylation when sperm were incubated under PKA-inhibitory conditions (i.e. in the absence of bicarbonate or in the presence of PKA inhibitor). Moreover, the increase in PKA activity by exposure to a cAMP analog and a phosphodiesterase inhibitor did not overcome the inhibition produced by SKI606. Whereas the presence of SKI606 during capacitation produced a negative effect (P < 0.05) on sperm motility, progesterone-induced AR and fertilizing ability, none of these inhibitions were observed when sperm were exposed to SKI606 and OA. Interestingly, different concentrations of inhibitors were required to modulate human and mouse capacitation revealing the species specificity of the molecular mechanisms underlying this process. In conclusion, our results describe for the first time the involvement of both PKA activation and Ser/Thr phosphatase down-regulation in functional human sperm capacitation and provide convincing evidence that early PKA-dependent phosphorylation is the convergent regulatory point between these two signaling pathways.
1 aBattistone, M, A1 aDa Ros, V, G1 aSalicioni, A, M1 aNavarrete, F, A1 aKrapf, D1 aVisconti, P, E1 aCuasnicú, P, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/functional-human-sperm-capacitation-requires-both-bicarbonate01705nas a2200241 4500008004100000245013300041210006900174260001600243300001100259490000700270520084300277100002001120700002401140700002001164700002201184700001701206700001801223700002001241700001801261700001701279700002001296856014701316 2013 eng d00aA Genome-wide siRNA Screen Identifies Proteasome Addiction as a Vulnerability of Basal-like Triple-Negative Breast Cancer Cells.0 aGenomewide siRNA Screen Identifies Proteasome Addiction as a Vul c2013 Aug 12 a182-960 v243 aBasal-like triple-negative breast cancers (TNBCs) have poor prognosis. To identify basal-like TNBC dependencies, a genome-wide siRNA lethality screen compared two human breast epithelial cell lines transformed with the same genes: basal-like BPLER and myoepithelial HMLER. Expression of the screen's 154 BPLER dependency genes correlated with poor prognosis in breast, but not lung or colon, cancer. Proteasome genes were overrepresented hits. Basal-like TNBC lines were selectively sensitive to proteasome inhibitor drugs relative to normal epithelial, luminal, and mesenchymal TNBC lines. Proteasome inhibition reduced growth of established basal-like TNBC tumors in mice and blocked tumor-initiating cell function and macrometastasis. Proteasome addiction in basal-like TNBCs was mediated by NOXA and linked to MCL-1 dependence.
1 aPetrocca, Fabio1 aAltschuler, Gabriel1 aTan, Shen, Mynn1 aMendillo, Marc, L1 aYan, Haoheng1 aJerry, Joseph1 aKung, Andrew, L1 aHide, Winston1 aInce, Tan, A1 aLieberman, Judy uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/a-genome-wide-sirna-screen-identifies-proteasome-addiction-as-a02812nas a2200385 4500008004100000245015800041210006900199260001500268300001300283490000800296520152000304653001201824653002701836653001401863653002101877653001401898653003001912653001101942653000901953653000901962653002501971653003701996653003002033653003702063653001602100653001502116653001902131100001702150700002302167700001502190700002202205700002402227700002302251856015202274 2013 eng d00aHeat shock protein 90 functions to stabilize and activate the testis-specific serine/threonine kinases, a family of kinases essential for male fertility.0 aHeat shock protein 90 functions to stabilize and activate the te c2013 Jun 7 a16308-200 v2883 aSpermiogenesis is characterized by a profound morphological differentiation of the haploid spermatid into spermatozoa. The testis-specific serine/threonine kinases (TSSKs) comprise a family of post-meiotic kinases expressed in spermatids, are critical to spermiogenesis, and are required for male fertility in mammals. To explore the role of heat shock protein 90 (HSP90) in regulation of TSSKs, the stability and catalytic activity of epitope-tagged murine TSSKs were assessed in 293T and COS-7 cells. TSSK1, -2, -4, and -6 (small serine/threonine kinase) were all found to associate with HSP90, and pharmacological inhibition of HSP90 function using the highly specific drugs 17-AAG, SNX-5422, or NVP-AUY922 reduced TSSK protein levels in cells. The attenuation of HSP90 function abolished the catalytic activities of TSSK4 and -6 but did not significantly alter the specific activities of TSSK1 and -2. Inhibition of HSP90 resulted in increased TSSK ubiquitination and proteasomal degradation, indicating that HSP90 acts to control ubiquitin-mediated catabolism of the TSSKs. To study HSP90 and TSSKs in germ cells, a mouse primary spermatid culture model was developed and characterized. Using specific antibodies against murine TSSK2 and -6, it was demonstrated that HSP90 inhibition resulted in a marked decrease of the endogenous kinases in spermatids. Together, our findings demonstrate that HSP90 plays a broad and critical role in stabilization and activation of the TSSK family of protein kinases.
10aAnimals10aCercopithecus aethiops10aCOS Cells10aEnzyme Stability10aFertility10aHSP90 Heat-Shock Proteins10aHumans10aMale10aMice10aMice, Mutant Strains10aProteasome Endopeptidase Complex10aProtein Kinase Inhibitors10aProtein-Serine-Threonine Kinases10aProteolysis10aSpermatids10aUbiquitination1 aJha, Kula, N1 aColeman, Alyssa, R1 aWong, Lily1 aSalicioni, Ana, M1 aHowcroft, Elizabeth1 aJohnson, Gibbes, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/heat-shock-protein-90-functions-to-stabilize-and-activate-the-testis00712nas a2200205 4500008004100000245009600041210006900137260002300206300001400229490000700243100003100250700002600281700002300307700001600330700002000346700002300366700003500389700002500424856005700449 2013 eng d00aIdentification of phospholipase C zeta in normospermic and teratospermic domestic cat sperm0 aIdentification of phospholipase C zeta in normospermic and terat bElsevier {BV}coct a722–7290 v801 aVillaverde, Ana, Izabel S.1 aFioratti, Eduardo, G.1 aFissore, Rafael, A1 aHe, Changli1 aLee, Hoi, Chang1 aSouza, Fabiana, F.1 aLandim-Alvarenga, Fernanda, C.1 aLopes, Maria, Denise uhttps://doi.org/10.1016/j.theriogenology.2013.06.00502191nas a2200397 4500008004100000245013800041210007100179260001300250300001100263490000700274520088300281653001201164653003101176653001501207653002301222653001801245653003101263653001601294653002101310653001801331653002601349653000901375653002101384653003101405653002401436653001701460100002201477700002601499700002201525700001901547700001901566700002001585700001701605700001901622856015201641 2013 eng d00aIL-7 signaling must be intermittent, not continuous, during CD8⁺ T cell homeostasis to promote cell survival instead of cell death.0 aIL7 signaling must be intermittent not continuous during CD8⁺ T c2013 Feb a143-510 v143 aThe maintenance of naive CD8(+) T cells is necessary for lifelong immunocompetence but for unknown reasons requires signaling via both interleukin 7 (IL-7) and the T cell antigen receptor (TCR). We now report that naive CD8(+) T cells required IL-7 signaling to be intermittent, not continuous, because prolonged IL-7 signaling induced naive CD8(+) T cells to proliferate, produce interferon-γ (IFN-γ) and undergo IFN-γ-triggered cell death. Homeostatic engagement of the TCR interrupted IL-7 signaling and thereby supported the survival and quiescence of CD8(+) T cells. However, CD8(+) T cells with insufficient TCR affinity for self ligands received prolonged IL-7 signaling and died during homeostasis. In this study we identified regulation of the duration of IL-7 signaling by homeostatic engagement of the TCR as the basis for in vivo CD8(+) T cell homeostasis.
10aAnimals10aCD8-Positive T-Lymphocytes10aCell Death10aCell Proliferation10aCell Survival10aGene Expression Regulation10aHomeostasis10aInterferon-gamma10aInterleukin-710aLymphocyte Activation10aMice10aMice, Transgenic10aReceptors, Antigen, T-Cell10aSignal Transduction10aTime Factors1 aKimura, Motoko, Y1 aPobezinsky, Leonid, A1 aGuinter, Terry, I1 aThomas, Julien1 aAdams, Anthony1 aPark, Jung-Hyun1 aTai, Xuguang1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/il-7-signaling-must-be-intermittent-not-continuous-during-cd8-t-cell00737nas a2200217 4500008004100000245011900041210006900160260004400229300001100273490000600284100001300290700002200303700002300325700002200348700001700370700001800387700001900405700002100424700002500445856004900470 2013 eng d00aImpact of Laminitis on the Canonical Wnt Signaling Pathway in Basal Epithelial Cells of the Equine Digital Laminae0 aImpact of Laminitis on the Canonical Wnt Signaling Pathway in Ba bPublic Library of Science ({PLoS})cfeb ae560250 v81 aWang, Le1 aPawlak, Erica, A.1 aJohnson, Philip, J1 aBelknap, James, K1 aEades, Susan1 aStack, Sharon1 aCousin, Helene1 aBlack, Samuel, J1 aLaird, Elizabeth, G. uhttps://doi.org/10.1371/journal.pone.005602502095nas a2200301 4500008004100000245006600041210006500107260001300172300001000185490000700195520112900202653001201331653002101343653003101364653002701395653002701422653001101449653001101460653002701471653002601498653001701524653003001541100002101571700002501592700001801617700001401635856014401649 2013 eng d00aImpaired mitochondrial metabolism and mammary carcinogenesis.0 aImpaired mitochondrial metabolism and mammary carcinogenesis c2013 Mar a75-870 v183 aMitochondrial oxidative metabolism plays a key role in meeting energetic demands of cells by oxidative phosphorylation (OxPhos). Here, we have briefly discussed (a) the dynamic relationship that exists among glycolysis, the tricarboxylic acid (TCA) cycle, and OxPhos; (b) the evidence of impaired OxPhos (i.e. mitochondrial dysfunction) in breast cancer; (c) the mechanisms by which mitochondrial dysfunction can predispose to cancer; and (d) the effects of host and environmental factors that can negatively affect mitochondrial function. We propose that impaired OxPhos could increase susceptibility to breast cancer via suppression of the p53 pathway, which plays a critical role in preventing tumorigenesis. OxPhos is sensitive to a large number of factors intrinsic to the host (e.g. inflammation) as well as environmental exposures (e.g. pesticides, herbicides and other compounds). Polymorphisms in over 143 genes can also influence the OxPhos system. Therefore, declining mitochondrial oxidative metabolism with age due to host and environmental exposures could be a common mechanism predisposing to cancer.
10aAnimals10aBreast Neoplasms10aCarcinogens, Environmental10aDisease Susceptibility10aEnvironmental Exposure10aFemale10aHumans10aMammary Glands, Animal10aMammary Glands, Human10aMitochondria10aOxidative Phosphorylation1 aYadava, Nagendra1 aSchneider, Sallie, S1 aJerry, Joseph1 aKim, Chul uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/impaired-mitochondrial-metabolism-and-mammary-carcinogenesis00618nas a2200157 4500008004100000245015100041210006900192260004400261300001100305490000600316100002000322700002100342700002600363700002200389856004900411 2013 eng d00aAn In Vitro Model of the Horse Gut Microbiome Enables Identification of Lactate-Utilizing Bacteria that Differentially Respond to Starch Induction0 aIn Vitro Model of the Horse Gut Microbiome Enables Identificatio bPublic Library of Science ({PLoS})coct ae775990 v81 aBiddle, Amy, S.1 aBlack, Samuel, J1 aBlanchard, Jeffrey, L1 aBereswill, Stefan uhttps://doi.org/10.1371/journal.pone.007759900929nas a2200277 4500008004100000245012000041210006900161260004400230300001100274490000600285100001600291700001700307700002500324700002400349700001800373700001900391700002300410700001600433700003100449700001900480700002300499700002400522700002600546700003000572856004900602 2013 eng d00aInvolvement of a Bacterial Microcompartment in the Metabolism of Fucose and Rhamnose by Clostridium phytofermentans0 aInvolvement of a Bacterial Microcompartment in the Metabolism of bPublic Library of Science ({PLoS})cjan ae543370 v81 aPetit, Elsa1 aLaTouf, Greg1 aCoppi, Maddalena, V.1 aWarnick, Thomas, A.1 aCurrie, Devin1 aRomashko, Igor1 aDeshpande, Supriya1 aHaas, Kelly1 aAlvelo-Maurosa, Jesús, G.1 aWardman, Colin1 aSchnell, Danny, J.1 aLeschine, Susan, B.1 aBlanchard, Jeffrey, L1 ade Crécy-Lagard, Valerie uhttps://doi.org/10.1371/journal.pone.005433702430nas a2200385 4500008004100000245011000041210006900151260001600220300001200236490000800248520111500256653001201371653005701383653003701440653000901477653004301486653002101529653002401550653001801574653001501592653001701607100002601624700002801650700002601678700001701704700002201721700002201743700002201765700001901787700002201806700002301828700002301851700001901874856015101893 2013 eng d00aLck availability during thymic selection determines the recognition specificity of the T cell repertoire.0 aLck availability during thymic selection determines the recognit c2013 Sep 12 a1326-410 v1543 aThymic selection requires signaling by the protein tyrosine kinase Lck to generate T cells expressing αβ T cell antigen receptors (TCR). For reasons not understood, the thymus selects only αβTCR that are restricted by major histocompatibility complex (MHC)-encoded determinants. Here, we report that Lck proteins that were coreceptor associated promoted thymic selection of conventionally MHC-restricted TCR, but Lck proteins that were coreceptor free promoted thymic selection of MHC-independent TCR. Transgenic TCR with MHC-independent specificity for CD155 utilized coreceptor-free Lck to signal thymic selection in the absence of MHC, unlike any transgenic TCR previously described. Thus, the thymus can select either MHC-restricted or MHC-independent αβTCR depending on whether Lck is coreceptor associated or coreceptor free. We conclude that the intracellular state of Lck determines the specificity of thymic selection and that Lck association with coreceptor proteins during thymic selection is the mechanism by which MHC restriction is imposed on a randomly generated αβTCR repertoire.
10aAnimals10aLymphocyte Specific Protein Tyrosine Kinase p56(lck)10aMajor Histocompatibility Complex10aMice10aReceptors, Antigen, T-Cell, alpha-beta10aReceptors, Virus10aSignal Transduction10aT-Lymphocytes10aThymocytes10aThymus Gland1 aVan Laethem, Francois1 aTikhonova, Anastasia, N1 aPobezinsky, Leonid, A1 aTai, Xuguang1 aKimura, Motoko, Y1 aLe Saout, Cécile1 aGuinter, Terry, I1 aAdams, Anthony1 aSharrow, Susan, O1 aBernhardt, Günter1 aFeigenbaum, Lionel1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/lck-availability-during-thymic-selection-determines-the-recognition00582nas a2200169 4500008004100000245013400041210006900175260001500244300001400259490000700273100001800280700001900298700002300317700001500340700001900355856003800374 2013 eng d00aManual corneal thickness measurements of healthy equine eyes using a portable spectral-domain optical coherence tomography device0 aManual corneal thickness measurements of healthy equine eyes usi bWileycdec a631–6340 v461 aPirie, C., G.1 aAlario, A., F.1 aBarysauskas, C., M1 aGradil, C.1 aUricchio, C.K. uhttps://doi.org/10.1111/evj.1219800716nas a2200193 4500008004100000245017700041210006900218260004600287490000700333100001500340700001300355700001300368700002000381700001800401700001800419700001800437700001700455856005000472 2013 eng d00aMechanisms by which a Lack of Germinal Vesicle (GV) Material Causes Oocyte Meiotic Defects: A Study Using Oocytes Manipulated to Replace GV with Primary Spermatocyte Nuclei0 aMechanisms by which a Lack of Germinal Vesicle GV Material Cause bOxford University Press ({OUP})coct/20130 v891 aZhang, Jie1 aCui, Wei1 aLi, Qing1 aWang, Tian-Yang1 aSui, Hong-Shu1 aWang, Jun-Zuo1 aLuo, Ming-Jiu1 aTan, Jing-He uhttps://doi.org/10.1095/biolreprod.113.11150003114nas a2200433 4500008004100000245017500041210006900216260000900285300000900294490000600303520168300309653001201992653002502004653001402029653003802043653003602081653002502117653001102142653002702153653002602180653001602206653000902222653001402231653002602245653002102271653002102292653002702313653001502340653002602355653003002381100001902411700001702430700001802447700001802465700001702483700001602500700001502516856014902531 2013 eng d00aMiR203 mediates subversion of stem cell properties during mammary epithelial differentiation via repression of ΔNP63α and promotes mesenchymal-to-epithelial transition.0 aMiR203 mediates subversion of stem cell properties during mammar c2013 ae5140 v43 aDuring reproductive life, the mammary epithelium undergoes consecutive cycles of proliferation, differentiation and apoptosis. Doing so relies on the retained proliferative capacity, prolonged lifespan and developmental potency of mammary stem cells (MaSCs). ΔNp63α, the predominant TP63 isoform in mammary epithelia, is robustly expressed in MaSCs and is required for preservation of self-renewing capacity in diverse epithelial structures. However, the mechanism(s) underlying subversion of this activity during forfeiture of self-renewing capacity are poorly understood. MicroRNAs (miRNAs) govern critical cellular functions including stem cell maintenance, development, cell cycle regulation and differentiation by disrupting translation of target mRNAs. Data presented here indicate that expression of miR203, a miRNA that targets ΔNp63α and ΔNp63β is activated during luminal epithelial differentiation and that this pattern is observed in the murine mammary hierarchy. In addition, we present evidence that the transcription factor Zeb1 represses miR203 expression, thus enhancing ΔNp63α protein levels. Furthermore, ectopic miR203 suppresses ΔNp63α expression, proliferation and colony formation. The anti-clonogenic effects mediated by miR203 require suppression of ΔNp63α. In addition, ectopic miR203 promotes mesenchymal-to-epithelial transition and disrupts activities associated with epithelial stem cells. These studies support a model in which induction of miR203 mediates forfeiture of self-renewing capacity via suppression of ΔNp63α and may also have anti-tumorigenic activity through its reduction of EMT and cancer stem cell populations.
10aAnimals10aCell Differentiation10aCell Line10aEpithelial-Mesenchymal Transition10aG1 Phase Cell Cycle Checkpoints10aHomeodomain Proteins10aHumans10aMammary Glands, Animal10aMammary Glands, Human10aMCF-7 Cells10aMice10aMicroRNAs10aNeoplastic Stem Cells10aProtein Isoforms10aRNA Interference10aRNA, Small Interfering10aStem Cells10aTranscription Factors10aTumor Suppressor Proteins1 aDeCastro, A, J1 aDunphy, K, A1 aHutchinson, J1 aBalboni, A, L1 aCherukuri, P1 aJerry, D, J1 aDiRenzo, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mir203-mediates-subversion-of-stem-cell-properties-during-mammary00591nas a2200193 4500008004100000245007000041210006800111260002500179300001400204490000700218100001500225700001800240700002000258700002000278700002300298700001500321700002300336856003800359 2013 eng d00aMolecular characteristics of horse phospholipase C zeta (PLCzeta)0 aMolecular characteristics of horse phospholipase C zeta PLCzeta bWiley-Blackwellcfeb a359–3680 v841 aSato, Kana1 aWakai, Takuya1 aSeita, Yasunari1 aTakizawa, Akiko1 aFissore, Rafael, A1 aIto, Junya1 aKashiwazaki, Naomi uhttps://doi.org/10.1111/asj.1204401992nas a2200157 4500008004100000245006100041210005900102260001300161300001200174490000800186520143800194100002501632700002101657700001701678856013901695 2013 eng d00aMorphogenetic analysis of peri-implantation development.0 aMorphogenetic analysis of periimplantation development c2013 Sep a1110-200 v2423 aBACKGROUND: Although successful implantation is required for development in placental mammals, the molecular and morphogenetic events that define peri-implantation remain largely unexplored. RESULTS: Here we present detailed morphological and immunohistochemical analysis of mouse embryos between embryonic day 3.75 and 5.25 of gestation, during the implantation process in vivo. We examined expression patterns of key transcription factors (Sox2, Oct4, Nanog, Cdx2, Gata6, Sox17, and Yy1) during pre- and postimplantation development. Additionally, we examined morphogenetic changes through analysis of ZO-1, Laminin, and E-Cadherin localization. The results presented reveal novel changes in gene expression and morphogenetic events during peri-implantation in utero. Here we show: (1) molecular and morphological changes in primitive endoderm cells as they transition from a salt and pepper distribution to a sheet covering the inner cell mass; (2) tissue-specific GATA6 levels; and (3) a striking pattern of SOX17 that is suggestive of a functional role either directing or permitting implantation at specific sites in the uterine epithelium. CONCLUSIONS: A growing number of knockout mice display peri-implantation lethality, and the data presented herein identify key morphogenetic landmarks that can be used to characterize mutant phenotypes, as well as further our basic understanding of peri-implantation development.
1 aWallingford, Mary, C1 aAngelo, Jesse, R1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/morphogenetic-analysis-of-peri-implantation-development01998nas a2200253 4500008004100000245007400041210006900115260001300184300001300197490000700210520115200217653002701369653001101396653001701407653002801424653002401452100001901476700002101495700002301516700001701539700002001556700001601576856015201592 2013 eng d00aNonclassical progesterone signalling molecules in the nervous system.0 aNonclassical progesterone signalling molecules in the nervous sy c2013 Nov a991-10010 v253 aProgesterone (P4) regulates a wide range of cognitive, neuroendocrine, neuroimmune and neuroprotective functions. Therefore, it is not surprising that this ovarian hormone acts through multiple receptors. Ever since the 1980s, studies investigating the neural effects of P4 have focused mainly on genomic and nongenomic actions of the classical progestin receptor (PGR). More recently, two groups of nonclassical P4 signalling molecules have been identified: (i) the class II progestin and adipoQ receptor (PAQR) family, which includes PAQR 5, 6, 7, 8 and 9, also called membrane progestin receptor α (mPRα; PAQR7), mPRβ (PAQR8), mPRγ (PAQR5), mPRδ (PAQR6) and mPRε (PAQR9), and (ii) the b5-like haeme/steroid-binding protein family, which includes progesterone receptor membrane component 1 (Pgrmc1), Pgrmc2, neudesin and neuferricin. In this review, we describe the structures, neuroanatomical localisation and signalling mechanisms of these molecules. We also discuss gonadotrophin-releasing hormone regulation as an example of a physiological function regulated by multiple progesterone receptors but through different mechanisms.
10aCentral Nervous System10aHumans10aProgesterone10aReceptors, Progesterone10aSignal Transduction1 aPetersen, S, L1 aIntlekofer, K, A1 aMoura-Conlon, P, J1 aBrewer, D, N1 aSans, Del, Pino1 aLopez, J, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/nonclassical-progesterone-signalling-molecules-in-the-nervous-system02054nas a2200133 4500008004100000245007900041210006900120260000900189300001100198490000600209520153500215100002001750856015001770 2013 eng d00aNOTCH signaling in immune-mediated bone marrow failure of aplastic anemia.0 aNOTCH signaling in immunemediated bone marrow failure of aplasti c2013 ae267640 v13 aSevere aplastic anemia is a rare bone marrow failure disease with the majority of cases caused by aberrant immune destruction of blood progenitors. Although the Th1-mediated pathology of aplastic anemia is well-described, the molecular mechanisms that drive disease progression remain ill-defined. The NOTCH signaling pathway mediates Th1 differentiation in the presence of polarizing cytokines, an action requiring enzymatic processing of NOTCH receptors by γ- secretase. We used a mouse model of aplastic anemia to demonstrate that expression both of intracellular NOTCH1 (NOTCH1(IC)) and T-BET, a key transcription factor regulating Th1 differentiation, were increased in T cells in the spleen and bone marrow during active disease. Conditionally deleting NOTCH1 or administering γ-secretase inhibitors (GSI) in vivo, attenuated disease and rescued mice from lethal bone marrow failure. In peripheral T cells from patients with untreated aplastic anemia, NOTCH1(IC) was significantly elevated and was detected at the TBX21 promoter, showing NOTCH1 directly regulates the gene encoding T-BET. Treating patients' cells ex vivo with GSI lowered NOTCH1(IC) levels, decreased the level of NOTCH1 detectable at the TBX21 promoter, and also decreased T-BET expression, indicating NOTCH1 signaling is responsive to GSI during active disease. Collectively, these results identify NOTCH1 signaling as a primary driver of Th1-mediated pathogenesis in aplastic anemia and may represent a novel target for therapeutic intervention.
1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch-signaling-in-immune-mediated-bone-marrow-failure-of-aplastic02056nas a2200301 4500008004100000245012700041210006900168260001300237300001000250490000700260520099000267653003101257653002501288653003001313653001101343653001701354653002101371653002101392653002701413653002601440100002001466700002901486700002201515700002401537700002001561700002001581856015301601 2013 eng d00aNovel protein transduction domain mimics as nonviral delivery vectors for siRNA targeting NOTCH1 in primary human T cells.0 aNovel protein transduction domain mimics as nonviral delivery ve c2013 Jan a201-90 v213 aRNA interference technology has recently been highlighted as a powerful research method as well as a potential therapeutic treatment for several diseases. However, the delivery of small interfering RNA (siRNA) into T cell lines and primary blood cells is exceedingly challenging, as they are resistant to transfection by conventional reagents. As a result, there is an unmet need for nonviral, efficient, and easily prepared carriers for siRNA delivery into hard-to-transfect cell types. Here, we report a novel system based on protein transduction domain mimics (PTDMs), generated by ring opening metathesis polymerization, for intracellular delivery of siRNA molecules. PTDM-based siRNA delivery induced efficient NOTCH1 knockdown in Jurkat T cells and human peripheral blood mononuclear cells without any measured toxicity. Furthermore, delivering siRNA to NOTCH1 in human peripheral blood cells modulated cell proliferation and differentiation of T cells into T(H)1 cells.
10aCD4-Positive T-Lymphocytes10aCell Differentiation10aGene Knockdown Techniques10aHumans10aJurkat Cells10aReceptor, Notch110aRNA Interference10aRNA, Small Interfering10aTransduction, Genetic1 aTezgel, Özgül1 aGonzalez-Perez, Gabriela1 aTelfer, Janice, C1 aOsborne, Barbara, A1 aMinter, Lisa, M1 aTew, Gregory, N uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/novel-protein-transduction-domain-mimics-as-nonviral-delivery-vectors02700nas a2200277 4500008004100000245013700041210006900178260001600247300000700263490000700270520171600277100002101993700001802014700001902032700002002051700001502071700001902086700002302105700002302128700001902151700003202170700002102202700002902223700001802252856015202270 2013 ENG d00aOncogenic transformation of mammary epithelial cells by transforming growth factor beta independent of mammary stem cell regulation.0 aOncogenic transformation of mammary epithelial cells by transfor c2013 Jul 25 a740 v133 aBACKGROUND: Transforming growth factor beta (TGFbeta) is transiently increased in the mammary gland during involution and by radiation. While TGFbeta normally has a tumour suppressor role, prolonged exposure to TGFbeta can induce an oncogenic epithelial to mesenchymal transition (EMT) program in permissive cells and initiate the generation of cancer stem cells. Our objective is to mimic the transient exposure to TGFbeta during involution to determine the persistent effects on premalignant mammary epithelium. METHOD: CDbetaGeo cells, a transplantable mouse mammary epithelial cell line, were treated in vitro for 14 days with TGFbeta (5 ng/ml). The cells were passaged for an additional 14 days in media without TGFbeta and then assessed for markers of EMT and transformation. RESULTS: The 14-day exposure to TGFbeta induced EMT and transdifferentiation in vitro that persists after withdrawal of TGFbeta. TGFbeta-treated cells are highly tumorigenic in vivo, producing invasive solid de-differentiated tumours (100%; latency 6.7 weeks) compared to control (43%; latency 32.7 weeks). Although the TGFbeta-treated cells have initiated a persistent EMT program, the stem cell population was unchanged relative to the controls. The gene expression profiles of TGFbeta-treated cells demonstrate de-differentiation with decreases in the expression of genes that define luminal, basal and stem cells. Additionally, the gene expression profiles demonstrate increases in markers of EMT, growth factor signalling, TGFbeta2 and changes in extra cellular matrix. CONCLUSION: This model demonstrates full oncogenic EMT without an increase in stem cells, serving to separate EMT markers from stem cell markers.
1 aDunphy, Karen, A1 aSeo, Jae-Hong1 aKim, Daniel, J1 aRoberts, Amy, L1 aTao, Luwei1 aDirenzo, James1 aBalboni, Amanda, L1 aCrisi, Giovanna, M1 aHagen, Mary, J1 aChandrasekaran, Thiruppavai1 aGauger, Kelly, J1 aSchneider, Sallie, Smith1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/oncogenic-transformation-of-mammary-epithelial-cells-by-transforming02242nas a2200241 4500008004100000245012100041210006900162260001600231520132700247100003001574700002401604700002001628700002201648700002301670700002501693700002201718700002101740700002001761700002201781700002501803700002101828856015101849 2013 ENG d00aPostpartum Remodeling, Lactation, and Breast Cancer Risk: Summary of a National Cancer Institute-Sponsored Workshop.0 aPostpartum Remodeling Lactation and Breast Cancer Risk Summary o c2013 Nov 133 aThe pregnancy-lactation cycle (PLC) is a period in which the breast is transformed from a less-developed, nonfunctional organ into a mature, milk-producing gland that has evolved to meet the nutritional, developmental, and immune protection needs of the newborn. Cessation of lactation initiates a process whereby the breast reverts to a resting state until the next pregnancy. Changes during this period permanently alter the morphology and molecular characteristics of the breast (molecular histology) and produce important, yet poorly understood, effects on breast cancer risk. To provide a state-of-the-science summary of this topic, the National Cancer Institute invited a multidisciplinary group of experts to participate in a workshop in Rockville, Maryland, on March 2, 2012. Topics discussed included: 1) the epidemiology of the PLC in relation to breast cancer risk, 2) breast milk as a biospecimen for molecular epidemiological and translational research, and 3) use of animal models to gain mechanistic insights into the effects of the PLC on breast carcinogenesis. This report summarizes conclusions of the workshop, proposes avenues for future research on the PLC and its relationship with breast cancer risk, and identifies opportunities to translate this knowledge to improve breast cancer outcomes.
1 aFaupel-Badger, Jessica, M1 aArcaro, Kathleen, F1 aBalkam, Jane, J1 aEliassen, Heather1 aHassiotou, Foteini1 aLebrilla, Carlito, B1 aMichels, Karin, B1 aPalmer, Julie, R1 aSchedin, Pepper1 aStuebe, Alison, M1 aWatson, Christine, J1 aSherman, Mark, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/postpartum-remodeling-lactation-and-breast-cancer-risk-summary-of-a01416nas a2200289 4500008004100000245008800041210006900129260000900198300000800207490000700215520046800222653002100690653001700711653004200728653001100770653003000781653001100811653002600822653001100848653001400859653001500873100001800888700002500906700002300931700002100954856015100975 2013 eng d00aPregnancy offers new insights into mechanisms of breast cancer risk and resistance.0 aPregnancy offers new insights into mechanisms of breast cancer r c2013 a3120 v153 aPregnancy induces long-lasting changes in gene expression that are associated with a reduction in breast cancer risk. Although several mechanisms have been proposed to mediate the reduction in breast cancer risk among parous women, recent studies focus attention on progenitor cells as major targets. The results suggest new biomarkers that may improve risk prediction and provide endpoints for assessment of clinical responses to prophylactic therapies.
10aBreast Neoplasms10aCell Lineage10aCyclin-Dependent Kinase Inhibitor p2710aFemale10aGene Expression Profiling10aHumans10aMammary Glands, Human10aParity10aPregnancy10aStem Cells1 aJerry, Joseph1 aMakari-Judson, Grace1 aCrisi, Giovanna, M1 aDunphy, Karen, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pregnancy-offers-new-insights-into-mechanisms-of-breast-cancer-risk02630nas a2200181 4500008004100000245015800041210006900199260001300268520184100281100002902122700002902151700002402180700002202204700002502226700002102251700002402272856015202296 2013 ENG d00aThe ras homolog gene family, member A (RhoA) pathway mediates MMP-2 and MMP-9-independent invasive behavior in a triple-negative breast cancer cell line.0 aras homolog gene family member A RhoA pathway mediates MMP2 and c2013 Jun3 aBreast cancer is a heterogeneous disease that varies in its biology and response to therapy. A foremost threat to patients is tumor invasion and metastasis, with the greatest risk among patients diagnosed with triple-negative and/or basal-like breast cancers. A greater understanding of the molecular mechanisms underlying cancer cell spreading is needed as 90% of cancer-associated deaths result from metastasis. We previously demonstrated that the tamoxifen-selected, MCF-7 derivative, TMX2-28, lacks expression of estrogen receptor α (ERα) and is highly invasive, yet maintains an epithelial morphology. The present study was designed to further characterize TMX2-28 cells and elucidate their invasion mechanism. We found that TMX2-28 cells do not express human epidermal growth factor receptor 2 (HER2) and progesterone receptor (PR), in addition to lacking ERα, making the cells triple-negative. We then determined that TMX2-28 cells lack expression of active matrix metalloproteinases (MMPs) -1, MMP-2, MMP-9, and other genes involved in epithelial-mesenchymal transition (EMT) suggesting that TMX2-28 may not utilize mesenchymal invasion. In contrast, TMX2-28 cells have high expression of RhoA, a protein known to play a critical role in amoeboid invasion. Blocking RhoA activity with the RhoA pathway specific inhibitor H-1152, or a RhoA specific siRNA, resulted in inhibition of invasive behavior. Collectively, these results suggest that TMX2-28 breast cancer cells exploit a RhoA-dependent, proteolytic-independent invasion mechanism. Targeting the RhoA pathway in triple-negative, basal-like breast cancers that have a proteolytic-independent invasion mechanism may provide therapeutic strategies for the treatment of patients with increased risk of metastasis. J. Cell. Biochem. © 2012 Wiley Periodicals, Inc.
1 aFagan-Solis, Katerina, D1 aSchneider, Sallie, Smith1 aPentecost, Brian, T1 aBentley, Brook, A1 aOtis, Christopher, N1 aGierthy, John, F1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-ras-homolog-gene-family-member-a-rhoa-pathway-mediates-mmp-2-and00914nas a2200301 4500008004100000245009600041210006900137260005200206300001600258490000800274100002400282700002100306700001700327700001500344700001800359700001500377700002000392700001600412700001700428700001600445700001700461700002000478700001800498700001800516700002000534700001400554856004400568 2013 eng d00aRecognition of microbial and mammalian phospholipid antigens by NKT cells with diverse TCRs0 aRecognition of microbial and mammalian phospholipid antigens by bProceedings of the National Academy of Sciences a1827–18320 v1101 aTatituri, R., V. V.1 aWatts, G., F. M.1 aBhowruth, V.1 aBarton, N.1 aRothchild, A.1 aHsu, F.-F.1 aAlmeida, C., F.1 aCox, L., R.1 aEggeling, L.1 aCardell, S.1 aRossjohn, J.1 aGodfrey, D., I.1 aBehar, S., M.1 aBesra, G., S.1 aBrenner, M., B.1 aBrigl, M. uhttps://doi.org/10.1073/pnas.122060111000521nas a2200157 4500008004100000245007700041210006900118260003500187300001600222490000800238100001800246700001500264700002200279700002300301856003900324 2013 eng d00aRegulation of endoplasmic reticulum Ca 2+ oscillations in mammalian eggs0 aRegulation of endoplasmic reticulum Ca 2 oscillations in mammali bThe Company of Biologistscoct a5714–57240 v1261 aWakai, Takuya1 aZhang, Nan1 aVangheluwe, Peter1 aFissore, Rafael, A uhttps://doi.org/10.1242/jcs.13654900623nas a2200181 4500008004100000245013900041210006900180260002500249300001400274490000700288100001900295700001700314700001800331700001600349700002000365700001800385856003800403 2013 eng d00aRegulation of hypoxia-inducible factor-1-alpha and HIF-1 alpha-related genes in equine digital lamellae and in cultured keratinocytes0 aRegulation of hypoxiainducible factor1alpha and HIF1 alpharelate bWiley-Blackwellcjul a203–2090 v461 aPawlak, E., A.1 aGeor, R., J.1 aWatts, M., R.1 aBlack, S, J1 aJohnson, P., J.1 aBelknap, J, K uhttps://doi.org/10.1111/evj.1209202902nas a2200457 4500008004100000245011300041210006900154260001500223300001200238490000800250520142900258653004101687653002101728653001201749653001601761653002701777653002201804653001101826653004401837653001101881653000901892653000901901653002401910653002301934653001901957653002101976653002101997653002402018100002502042700002902067700003002096700002002126700002202146700002302168700002602191700001902217700001902236700001702255700002002272856015202292 2013 eng d00aTherapeutic targeting of NOTCH signaling ameliorates immune-mediated bone marrow failure of aplastic anemia.0 aTherapeutic targeting of NOTCH signaling ameliorates immunemedia c2013 Jul 1 a1311-290 v2103 aSevere aplastic anemia (AA) is a bone marrow (BM) failure (BMF) disease frequently caused by aberrant immune destruction of blood progenitors. Although a Th1-mediated pathology is well described for AA, molecular mechanisms driving disease progression remain ill defined. The NOTCH signaling pathway mediates Th1 cell differentiation in the presence of polarizing cytokines, an action requiring enzymatic processing of NOTCH receptors by γ-secretase. Using a mouse model of AA, we demonstrate that expression of both intracellular NOTCH1(IC) and T-BET, a key transcription factor regulating Th1 cell differentiation, was increased in spleen and BM-infiltrating T cells during active disease. Conditionally deleting Notch1 or administering γ-secretase inhibitors (GSIs) in vivo attenuated disease and rescued mice from lethal BMF. In peripheral T cells from patients with untreated AA, NOTCH1(IC) was significantly elevated and bound to the TBX21 promoter, showing NOTCH1 directly regulates the gene encoding T-BET. Treating patient cells with GSIs in vitro lowered NOTCH1(IC) levels, decreased NOTCH1 detectable at the TBX21 promoter, and decreased T-BET expression, indicating that NOTCH1 signaling is responsive to GSIs during active disease. Collectively, these results identify NOTCH signaling as a primary driver of Th1-mediated pathogenesis in AA and may represent a novel target for therapeutic intervention.
10aAmyloid Precursor Protein Secretases10aAnemia, Aplastic10aAnimals10aBone Marrow10aDisease Models, Animal10aEnzyme Inhibitors10aFemale10aHematopoietic Stem Cell Transplantation10aHumans10aMale10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aMice, Knockout10aMice, Transgenic10aReceptor, Notch110aSignal Transduction1 aRoderick, Justine, E1 aGonzalez-Perez, Gabriela1 aKuksin, Christina, Arieta1 aDongre, Anushka1 aRoberts, Emily, R1 aSrinivasan, Janani1 aAndrzejewski, Chester1 aFauq, Abdul, H1 aGolde, Todd, E1 aMiele, Lucio1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/therapeutic-targeting-of-notch-signaling-ameliorates-immune-mediated00840nas a2200265 4500008004100000245008300041210006900124260001800193300001600211490000800227100002200235700002200257700002400279700002600303700002300329700002400352700002000376700001900396700001900415700002400434700002700458700002200485700002000507856004700527 2013 eng d00aTryptophan Biosynthesis Protects Mycobacteria from CD4 T-Cell-Mediated Killing0 aTryptophan Biosynthesis Protects Mycobacteria from CD4 TCellMedi bElsevier {BV} a1296–13080 v1551 aZhang, Yanjia, J.1 aReddy, Manchi, C.1 aIoerger, Thomas, R.1 aRothchild, Alissa, C.1 aDartois, Veronique1 aSchuster, Brian, M.1 aTrauner, Andrej1 aWallis, Deeann1 aGalaviz, Stacy1 aHuttenhower, Curtis1 aSacchettini, James, C.1 aBehar, Samuel, M.1 aRubin, Eric, J. uhttps://doi.org/10.1016/j.cell.2013.10.04502767nas a2200349 4500008004100000245011200041210006900153260000900222300001100231490000600242520162700248653001201875653001501887653002301902653001301925653001101938653002001949653002901969653000901998653001302007653002402020653004102044653001302085653002902098100001702127700003002144700002502174700002402199700001702223700002602240856015102266 2013 eng d00aVisceral endoderm expression of Yin-Yang1 (YY1) is required for VEGFA maintenance and yolk sac development.0 aVisceral endoderm expression of YinYang1 YY1 is required for VEG c2013 ae588280 v83 aMouse embryos lacking the polycomb group gene member Yin-Yang1 (YY1) die during the peri-implantation stage. To assess the post-gastrulation role of YY1, a conditional knock-out (cKO) strategy was used to delete YY1 from the visceral endoderm of the yolk sac and the definitive endoderm of the embryo. cKO embryos display profound yolk sac defects at 9.5 days post coitum (dpc), including disrupted angiogenesis in mesoderm derivatives and altered epithelial characteristics in the visceral endoderm. Significant changes in both cell death and proliferation were confined to the YY1-expressing yolk sac mesoderm indicating that loss of YY1 in the visceral endoderm causes defects in the adjacent yolk sac mesoderm. Production of Vascular Endothelial Growth Factor A (VEGFA) by the visceral endoderm is essential for normal growth and development of the yolk sac vasculature. Reduced levels of VEGFA are observed in the cKO yolk sac, suggesting a cause for the angiogenesis defects. Ex vivo culture with exogenous VEGF not only rescued angiogenesis and apoptosis in the cKO yolk sac mesoderm, but also restored the epithelial defects observed in the cKO visceral endoderm. Intriguingly, blocking the activity of the mesoderm-localized VEGF receptor, FLK1, recapitulates both the mesoderm and visceral endoderm defects observed in the cKO yolk sac. Taken together, these results demonstrate that YY1 is responsible for maintaining VEGF in the developing visceral endoderm and that a VEGF-responsive paracrine signal, originating in the yolk sac mesoderm, is required to promote normal visceral endoderm development.
10aAnimals10aCell Death10aCell Proliferation10aEndoderm10aFemale10aGene Expression10aGene Knockout Techniques10aMale10aMesoderm10aSignal Transduction10aVascular Endothelial Growth Factor A10aYolk Sac10aYY1 Transcription Factor1 aRhee, Siyeon1 aGuerrero-Zayas, Mara-Isel1 aWallingford, Mary, C1 aOrtiz-Pineda, Pablo1 aMager, Jesse1 aTremblay, Kimberly, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/visceral-endoderm-expression-of-yin-yang1-yy1-is-required-for-vegfa01985nas a2200145 4500008004100000245013700041210006900178260001500247520132700262100002101589700002601610700002401636700002501660856015401685 2012 ENG d00aADAM13 function is required in the 3 dimensional context of the embryo during cranial neural crest cell migration in Xenopus laevis.0 aADAM13 function is required in the 3 dimensional context of the c2012 Jun 73 aThe cranial neural crest (CNC) is a population of cells that arises from the lateral part of the developing brain, migrates ventrally and coordinates the entire craniofacial development of vertebrates. Many molecules are involved in CNC migration including the transmembrane metalloproteases ADAM13 and 19. We have previously shown that these ADAMs cleave a number of extracellular proteins and modify the transcription of a number of genes, and that both of these activities are important for cell migration. Here we show that the knock down of ADAM13 inhibits CNC migration in vivo but not in vitro, indicating that ADAM13 function is required in the 3-dimentional context of the embryo. We further show that the migration of CNC that do not express ADAM13 and ADAM19 can be rescued in vivo by co-grafting wild type CNC. Furthermore, the migration of CNC lacking ADAM13 can be rescued by mechanically separating the CNC from the surrounding ectoderm and mesoderm. Finally, we show that ADAM13 function is autonomous to CNC tissue, as the migration of morphant CNC can only be rescued by ADAM13 expression in the CNC and not the surrounding tissues. Together our results suggest that ADAM13 changes CNC interaction with the extracellular environment and that this change is necessary for their migration in vivo.
1 aCousin, Hélène1 aAbbruzzese, Genevieve1 aMcCusker, Catherine1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/adam13-function-is-required-in-the-3-dimensional-context-of-the-embryo02040nas a2200145 4500008004100000245005100041210005000092260000900142300001000151490000800161520155100169100002001720700002501740856012901765 2012 eng d00aAdvances in the mode of action of pyrethroids.0 aAdvances in the mode of action of pyrethroids c2012 a49-720 v3143 aThe ability to clone, express, and electrophysiologically measure currents carried by voltage-gated ion channels has allowed a detailed assessment of the action of pyrethroids on various target proteins.Recently, the heterologous expression of various rat brain voltage-gated sodium channel isoforms in Xenopus laevis oocytes has determined a wide range of sensitivities to the pyrethroids, with some channels virtually insensitive and others highly sensitive. Furthermore, some isoforms show selective sensitivity to certain pyrethroids and this selectivity can be altered in a state-dependent manner. Additionally, some rat brain isoforms are apparently more sensitive to pyrethroids than the corresponding human isoform. These finding may have significant relevance in judging the merit and value of assessing the risk of pyrethroid exposures to humans using toxicological studies done in rat.Other target sites for certain pyrethroids include the voltage-gated calcium and chloride channels. Of particular interest is the increased effect of Type II pyrethroids on certain phosphoforms of the N-type Ca(v)2.2 calcium channel following post-translational modification and its relationship to enhanced neurotransmitter release seen in vivo.Lastly, parallel neurobehavioral and mechanistic studies on three target sites suggest that a fundamental difference exists between the action of Types I and II pyrethroids, both on a functional and molecular level. These differences should be considered in any future risk evaluation of the pyrethroids.1 aClark, Marshall1 aSymington, Steven, B uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/advances-in-the-mode-of-action-of-pyrethroids02750nas a2200385 4500008004100000245009100041210006900132260001500201300000900216490000800225520155000233653001201783653002601795653001801821653002101839653001601860653003101876653002001907653001401927653002101941653001401962653001601976653000901992653002402001653001402025653001702039653001502056653001302071653003702084100002002121700002602141700002102167700002402188856015202212 2012 eng d00aB Lymphocytes provide an infection niche for intracellular bacterium Brucella abortus.0 aB Lymphocytes provide an infection niche for intracellular bacte c2012 Jul 1 a91-80 v2063 aBACKGROUND: Brucella spp. are intracellular bacteria that establish lifelong infections whose mechanisms of chronicity are poorly understood. Notably, B cells facilitate the establishment of the high infection plateau that persists for months. METHODS: We evaluated the contribution of murine B cells toward providing infection niches for Brucella by using flow cytometry and microscopy and by determining live bacterial counts associated with B cells both in vivo and in vitro. RESULTS: Herein we demonstrate that immunoglobulin M and complement-opsonized Brucella abortus infects and survives inside primary murine B cells protected from bactericidal effects of gentamicin. The entry was dependent on microfilaments for internalization and subsequently brucellae reside in a late endosomal/lysosomal compartment. Throughout the infection, 10% of colony-forming units from infected mice was associated with B cells, and these cells transferred disease to naive hosts. Furthermore, Brucella-positive cells were positive for transforming growth factor (TGF) β1, and about 10% of such cells were B cells, similar to rates found for other intracellular pathogens that induce their hosts cells to produce TGF-β1. CONCLUSIONS: To conclude, infected B cells contribute to chronic bacterial infections by providing an intracellular niche that may exert an immunoregulatory role. Although professional phagocytic cells harbor intracellular bacteria including Brucella, infection of lymphocytes by bacteria has not been previously appreciated.
10aAnimals10aAntibodies, Bacterial10aB-Lymphocytes10aBrucella abortus10aBrucellosis10aComplement System Proteins10aDNA Replication10aEndosomes10aImmunoglobulin M10aLysosomes10aMacrophages10aMice10aMice, Inbred BALB C10aMonocytes10aPhagocytosis10aStem Cells10aSurvival10aTransforming Growth Factor beta11 aGoenka, Radhika1 aGuirnalda, Patrick, D1 aBlack, Samuel, J1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/b-lymphocytes-provide-an-infection-niche-for-intracellular-bacterium02662nas a2200397 4500008004100000245007600041210006900117260001600186300001200202490000800214520141300222653001201635653001801647653001801665653001901683653002001702653002601722653000901748653001901757653003101776653002401807653003001831100001701861700002601878700002301904700001901927700002201946700001901968700001901987700002102006700002102027700002302048700002302071700001902094856015102113 2012 eng d00aBasis of CTLA-4 function in regulatory and conventional CD4(+) T cells.0 aBasis of CTLA4 function in regulatory and conventional CD4 T cel c2012 May 31 a5155-630 v1193 aCTLA-4 proteins contribute to the suppressor function of regulatory T cells (Tregs), but the mechanism by which they do so remains incompletely understood. In the present study, we assessed CTLA-4 protein function in both Tregs and conventional (Tconv) CD4(+) T cells. We report that CTLA-4 proteins are responsible for all 3 characteristic Treg functions of suppression, TCR hyposignaling, and anergy. However, Treg suppression and anergy only required the external domain of CTLA-4, whereas TCR hyposignaling required its internal domain. Surprisingly, TCR hyposignaling was neither required for Treg suppression nor anergy because costimulatory blockade by the external domain of CTLA-4 was sufficient for both functions. We also report that CTLA-4 proteins were localized in Tregs in submembrane vesicles that rapidly recycled to/from the cell surface, whereas CTLA-4 proteins in naive Tconv cells were retained in Golgi vesicles away from the cell membrane and had no effect on Tconv cell function. However, TCR signaling of Tconv cells released CTLA-4 proteins from Golgi retention and caused activated Tconv cells to acquire suppressor function. Therefore, the results of this study demonstrate the importance of intracellular localization for CTLA-4 protein function and reveal that CTLA-4 protein externalization imparts suppressor function to both regulatory and conventional CD4(+) T cells.
10aAnimals10aCell Membrane10aClonal Anergy10aCTLA-4 Antigen10aGolgi Apparatus10aLymphocyte Activation10aMice10aMice, Knockout10aReceptors, Antigen, T-Cell10aSignal Transduction10aT-Lymphocytes, Regulatory1 aTai, Xuguang1 aVan Laethem, Francois1 aPobezinsky, Leonid1 aGuinter, Terry1 aSharrow, Susan, O1 aAdams, Anthony1 aGranger, Larry1 aKruhlak, Michael1 aLindsten, Tullia1 aThompson, Craig, B1 aFeigenbaum, Lionel1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/basis-of-ctla-4-function-in-regulatory-and-conventional-cd4-t-cells00892nas a2200277 4500008004100000245010200041210006900143260002000212300000600232490000600238100001900244700002400263700002500287700003100312700002800343700002300371700002000394700002000414700002300434700002300457700002100480700002500501700002400526700002200550856004200572 2012 eng d00aBiological conversion assay using Clostridium phytofermentans to estimate plant feedstock quality0 aBiological conversion assay using Clostridium phytofermentans to bSpringer Nature a50 v51 aLee, Scott, J.1 aWarnick, Thomas, A.1 aPattathil, Sivakumar1 aAlvelo-Maurosa, Jesús, G.1 aSerapiglia, Michelle, J1 aMcCormick, Heather1 aBrown, Virginia1 aYoung, Naomi, F1 aSchnell, Danny, J.1 aSmart, Lawrence, B1 aHahn, Michael, G1 aPedersen, Jeffrey, F1 aLeschine, Susan, B.1 aHazen, Samuel, P. uhttps://doi.org/10.1186/1754-6834-5-501410nas a2200121 4500008004100000245006700041210006200108260001600170520091600186100002001102700002401122856014201146 2012 ENG d00aCanonical and Non-Canonical Notch Signaling in CD4(+) T Cells.0 aCanonical and NonCanonical Notch Signaling in CD4 T Cells c2012 Jun 153 aFor T cells to become fully activated, they must integrate a myriad of signals, both extrinsic and intrinsic. External stimuli accrued through various cell surface receptors are transduced and amplified through a coordinated circuitry of signaling cascades that ultimately result in the transcription of new genes. Along the way, extracellular and intracellular signaling components function to impart a fully activated state. Evidence is accumulating to show that the Notch family of cell surface receptors, long known to function as transcriptional regulators through their interactions with the canonical nuclear binding protein CSL/RBP-J, may also be playing an as-yet-unappreciated role in T cell activation by virtue of its signaling via non-canonical as well as nonnuclear mechanisms. In this review we will discuss these and other better-known means by which Notch signaling influences T cell responses.1 aMinter, Lisa, M1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/canonical-and-non-canonical-notch-signaling-in-cd4-t-cells02389nas a2200421 4500008004100000245009300041210006900134260001300203300001100216490000700227520107000234653001201304653001901316653001801335653001801353653002501371653002001396653001901416653002101435653000901456653002401465653002301489653001901512653002101531653003101552653002401583653001501607653001701622100002601639700002301665700001701688700002201705700002601727700002301753700002001776700001901796856015201815 2012 eng d00aClonal deletion and the fate of autoreactive thymocytes that survive negative selection.0 aClonal deletion and the fate of autoreactive thymocytes that sur c2012 Jun a569-780 v133 aClonal deletion of autoreactive thymocytes is important for self-tolerance, but the intrathymic signals that induce clonal deletion have not been clearly identified. We now report that clonal deletion during negative selection required CD28-mediated costimulation of autoreactive thymocytes at the CD4(+)CD8(lo) intermediate stage of differentiation. Autoreactive thymocytes were prevented from undergoing clonal deletion by either a lack of CD28 costimulation or transgenic overexpression of the antiapoptotic factors Bcl-2 or Mcl-1, with surviving thymocytes differentiating into anergic CD4(-)CD8(-) double-negative thymocytes positive for the T cell antigen receptor αβ subtype (TCRαβ) that 'preferentially' migrated to the intestine, where they re-expressed CD8α and were sequestered as CD8αα(+) intraepithelial lymphocytes (IELs). Our study identifies costimulation by CD28 as the intrathymic signal required for clonal deletion and identifies CD8αα(+) IELs as the developmental fate of autoreactive thymocytes that survive negative selection.
10aAnimals10aAntigens, CD2810aAntigens, CD410aAntigens, CD810aCell Differentiation10aClonal Deletion10aFlow Cytometry10aImmune Tolerance10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aMice, Knockout10aMice, Transgenic10aReceptors, Antigen, T-Cell10aSignal Transduction10aThymocytes10aThymus Gland1 aPobezinsky, Leonid, A1 aAngelov, Georgi, S1 aTai, Xuguang1 aJeurling, Susanna1 aVan Laethem, Francois1 aFeigenbaum, Lionel1 aPark, Jung-Hyun1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/clonal-deletion-and-the-fate-of-autoreactive-thymocytes-that-survive02512nas a2200241 4500008004100000245010700041210006900148260001500217300001000232490000800242520165500250100001701905700001901922700002301941700002501964700002001989700002002009700002302029700002302052700001902075700002302094856015302117 2012 eng d00acSrc is necessary for epididymal development and is incorporated into sperm during epididymal transit.0 acSrc is necessary for epididymal development and is incorporated c2012 Sep 1 a43-530 v3693 aChanges that occur to mammalian sperm upon epididymal transit and maturation render these cells capable of moving progressively and capacitating. Signaling events leading to mammalian sperm capacitation depend on the modulation of proteins by phosphorylation and dephosphorylation cascades. Recent experiments have demonstrated that the Src family of kinases plays an important role in the regulation of these events. However, sperm from cSrc null mice display normal tyrosine phosphorylation associated with capacitation. We report here that, despite normal phosphorylation, sperm from cSrc null mice display a severe reduction in forward motility, and are unable to fertilize in vitro. Histological analysis of seminiferous tubules in the testes, caput and corpus epididymis do not reveal obvious defects. However, the cauda epididymis is significantly smaller, and expression of key transport proteins in the epithelial cells lining this region is reduced in cSrc null mice compared to wild type littermates. Although previously, we and others have shown the presence of cSrc in mature sperm from cauda epididymis, a closer evaluation indicates that this tyrosine kinase is not present in sperm from the caput epididymis, suggesting that this protein is acquired by sperm later during epididymal maturation. Consistent with this observation, cSrc is enriched in vesicles released by the epididymal epithelium known as epididymosomes. Altogether, these observations indicate that cSrc is essential for cauda epididymal development and suggest an essential role of this kinase in epididymal sperm maturation involving cSrc extracellular trafficking.1 aKrapf, Dario1 aRuan, Ye, Chun1 aWertheimer, Eva, V1 aBattistone, Maria, A1 aPawlak, John, B1 aSanjay, Archana1 aPilder, Stephen, H1 aCuasnicu, Patricia1 aBreton, Sylvie1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/csrc-is-necessary-for-epididymal-development-and-is-incorporated-into01548nas a2200145 4500008004100000245008100041210006900122260001500191520097500206100001501181700001901196700002501215700001701240856014501257 2012 ENG d00aDepletion of Suds3 reveals an essential role in early lineage specification.0 aDepletion of Suds3 reveals an essential role in early lineage sp c2012 Nov 13 aPreimplantation development culminates with the emergence of three distinct populations: the inner cell mass, primitive endoderm and trophectoderm. Here, we define the mechanisms underlying the requirement of Suds3 in pre/peri-implantation development. Suds3 knockdown blastocysts exhibit a failure of both trophectoderm proliferation as well as a conspicuous lack of primitive endoderm. Expression of essential lineage factors Nanog, Sox2, Cdx2, Eomes, Elf5 and Sox17 are severely reduced in the absence of Suds3. Importantly, we document deficient FGF4/ERK signaling and show that exogenous FGF4 rescues primitive endoderm formation and trophectoderm proliferation in Suds3 knockdown blastocysts. We also show that Hdac1 knockdown reduces Sox2/FGF4/ERK signaling in blastocysts. Collectively, these data define a role for Suds3 in activation of FGF4/ERK signaling and determine an essential molecular role of Suds3/Sin3/HDAC complexes in lineage specification in vivo.1 aZhang, Kun1 aDai, Xiangpeng1 aWallingford, Mary, C1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/depletion-of-suds3-reveals-an-essential-role-in-early-lineage02858nas a2200193 4500008004100000245013600041210006900177260001500246300001200261490000600273520209400279100002302373700002602396700002402422700001902446700002302465700002402488856015202512 2012 eng d00aDetection of promoter methylation of tumor suppressor genes in serum DNA of breast cancer cases and benign breast disease controls.0 aDetection of promoter methylation of tumor suppressor genes in s c2012 Nov 1 a1258-670 v73 aTumors are capable of shedding DNA into the blood stream. This shed DNA may be recovered from serum or plasma. The objective of this study was to evaluate whether pyrosequencing promoter DNA in a panel of 12 breast cancer-related genes (APC, BRCA1, CCND2, CDH1, ESR1, GSTP1, HIN1, P16, RARβ, RASSF1, SFRP1 and TWIST) to measure the degree of methylation would lead to a useful serum-based marker of breast cancer. Serum was obtained from women who were about to undergo a breast biopsy or mastectomy at three hospitals from 1977 to 1987 in Grand Rapids, MI USA. We compared the methylation status of 12 genes in serum DNA obtained from three groups of postmenopausal women (mean age at blood collection: 63.0 y; SD 9.9; range 35-91): breast cancer cases with lymph node-positive disease (n = 241); breast cancer cases with lymph node-negative disease (n = 63); and benign breast disease control subjects (n = 234). Overall, median levels of promoter methylation were low, typically below 5%, for all genes in all study groups. For all genes, median levels of methylation were higher (by 3.3 to 47.6%) in lymph node-positive breast cancer cases than in the controls. Comparing mean methylation level between lymph-node positive cases and controls, the most statistically significant findings, after adjustment of the false-positive rate (q-value), were for TWIST (p = 0.04), SFRP1 (p = 0.16), ESR1 (p = 0.17), P16 (p = 0.19) and APC (p = 0.19). For two of these four genes (TWIST, P16), the median methylation level was also highest in lymph-node positive cases, intermediate in lymph node-negative cases and lowest in the controls. The percent of study subjects with mean methylation scores ≥ 5% was higher among lymph node-positive cases than controls for ten genes, and significantly higher for HIN1 and TWIST (22.0 vs. 12.2%, p = 0.04 and 37.9 vs. 24.5%, p = 0.004, respectively). Despite relatively consistent variation in methylation patterns among groups, these modest differences did not provide sufficient ability to distinguish between cases and controls in a clinical setting.1 aSturgeon, Susan, R1 aBalasubramanian, Raji1 aSchairer, Catherine1 aMuss, Hyman, B1 aZiegler, Regina, G1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/detection-of-promoter-methylation-of-tumor-suppressor-genes-in-serum02447nas a2200205 4500008004100000245011600041210006900157260000900226300000800235490000700243520170100250100001501951700001401966700002201980700002202002700001902024700002402043700002202067856015202089 2012 eng d00aDifferential expression of cancer associated proteins in breast milk based on age at first full term pregnancy.0 aDifferential expression of cancer associated proteins in breast c2012 a1000 v123 aBACKGROUND: First full term pregnancy (FFTP) completed at a young age has been linked to low long term breast cancer risk, whereas late FFTP pregnancy age confers high long term risk, compared to nulliparity. Our hypothesis was that proteins linked to breast cancer would be differentially expressed in human milk collected at three time points during lactation based on age at FFTP. METHODS: We analyzed breast milk from 72 lactating women. Samples were collected within 10 days of the onset of lactation (baseline-BL), two months after lactation started and during breast weaning (W). We measured 16 proteins (11 kallikreins (KLKs), basic fibroblast growth factor, YKL-40, neutrophil gelatinase-associated lipocalin and transforming growth factor (TGF) β-1 and -2) associated with breast cancer, most known to be secreted into milk. RESULTS: During lactation there was a significant change in the expression of 14 proteins in women < 26 years old and 9 proteins in women > = 26 at FFTP. The most significant (p < .001) changes from BL to W in women divided by FFTP age (< 26 vs. > = 26) were in KLK3,6, 8, and TGFβ2 in women < 26; and KLK6, 8, and TGFβ2 in women > = 26. There was a significant increase (p = .022) in KLK8 expression from BL to W depending on FFTP age. Examination of DNA methylation in the promoter region of KLK6 revealed high levels of methylation that did not explain the observed changes in protein levels. On the other hand, KLK6 and TGFβ1 expression were significantly associated (r2 = .43, p = .0050). CONCLUSIONS: The expression profile of milk proteins linked to breast cancer is influenced by age at FFTP. These proteins may play a role in future cancer risk.1 aQin, Wenyi1 aZhang, Ke1 aKliethermes, Beth1 aRuhlen, Rachel, L1 aBrowne, Eva, P1 aArcaro, Kathleen, F1 aSauter, Edward, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/differential-expression-of-cancer-associated-proteins-in-breast-milk01990nas a2200193 4500008004100000245011600041210006900157260001300226300001000239490000700249520127000256100002401526700001901550700001501569700001401584700002501598700002201623856015101645 2012 eng d00aDifferential expression of cancer-related proteins in paired breast milk samples from women with breast cancer.0 aDifferential expression of cancerrelated proteins in paired brea c2012 Nov a543-60 v283 aBackground: Breast cancer risk increases during pregnancy and remains elevated for a number of years thereafter. Cancer-associated proteins that are secreted into breast milk may provide a means to detect cancer in the lactating breast or to assess future breast cancer risk. Objective: To determine whether proteins linked to breast cancer would be differentially expressed in matched (both breasts from each participant) human milk samples collected from women with unilateral breast cancer. Methods: Five cancer-associated proteins (basic fibroblast growth factor [bFGF], YKL-40, neutrophil gelatinase-associated lipocalin, and transforming growth factor β1 and β2) were analyzed in milk provided by 5 lactating women, 4 of whom were known to have cancer in 1 breast (and the opposite breast clinically disease free) at the time of milk collection and 1 who developed breast cancer 2 years after milk collection. Results: Expression was significantly higher for TGFβ2 (P = .03) and bFGF (P =.03) in the breasts with cancer. Conclusion: These proteins may play a role in assessing a woman's risk of pregnancy-associated breast cancer. Because of variable protein concentration among patients and the limited sample size, the results are considered preliminary.1 aArcaro, Kathleen, F1 aBrowne, Eva, P1 aQin, Wenyi1 aZhang, Ke1 aAnderton, Douglas, L1 aSauter, Edward, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/differential-expression-of-cancer-related-proteins-in-paired-breast02535nas a2200217 4500008004100000245016300041210006900204260001300273300001200286490000700298520170600305100001802011700001302029700002302042700001802065700001602083700002202099700002502121700002102146856015002167 2012 eng d00aDistribution and processing of a disintegrin and metalloproteinase with thrombospondin motifs-4, aggrecan, versican, and hyaluronan in equine digital laminae.0 aDistribution and processing of a disintegrin and metalloproteina c2012 Jul a1035-460 v733 aOBJECTIVE: To determine the expression and distribution of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), its substrates aggrecan and versican, and their binding partner hyaluronan in laminae of healthy horses. SAMPLE: Laminae from the forelimb hooves of 8 healthy horses. PROCEDURES: Real-time quantitative PCR assay was used for gene expression analysis. Hyaluronidase, chondroitinase, and keratanase digestion of lamina extracts combined with SDS-PAGE and western blotting were used for protein and proteoglycan analysis. Immunofluorescent and immunohistochemical staining of tissue sections were used for protein and hyaluronan localization. RESULTS: Genes encoding ADAMTS-4, aggrecan, versican, and hyaluronan synthase II were expressed in laminae. The ADAMTS-4 was predominantly evident as a 51-kDa protein bearing a catalytic site neoepitope indicative of active enzyme and in situ activity, which was confirmed by the presence of aggrecan and versican fragments bearing ADAMTS-4 cleavage neoepitopes in laminar protein extracts. Aggrecan, versican, and hyaluronan were localized to basal epithelial cells within the secondary epidermal laminae. The ADAMTS-4 localized to these cells but was also present in some cells in the dermal laminae. CONCLUSIONS AND CLINICAL RELEVANCE: Within digital laminae, versican exclusively and aggrecan primarily localized within basal epithelial cells and both were constitutively cleaved by ADAMTS-4, which therefore contributed to their turnover. On the basis of known properties of these proteoglycans, it is possible that they can protect the basal epithelial cells of horses from biomechanical and concussive stress.
1 aPawlak, Erica1 aWang, Le1 aJohnson, Philip, J1 aNuovo, Gerard1 aTaye, Almaz1 aBelknap, James, K1 aAlfandari, Dominique1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/distribution-and-processing-of-a-disintegrin-and-metalloproteinase02550nas a2200193 4500008004100000245022600041210006900267260001300336300001200349490000700361520171900368100001302087700001802100700002302118700002202141700002502163700002102188856014702209 2012 eng d00aEffects of cleavage by a disintegrin and metalloproteinase with thrombospondin motifs-4 on gene expression and protein content of versican and aggrecan in the digital laminae of horses with starch gruel-induced laminitis.0 aEffects of cleavage by a disintegrin and metalloproteinase with c2012 Jul a1047-560 v733 aOBJECTIVE: To determine whether increased gene expression of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) in laminae of horses with starch gruel-induced laminitis was accompanied by increased enzyme activity and substrate degradation. SAMPLE: Laminae from the forelimb hooves of 8 healthy horses and 17 horses with starch gruel-induced laminitis (6 at onset of fever, 6 at onset of Obel grade 1 lameness, and 5 at onset of Obel grade 3 lameness). PROCEDURES: Gene expression was determined by use of cDNA and real-time quantitative PCR assay. Protein expression and processing were determined via SDS-PAGE and quantitative western blotting. Protein distribution and abundance were determined via quantitative immunofluorescent staining. RESULTS: ADAMTS-4 gene expression was increased and that of versican decreased in laminitic laminae, compared with expression in healthy laminae. Catalytically active ADAMTS-4 also was increased in the tissue, as were ADAMTS-4-cleavage fragments of versican. Immunofluorescent analyses indicated that versican was depleted from the basal epithelia of laminae of horses at onset of Obel grade 3 lameness, compared with results for healthy laminae, and this was accompanied by regional separation of basal epithelial cells from the basement membrane. Aggrecan gene and protein expression were not significantly affected. CONCLUSIONS AND CLINICAL RELEVANCE: Changes in gene and protein expression of ADAMTS-4 and versican in the basal epithelium of laminitic laminae indicated a fundamental change in the physiology of basal epithelial cells. This was accompanied by and may have caused detachment of these cells from the basement membrane.
1 aWang, Le1 aPawlak, Erica1 aJohnson, Philip, J1 aBelknap, James, K1 aAlfandari, Dominique1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/effects-of-cleavage-by-a-disintegrin-and-metalloproteinase-with02428nas a2200205 4500008004100000245012900041210006900170260001600239520161000255100002901865700002501894700002101919700002301940700002901963700002201992700001802014700002002032700002102052856014902073 2012 eng d00aElectrophysiological evidence for the presence of cystic fibrosis transmembrane conductance regulator (CFTR) in mouse sperm.0 aElectrophysiological evidence for the presence of cystic fibrosi c2012 Jul 253 aMammalian sperm must undergo a maturational process, named capacitation, in the female reproductive tract to fertilize the egg. Sperm capacitation is regulated by a cAMP/PKA pathway and involves increases in intracellular Ca(2+) , pH, Cl(-) , protein tyrosine phosphorylation, and in mouse and some other mammals a membrane potential hyperpolarization. The cystic fibrosis transmembrane conductance regulator (CFTR), a Cl(-) channel modulated by cAMP/PKA and ATP, was detected in mammalian sperm and proposed to modulate capacitation. Our whole-cell patch-clamp recordings from testicular mouse sperm now reveal a Cl(-) selective component to membrane current that is ATP-dependent, stimulated by cAMP, cGMP and genistein (a CFTR agonist, at low concentrations), and inhibited by DPC and CFTR(inh) -172, two well-known CFTR antagonists. Furthermore, the Cl(-) current component activated by cAMP and inhibited by CFTR(inh) -172 is absent in recordings on testicular sperm from mice possessing the CFTR ▵F508 loss-of-function mutation, indicating that CFTR is responsible for this component. A Cl(-) selective like current component displaying CFTR characteristics was also found in wild type epididymal sperm bearing the cytoplasmatic droplet. Capacitated sperm treated with CFTR(inh) -172 undergo a shape change, suggesting that CFTR is involved in cell volume regulation. These findings indicate that functional CFTR channels are present in mouse sperm and their biophysical properties are consistent with their proposed participation in capacitation. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.1 aFierro, Dulce, Figueiras1 aAcevedo, Juan, José1 aMartínez, Pablo1 aEscoffier, Jessica1 aSepúlveda, Francisco, V1 aBalderas, Enrique1 aOrta, Gerardo1 aVisconti, Pablo1 aDarszon, Alberto uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/electrophysiological-evidence-for-the-presence-of-cystic-fibrosis02288nas a2200157 4500008004100000245009800041210006900139260001500208520165100223100002301874700001701897700002201914700002101936700002301957856015001980 2012 ENG d00aFlow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation.0 aFlow cytometry analysis reveals a decrease in intracellular sodi c2012 Feb 23 aMammalian sperm require time in the female tract in order to be able to fertilize an egg. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential. Recently, we presented evidence showing that epithelial Na(+) channels (ENaC) are present in mature sperm and that ENaCs are blocked during capacitation. In the present work, we used flow cytometry to analyze changes in intracellular Na(+) concentration ([Na(+)](i)) during capacitation in individual cells. Our results indicate that capacitated sperm have lower Na(+) concentrations. Using sperm with green fluorescent protein in their acrosomes, it was shown that the lower [Na(+)](i) concentration only occurs in sperm having intact acrosomes. ENaC inhibition has been shown in other cell types to depend on the activation of cystic fibrosis transmembrane conductance regulator (CFTR). In non-capacitated sperm, amiloride, an ENaC inhibitor, and genistein, a CFTR activator, caused a decrease in [Na(+)](i), suggesting that also in these cells [Na(+)](i) is dependent on the crosstalk between ENaC and CFTR. In addition, PKA inhibition blocked [Na(+)](i) decrease in capacitated sperm. Altogether, these data are consistent with the hypothesis that the capacitation-associated hyperpolarization involves a decrease in [Na(+)](i) mediated by inhibition of ENaC and regulated by PKA through activation of CFTR channels.1 aEscoffier, Jessica1 aKrapf, Dario1 aNavarrete, Felipe1 aDarszon, Alberto1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/flow-cytometry-analysis-reveals-a-decrease-in-intracellular-sodium03525nas a2200385 4500008004100000245010400041210006900145260000900214300000700223490000700230520224900237653001202486653001802498653001102516653002302527653002302550653001602573653001102589653002702600653002802627653002102655653001402676653002602690653003202716653004002748653004402788100001702832700002502849700002502874700001902899700002202918700002202940700002402962856015302986 2012 eng d00aGene number determination and genetic polymorphism of the gamma delta T cell co-receptor WC1 genes.0 aGene number determination and genetic polymorphism of the gamma c2012 a860 v133 aBACKGROUND: WC1 co-receptors belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. Expression of particular WC1 genes defines functional subpopulations of WC1(+) γδ T cells. We have previously identified partial or complete genomic sequences for thirteen different WC1 genes through annotation of the bovine genome Btau_3.1 build. We also identified two WC1 cDNA sequences from other cattle that did not correspond to sequences in the Btau_3.1 build. Their absence in the Btau_3.1 build may have reflected gaps in the genome assembly or polymorphisms among animals. Since the response of γδ T cells to bacterial challenge is determined by WC1 gene expression, it was critical to understand whether individual cattle or breeds differ in the number of WC1 genes or display polymorphisms. RESULTS: Real-time quantitative PCR using DNA from the animal whose genome was sequenced ("Dominette") and sixteen other animals representing ten breeds of cattle, showed that the number of genes coding for WC1 co-receptors is thirteen. The complete coding sequences of those thirteen WC1 genes is presented, including the correction of an error in the WC1-2 gene due to mis-assembly in the Btau_3.1 build. All other cDNA sequences were found to agree with the previous annotation of complete or partial WC1 genes. PCR amplification and sequencing of the most variable N-terminal SRCR domain (domain 1 which has the SRCR "a" pattern) of each of the thirteen WC1 genes showed that the sequences are highly conserved among individuals and breeds. Of 160 sequences of domain 1 from three breeds of cattle, no additional sequences beyond the thirteen described WC1 genes were found. Analysis of the complete WC1 cDNA sequences indicated that the thirteen WC1 genes code for three distinct WC1 molecular forms. CONCLUSION: The bovine WC1 multi-gene family is composed of thirteen genes coding for three structural forms whose sequences are highly conserved among individual cattle and breeds. The sequence diversity necessary for WC1 genes to function as a multi-genic pattern recognition receptor array is encoded in the genome, rather than generated by recombinatorial diversity or hypermutation.
10aAnimals10aBase Sequence10aCattle10aDatabases, Genetic10aDNA, Complementary10aGene Dosage10aGenome10aMembrane Glycoproteins10aMolecular Sequence Data10aMultigene Family10aPhylogeny10aPolymorphism, Genetic10aProtein Structure, Tertiary10aReal-Time Polymerase Chain Reaction10aReceptors, Antigen, T-Cell, gamma-delta1 aChen, Chuang1 aHerzig, Carolyn, T A1 aAlexander, Leeson, J1 aKeele, John, W1 aMcDaneld, Tara, G1 aTelfer, Janice, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/gene-number-determination-and-genetic-polymorphism-of-the-gamma-delta00562nas a2200133 4500008004100000245007500041210006900116490001300185100001900198700001900217700002200236700001700258856015300275 2012 eng d00aIdentification of 4 genes required for mammalian blastocyst formation.0 aIdentification of 4 genes required for mammalian blastocyst form0 vIn Press1 aMaserati, Marc1 aDai, Xiangpeng1 aWalentuk, Melanie1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-4-genes-required-for-mammalian-blastocyst-formation02816nas a2200277 4500008004100000245011700041210006900158260001300227300001100240490000700251520185200258100001902110700002902129700001802158700002302176700002002199700001902219700002202238700002402260700002102284700002302305700002102328700001702349700001802366856015402384 2012 eng d00aA maternally inherited autosomal point mutation in human phospholipase C zeta (PLCζ) leads to male infertility.0 amaternally inherited autosomal point mutation in human phospholi c2012 Jan a222-310 v273 aBACKGROUND: Male factor and idiopathic infertility contribute significantly to global infertility, with abnormal testicular gene expression considered to be a major cause. Certain types of male infertility are caused by failure of the sperm to activate the oocyte, a process normally regulated by calcium oscillations, thought to be induced by a sperm-specific phospholipase C, PLCzeta (PLCζ). Previously, we identified a point mutation in an infertile male resulting in the substitution of histidine for proline at position 398 of the protein sequence (PLCζ(H398P)), leading to abnormal PLCζ function and infertility. METHODS AND RESULTS: Here, using a combination of direct-sequencing and mini-sequencing of the PLCζ gene from the patient and his family, we report the identification of a second PLCζ mutation in the same patient resulting in a histidine to leucine substitution at position 233 (PLCζ(H233L)), which is predicted to disrupt local protein interactions in a manner similar to PLCζ(H398P) and was shown to exhibit abnormal calcium oscillatory ability following predictive 3D modelling and cRNA injection in mouse oocytes respectively. We show that PLCζ(H233L) and PLCζ(H398P) exist on distinct parental chromosomes, the former inherited from the patient's mother and the latter from his father. Neither mutation was detected utilizing custom-made single-nucleotide polymorphism assays in 100 fertile males and females, or 8 infertile males with characterized oocyte activation deficiency. CONCLUSIONS: Collectively, our findings provide further evidence regarding the importance of PLCζ at oocyte activation and forms of male infertility where this is deficient. Additionally, we show that the inheritance patterns underlying male infertility are more complex than previously thought and may involve maternal mechanisms.1 aKashir, Junaid1 aKonstantinidis, Michalis1 aJones, Celine1 aLemmon, Bernadette1 aLee, Hoi, Chang1 aHamer, Rebecca1 aHeindryckx, Bjorn1 aDeane, Charlotte, M1 aDe Sutter, Petra1 aFissore, Rafael, A1 aParrington, John1 aWells, Dagan1 aCoward, Kevin uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/a-maternally-inherited-autosomal-point-mutation-in-human-phospholipase03119nas a2200409 4500008004100000245011800041210006900159260001300228300001200241490000800253520184000261653001902101653001502120653001002135653002302145653002002168653001102188653002102199653001102220653003002231653003102261653001102292653001702303653001602320653001602336653001602352653001202368653001602380100001802396700003102414700001902445700001902464700001802483700002902501700002502530856015402555 2012 eng d00aNormal breast tissue of obese women is enriched for macrophage markers and macrophage-associated gene expression.0 aNormal breast tissue of obese women is enriched for macrophage m c2012 Feb a1003-120 v1313 aActivation of inflammatory pathways is one plausible mechanism underlying the association between obesity and increased breast cancer risk. However, macrophage infiltration and local biomarkers of inflammation in breast adipose tissue have seldom been studied in association with obesity. Gene expression profiles of normal breast tissue from reduction mammoplasty patients were evaluated by whole genome microarrays to identify patterns associated with obesity status (normal-weight, body mass index (BMI) <25; overweight, BMI 25-29.9; obese, BMI ≥30). The presence of macrophage-enriched inflammatory loci with immunopositivity for CD68 protein was evaluated by immunohistochemistry (IHC). After adjusting for confounding by age, 760 genes were differentially expressed (203 up and 557 down; FDR = 0.026) between normal-weight and obese women. Gene ontology analysis suggested significant enrichment for pathways involving IL-6, IL-8, CCR5 signaling in macrophages and RXRα and PPARα activation, consistent with a pro-inflammatory state and suggestive of macrophage infiltration. Gene set enrichment analysis also demonstrated that the genomic signatures of monocytes and macrophages were over-represented in the obese group with FDR of 0.08 and 0.13, respectively. Increased macrophage infiltration was confirmed by IHC, which showed that the breast adipose tissue of obese women had higher average macrophage counts (mean = 8.96 vs. 3.56 in normal-weight women) and inflammatory foci counts (mean = 4.91 vs. 2.67 in normal-weight women). Obesity is associated with local inflammation and macrophage infiltration in normal human breast adipose tissues. Given the role of macrophages in carcinogenesis, these findings have important implications for breast cancer etiology and progression.
10aAdipose Tissue10aAdolescent10aAdult10aBiological Markers10aBody Mass Index10aBreast10aCluster Analysis10aFemale10aGene Expression Profiling10aGene Expression Regulation10aHumans10aInflammation10aMacrophages10aMammaplasty10aMiddle Aged10aObesity10aYoung Adult1 aSun, Xuezheng1 aCasbas-Hernandez, Patricia1 aBigelow, Carol1 aMakowski, Liza1 aJerry, Joseph1 aSchneider, Sallie, Smith1 aTroester, Melissa, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/normal-breast-tissue-of-obese-women-is-enriched-for-macrophage-markers01775nas a2200145 4500008004100000245007400041210006900115260000900184300000900193490000600202520122600208100002001434700002401454856015101478 2012 eng d00aNotch and the survival of regulatory T cells: location is everything!0 aNotch and the survival of regulatory T cells location is everyth c2012 ape310 v53 aSignaling through the T cell receptor induces T lymphocytes to divide, differentiate, and perform numerous effector functions. Once activated, effector T cells are exquisitely sensitive to changes in available cytokines, particularly the survival cytokine interleukin-2 (IL-2). Removal of IL-2 rapidly initiates apoptosis in response to cytokine withdrawal. In contrast to effector T cells, regulatory T cells (T(regs)) are resistant to apoptosis induced by cytokine withdrawal. A study exploring the differences between these two T cell subsets reveals a role for Notch1 in protecting T(regs) from apoptosis. Protection from apoptosis induced by cytokine withdrawal correlated with Notch1 localization in the cytosol of T(regs) and its association with phosphatidylinositol 3-kinase and Rictor, a component of the mammalian target of rapamycin complex. Notch1 localization in the nucleus in effector T cells, on the other hand, was correlated with susceptibility to apoptosis induced by cytokine withdrawal. This study highlights how Notch1 can deliver opposing signals in different cellular contexts and suggests that localization of Notch1 can have a substantial influence on life-and-death decisions in T lymphocytes.1 aMinter, Lisa, M1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch-and-the-survival-of-regulatory-t-cells-location-is-everything02210nas a2200301 4500008004100000022001400041245011900055210006900174260001300243300001100256490000700267520126200274653001201536653002101548653002601569653001101595653000901606653001101615653001001626653000901636653001401645653001901659100002501678700001801703700002201721700002001743856014501763 2012 eng d a0022-258500aOvicidal response of NYDA formulations on the human head louse (Anoplura: Pediculidae) using a hair tuft bioassay.0 aOvicidal response of NYDA formulations on the human head louse A c2012 Mar a336-420 v493 aUsing the in vitro rearing system in conjunction with the hair tuft bioassay, NYDA and NYDA without fragrances formulations (92% wt:wt dimeticones) were 100% ovicidal (0% of treated eggs hatched) after an 8-h exposure of the eggs of the human head louse (Pediculus humanus capitis De Geer) following the manufacturer's instructions. Comparatively, 78 and 66% of eggs similarly exposed hatched after distilled deionized water or Nix (1% permethrin) treatments, respectively. NYDA and NYDA without fragrances formulations were also statistically and substantially more ovicidal than either distilled deionized water or Nix treatments after 10, 30 min, and 1 h exposures. Only the 10 min exposure of eggs to NYDA and NYDA without fragrances formulations resulted in hatched lice that survived to adulthood (5-8% survival). Of the lice that hatched from eggs exposed to NYDA formulations for 10 min, there were no significant differences in the time it took them to become adults, female fecundity or the viability of eggs laid by surviving females. The longevity of adults, however, was reduced after the 10 min treatments of eggs with NYDA and NYDA without fragrances formulations compared with either the distilled deionized water or Nix treatments.
10aAnimals10aBiological Assay10aDimethylpolysiloxanes10aFemale10aHair10aHumans10aLarva10aOvum10aPediculus10aToxicity Tests1 aStrycharz, Joseph, P1 aLao, Alice, R1 aAlves, Anna-Maria1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/ovicidal-response-nyda-formulations-human-head-louse-anoplura02436nas a2200253 4500008004100000245008100041210006900122260000900191300000700200490000700207520154900214100001901763700002901782700002501811700002201836700002801858700002301886700002501909700002301934700002501957700002501982700002402007856015102031 2012 eng d00aParalemmin-1 is over-expressed in estrogen-receptor positive breast cancers.0 aParalemmin1 is overexpressed in estrogenreceptor positive breast c2012 a170 v123 aUNLABELLED: ABSTRACT: BACKGROUND: Paralemmin-1 is a phosphoprotein lipid-anchored to the cytoplasmic face of membranes where it functions in membrane dynamics, maintenance of cell shape, and process formation. Expression of paralemmin-1 and its major splice variant (Δ exon 8) as well as the extent of posttranslational modifications are tissue- and development-specific. Paralemmin-1 expression in normal breast and breast cancer tissue has not been described previously. RESULTS: Paralemmin-1 mRNA and protein expression was evaluated in ten breast cell lines, 26 primary tumors, and 10 reduction mammoplasty (RM) tissues using real time RT-PCR. Paralemmin-1 splice variants were assessed in tumor and RM tissues using a series of primers and RT-PCR. Paralemmin-1 protein expression was examined in cell lines using Western Blots and in 31 ductal carcinomas in situ, 65 infiltrating ductal carcinomas, and 40 RM tissues using immunohistochemistry. Paralemmin-1 mRNA levels were higher in breast cancers than in RM tissue and estrogen receptor (ER)-positive tumors had higher transcript levels than ER-negative tumors. The Δ exon 8 splice variant was detected more frequently in tumor than in RM tissues. Protein expression was consistent with mRNA results showing higher paralemmin-1 expression in ER-positive tumors. CONCLUSIONS: The differential expression of paralemmin-1 in a subset of breast cancers suggests the existence of variation in membrane dynamics that may be exploited to improve diagnosis or provide a therapeutic target.1 aTurk, Casey, M1 aFagan-Solis, Katerina, D1 aWilliams, Kristin, E1 aGozgit, Joseph, M1 aSmith-Schneider, Sallie1 aMarconi, Sharon, A1 aOtis, Christopher, N1 aCrisi, Giovanna, M1 aAnderton, Douglas, L1 aKilimann, Manfred, W1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/paralemmin-1-is-over-expressed-in-estrogen-receptor-positive-breast02423nas a2200193 4500008004100000245015300041210006900194260000900263300000900272490000700281520164300288100002201931700003701953700002001990700002302010700002102033700002502054856015002079 2012 eng d00aParticipation of the Cl-/HCO3- Exchangers SLC26A3 and SLC26A6, the Cl- Channel CFTR, and the Regulatory Factor SLC9A3R1 in Mouse Sperm Capacitation.0 aParticipation of the ClHCO3 Exchangers SLC26A3 and SLC26A6 the C c2012 a1-140 v863 aSperm capacitation is required for fertilization and involves several ion permeability changes. Although Cl(-) and HCO(3)(-) are essential for capacitation, the molecular entities responsible for their transport are not fully known. During mouse sperm capacitation, the intracellular concentration of Cl(-) ([Cl(-)](i)) increases and membrane potential (Em) hyperpolarizes. As in noncapacitated sperm, the Cl(-) equilibrium potential appears to be close to the cell resting Em, opening of Cl(-) channels could not support the [Cl(-)](i) increase observed during capacitation. Alternatively, the [Cl(-)](i) increase might be mediated by anion exchangers. Among them, SLC26A3 and SLC26A6 are good candidates, since, in several cell types, they increase [Cl(-)](i) and interact with cystic fibrosis transmembrane conductance regulator (CFTR), a Cl(-) channel present in mouse and human sperm. This interaction is known to be mediated and probably regulated by the Na(+)/H(+) regulatory factor-1 (official symbol, SLC9A3R1). Our RT-PCR, immunocytochemistry, Western blot, and immunoprecipitation data indicate that SLC26A3, SLC26A6, and SLC9A3R1 are expressed in mouse sperm, localize to the midpiece, and interact between each other and with CFTR. Moreover, we present evidence indicating that CFTR and SLC26A3 are involved in the [Cl(-)](i) increase induced by db-cAMP in noncapacitated sperm. Furthermore, we found that inhibitors of SLC26A3 (Tenidap and 5099) interfere with the Em changes that accompany capacitation. Together, these findings indicate that a CFTR/SLC26A3 functional interaction is important for mouse sperm capacitation.1 aChávez, Julio, C1 aHernández-González, Enrique, O1 aWertheimer, Eva1 aVisconti, Pablo, E1 aDarszon, Alberto1 aTreviño, Claudia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/participation-of-the-cl-hco3-exchangers-slc26a3-and-slc26a6-the-cl02891nas a2200325 4500008004100000245009700041210006900138260001300207300001100220490000800231520180900239653001202048653002202060653004702082653001102129653003102140653004302171653000902214653001202223653002002235653002202255100001802277700002502295700002102320700001902341700001502360700001802375700002302393856014902416 2012 eng d00aRegulation of inositol 1,4,5-trisphosphate receptor function during mouse oocyte maturation.0 aRegulation of inositol 145trisphosphate receptor function during c2012 Feb a705-170 v2273 aAt the time of fertilization, an increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)) underlies egg activation and initiation of development in all species studied to date. The inositol 1,4,5-trisphosphate receptor (IP(3)R1), which is mostly located in the endoplasmic reticulum (ER) mediates the majority of this Ca(2+) release. The sensitivity of IP(3)R1, that is, its Ca(2+) releasing capability, is increased during oocyte maturation so that the optimum [Ca(2+)](i) response concurs with fertilization, which in mammals occurs at metaphase of second meiosis. Multiple IP(3)R1 modifications affect its sensitivity, including phosphorylation, sub-cellular localization, and ER Ca(2+) concentration ([Ca(2+)](ER)). Here, we evaluated using mouse oocytes how each of these factors affected IP(3)R1 sensitivity. The capacity for IP(3)-induced Ca(2+) release markedly increased at the germinal vesicle breakdown stage, although oocytes only acquire the ability to initiate fertilization-like oscillations at later stages of maturation. The increase in IP(3)R1 sensitivity was underpinned by an increase in [Ca(2+)](ER) and receptor phosphorylation(s) but not by changes in IP(3)R1 cellular distribution, as inhibition of the former factors reduced Ca(2+) release, whereas inhibition of the latter had no impact. Therefore, the results suggest that the regulation of [Ca(2+)](ER) and IP(3)R1 phosphorylation during maturation enhance IP(3)R1 sensitivity rendering oocytes competent to initiate oscillations at the expected time of fertilization. The temporal discrepancy between the initiation of changes in IP(3)R1 sensitivity and acquisition of mature oscillatory capacity suggest that other mechanisms that regulate Ca(2+) homeostasis also shape the pattern of oscillations in mammalian eggs.10aAnimals10aCalcium Signaling10aCyclin-Dependent Kinase Inhibitor Proteins10aFemale10aGene Expression Regulation10aInositol 1,4,5-Trisphosphate Receptors10aMice10aOocytes10aPhosphorylation10aProtein Transport1 aWakai, Takuya1 aVanderheyden, Veerle1 aYoon, Sook-Young1 aCheon, Banyoon1 aZhang, Nan1 aParys, Jan, B1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-inositol-145-trisphosphate-receptor-function-during01595nas a2200277 4500008004100000245005100041210004600092260001500138300001000153490000700163520059700170653001400767653001100781653003800792653001500830653002800845653005300873653004500926653002400971653004900995653004201044653005501086100002801141700001901169856012901188 2012 eng d00aThe role of TRADD in death receptor signaling.0 arole of TRADD in death receptor signaling c2012 Mar 1 a871-60 v113 aTRADD (TNFR1-associated death domain protein) was initially identified as an adaptor molecule that transduces the signal downstream of the TNFR1 (tumor necrosis factor receptor 1). TNFR1 belongs to the so-called death receptor (DR) family of receptors that depending on the context can induce either apoptosis or proliferation, as well as NF-κB and MAP kinase activation. The receptors of this group contain death domain (DD) that is necessary for the induction of apoptosis. This review summarizes the recent advances in the field of DR signaling and in particular the role of TRADD.
10aApoptosis10aHumans10aMitogen-Activated Protein Kinases10aNF-kappa B10aReceptors, Death Domain10aReceptors, TNF-Related Apoptosis-Inducing Ligand10aReceptors, Tumor Necrosis Factor, Type I10aSignal Transduction10aTNF Receptor-Associated Death Domain Protein10aTNF-Related Apoptosis-Inducing Ligand10aTumor Necrosis Factor Ligand Superfamily Member 151 aPobezinskaya, Yelena, L1 aLiu, Zhenggang uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-role-of-tradd-in-death-receptor-signaling00701nas a2200217 4500008004100000245010700041210006900148260004400217300001100261490000600272100001300278700001500291700001700306700001700323700001900340700002100359700001800380700001700398700001900415856004900434 2012 eng d00aRoles of MAPK and Spindle Assembly Checkpoint in Spontaneous Activation and MIII Arrest of Rat Oocytes0 aRoles of MAPK and Spindle Assembly Checkpoint in Spontaneous Act bPublic Library of Science ({PLoS})cfeb ae320440 v71 aCui, Wei1 aZhang, Jie1 aLian, Hua-Yu1 aWang, Hui-Li1 aMiao, De-Qiang1 aZhang, Chuan-Xin1 aLuo, Ming-Jiu1 aTan, Jing-He1 aBaltz, Jay, M. uhttps://doi.org/10.1371/journal.pone.003204400711nas a2200109 4500008004100000245003900041210003800080260001600118520033400134100001600468856011700484 2012 eng d00aSperm Bioenergetics in a Nutshell.0 aSperm Bioenergetics in a Nutshell c2012 Aug 223 aIn this issue of Biology of Reproduction, Goodson et al. used a battery of glycolysable and non-glycolysable metabolic substrates to analyze the contribution of glycolysis and oxidative phosphorylation in ATP formation, sperm motility, hyperactivation and the capacitation-associated increase in protein tyrosine phosphorylation.1 aVisconti, P uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sperm-bioenergetics-in-a-nutshell02183nas a2200289 4500008004100000245008000041210006900121260001300190300001100203490000600214520125900220653001801479653003101497653001101528653002201539653000901561653002201570653002001592653002001612653001501632653001601647100002301663700002201686700002301708700001901731856014301750 2012 eng d00aSperm phosphoproteomics: historical perspectives and current methodologies.0 aSperm phosphoproteomics historical perspectives and current meth c2012 Oct a533-480 v93 aMammalian sperm are differentiated germ cells that transfer genetic material from the male to the female. Owing to this essential role in the reproductive process, an understanding of the complex mechanisms that underlie sperm function has implications ranging from the development of novel contraceptives to the treatment of male infertility. While the importance of phosphorylation in sperm differentiation, maturation and fertilization has been well established, the ability to directly determine the sites of phosphorylation within sperm proteins and to quantitate the extent of phosphorylation at these sites is a recent development that has relied almost exclusively on advances in the field of proteomics. This review will summarize the work that has been carried out to date on sperm phosphoproteomics and discuss how the resulting qualitative and quantitative information has been used to provide insight into the manner in which protein phosphorylation events modulate sperm function. The authors also present the proteomics process as it is most often utilized for the elucidation of protein expression, with a particular emphasis on the way in which the process has been modified for the analysis of protein phosphorylation in sperm.
10aFertilization10aGene Expression Regulation10aHumans10aInfertility, Male10aMale10aMass Spectrometry10aPhosphoproteins10aPhosphorylation10aProteomics10aSpermatozoa1 aPorambo, James, R.1 aSalicioni, Ana, M1 aVisconti, Pablo, E1 aPlatt, Mark, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sperm-phosphoproteomics-historical-perspectives-and-current00553nas a2200133 4500008004100000245007900041210006900120260001200189490000700201100002200208700002200230700002300252856014400275 2012 eng d00aTestis-Specific Kinases in male Fertility and as Targets for Contraception0 aTestisSpecific Kinases in male Fertility and as Targets for Cont c09/20120 v151 aSalicioni, Ana, M1 aRomano, Fabian, B1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/testis-specific-kinases-in-male-fertility-and-as-targets-for02308nas a2200241 4500008004100000245013500041210007000176260001600246300000700262490000600269520143300275100001601708700002501724700001701749700002601766700002301792700001801815700002001833700001601853700002401869700001901893856015401912 2012 ENG d00aTransient pharmacologic lowering of Aß production prior to deposition results in sustained reduction of amyloid plaque pathology.0 aTransient pharmacologic lowering of Aß production prior to depos c2012 Aug 14 a390 v73 aABSTRACT: BACKGROUND: Alzheimer's disease (AD) is the leading cause of dementia among the elderly. Disease modifying therapies targeting Abeta that are in development have been proposed to be more effective if treatment was initiated prior to significant accumulation of Abeta in the brain, but optimal timing of treatment initiation has not been clearly established in the clinic. We compared the efficacy of transient pharmacologic reduction of brain Abeta with a gamma-secretase inhibitor (GSI ) for 1--3 months (M) treatment windows in APP Tg2576 mice and subsequent aging of the mice to either 15M or 18M. RESULTS: These data show that reducing Abeta production in a 2-3M windows both initiated and discontinued before detectable Abeta deposition has the most significant impact on Abeta loads up to 11M after treatment discontinuation. In contrast, initiation of treatment for 3M windows from 7-10M or 12-15M shows progressively decreasing efficacy. CONCLUSIONS: These data have major implications for clinical testing of therapeutics aimed at lowering Abeta production, indicating that; i) these therapies may have little efficacy unless tested as prophylactics or in the earliest preclinical stage of AD where there is no or minimal Abeta accumulation and ii) lowering Abeta production transiently during a critical pre-deposition window potentially provides long-lasting efficacy after discontinuation of the treatment.1 aDas, Pritam1 aVerbeeck, Christophe1 aMinter, Lisa1 aChakrabarty, Paramita1 aFelsenstein, Kevin1 aKukar, Thomas1 aMaharvi, Ghulam1 aFauq, Abdul1 aOsborne, Barbara, A1 aGolde, Todd, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transient-pharmacologic-lowering-of-ass-production-prior-to-deposition02285nas a2200157 4500008004100000245013200041210006900173260001600242300001100258490000800269520163800277100001901915700002601934700001701960856015001977 2012 eng d00aYin-Yang1 is required for epithelial-to-mesenchymal transition and regulation of Nodal signaling during mammalian gastrulation.0 aYinYang1 is required for epithelialtomesenchymal transition and c2012 Aug 15 a273-820 v3683 aThe ubiquitously expressed Polycomb Group protein Yin-Yang1 (YY1) is believed to regulate gene expression through direct binding to DNA elements found in promoters or enhancers of target loci. Additionally, YY1 contains diverse domains that enable a plethora of protein-protein interactions, including association with the Oct4/Sox2 pluripotency complex and Polycomb Group silencing complexes. To elucidate the in vivo role of YY1 during gastrulation, we generated embryos with an epiblast specific deletion of Yy1. Yy1 conditional knockout (cKO) embryos initiate gastrulation, but both primitive streak formation and ingression through the streak is severely impaired. These streak descendants fail to repress E-Cadherin and are unable to undergo an appropriate epithelial to mesenchymal transition (EMT). Intriguingly, overexpression of Nodal and concomitant reduction of Lefty2 are observed in Yy1 cKO embryos, suggesting that YY1 is normally required for proper Nodal regulation during gastrulation. Furthermore, definitive endoderm is specified but fails to properly integrate into the outer layer. Although anterior neuroectoderm is specified, mesoderm production is severely restricted. We show that YY1 directly binds to the Lefty2 locus in E7.5 embryos and that pharmacological inhibition of Nodal signaling partially restores mesoderm production in Yy1 cKO mutant embryos. Our results reveal critical requirements for YY1 during several important developmental processes, including EMT and regulation of Nodal signaling. These results are the first to elucidate the diverse role of YY1 during gastrulation in vivo.
1 aTrask, Mary, C1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/yin-yang1-is-required-for-epithelial-to-mesenchymal-transition-and02495nas a2200445 4500008004100000245015400041210007100195260001600266300001000282490000700292520100900299653002401308653001201332653002501344653002701369653001801396653001201414653003701426653000901463653002801472653002001500653004301520653002101563653001801584653001701602100002801619700002601647700002101673700001601694700002601710700001901736700002201755700002501777700002301802700002001825700001901845700001801864700001901882856014801901 2012 eng d00aαβ T cell receptors that do not undergo major histocompatibility complex-specific thymic selection possess antibody-like recognition specificities.0 aαβ T cell receptors that do not undergo major histocompatibility c2012 Jan 27 a79-910 v363 aMajor histocompatibility complex (MHC) restriction is the cardinal feature of T cell antigen recognition and is thought to be intrinsic to αβ T cell receptor (TCR) structure because of germline-encoded residues that impose MHC specificity. Here, we analyzed αβTCRs from T cells that had not undergone MHC-specific thymic selection. Instead of recognizing peptide-MHC complexes, the two αβTCRs studied here resembled antibodies in recognizing glycosylation-dependent conformational epitopes on a native self-protein, CD155, and they did so with high affinity independently of MHC molecules. Ligand recognition was via the αβTCR combining site and involved the identical germline-encoded residues that have been thought to uniquely impose MHC specificity, demonstrating that these residues do not only promote MHC binding. This study demonstrates that, without MHC-specific thymic selection, αβTCRs can resemble antibodies in recognizing conformational epitopes on MHC-independent ligands.
10aAmino Acid Sequence10aAnimals10aAntibody Specificity10aEpitopes, T-Lymphocyte10aGene Deletion10aLigands10aMajor Histocompatibility Complex10aMice10aMolecular Sequence Data10aProtein Binding10aReceptors, Antigen, T-Cell, alpha-beta10aReceptors, Virus10aT-Lymphocytes10aThymus Gland1 aTikhonova, Anastasia, N1 aVan Laethem, Francois1 aHanada, Ken-ichi1 aLu, Jinghua1 aPobezinsky, Leonid, A1 aHong, Changwan1 aGuinter, Terry, I1 aJeurling, Susanna, K1 aBernhardt, Günter1 aPark, Jung-Hyun1 aYang, James, C1 aSun, Peter, D1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/av-t-cell-receptors-that-do-not-undergo-major-histocompatibility02520nas a2200145 4500008004100000245021900041210007000260260001600330300001000346490000800356520181800364100002102182700001902203856015202222 2011 eng d00a17β-estradiol and progesterone regulate multiple progestin signaling molecules in the anteroventral periventricular nucleus, ventromedial nucleus and sexually dimorphic nucleus of the preoptic area in female rats.0 a17βestradiol and progesterone regulate multiple progestin signal c2011 Mar 10 a86-920 v1763 aRecent work identified novel progestin signaling molecules, including progesterone receptor membrane component 1 (Pgrmc1), Pgrmc2, serpine mRNA binding protein 1 (Serbp1), progestin and adiponectin receptors 7 (Paqr7) and Paqr8. These molecules mediate rapid progesterone (P(4)) effects in non-neural tissue and we recently mapped their expression in the brain. Many rapid effects of P(4) require 17β-estradiol (E(2)) and P(4) priming; therefore, we examined the effects of ovarian hormones on the expression of these non-classical progestin signaling molecules. We focused specifically on the anteroventral periventricular nucleus (AVPV), the sexually dimorphic nucleus of the preoptic area (SDN-POA) and the ventrolateral portion of the ventromedial nucleus (VMNvl). These brain nuclei are important for female reproduction. Ovariectomized adult female rats were implanted with capsules containing sesame oil or E(2), and injected 48 h later with sesame oil or P(4). Brains were collected 8 h later and RNA was isolated from the AVPV, SDN-POA and VMNvl. We assessed the effects of ovarian hormones on mRNA levels using quantitative polymerase chain reaction (QPCR). In the AVPV, Serbp1 mRNA levels were increased by P(4) in the presence of E(2), and Paqr8 was downregulated by P(4) alone. In the SDN-POA, combined E(2) and P(4) increased Pgrmc1 and Serbp1 mRNA levels, and E(2) alone increased Paqr8 mRNA levels. Finally, in the VMNvl, P(4) increased mRNA levels encoding Pgrmc1, Pgrmc2 and Serbp1, and the combination of E(2) and P(4) increased Pgrmc1 and Serbp1 mRNA levels. Paqr7 was not regulated by E(2) or P(4) in any brain region examined. In summary, we showed that ovarian hormones regulate novel progestin signaling molecules in brain regions important for the neuroendocrine control of reproduction.1 aIntlekofer, K, A1 aPetersen, S, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/17v-estradiol-and-progesterone-regulate-multiple-progestin-signaling00550nas a2200133 4500008004100000245007500041210006900116260001300185300001200198490000700210100002100217700002500238856015300263 2011 fre d00aADAM and cell migration: the unexpected role of the cytoplasmic domain0 aADAM and cell migration the unexpected role of the cytoplasmic d c2011 Dec a1069-710 v271 aCousin, Hélène1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/adam-and-cell-migration-the-unexpected-role-of-the-cytoplasmic-domain02356nas a2200349 4500008004100000245009400041210006900135260001500204300001100219490000800230520112600238653001201364653002201376653002001398653004101418653001901459653002401478653002601502653000901528653001901537653004801556653002401604653001801628653004901646653005501695100002801750700001801778700002301796700001601819700002001835856015101855 2011 eng d00aThe adaptor protein TRADD is essential for TNF-like ligand 1A/death receptor 3 signaling.0 aadaptor protein TRADD is essential for TNFlike ligand 1Adeath re c2011 May 1 a5212-60 v1863 aTNFR-associated death domain protein (TRADD) is a key effector protein of TNFR1 signaling. However, the role of TRADD in other death receptor (DR) signaling pathways, including DR3, has not been completely characterized. Previous studies using overexpression systems suggested that TRADD is recruited to the DR3 complex in response to the DR3 ligand, TNF-like ligand 1A (TL1A), indicating a possible role in DR3 signaling. Using T cells from TRADD knockout mice, we demonstrate in this study that the response of both CD4(+) and CD8(+) T cells to TL1A is dependent upon the presence of TRADD. TRADD knockout T cells therefore lack the appropriate proliferative response to TL1A. Moreover, in the absence of TRADD, both the stimulation of MAPK signaling and activation of NF-κB in response to TL1A are dramatically reduced. Unsurprisingly, TRADD is required for recruitment of receptor interacting protein 1 and TNFR-associated factor 2 to the DR3 signaling complex and for the ubiquitination of receptor interacting protein 1. Thus, our findings definitively establish an essential role of TRADD in DR3 signaling.
10aAnimals10aBlotting, Western10aCell Separation10aElectrophoretic Mobility Shift Assay10aFlow Cytometry10aImmunoprecipitation10aLymphocyte Activation10aMice10aMice, Knockout10aReceptors, Tumor Necrosis Factor, Member 2510aSignal Transduction10aT-Lymphocytes10aTNF Receptor-Associated Death Domain Protein10aTumor Necrosis Factor Ligand Superfamily Member 151 aPobezinskaya, Yelena, L1 aChoksi, Swati1 aMorgan, Michael, J1 aCao, Xiumei1 aLiu, Zheng-Gang uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-adaptor-protein-tradd-is-essential-for-tnf-like-ligand-1a-death03114nas a2200385 4500008004100000245013200041210006900173260001300242300001100255490000800266520185100274653001202125653002202137653002602159653002302185653002602208653002002234653001102254653002702265653004302292653000902335653000902344653000902353653003902362653001802401100001802419700002102437700002102458700001902479700002302498700001802521700001802539700002002557856015102577 2011 eng d00aAlterations in calcium oscillatory activity in vitrified mouse eggs impact on egg quality and subsequent embryonic development.0 aAlterations in calcium oscillatory activity in vitrified mouse e c2011 May a515-260 v4613 aCryopreservation of mature eggs is a useful technique that can be applied in assisted reproductive technology. However, the method has some limitations, such as cryodamage induced by biophysical modifications during the cryopreservation process. To assess these biophysical damage, we analyzed the relationship between intracellular calcium ([Ca2+]i) oscillatory activity via type 1 inositol 1,4,5-trisphosphate receptor (IP(3)R1) distribution after vitrification and efficiency of cryopreservation according to cryoprotectant (CPA) composition. In immunostaining, results of IP(3)R1 with eggs after the vitrification performed using ethylene glycol (EG) alone or EG + dimethylsulfoxide (DMSO) as CPAs, CPA-treated, and fresh eggs displayed a homogeneous IP(3)R1 distribution which is spread uniformly throughout cytoplasm with clusters on the cortex. However, after vitrification and warming process, more than 60% of eggs displayed a heterogeneous distribution which is non-uniform distribution with patches and disconnection of IP(3)R1. In 90-min incubation for recovery from cryodamage, eggs from the EG + DMSO group recovered from with a heterogeneous IP(3)R1 distribution to the homogeneous distribution, but not in EG alone group. In ICSI experiments, vitrified eggs in the EG-alone group presented significantly low blastocyst formation compared to those of the fresh and EG + DMSO groups. These results suggest that the vitrification process influences IP(3)R1 distribution, and subsequently, [Ca2+]i oscillatory activity and embryonic development. Accordingly, we propose that IP(3)R1 distribution and [Ca2+]i oscillatory activity are correlated with egg quality and developmental potential after vitrification, and may thus be applied as an effective indicator to evaluate the efficiency of oocyte cryopreservation methods.10aAnimals10aCalcium Signaling10aCryoprotective Agents10aDimethyl Sulfoxide10aEmbryonic Development10aEthylene Glycol10aFemale10aFertilization in Vitro10aInositol 1,4,5-Trisphosphate Receptors10aMale10aMice10aOvum10aSperm Injections, Intracytoplasmic10aVitrification1 aKim, Bo, Yeun1 aYoon, Sook-Young1 aCha, Soo, Kyoung1 aKwak, Ki, Hoon1 aFissore, Rafael, A1 aParys, Jan, B1 aYoon, Tae, Ki1 aLee, Dong, Ryul uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/alterations-in-calcium-oscillatory-activity-in-vitrified-mouse-eggs00782nas a2200169 4500008004100000245011200041210006900153260001600222300001200238490000800250520011600258100002000374700002400394700002100418700002400439856014900463 2011 eng d00aB cell-deficient mice display markedly enhanced resistance to the intracellular bacterium Brucella abortus.0 aB celldeficient mice display markedly enhanced resistance to the c2011 Apr 15 a1136-460 v2033 aBrucella species are facultative intracellular bacteria that cause lifelong infections in humans and livestock.1 aGoenka, Radhika1 aParent, Michelle, A1 aElzer, Philip, H1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/b-cell-deficient-mice-display-markedly-enhanced-resistance-to-the01973nas a2200229 4500008004100000245013500041210006900176260001300245300001100258490000700269520117200276100001501448700002001463700001501483700001101498700001401509700001501523700002201538700001401560700001601574856015301590 2011 eng d00aBrief exposures of human body lice to sublethal amounts of ivermectin over-transcribes detoxification genes involved in tolerance.0 aBrief exposures of human body lice to sublethal amounts of iverm c2011 Dec a687-990 v203 aTranscriptional profiling results, using our non-invasive induction assay {short exposure intervals (2-5 h) to sublethal amounts of insecticides [Members of the testis-specific serine/threonine kinases (Tssk) family may have a role in sperm differentiation in the testis and/or fertilization. To gain insight into the functional relevance of these kinases, their expression was examined both at the mRNA and protein levels. Quantitative PCR analysis confirmed that all five Tssk mRNAs are almost exclusively expressed postmeiotically in the testis. Recombinant mouse and human Tssks were cloned and used for validation of an array of commercial and custom-made antibodies against Tssks. Immunolocalization in mouse testis, and in mouse and human sperm, showed that Tssk1, Tssk2, Tssk4 and Tssk6, but not Tssk3, were present in mouse sperm and in germ cells from mouse testis. TSSK1, TSSK2 and TSSK6 were also detected in human sperm, while TSSK3 was absent. In both mouse and human sperm, Tssk1 was partially soluble, while Tssk2, Tssk4 and Tssk6 were insoluble in non-ionic detergents. In vitro recombinant TSSK2 activity assays showed maximum enzymatic activity at 5 mM Mg(2+) and a Km for ATP of ∼10 µM. These, observations together with findings that the Tssk1/Tssk2 double knock-out as well as the Tssk6 null mice are sterile without presenting other detectable defects, suggest that these kinases could be used as targets for male contraception.
1 aLi, Y1 aSosnik, J1 aBrassard, L1 aReese, M1 aSpiridonov, NA1 aBates, TC1 aJohnson, GR1 aAnguita, J1 aVisconti, P, E1 aSalicioni, A, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/expression-and-localization-of-five-members-of-the-testis-specific03236nas a2200349 4500008004100000245015100041210006900192260001300261300001100274490000700285520202100292653001302313653001202326653001502338653005902353653003902412653001502451653004002466653001102506653000902517653002402526653000902550653002002559653002902579653002402608653002302632653002002655100002002675700001902695700002402714856014802738 2011 eng d00aGuanine-nucleotide exchange factors (RAPGEF3/RAPGEF4) induce sperm membrane depolarization and acrosomal exocytosis in capacitated stallion sperm.0 aGuaninenucleotide exchange factors RAPGEF3RAPGEF4 induce sperm m c2011 Jul a179-880 v853 aCapacitation encompasses the molecular changes sperm undergo to fertilize an oocyte, some of which are postulated to occur via a cAMP-PRKACA (protein kinase A)-mediated pathway. Due to the recent discovery of cAMP-activated guanine nucleotide exchange factors RAPGEF3 and RAPGEF4, we sought to investigate the separate roles of PRKACA and RAPGEF3/RAPGEF4 in modulating capacitation and acrosomal exocytosis. Indirect immunofluorescence localized RAPGEF3 to the acrosome and subacrosomal ring and RAPGEF4 to the midpiece in equine sperm. Addition of the RAPGEF3/RAPGEF4-specific cAMP analogue 8-(p-chlorophenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate (8pCPT) to sperm incubated under both noncapacitating and capacitating conditions had no effect on protein tyrosine phosphorylation, thus supporting a PRKACA-mediated event. Conversely, activation of RAPGEF3/RAPGEF4 with 8pCPT induced acrosomal exocytosis in capacitated equine sperm at rates (34%) similar (P > 0.05) to those obtained in progesterone- and calcium ionophore-treated sperm. In the mouse, capacitation-dependent hyperpolarization of the sperm plasma membrane has been shown to recruit low voltage-activated T-type Ca(2+) channels, which later open in response to zona pellucida-induced membrane depolarization. We hypothesized that RAPGEF3 may be inducing acrosomal exocytosis via depolarization-dependent Ca(2+) influx, as RAPGEF3/RAPGEF4 have been demonstrated to play a role in the regulation of ion channels in somatic cells. We first compared the membrane potential (E(m)) of noncapacitated (-37.11 mV) and capacitated (-53.74 mV; P = 0.002) equine sperm. Interestingly, when sperm were incubated (6 h) under capacitating conditions in the presence of 8pCPT, E(m) remained depolarized (-32.06 mV). Altogether, these experiments support the hypothesis that RAPGEF3/RAPGEF4 activation regulates acrosomal exocytosis via its modulation of E(m), a novel role for RAPGEF3/RAPGEF4 in the series of events required to achieve fertilization.10aAcrosome10aAnimals10aCyclic AMP10aCyclic AMP-Dependent Protein Kinase Catalytic Subunits10aDichlororibofuranosylbenzimidazole10aExocytosis10aGuanine Nucleotide Exchange Factors10aHorses10aMale10aMembrane Potentials10aMice10aPhosphorylation10aProtein-Tyrosine Kinases10aSignal Transduction10aSperm Capacitation10aThionucleotides1 aMcPartlin, L, A1 aVisconti, P, E1 aBedford-Guaus, S, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/guanine-nucleotide-exchange-factors-rapgef3-rapgef4-induce-sperm02320nas a2200433 4500008004100000245013500041210006900176260001300245300001000258490000700268520098200275653001201257653001801269653001201287653001301299653001101312653001801323653004401341653001101385653000901396653000901405653002801414653002201442653001001464653000901474653003701483653002401520653001301544100001801557700002201575700001501597700001801612700002101630700002001651700001901671700002301690700002101713856015201734 2011 eng d00aIdentification and functional analysis of an ovarian form of the egg activation factor phospholipase C zeta (PLCζ) in pufferfish.0 aIdentification and functional analysis of an ovarian form of the c2011 Jan a48-560 v783 aRecent studies suggest that egg activation in mammals is triggered by a sperm-specific phospholipase C, PLCzeta. In other vertebrate species such as medaka fish, chickens, and quail, PLCzeta is also expressed as a testis-specific mRNA. Functional studies suggest that PLCzeta plays a similar role as a trigger of egg activation in these species. Here, we report the identification of PLCzeta orthologues in pufferfish species Takifugu rubripes (Fugu) and Tetraodon nigroviridis (Tetraodon). Unexpectedly in these species PLCzeta is expressed not in the testis, but in ovary and brain. Injection of pufferfish PLCzeta copy ribonucleic acid (cRNA) into mouse eggs failed to trigger calcium oscillations, unlike medaka PLCzeta cRNA. Our findings provide the first evidence that PLCzeta may be expressed in the egg, rather than the sperm, in some vertebrate species, and that its mechanism of action and physiologic role at fertilization may differ in different vertebrate species.10aAnimals10aBase Sequence10aCalcium10aChickens10aFemale10aFish Proteins10aGene Expression Regulation, Enzymologic10aHumans10aMale10aMice10aMolecular Sequence Data10aOrgan Specificity10aOvary10aOvum10aPhosphoinositide Phospholipase C10aSpecies Specificity10aTakifugu1 aCoward, Kevin1 aPonting, Chris, P1 aZhang, Nan1 aYoung, Claire1 aHuang, Chang-Jen1 aChou, Chih-Ming1 aKashir, Junaid1 aFissore, Rafael, A1 aParrington, John uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-and-functional-analysis-of-an-ovarian-form-of-the-egg02256nas a2200193 4500008004100000245011300041210006900154260001500223300001200238490000600250520151500256100001901771700002501790700002101815700002501836700002501861700002401886856015201910 2011 eng d00aIncreased promoter methylation in exfoliated breast epithelial cells in women with a previous breast biopsy.0 aIncreased promoter methylation in exfoliated breast epithelial c c2011 Dec 1 a1425-350 v63 aAccurately identifying women at increased risk of developing breast cancer will provide greater opportunity for early detection and prevention. DNA promoter methylation is a promising biomarker for assessing breast cancer risk. Breast milk contains large numbers of exfoliated epithelial cells that are ideal for methylation analyses. Exfoliated epithelial cells were isolated from the milk obtained from each breast of 134 women with a history of a non-proliferative benign breast biopsy (Biopsy Group). Promoter methylation of three tumor suppressor genes, RASSF1, SFRP1 and GSTP1, was assessed by pyrosequencing of bisulfite-modified DNA. Methylation scores from the milk of the 134 women in the Biopsy Group were compared to scores from 102 women for whom a breast biopsy was not a recruitment requirement (Reference Group). Mean methylation scores for RASSF1 and GSTP1 were significantly higher in the Biopsy than in the Reference Group. For all three genes the percentage of outlier scores was greater in the Biopsy than in the Reference Group but reached statistical significance only for GSTP1. A comparison between the biopsied and non-biopsied breasts of the Biopsy Group revealed higher mean methylation and a greater number of outlier scores in the biopsied breast for both SFRP1 and RASSF1, but not for GSTP1. This is the first evidence of CpG island methylation in tumor suppressor genes of women who may be at increased risk of developing breast cancer based on having had a prior breast biopsy.1 aBrowne, Eva, P1 aPunska, Elizabeth, C1 aLenington, Sarah1 aOtis, Christopher, N1 aAnderton, Douglas, L1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/increased-promoter-methylation-in-exfoliated-breast-epithelial-cells01730nas a2200133 4500008004100000245008500041210006900126260001300195300001200208490000800220520118800228100002601416856015401442 2011 eng d00aInducing the liver: understanding the signals that promote murine liver budding.0 aInducing the liver understanding the signals that promote murine c2011 Jul a1727-310 v2263 aThe endoderm emerges as an epithelial sheet that covers the surface of the developing murine embryo. This tissue will produce the entire gut tube as well as associated digestive and respiratory organs including the thyroid, thymus, lung, liver, and pancreas. The emergence of each endodermal organ occurs in a temporally distinct manner that is dependant upon reciprocal inductive interactions between the endoderm and the underlying mesoderm. The emergence of the hepatic endoderm, which occurs using a morphological process termed liver budding, initiates during early somitogenesis in the mouse at approximately 8.25 days post-coitum (dpc). Explant and transplant studies performed in chicken and mouse have demonstrated that secreted signals from adjacent mesodermal tissues initiate the hepatic gene program from ventral-fated endoderm. Here, we review the data in support of the roles of members of the fibroblast growth factor (FGF), bone morphogenetic protein (BMP), and Wnt signaling pathways in liver budding and discover that little is known about the precise endogenous signals involved in the molecular and morphological induction of liver budding in the mouse.
1 aTremblay, Kimberly, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/inducing-the-liver-understanding-the-signals-that-promote-murine-liver02038nas a2200181 4500008004100000245006800041210006600109260001300175300001200188490000700200520138300207100002301590700001701613700003501630700002501665700002101690856014501711 2011 eng d00aIon channels, phosphorylation and mammalian sperm capacitation.0 aIon channels phosphorylation and mammalian sperm capacitation c2011 May a395-4050 v133 aSexually reproducing animals require an orchestrated communication between spermatozoa and the egg to generate a new individual. Capacitation, a maturational complex phenomenon that occurs in the female reproductive tract, renders spermatozoa capable of binding and fusing with the oocyte, and it is a requirement for mammalian fertilization. Capacitation encompasses plasma membrane reorganization, ion permeability regulation, cholesterol loss and changes in the phosphorylation state of many proteins. Novel tools to study sperm ion channels, image intracellular ionic changes and proteins with better spatial and temporal resolution, are unraveling how modifications in sperm ion transport and phosphorylation states lead to capacitation. Recent evidence indicates that two parallel pathways regulate phosphorylation events leading to capacitation, one of them requiring activation of protein kinase A and the second one involving inactivation of ser/thr phosphatases. This review examines the involvement of ion transporters and phosphorylation signaling processes needed for spermatozoa to achieve capacitation. Understanding the molecular mechanisms leading to fertilization is central for societies to deal with rising male infertility rates, to develop safe male gamete-based contraceptives and to preserve biodiversity through better assisted fertilization strategies.1 aVisconti, Pablo, E1 aKrapf, Dario1 aDe la Vega-Beltran, Jose, Luis1 aAcevedo, Juan, José1 aDarszon, Alberto uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/ion-channels-phosphorylation-and-mammalian-sperm-capacitation02030nas a2200169 4500008004100000245015000041210006900191260001300260300001200273490000700285520132800292100002501620700001901645700002201664700002001686856015401706 2011 eng d00aIvermectin acts as a posteclosion nymphicide by reducing blood feeding of human head lice (Anoplura: Pediculidae) that hatched from treated eggs.0 aIvermectin acts as a posteclosion nymphicide by reducing blood f c2011 Nov a1174-820 v483 aThe 0.5% ivermectin topical cream formulation was not directly ovicidal to treated eggs of head lice, as hatchability was not decreased. Nevertheless, the percent of hatched lice from treated eggs that took a blood meal significantly decreased (80-95%) compared with lice that hatched from untreated eggs and all treated lice died within 48 h of hatching, including those that fed. Dilutions of ivermectin formulation of 0.15 and 0.2 microg/ml, which were topically applied to 0-8 d old eggs, were not lethal to lice at 24 h posteclosion. However, 9 and 16% less lice fed when hatched from these treated eggs, respectively. Total [3H] inulin ingested by untreated first instars significantly increased over a 48 h feeding interval but was significantly less in instars that hatched from eggs receiving the 0.15 (36% less) and 0.2 (55% less) microg/ml ivermectin treatments compared with placebo. The reduced feeding that occurred after the 0.15 and 0.2 microg/ml ivermectin treatments occurred in the absence of mortality and suggests a unique mode of action of ivermectin on feeding that is separate from the mode of action of ivermectin leading to mortality. Failure of hatched instars to take a blood meal after egg treatments with formulated ivermectin is likely responsible for its action as a posteclosion nymphicide.1 aStrycharz, Joseph, P1 aBerge, Noah, M1 aAlves, Anna-Maria1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/ivermectin-acts-as-a-posteclosion-nymphicide-by-reducing-blood-feeding03206nas a2200289 4500008004100000245014800041210007000189260001300259300001200272490000700284520219100291100001902482700001802501700002002519700002002539700002402559700002002583700002002603700001902623700002202642700002002664700002102684700002302705700002102728700001802749856014902767 2011 eng d00aLoss of activity mutations in phospholipase C zeta (PLCζ) abolishes calcium oscillatory ability of human recombinant protein in mouse oocytes.0 aLoss of activity mutations in phospholipase C zeta PLCζ abolishe c2011 Dec a3372-870 v263 aBACKGROUND: Mammalian oocyte activation occurs via a series of intracellular calcium (Ca(2+)) oscillations thought to be induced by a sperm-specific phospholipase C zeta (PLCζ). There is now strong evidence to indicate that certain types of human male infertility are caused by failure of the sperm to activate the oocyte in an appropriate manner. Molecular analysis of the PLCζ gene of a male patient with oocyte activation deficiency has previously identified a point mutation causing a histidine to proline substitution at PLCζ residue 398 (PLCζ(H398P)), leading to abnormal Ca(2+) release profiles and reduced oocyte activation efficiency. METHODS AND RESULTS: In the present study, we used HEK293T cells to produce recombinant human wild-type PLCζ (PLCζ(WT)) protein which, upon microinjection into mouse oocytes, induced Ca(2+) oscillations characteristic of oocyte activation. Injection of recombinant PLCζ(H398P) was unable to elicit Ca(2+) oscillations in mouse oocytes. Loss of activity mutations, such as PLCζ(H398P) and an artificially induced frameshift mutation (PLCζ(ΔYC2)) did not affect Ca(2+) release when over-expressed in HEK293T cells, whereas PLCζ(WT) inhibited adenosine triphosphate-activated Ca(2+) release. Confocal imaging of fluorescently tagged PLCζ isoforms in HEK293T cells suggested a cytoplasmic pattern of localization, while quantitative analysis of fluorescence levels showed that PLCζ(WT) > PLCζ(H398P) > PLCζ(ΔYC2), indicating that loss of activity mutations may lead to protein instability. This was further indicated by the low proportion of sperm and the lower levels of total PLCζ immunofluorescence from the patient exhibiting PLCζ(H398P) compared with fertile controls. CONCLUSIONS: We demonstrate, for the first time, the production of active recombinant human PLCζ protein which retained the ability to elicit characteristic Ca(2+) oscillations in mouse oocytes, an ability which was eliminated by an infertility-linked mutation. These findings advance our understanding of PLCζ, and provide a critical step forward in obtaining purified PLCζ protein as a potential therapeutic agent for oocyte activation deficiency.1 aKashir, Junaid1 aJones, Celine1 aLee, Hoi, Chang1 aRietdorf, Katja1 aNikiforaki, Dimitra1 aDurrans, Claire1 aRuas, Margarida1 aTee, Sze, Tian1 aHeindryckx, Bjorn1 aGalione, Antony1 aDe Sutter, Petra1 aFissore, Rafael, A1 aParrington, John1 aCoward, Kevin uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/loss-of-activity-mutations-in-phospholipase-c-zeta-plcz-abolishes02549nas a2200241 4500008004100000245010100041210006900142260001300211300001000224490000700234520170600241653001201947653002101959653001101980653003801991653001102029653003002040653000902070653003302079100002502112700001802137856015202155 2011 eng d00aMap making in the 21st century: charting breast cancer susceptibility pathways in rodent models.0 aMap making in the 21st century charting breast cancer susceptibi c2011 Apr a57-640 v163 aGenetic factors play an important role in determining risk and resistance to increased breast cancer. Recent technological advances have made it possible to analyze hundreds of thousands of single nucleotide polymorphisms in large-scale association studies in humans and have resulted in identification of alleles in over 20 genes that influence breast cancer risk. Despite these advances, the challenge remains in identifying what the functional polymorphisms are that confer the increased risk, and how these genetic variants interact with each other and with environmental factors. In rodents, the incidence of mammary tumors varies among strains, such that they can provide alternate ideas for candidate pathways involved in humans. Mapping studies in animals have unearthed numerous loci for breast cancer susceptibility that have been validated in human populations. In a reciprocal manner, knockin and knockout mice have been used to validate the tumorigenicity of risk alleles found in population studies. Rodent studies also underscore the complexity of interactions among alleles. The fact that genes affecting risk and resistance to mammary tumors in rodents depend greatly upon the carcinogenic challenge emphasizes the importance of gene x environment interactions. The challenge to rodent geneticists now is to capitalize on the ability to control the genetics and environment in rodent models of tumorigenesis to better understand the biology of breast cancer development, to identify those polymorphisms most relevant to human susceptibility and to identify compensatory pathways that can be targeted for improved prevention in women at highest risk of developing breast cancer.
10aAnimals10aBreast Neoplasms10aFemale10aGenetic Predisposition to Disease10aHumans10aMammary Neoplasms, Animal10aMice10aTumor Suppressor Protein p531 aBlackburn, Anneke, C1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/map-making-in-the-21st-century-charting-breast-cancer-susceptibility02349nas a2200301 4500008004100000245008500041210006900126260001600195300001300211490000800224520137300232653001201605653001401617653001301631653001401644653002001658653003101678653000901709653002901718653001801747653001601765100002601781700001301807700002601820700002601846700002101872856015401893 2011 eng d00aMechanism of a genetically encoded dark-to-bright reporter for caspase activity.0 aMechanism of a genetically encoded darktobright reporter for cas c2011 Jul 15 a24977-860 v2863 aFluorescent proteins have revolutionized modern biology with their ability to report the presence of tagged proteins in living systems. Although several fluorescent proteins have been described in which the excitation and emission properties can be modulated by external triggers, no fluorescent proteins have been described that can be activated from a silent dark state to a bright fluorescent state directly by the activity of an enzyme. We have developed a version of GFP in which fluorescence is completely quenched by appendage of a hydrophobic quenching peptide that tetramerizes GFP and prevents maturation of the chromophore. The fluorescence can be fully restored by catalytic removal of the quenching peptide, making it a robust reporter of proteolysis. We have demonstrated the utility of this uniquely dark state of GFP as a genetically encoded apoptosis reporter that monitors the function of caspases, which catalyze the fate-determining step in programmed cell death. Caspase Activatable-GFP (CA-GFP) can be activated both in vitro and in vivo, resulting in up to a 45-fold increase in fluorescent signal in bacteria and a 3-fold increase in mammalian cells. We used CA-GFP successfully to monitor real-time apoptosis in mammalian cells. This dark state of GFP may ultimately serve as a useful platform for probes of other enzymatic processes.
10aAnimals10aApoptosis10aCaspases10aCatalysis10aGenes, Reporter10aGreen Fluorescent Proteins10aMice10aMicroscopy, Fluorescence10aNIH 3T3 Cells10aProteolysis1 aNicholls, Samantha, B1 aChu, Jun1 aAbbruzzese, Genevieve1 aTremblay, Kimberly, D1 aHardy, Jeanne, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mechanism-of-a-genetically-encoded-dark-to-bright-reporter-for-caspase02966nas a2200409 4500008004100000245008000041210006900121260001600190300001400206490000800220520168500228653001201913653001801925653001401943653003301957653001501990653003102005653000902036653001902045653001702064653002802081653001302109653001402122653003002136653002302166653003302189100002102222700001402243700002402257700002202281700002302303700002002326700001802346700002202364700002102386856014902407 2011 eng d00aMitochondrial dysfunction impairs tumor suppressor p53 expression/function.0 aMitochondrial dysfunction impairs tumor suppressor p53 expressio c2011 Jun 10 a20297-3120 v2863 aRecently, mitochondria have been suggested to act in tumor suppression. However, the underlying mechanisms by which mitochondria suppress tumorigenesis are far from being clear. In this study, we have investigated the link between mitochondrial dysfunction and the tumor suppressor protein p53 using a set of respiration-deficient (Res(-)) mammalian cell mutants with impaired assembly of the oxidative phosphorylation machinery. Our data suggest that normal mitochondrial function is required for γ-irradiation (γIR)-induced cell death, which is mainly a p53-dependent process. The Res(-) cells are protected against γIR-induced cell death due to impaired p53 expression/function. We find that the loss of complex I biogenesis in the absence of the MWFE subunit reduces the steady-state level of the p53 protein, although there is no effect on the p53 protein level in the absence of the ESSS subunit that is also essential for complex I assembly. The p53 protein level was also reduced to undetectable levels in Res(-) cells with severely impaired mitochondrial protein synthesis. This suggests that p53 protein expression is differentially regulated depending upon the type of electron transport chain/respiratory chain deficiency. Moreover, irrespective of the differences in the p53 protein expression profile, γIR-induced p53 activity is compromised in all Res(-) cells. Using two different conditional systems for complex I assembly, we also show that the effect of mitochondrial dysfunction on p53 expression/function is a reversible phenomenon. We believe that these findings will have major implications in the understanding of cancer development and therapy.
10aAnimals10aBase Sequence10aCell Line10aElectron Transport Complex I10aGamma Rays10aGene Expression Regulation10aMice10aMice, Knockout10aMitochondria10aMolecular Sequence Data10aMutation10aNeoplasms10aOxidative Phosphorylation10aOxygen Consumption10aTumor Suppressor Protein p531 aCompton, Shannon1 aKim, Chul1 aGriner, Nicholas, B1 aPotluri, Prasanth1 aScheffler, Immo, E1 aSen, Sabyasachi1 aJerry, Joseph1 aSchneider, Sallie1 aYadava, Nagendra uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mitochondrial-dysfunction-impairs-tumor-suppressor-p53-expression00800nas a2200253 4500008004100000245006800041210006700109260001200176300001200188490000800200100002000208700001600228700001400244700001600258700001300274700001400287700001500301700001800316700001800334700001700352700001300369700001800382856014600400 2011 eng d00aNotch signaling regulates mouse and human Th17 differentiation.0 aNotch signaling regulates mouse and human Th17 differentiation c07/2011 a692-7010 v1871 aKeerthivasan, S1 aSuleiman, R1 aLawlor, R1 aRoderick, J1 aBates, T1 aMinter, L1 aAnguita, J1 aJuncadella, I1 aNickoloff, BJ1 aLe Poole, IC1 aMiele, L1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch-signaling-regulates-mouse-and-human-th17-differentiation01681nas a2200157 4500008004100000245006700041210006600108260001700174300001100191490000700202520111100209100001501320700002101335700002301356856014401379 2011 eng d00aPLCζ and its role as a trigger of development in vertebrates.0 aPLCζ and its role as a trigger of development in vertebrates c2011 Oct-Nov a846-530 v783 aA major unresolved issue in developmental biology is the precise mechanism whereby the sperm activates the oocyte. With the discovery that calcium signals are the primary trigger for oocyte activation, a key remaining question became the identification of the signaling protein that mediates such calcium signals at fertilization. A major step forward came in 2002 with the discovery of a sperm-specific mammalian phospholipase C called phospholipase C zeta (PLCζ), which had the expected properties of the mammalian oocyte activation factor and was subsequently identified in other vertebrate groups. Most recently, defects in PLCζ have been shown to be linked to certain types of male infertility in humans. Despite these advances, many questions remain about the precise mechanism of action of PLCζ and the extent of its role during oocyte activation in the vertebrate kingdom. In this review, we will look at the current state of understanding of PLCζ's mechanism of action and physiological role in mammals and other vertebrates, and identify areas of uncertainty that still remain to be resolved.1 aIto, Junya1 aParrington, John1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/plcz-and-its-role-as-a-trigger-of-development-in-vertebrates02181nas a2200301 4500008004100000022001400041245009800055210006900153260001600222300001200238490000700250520120000257653001201457653001601469653002301485653001701508653001701525653001901542653001401561653001501575653000901590653001801599653005001617100002101667700002401688700001901712856014801731 2011 eng d a1520-511800aPyrethroid mode(s) of action in the context of Food Quality Protection Act (FQPA) regulation.0 aPyrethroid modes of action in the context of Food Quality Protec c2011 Apr 13 a2773-850 v593 aA Scientific Advisory Panel (SAP) in June 2009 concluded that a common mode of action existed for pyrethroids, with two subgroups. The purpose of this SAP was to advise the U.S. Environmental Protection Agency on the validity of regulation of pyrethroids as a single class under the Food Quality Protection Act of 1996. Two types of pyrethroid action were first described for clinical signs in the rat and clinical signs/nerve effects in the cockroach. In insects, Type I clinical signs correlate with repetitive firing in nerve axons, especially fine sensory axons. The Na(+) inward current is via a TTX-sensitive voltage-gated sodium channel (VGSC). Type II (α-CN) effects on VGSCs do not include repetitive firing following stimulation in these axons. Instead, Type II effects on VGSCs include prolonged Na(+) tail currents along with depolarization of nerve membrane. Other Type II effects have been measured on VG Ca(2+) and K(+) channels and VG and GABA-activated Cl(-) channels. In conclusion, in vivo pyrethroid effects in mammals should be linked with specific channel effects, allowing the use of specific clinical signs or ion channel effects for pyrethroid risk assessment.
10aAnimals10aCockroaches10aFood Contamination10aInsecticides10aIon Channels10aNervous System10aPoisoning10aPyrethrins10aRats10aUnited States10aUnited States Environmental Protection Agency1 aGammon, Derek, W1 aLeggett, Michael, F1 aClark, John, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pyrethroid-modes-action-context-food-quality-protection-act-fqpa02908nas a2200577 4500008004100000245015000041210006900191260001600260300001100276490000700287520096800294653001201262653002101274653003601295653004201331653002101373653001101394653003001405653004301435653002901478653002701507653000901534653002401543653001901567653003301586653002201619653001801641653002401659653001701683653003701700653001701737653002701754653003301781653002701814100002101841700002801862700002501890700002101915700002501936700001801961700002401979700001802003700002702021700001802048700002102066700001902087700001902106700001902125700003202144856015402176 2011 eng d00aRadiation acts on the microenvironment to affect breast carcinogenesis by distinct mechanisms that decrease cancer latency and affect tumor type.0 aRadiation acts on the microenvironment to affect breast carcinog c2011 May 17 a640-510 v193 aTissue microenvironment is an important determinant of carcinogenesis. We demonstrate that ionizing radiation, a known carcinogen, affects cancer frequency and characteristics by acting on the microenvironment. Using a mammary chimera model in which an irradiated host is transplanted with oncogenic Trp53 null epithelium, we show accelerated development of aggressive tumors whose molecular signatures were distinct from tumors arising in nonirradiated hosts. Molecular and genetic approaches show that TGFβ mediated tumor acceleration. Tumor molecular signatures implicated TGFβ, and genetically reducing TGFβ abrogated the effect on latency. Surprisingly, tumors from irradiated hosts were predominantly estrogen receptor negative. This effect was TGFβ independent and linked to mammary stem cell activity. Thus, the irradiated microenvironment affects latency and clinically relevant features of cancer through distinct and unexpected mechanisms.
10aAnimals10aBreast Neoplasms10aCell Transformation, Neoplastic10aDose-Response Relationship, Radiation10aEpithelial Cells10aFemale10aGene Expression Profiling10aGene Expression Regulation, Neoplastic10aGene Regulatory Networks10aMammary Glands, Animal10aMice10aMice, Inbred BALB C10aMice, Knockout10aNeoplasms, Radiation-Induced10aRadiation Chimera10aReaction Time10aReceptors, Estrogen10aTime Factors10aTransforming Growth Factor beta110aTumor Burden10aTumor Microenvironment10aTumor Suppressor Protein p5310aWhole-Body Irradiation1 aNguyen, David, H1 aOketch-Rabah, Hellen, A1 aIlla-Bochaca, Irineu1 aGeyer, Felipe, C1 aReis-Filho, Jorge, S1 aMao, Jian-Hua1 aRavani, Shraddha, A1 aZavadil, Jiri1 aBorowsky, Alexander, D1 aJerry, Joseph1 aDunphy, Karen, A1 aSeo, Jae, Hong1 aHaslam, Sandra1 aMedina, Daniel1 aBarcellos-Hoff, Mary, Helen uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/radiation-acts-on-the-microenvironment-to-affect-breast-carcinogenesis02194nas a2200193 4500008004100000245011500041210006900156260001300225300001100238490000700249520148500256100001501741700002001756700002101776700001901797700001701816700001801833856014901851 2011 eng d00aRepression of mammary stem/progenitor cells by p53 is mediated by Notch and separable from apoptotic activity.0 aRepression of mammary stemprogenitor cells by p53 is mediated by c2011 Jan a119-270 v293 aBreast cancer is the most common tumor among women with inherited mutations in the p53 gene (Li-Fraumeni syndrome). The tumors represent the basal-like subtype, which has been suggested to originate from mammary stem/progenitor cells. In mouse mammary epithelium, mammosphere-forming potential was increased with decreased dosage of the gene encoding the p53 tumor suppressor protein (Trp53). Limiting dilution transplantation also showed a 3.3-fold increase in the frequency of long-term regenerative mammary stem cells in Trp53-/- mice. The repression of mammospheres by p53 was apparent despite the absence of apoptotic responses to radiation indicating a dissociation of these two activities of p53. The effects of p53 on progenitor cells were also observed in TM40A cells using both mammosphere-forming assays and the DsRed-let7c-sensor. The frequency of long-term label-retaining epithelial cells was decreased in Trp53-/- mammary glands indicating that asymmetric segregation of DNA is diminished and contributes to the expansion of the mammary stem cells. Treatment with an inhibitor of γ-secretase (N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester) reduced the number of Trp53-/- mammospheres to the level found in Trp53+/+ cells. These results demonstrate that basal levels of p53 restrict mammary stem/progenitor cells through Notch and that the Notch pathway is a therapeutic target to prevent expansion of this vulnerable pool of cells.
1 aTao, Luwei1 aRoberts, Amy, L1 aDunphy, Karen, A1 aBigelow, Carol1 aYan, Haoheng1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/repression-of-mammary-stem-progenitor-cells-by-p53-is-mediated-by02239nas a2200289 4500008004100000245007000041210006500111260001300176300001100189490000700200520129500207653001301502653001201515653002101527653001101548653001101559653002701570653002601597653003601623653002401659653003601683100002101719700002201740700001901762700001801781856015001799 2011 eng d00aThe role of activin in mammary gland development and oncogenesis.0 arole of activin in mammary gland development and oncogenesis c2011 Jun a117-260 v163 aTGFβ contributes to mammary gland development and has paradoxical roles in breast cancer because it has both tumor suppressor and tumor promoter activity. Another member of the TGFβ superfamily, activin, also has roles in the developing mammary gland, but these functions, and the role of activin in breast cancer, are not well characterized. TGFβ and activin share the same intracellular signaling pathways, but divergence in their signaling pathways are suggested. The purpose of this review is to compare the spatial and temporal expression of TGFβ and activin during mammary gland development, with consideration given to their functions during each developmental period. We also review the contributions of TGFβ and activin to breast cancer resistance and susceptibility. Finally, we consider the systemic contributions of activin in regulating obesity and diabetes; and the impact this regulation has on breast cancer. Elevated levels of activin in serum during pregnancy and its influence on pregnancy associated breast cancer are also considered. We conclude that evidence demonstrates that activin has tumor suppressing potential, without definitive indication of tumor promoting activity in the mammary gland, making it a good target for development of therapeutics.
10aActivins10aAnimals10aBreast Neoplasms10aFemale10aHumans10aMammary Glands, Animal10aMammary Glands, Human10aMammary Neoplasms, Experimental10aSignal Transduction10aTransforming Growth Factor beta1 aDunphy, Karen, A1 aSchneyer, Alan, L1 aHagen, Mary, J1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-role-of-activin-in-mammary-gland-development-and-oncogenesis-001900nas a2200169 4500008004100000245007000041210006500111260001300176300001100189490000700200520129500207100002101502700002201523700001901545700001801564856014801582 2011 eng d00aThe role of activin in mammary gland development and oncogenesis.0 arole of activin in mammary gland development and oncogenesis c2011 Jun a117-260 v163 aTGFβ contributes to mammary gland development and has paradoxical roles in breast cancer because it has both tumor suppressor and tumor promoter activity. Another member of the TGFβ superfamily, activin, also has roles in the developing mammary gland, but these functions, and the role of activin in breast cancer, are not well characterized. TGFβ and activin share the same intracellular signaling pathways, but divergence in their signaling pathways are suggested. The purpose of this review is to compare the spatial and temporal expression of TGFβ and activin during mammary gland development, with consideration given to their functions during each developmental period. We also review the contributions of TGFβ and activin to breast cancer resistance and susceptibility. Finally, we consider the systemic contributions of activin in regulating obesity and diabetes; and the impact this regulation has on breast cancer. Elevated levels of activin in serum during pregnancy and its influence on pregnancy associated breast cancer are also considered. We conclude that evidence demonstrates that activin has tumor suppressing potential, without definitive indication of tumor promoting activity in the mammary gland, making it a good target for development of therapeutics.
1 aDunphy, Karen, A1 aSchneyer, Alan, L1 aHagen, Mary, J1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-role-of-activin-in-mammary-gland-development-and-oncogenesis02640nas a2200289 4500008004100000245006400041210005800105260001300163300001100176490000700187520164200194653001201836653001401848653004001862653003101902653001101933653000901944653005301953653002402006653004902030653004202079100001602121700002802137700002302165700002002188856014202208 2011 eng d00aThe role of TRADD in TRAIL-induced apoptosis and signaling.0 arole of TRADD in TRAILinduced apoptosis and signaling c2011 Apr a1353-80 v253 aTumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily. TRAIL is promising for anticancer therapy because it induces apoptosis in cancer cells with little or no toxicity to normal cells; hence, TRAIL-receptor agonists are currently undergoing clinical trials for cancer treatment. However, many molecular signaling mechanisms in TRAIL signaling are not completely characterized. The functions of adaptor proteins, including TNF-receptor-associated death domain protein (TRADD) and receptor-interacting protein-1 (RIP1) in TRAIL signaling have been controversial. We demonstrate that while wild-type mouse embryonic fibroblasts (MEFs) are completely resistant to TRAIL-induced apoptosis, MEFs derived from Tradd(-/-) mice are hypersensitive to TRAIL (IC(50)~0.5 nM rmTRAIL, 24 h), an effect also seen in primary keratinocytes treated with TRAIL/CHX. Restoration of TRADD in Tradd(-/-) MEFs restores TRAIL resistance, indicating that TRADD plays a survival role in TRAIL signaling. We show that TRADD is recruited to the TRAIL-receptor complex, and RIP1 recruitment is mediated by TRADD. While early activation of the MAP kinase ERK is deficient in Tradd(-/-) cells, the main mechanism for enhanced TRAIL sensitivity is likely due to increased recruitment of FADD to the receptor complex, indicating that TRADD may limit FADD binding within the receptor complex and also mediate RIP1-dependent nonapoptotic signaling events, thus reducing caspase activation and subsequent apoptosis. These novel findings have potential implications for cancer therapy using TRAIL-receptor agonists.
10aAnimals10aApoptosis10aFas-Associated Death Domain Protein10aGTPase-Activating Proteins10aHumans10aMice10aReceptors, TNF-Related Apoptosis-Inducing Ligand10aSignal Transduction10aTNF Receptor-Associated Death Domain Protein10aTNF-Related Apoptosis-Inducing Ligand1 aCao, Xiumei1 aPobezinskaya, Yelena, L1 aMorgan, Michael, J1 aLiu, Zheng-Gang uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-role-of-tradd-in-trail-induced-apoptosis-and-signaling01698nas a2200193 4500008004100000245008200041210007100123260001300194300001000207490000700217520101400224100001401238700002501252700001701277700001701294700002401311700002201335856014701357 2011 eng d00aScavenger receptor WC1 contributes to the γδ T cell response to Leptospira.0 aScavenger receptor WC1 contributes to the γδ T cell response to c2011 Mar a801-90 v483 aWC1 molecules are exclusively expressed on the surface of γδ T cells. They belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. WC1 molecules have been grouped on the basis of antibody reactivity. The expression of WC1 molecules from these serologically defined groups is correlated with differences in γδ T cell responses. The expression of receptors within the WC1.1 group correlates with the capacity of γδ T cells to respond to Leptospira antigen. In this study, we used RNA interference to directly investigate the role of WC1 expression in the response to Leptospira borgpetersenii. We found that when three out of thirteen WC1 gene products were downregulated by RNA interference, γδ T cell proliferation and IFN-γ production in response to Leptospira antigen was significantly reduced. Our data demonstrate that specific receptors in the WC1 family directly participate in Leptospira recognition and/or activation of γδ T cells.
1 aWang, Fei1 aHerzig, Carolyn, T A1 aChen, Chuang1 aHsu, Haoting1 aBaldwin, Cynthia, L1 aTelfer, Janice, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/scavenger-receptor-wc1-contributes-to-the-gd-t-cell-response-to02019nas a2200253 4500008004100000022001400041245012100055210006900176260001300245300001100258490000600269520112600275100002601401700002201427700002901449700002001478700002501498700001901523700001601542700001901558700002001577700001901597856014901616 2011 eng d a1942-088900aSimplify, simplify: Lifestyle and compact genome of the body louse provide a unique functional genomics opportunity.0 aSimplify simplify Lifestyle and compact genome of the body louse c2011 Mar a188-910 v43 aThe body louse, with its recently sequenced genome, is now primed to serve as a powerful model organism for addressing fundamental questions relating to how insects interact with their environment. One characteristic of the body louse that facilitates this research is the size of its genome-the smallest insect genome sequenced to date. This diminutive genome must nonetheless control an organism that senses and responds to its environment, reacting to threats of corporal and genomic integrity. Additionally, the body louse transmits several important human diseases compared to its very close relative, the head louse, which does not. Therefore, these two organisms comprise an excellent model system for studying molecular mechanisms associated with vector competence. To understand more fully the development of vector/pathogen interactions, we have developed an in vitro bioassay system and determined that the body louse genome appears to contain the genes necessary for RNAi. The body louse will therefore be useful for determining the set of conditions permissive to the evolution of vector competence.
1 aPittendrigh, Barry, R1 aBerenbaum, May, R1 aSeufferheld, Manfredo, J1 aMargam, Venu, M1 aStrycharz, Joseph, P1 aYoon, Kyong, S1 aSun, Weilin1 aReenan, Robert1 aLee, Si, Hyeock1 aClark, John, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/simplify-simplify-lifestyle-and-compact-genome-body-louse-provide01983nas a2200241 4500008004100000245012100041210006900162260001300231300001100244490000600255520111700261100002601378700002201404700002901426700002001455700002501475700001901500700001601519700001901535700002001554700001901574856014801593 2011 eng d00aSimplify, simplify: Lifestyle and compact genome of the body louse provide a unique functional genomics opportunity.0 aSimplify simplify Lifestyle and compact genome of the body louse c2011 Mar a188-910 v43 aThe body louse, with its recently sequenced genome, is now primed to serve as a powerful model organism for addressing fundamental questions relating to how insects interact with their environment. One characteristic of the body louse that facilitates this research is the size of its genome-the smallest insect genome sequenced to date. This diminutive genome must nonetheless control an organism that senses and responds to its environment, reacting to threats of corporal and genomic integrity. Additionally, the body louse transmits several important human diseases compared to its very close relative, the head louse, which does not. Therefore, these two organisms comprise an excellent model system for studying molecular mechanisms associated with vector competence. To understand more fully the development of vector/pathogen interactions, we have developed an in vitro bioassay system and determined that the body louse genome appears to contain the genes necessary for RNAi. The body louse will therefore be useful for determining the set of conditions permissive to the evolution of vector competence.1 aPittendrigh, Barry, R1 aBerenbaum, May, R1 aSeufferheld, Manfredo, J1 aMargam, Venu, M1 aStrycharz, Joseph, P1 aYoon, Kyong, S1 aSun, Weilin1 aReenan, Robert1 aLee, Si, Hyeock1 aClark, John, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/simplify-simplify-lifestyle-and-compact-genome-of-the-body-louse02716nas a2200229 4500008004100000245015100041210006900192260001300261300001300274490000600287520186300293100001902156700002302175700001602198700002002214700002202234700002102256700002502277700001802302700002102320856014502341 2011 eng d00aT. brucei infection reduces B lymphopoiesis in bone marrow and truncates compensatory splenic lymphopoiesis through transitional B-cell apoptosis.0 aT brucei infection reduces B lymphopoiesis in bone marrow and tr c2011 Jun ae10020890 v73 aAfrican trypanosomes of the Trypanosoma brucei species are extracellular protozoan parasites that cause the deadly disease African trypanosomiasis in humans and contribute to the animal counterpart, Nagana. Trypanosome clearance from the bloodstream is mediated by antibodies specific for their Variant Surface Glycoprotein (VSG) coat antigens. However, T. brucei infection induces polyclonal B cell activation, B cell clonal exhaustion, sustained depletion of mature splenic Marginal Zone B (MZB) and Follicular B (FoB) cells, and destruction of the B-cell memory compartment. To determine how trypanosome infection compromises the humoral immune defense system we used a C57BL/6 T. brucei AnTat 1.1 mouse model and multicolor flow cytometry to document B cell development and maturation during infection. Our results show a more than 95% reduction in B cell precursor numbers from the CLP, pre-pro-B, pro-B, pre-B and immature B cell stages in the bone marrow. In the spleen, T. brucei induces extramedullary B lymphopoiesis as evidenced by significant increases in HSC-LMPP, CLP, pre-pro-B, pro-B and pre-B cell populations. However, final B cell maturation is abrogated by infection-induced apoptosis of transitional B cells of both the T1 and T2 populations which is not uniquely dependent on TNF-, Fas-, or prostaglandin-dependent death pathways. Results obtained from ex vivo co-cultures of living bloodstream form trypanosomes and splenocytes demonstrate that trypanosome surface coat-dependent contact with T1/2 B cells triggers their deletion. We conclude that infection-induced and possibly parasite-contact dependent deletion of transitional B cells prevents replenishment of mature B cell compartments during infection thus contributing to a loss of the host's capacity to sustain antibody responses against recurring parasitemic waves.
1 aBockstal, Viki1 aGuirnalda, Patrick1 aCaljon, Guy1 aGoenka, Radhika1 aTelfer, Janice, C1 aFrenkel, Deborah1 aRadwanska, Magdalena1 aMagez, Stefan1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/t-brucei-infection-reduces-b-lymphopoiesis-in-bone-marrow-and02443nas a2200421 4500008004100000245014900041210006900190260001600259300001100275490000700286520102500293653001801318653004101336653001201377653001201389653001401401653001801415653001701433653002301450653002501473653002501498653004601523653001101569653002201580653001701602653003201619653002201651653001001673653003601683653001901719653002101738100002101759700002601780700001901806700002001825700002501845856015101870 2011 eng d00aTranslocation of the cytoplasmic domain of ADAM13 to the nucleus is essential for Calpain8-a expression and cranial neural crest cell migration.0 aTranslocation of the cytoplasmic domain of ADAM13 to the nucleus c2011 Feb 15 a256-630 v203 aADAMs are transmembrane metalloproteases that control cell behavior by cleaving both cell adhesion and signaling molecules. The cytoplasmic domain of ADAMs can regulate the proteolytic activity by controlling the subcellular localization and/or the activation of the protease domain. Here, we show that the cytoplasmic domain of ADAM13 is cleaved and translocates into the nucleus. Preventing this translocation renders the protein incapable of promoting cranial neural crest (CNC) cell migration in vivo, without affecting its proteolytic activity. In addition, the cytoplasmic domain of ADAM13 regulates the expression of multiple genes in CNC, including the protease Calpain8-a. Restoring the expression of Calpain8-a is sufficient to rescue CNC migration in the absence of the ADAM13 cytoplasmic domain. This study shows that the cytoplasmic domain of ADAM metalloproteases can perform essential functions in the nucleus of cells and may contribute substantially to the overall function of the protein.
10aADAM Proteins10aAmyloid Precursor Protein Secretases10aAnimals10aCalpain10aCell Line10aCell Movement10aCell Nucleus10aConserved Sequence10aEmbryo, Nonmammalian10aEvolution, Molecular10aGene Expression Regulation, Developmental10aHumans10aMembrane Proteins10aNeural Crest10aProtein Structure, Tertiary10aProtein Transport10aSkull10aStructure-Activity Relationship10aXenopus laevis10aXenopus Proteins1 aCousin, Hélène1 aAbbruzzese, Genevieve1 aKerdavid, Erin1 aGaultier, Alban1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/translocation-of-the-cytoplasmic-domain-of-adam13-to-the-nucleus-is02101nas a2200325 4500008004100000245010700041210006900148260001500217300000900232490000800241520103500249653002201284653001201306653001801318653001801336653001501354653004101369653002201410653002401432653000901456653002001465653001901485653001601504100002101520700001701541700002301558700002301581700002201604856014901626 2011 eng d00aTransmembrane adenylyl cyclase regulates amphibian sperm motility through protein kinase A activation.0 aTransmembrane adenylyl cyclase regulates amphibian sperm motilit c2011 Feb 1 a80-80 v3503 aSperm motility is essential for achieving fertilization. In animals with external fertilization as amphibians, spermatozoa are stored in a quiescent state in the testis. Spermiation to hypotonic fertilization media triggers activation of sperm motility. Bufo arenarum sperm are immotile in artificial seminal plasma (ASP) but acquire in situ flagellar beating upon dilution. In addition to the effect of low osmolarity on sperm motility activation, we report that diffusible factors of the egg jelly coat (EW) regulate motility patterns, switching from in situ to progressive movement. The signal transduction pathway involved in amphibian sperm motility activation is mostly unknown. In the present study, we show a correlation between motility activation triggered by low osmotic pressure and activation of protein kinase A (PKA). Moreover, this is the first study to present strong evidences that point toward a role of a transmembrane adenyl-cyclase (tmAC) in the regulation of amphibian sperm motility through PKA activation.10aAdenylate Cyclase10aAnimals10aBufo arenarum10aCell Membrane10aCyclic AMP10aCyclic AMP-Dependent Protein Kinases10aEnzyme Activation10aHypotonic Solutions10aMale10aPhosphorylation10aSperm Motility10aSpermatozoa1 aO'Brien, Emma, D1 aKrapf, Dario1 aCabada, Marcelo, O1 aVisconti, Pablo, E1 aArranz, Silvia, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transmembrane-adenylyl-cyclase-regulates-amphibian-sperm-motility01968nas a2200337 4500008004100000245006100041210006000102260000900162300001100171490000600182520095900188653001201147653001401159653001501173653002501188653001101213653004601224653001501270653000901285653000901294653001301303653002501316653003301341100001901374700002201393700001901415700002001434700002001454700001701474856013901491 2011 eng d00aWdr74 is required for blastocyst formation in the mouse.0 aWdr74 is required for blastocyst formation in the mouse c2011 ae225160 v63 aPreimplantation is a dynamic developmental period during which a combination of maternal and zygotic factors program the early embryo resulting in lineage specification and implantation. A reverse genetic RNAi screen in mouse embryos identified the WD Repeat Domain 74 gene (Wdr74) as being required for these critical first steps of mammalian development. Knockdown of Wdr74 results in embryos that develop normally until the morula stage but fail to form blastocysts or properly specify the inner cell mass and trophectoderm. In Wdr74-deficient embryos, we find activated Trp53-dependent apoptosis as well as a global reduction of RNA polymerase I, II and III transcripts. In Wdr74-deficient embryos blocking Trp53 function rescues blastocyst formation and lineage differentiation. These results indicate that Wdr74 is required for RNA transcription, processing and/or stability during preimplantation development and is an essential gene in the mouse.10aAnimals10aApoptosis10aBlastocyst10aCell Differentiation10aFemale10aGene Expression Regulation, Developmental10aInjections10aMale10aMice10aProteins10aRNA, Double-Stranded10aTumor Suppressor Protein p531 aMaserati, Marc1 aWalentuk, Melanie1 aDai, Xiangpeng1 aHolston, Olivia1 aAdams, Danielle1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/wdr74-is-required-for-blastocyst-formation-in-the-mouse02292nas a2200397 4500008004100000245007400041210006900115260001300184300001100197490000700208520103200215653001201247653001801259653003401277653002301311653001101334653004601345653002001391653003601411653000901447653002301456653001901479653002301498653001201521653001401533653001901547653002901566100002501595700001901620700001801639700002201657700002001679700002601699700001701725856015201742 2011 eng d00aYin-yang1 is required in the mammalian oocyte for follicle expansion.0 aYinyang1 is required in the mammalian oocyte for follicle expans c2011 Apr a654-630 v843 aThe multifaceted polycomb group gene Yin-Yang1 (Yy1) has been implicated in a variety of transcriptional regulatory roles both as an activator and silencer of gene expression. Here we examine the role of Yy1 during oocyte growth by conditional deletion of the locus in the growing oocyte. Our results indicate that YY1 is required for oocyte maturation and granulosa cell expansion. In mutant oocytes, we observe severely reduced expression of both Gdf9 and Bmp15, suggesting a mechanism underlying the failure of granulosa cell expansion. Consequently, we observe infertility, failure of estrus cycling, and altered reproductive hormone levels in mutant females. Additionally, we find that YY1-deficient oocytes exhibit altered levels of several oocyte-specific factors, including Pou5f1, Figla, Lhx8, Oosp1, and Sohlh2. These results document YY1's involvement in folliculogenesis and ovarian function in the mouse and indicate that YY1 is required specifically in the oocyte for oocyte-granulosa cell communication.
10aAnimals10aBase Sequence10aBone Morphogenetic Protein 1510aCell Communication10aFemale10aGene Expression Regulation, Developmental10aGranulosa Cells10aGrowth Differentiation Factor 910aMice10aMice, Inbred C57BL10aMice, Knockout10aModels, Biological10aOocytes10aOogenesis10aRNA, Messenger10aYY1 Transcription Factor1 aGriffith, Gillian, J1 aTrask, Mary, C1 aHiller, Jacob1 aWalentuk, Melanie1 aPawlak, John, B1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/yin-yang1-is-required-in-the-mammalian-oocyte-for-follicle-expansion02018nas a2200157 4500008004100000245010100041210006900142260001300211300001100224490000800235520140100243100001901644700002401663700002301687856015001710 2010 eng d00aAnalysis of CAPZA3 localization reveals temporally discrete events during the acrosome reaction.0 aAnalysis of CAPZA3 localization reveals temporally discrete even c2010 Sep a575-800 v2243 aIn mammals, the starting point of development is the fusion between sperm and egg. It is well established that sperm fuse with the egg through the equatorial/post-acrosomal region. Apart from this observation and the requirement of two proteins (CD9 in the egg and IZUMO1 in the sperm) very little is known about this fundamental process. Actin polymerization correlates with sperm capacitation in different mammalian species and it has been proposed that F-actin breakdown is needed during the acrosome reaction. Recently, we have presented evidence that actin polymerization inhibitors block the movement of IZUMO1 that accompany the acrosome reaction. These results suggest that actin dynamics play a role in the observed changes in IZUMO1 localization. This finding is significant because IZUMO1 localization in acrosome-intact sperm is not compatible with the known location of the initiation of the fusion between the sperm and the egg. To further understand the actin-mediated changes in protein localization during the acrosome reaction, the distribution of the sperm-specific plus-end actin capping protein CAPZA3 was analyzed. Like IZUMO1, CAPZA3 shows a dynamic pattern of localization; however, these movements follow a different temporal pattern than the changes observed with IZUMO1. In addition, the actin polymerization inhibitor latrunculin A was unable to alter CAPZA3 movement.1 aSosnik, Julian1 aBuffone, Mariano, G1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/analysis-of-capza3-localization-reveals-temporally-discrete-events01215nas a2200157 4500008004100000245007700041210006900118260000900187300000800196490000700204520062300211100002500834700002500859700002400884856014900908 2010 eng d00aAnnotation and classification of the bovine T cell receptor delta genes.0 aAnnotation and classification of the bovine T cell receptor delt c2010 a1000 v113 agammadelta T cells differ from alphabeta T cells with regard to the types of antigen with which their T cell receptors interact; gammadelta T cell antigens are not necessarily peptides nor are they presented on MHC. Cattle are considered a "gammadelta T cell high" species indicating they have an increased proportion of gammadelta T cells in circulation relative to that in "gammadelta T cell low" species such as humans and mice. Prior to the onset of the studies described here, there was limited information regarding the genes that code for the T cell receptor delta chains of this gammadelta T cell high species.1 aHerzig, Carolyn, T A1 aLefranc, Marie-Paule1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/annotation-and-classification-of-the-bovine-t-cell-receptor-delta01745nas a2200361 4500008004100000245010600041210006900147260001600216300001200232490000800244520060300252653001200855653002600867653001400893653002600907653001700933653001500950653001100965653000900976653002400985653002401009653001401033653001301047653003201060100001901092700002201111700002101133700001601154700001801170700001901188700002401207856015201231 2010 eng d00aArray-based sensing of normal, cancerous, and metastatic cells using conjugated fluorescent polymers.0 aArraybased sensing of normal cancerous and metastatic cells usin c2010 Jan 27 a1018-220 v1323 aA family of conjugated fluorescent polymers was used to create an array for cell sensing. Fluorescent conjugated polymers with pendant charged residues provided multivalent interactions with cell membranes, allowing the detection of subtle differences between different cell types on the basis of cell surface features. Highly reproducible characteristic patterns were obtained from different cell types as well as from isogenic cell lines, enabling the identification of the cell type as well differentiating between normal, cancerous, and metastatic isogenic cell types with high accuracy.
10aAnimals10aBiosensing Techniques10aCell Line10aDiscriminant Analysis10aFluorescence10aHeLa Cells10aHumans10aMice10aMice, Inbred BALB C10aMolecular Structure10aNeoplasms10aPolymers10aSensitivity and Specificity1 aBajaj, Avinash1 aMiranda, Oscar, R1 aPhillips, Ronnie1 aKim, Ik-Bum1 aJerry, Joseph1 aBunz, Uwe, H F1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/array-based-sensing-of-normal-cancerous-and-metastatic-cells-using-001343nas a2200205 4500008004100000245010600041210006900147260001600216300001200232490000800244520059600252100001900848700002200867700002100889700001600910700001800926700001900944700002400963856015000987 2010 eng d00aArray-based sensing of normal, cancerous, and metastatic cells using conjugated fluorescent polymers.0 aArraybased sensing of normal cancerous and metastatic cells usin c2010 Jan 27 a1018-220 v1323 aA family of conjugated fluorescent polymers was used to create an array for cell sensing. Fluorescent conjugated polymers with pendant charged residues provided multivalent interactions with cell membranes, allowing the detection of subtle differences between different cell types on the basis of cell surface features. Highly reproducible characteristic patterns were obtained from different cell types as well as from isogenic cell lines, enabling the identification of the cell type as well differentiating between normal, cancerous, and metastatic isogenic cell types with high accuracy.1 aBajaj, Avinash1 aMiranda, Oscar, R1 aPhillips, Ronnie1 aKim, Ik-Bum1 aJerry, Joseph1 aBunz, Uwe, H F1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/array-based-sensing-of-normal-cancerous-and-metastatic-cells-using01142nas a2200205 4500008004100000245015700041210006900198260001300267300001200280490000700292520033300299100002500632700002100657700002400678700002000702700002400722700002000746700002000766856015000786 2010 eng d00aDetermination of knockdown resistance allele frequencies in global human head louse populations using the serial invasive signal amplification reaction.0 aDetermination of knockdown resistance allele frequencies in glob c2010 Sep a1031-400 v663 aPediculosis is the most prevalent parasitic infestation of humans. Resistance to pyrethrin- and pyrethroid-based pediculicides is due to knockdown (kdr)-type point mutations in the voltage-sensitive sodium channel alpha-subunit gene. Early detection of resistance is crucial for the selection of effective management strategies.1 aHodgdon, Hilliary, E1 aYoon, Kyong, Sup1 aPrevite, Domenic, J1 aKim, Hyo, Jeong1 aAboelghar, Gamal, E1 aLee, Si, Hyeock1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/determination-of-knockdown-resistance-allele-frequencies-in-global02722nas a2200469 4500008004100000245006600041210006400107260001300171300001200184490000700196520123700203653001201440653001201452653001801464653004901482653002101531653004001552653002001592653001601612653001101628653004601639653002301685653003401708653002801742653003901770653000901809653000901818653002301827653002501850653002101875653002001896653001301916653001401929653003201943100002101975700002201996700002302018700001702041700002402058700002602082856014402108 2010 eng d00aDomain-specific response of imprinted genes to reduced DNMT1.0 aDomainspecific response of imprinted genes to reduced DNMT1 c2010 Aug a3916-280 v303 aImprinted genes are expressed in a monoallelic, parent-of-origin-specific manner. Clusters of imprinted genes are regulated by imprinting control regions (ICRs) characterized by DNA methylation of one allele. This methylation is critical for imprinting; a reduction in the DNA methyltransferase DNMT1 causes a widespread loss of imprinting. To better understand the role of DNA methylation in the regulation of imprinting, we characterized the effects of Dnmt1 mutations on the expression of a panel of imprinted genes in the embryo and placenta. We found striking differences among imprinted domains. The Igf2 and Peg3 domains showed imprinting perturbations with both null and partial loss-of-function mutations, and both domains had pairs of coordinately regulated genes with opposite responses to loss of DNMT1 function, suggesting these domains employ similar regulatory mechanisms. Genes in the Kcnq1 domain were less sensitive to the absence of DNMT1. Cdkn1c exhibited imprinting perturbations only in null mutants, while Kcnq1 and Ascl2 were largely unaffected by a loss of DNMT1 function. These results emphasize the critical role for DNA methylation in imprinting and reveal the different ways it controls gene expression.10aAlleles10aAnimals10aBase Sequence10aBasic Helix-Loop-Helix Transcription Factors10aCrosses, Genetic10aDNA (Cytosine-5-)-Methyltransferase10aDNA Methylation10aDNA Primers10aFemale10aGene Expression Regulation, Developmental10aGenomic Imprinting10aInsulin-Like Growth Factor II10aKCNQ1 Potassium Channel10aKruppel-Like Transcription Factors10aMale10aMice10aMice, Inbred C57BL10aMice, Mutant Strains10aMultigene Family10aMutant Proteins10aPlacenta10aPregnancy10aProtein Structure, Tertiary1 aWeaver, Jamie, R1 aSarkisian, Garnik1 aKrapp, Christopher1 aMager, Jesse1 aMann, Mellissa, R W1 aBartolomei, Marisa, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/domain-specific-response-of-imprinted-genes-to-reduced-dnmt102202nas a2200181 4500008004100000245016600041210006900207260001300276300001200289490000700301520144600308100002301754700002001777700002301797700002201820700002401842856015401866 2010 eng d00aEffects of exposure water volume, depuration time, and feeding status on vitellogenin mRNA induction in male medaka (Oryzias latipes) exposed to 17 β-estradiol.0 aEffects of exposure water volume depuration time and feeding sta c2010 Nov a1835-410 v733 aBioassays measuring the induction of vitellogenin gene expression in male fish are widely used for revealing estrogenic activity in water samples. Measuring induction of vitellogenin mRNA in males by means of RT-PCR analysis is a sensitive way to detect exposure to estrogenic chemicals. To date, little work has been done to examine variations in exposure conditions for assessing estrogenic activity in water samples using this model system. Here we report the results of experiments investigating the effects of volume of treatment water, time since removal from treatment water (depuration), and short-term food deprivation on vitellogenin mRNA induction in male Japanese medaka (Oryzias latipes). Fish exposed to a single concentration of E(2) while volume was manipulated were found to have similar levels of vitellogenin mRNA, though more E(2) was present at larger volumes. Removal of fish from E(2)-treated-water to clean water after exposures reduced vitellogenin levels in as little 24h, however, the vitellogenin levels of the fish transferred to the clean water remained above those of the control fish for at least 72 h. Depriving fish of food for up to 72 h during exposure to E(2) did not significantly reduce vitellogenin induction. Together these results support the conclusion that real time RT-PCR measurement of vitellogenin in male fish can be used as a robust indicator of exposure to estrogenic contaminants in water.1 aMoffatt, Lauren, T1 aMay, Chelsea, L1 aStuder, Kirsten, E1 aReckhow, David, A1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/effects-of-exposure-water-volume-depuration-time-and-feeding-status-on01790nas a2200265 4500008004100000245007400041210006900115260001300184300001200197490000700209520090400216653002101120653003901141653002401180653001401204653001101218653001101229653002401240653001701264653003301281100001801314700002101332700001901353856015201372 2010 eng d00aEstrogens, regulation of p53 and breast cancer risk: a balancing act.0 aEstrogens regulation of p53 and breast cancer risk a balancing a c2010 Apr a1017-230 v673 aThe paradoxical effects of ovarian hormones in both the promotion and prevention of breast cancer have been debated for over 30 years. Genetic studies have demonstrated that ovarian hormones act through NF-kappaB to stimulate proliferation and ductal elongation, whereas the p53 tumor suppressor protein plays a central role in rendering the mammary epithelium resistant to tumorigenesis. Transcriptional profiles now suggest that ovarian hormones stimulate a constellation of genes that interact with NF-kappaB and p53 to arbitrate the competing demands for proliferation and surveillance. Genes that participate in chromatin remodeling are among the acute transcriptional responses to estrogens and progestins. These genes are proposed to initiate epigenetic programs that influence the balance between proliferation and surveillance, and render the breast epithelium resistant to tumors.
10aBreast Neoplasms10aChromatin Assembly and Disassembly10aEpigenesis, Genetic10aEstrogens10aFemale10aHumans10aReceptors, Estrogen10aRisk Factors10aTumor Suppressor Protein p531 aJerry, Joseph1 aDunphy, Karen, A1 aHagen, Mary, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/estrogens-regulation-of-p53-and-breast-cancer-risk-a-balancing-act-001486nas a2200157 4500008004100000245007400041210006900115260001300184300001200197490000700209520090400216100001801120700002101138700001901159856015001178 2010 eng d00aEstrogens, regulation of p53 and breast cancer risk: a balancing act.0 aEstrogens regulation of p53 and breast cancer risk a balancing a c2010 Apr a1017-230 v673 aThe paradoxical effects of ovarian hormones in both the promotion and prevention of breast cancer have been debated for over 30 years. Genetic studies have demonstrated that ovarian hormones act through NF-kappaB to stimulate proliferation and ductal elongation, whereas the p53 tumor suppressor protein plays a central role in rendering the mammary epithelium resistant to tumorigenesis. Transcriptional profiles now suggest that ovarian hormones stimulate a constellation of genes that interact with NF-kappaB and p53 to arbitrate the competing demands for proliferation and surveillance. Genes that participate in chromatin remodeling are among the acute transcriptional responses to estrogens and progestins. These genes are proposed to initiate epigenetic programs that influence the balance between proliferation and surveillance, and render the breast epithelium resistant to tumors.
1 aJerry, Joseph1 aDunphy, Karen, A1 aHagen, Mary, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/estrogens-regulation-of-p53-and-breast-cancer-risk-a-balancing-act01235nas a2200169 4500008004100000245010500041210006900146260000900215300000800224490000700232520058200239100002500821700001900846700002400865700002200889856015400911 2010 eng d00aEvolution of the CD163 family and its relationship to the bovine gamma delta T cell co-receptor WC1.0 aEvolution of the CD163 family and its relationship to the bovine c2010 a1810 v103 aThe scavenger receptor cysteine rich (SRCR) domain is an ancient and conserved protein domain. CD163 and WC1 molecules are classed together as group B SRCR superfamily members, along with Spalpha, CD5 and CD6, all of which are expressed by immune system cells. There are three known types of CD163 molecules in mammals, CD163A (M130, coded for by CD163), CD163b (M160, coded for by CD163L1) and CD163c-alpha (CD163L1 or SCART), while their nearest relative, WC1, is encoded by a multigene family so far identified in the artiodactyl species of cattle, sheep, and pigs.
1 aHerzig, Carolyn, T A1 aWaters, Ray, W1 aBaldwin, Cynthia, L1 aTelfer, Janice, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evolution-of-the-cd163-family-and-its-relationship-to-the-bovine-gamma01643nas a2200169 4500008004100000245008800041210006900129260001600198300001100214490000800225520100200233100002401235700002001259700001901279700002401298856015101322 2010 eng d00aExpressed gene sequences of the equine cytokines interleukin-17 and interleukin-23.0 aExpressed gene sequences of the equine cytokines interleukin17 a c2010 Feb 15 a309-130 v1333 aThis report describes the initial cloning and characterization of the equine interleukin-17 (IL-17) expressed gene sequence from mRNA obtained from equine intestinal tissue and interleukin-23 (IL-23) expressed gene sequence from mRNA obtained from equine peripheral blood mononuclear cells. Equine IL-17 has 462 nucleotides in the translated region, determined by homology with known human and mouse sequences, and shares 84% and 75% identity, respectively. For the deduced amino acid sequences, the identity with human and mouse is 76% and 70%. Equine IL-23 has 579 nucleotides in the translated region. Homology with known human and mouse sequences was determined to be 89% and 77%. Deduced amino acid identities are 89% with the human sequence and 70% with the mouse sequence. The gene sequences were identified as part of the U.S. Veterinary Immune Reagent Network with a goal of developing reagents in order to aid veterinary researchers in the investigation of diseases in livestock species.1 aTompkins, Dannielle1 aHudgens, Edward1 aHorohov, David1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/expressed-gene-sequences-of-the-equine-cytokines-interleukin-17-and01681nas a2200217 4500008004100000245008500041210006900126260000900195300000900204490000700213520097400220653001201194653001001206653001701216653001301233653001101246653000901257653001801266100002601284856015301310 2010 eng d00aFormation of the murine endoderm: lessons from the mouse, frog, fish, and chick.0 aFormation of the murine endoderm lessons from the mouse frog fis c2010 a1-340 v963 aThe mammalian definitive endoderm arises as a simple epithelial sheet. This sheet of cells will eventually produce the innermost tube that comprises the entire digestive tract from the esophagus to the colon as well as the epithelial component of the digestive and respiratory organs including the thymus, thyroid, lung, liver, gallbladder, and pancreas. Thus a wide array of tissue types are derived from the early endodermal sheet, and understanding the morphological and molecular mechanisms used to produce this tissue is integral to understanding the development of all these organs. The goal of this chapter is to summarize what is known about the morphological and molecular mechanisms used to produce this embryonic germ layer. Although this chapter mainly focuses on the mechanisms used to generate the murine endoderm, supportive or suggestive data from other species, including chick, frog (Xenopus laevis), and the Zebrafish (Danio rerio) are also examined.10aAnimals10aAnura10aChick Embryo10aEndoderm10aFishes10aMice10aMorphogenesis1 aTremblay, Kimberly, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/formation-of-the-murine-endoderm-lessons-from-the-mouse-frog-fish-and05071nas a2201141 4500008004100000245013100041210006900172260001500241300001300256490000800269520171200277653001201989653002302001653002102024653001802045653002202063653001902085653001302104653001102117653002202128653002802150653001402178653002702192653001402219100002202233700001902255700001602274700001902290700002302309700001902332700002002351700002302371700002102394700001702415700002002432700002802452700002402480700001502504700002302519700002102542700001802563700002802581700002402609700001702633700002202650700002102672700002402693700001602717700001902733700001902752700002402771700001702795700001802812700002602830700002002856700001702876700002002893700002102913700002802934700002602962700002502988700002203013700002403035700002203059700002003081700001903101700002503120700002003145700002103165700001803186700002903204700001803233700002203251700002703273700001603300700002203316700002003338700002003358700002003378700001603398700002303414700002103437700002403458700002003482700002003502700002503522700002403547700002303571700001903594700002603613700001803639700002603657700002203683700002203705700002303727700002603750856015303776 2010 eng d00aGenome sequences of the human body louse and its primary endosymbiont provide insights into the permanent parasitic lifestyle.0 aGenome sequences of the human body louse and its primary endosym c2010 Jul 6 a12168-730 v1073 aAs an obligatory parasite of humans, the body louse (Pediculus humanus humanus) is an important vector for human diseases, including epidemic typhus, relapsing fever, and trench fever. Here, we present genome sequences of the body louse and its primary bacterial endosymbiont Candidatus Riesia pediculicola. The body louse has the smallest known insect genome, spanning 108 Mb. Despite its status as an obligate parasite, it retains a remarkably complete basal insect repertoire of 10,773 protein-coding genes and 57 microRNAs. Representing hemimetabolous insects, the genome of the body louse thus provides a reference for studies of holometabolous insects. Compared with other insect genomes, the body louse genome contains significantly fewer genes associated with environmental sensing and response, including odorant and gustatory receptors and detoxifying enzymes. The unique architecture of the 18 minicircular mitochondrial chromosomes of the body louse may be linked to the loss of the gene encoding the mitochondrial single-stranded DNA binding protein. The genome of the obligatory louse endosymbiont Candidatus Riesia pediculicola encodes less than 600 genes on a short, linear chromosome and a circular plasmid. The plasmid harbors a unique arrangement of genes required for the synthesis of pantothenate, an essential vitamin deficient in the louse diet. The human body louse, its primary endosymbiont, and the bacterial pathogens that it vectors all possess genomes reduced in size compared with their free-living close relatives. Thus, the body louse genome project offers unique information and tools to use in advancing understanding of coevolution among vectors, symbionts, and pathogens.10aAnimals10aEnterobacteriaceae10aGenes, Bacterial10aGenes, Insect10aGenome, Bacterial10aGenome, Insect10aGenomics10aHumans10aLice Infestations10aMolecular Sequence Data10aPediculus10aSequence Analysis, DNA10aSymbiosis1 aKirkness, Ewen, F1 aHaas, Brian, J1 aSun, Weilin1 aBraig, Henk, R1 aPerotti, Alejandra1 aClark, John, M1 aLee, Si, Hyeock1 aRobertson, Hugh, M1 aKennedy, Ryan, C1 aElhaik, Eran1 aGerlach, Daniel1 aKriventseva, Evgenia, V1 aElsik, Christine, G1 aGraur, Dan1 aHill, Catherine, A1 aVeenstra, Jan, A1 aWalenz, Brian1 aTubío, José, Manuel C1 aRibeiro, José, M C1 aRozas, Julio1 aJohnston, Spencer1 aReese, Justin, T1 aPopadic, Aleksandar1 aTojo, Marta1 aRaoult, Didier1 aReed, David, L1 aTomoyasu, Yoshinori1 aKraus, Emily1 aKrause, Emily1 aMittapalli, Omprakash1 aMargam, Venu, M1 aLi, Hong-Mei1 aMeyer, Jason, M1 aJohnson, Reed, M1 aRomero-Severson, Jeanne1 aVanzee, Janice, Pagel1 aAlvarez-Ponce, David1 aVieira, Filipe, G1 aAguadé, Montserrat1 aGuirao-Rico, Sara1 aAnzola, Juan, M1 aYoon, Kyong, S1 aStrycharz, Joseph, P1 aUnger, Maria, F1 aChristley, Scott1 aLobo, Neil, F1 aSeufferheld, Manfredo, J1 aWang, Naikuan1 aDasch, Gregory, A1 aStruchiner, Claudio, J1 aMadey, Greg1 aHannick, Linda, I1 aBidwell, Shelby1 aJoardar, Vinita1 aCaler, Elisabet1 aShao, Renfu1 aBarker, Stephen, C1 aCameron, Stephen1 aBruggner, Robert, V1 aRegier, Allison1 aJohnson, Justin1 aViswanathan, Lakshmi1 aUtterback, Terry, R1 aSutton, Granger, G1 aLawson, Daniel1 aWaterhouse, Robert, M1 aVenter, Craig1 aStrausberg, Robert, L1 aBerenbaum, May, R1 aCollins, Frank, H1 aZdobnov, Evgeny, M1 aPittendrigh, Barry, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genome-sequences-of-the-human-body-louse-and-its-primary-endosymbiont01823nas a2200205 4500008004100000245013500041210006900176260001600245520102500261100001801286700002201304700001501326700001801341700002101359700002001380700001901400700002301419700002101442856015401463 2010 ENG d00aIdentification and functional analysis of an ovarian form of the egg activation factor phospholipase C Zeta (PLCζ) in pufferfish.0 aIdentification and functional analysis of an ovarian form of the c2010 Dec 103 aRecent studies suggest that egg activation in mammals is triggered by a sperm-specific phospholipase C, PLCzeta. In other vertebrate species such as medaka fish, chickens, and quail, PLCzeta is also expressed as a testis-specific mRNA. Functional studies suggest that PLCzeta plays a similar role as a trigger of egg activation in these species. Here, we report the identification of PLCzeta orthologues in pufferfish species Takifugu rubripes (Fugu) and Tetraodon nigroviridis (Tetraodon). Unexpectedly in these species PLCzeta is expressed not in the testis, but in ovary and brain. Injection of pufferfish PLCzeta copy ribonucleic acid (cRNA) into mouse eggs failed to trigger calcium oscillations, unlike medaka PLCzeta cRNA. Our findings provide the first evidence that PLCzeta may be expressed in the egg, rather than the sperm, in some vertebrate species, and that its mechanism of action and physiologic role at fertilization may differ in different vertebrate species. Mol. Reprod. Dev. © 2010 Wiley-Liss, Inc.1 aCoward, Kevin1 aPonting, Chris, P1 aZhang, Nan1 aYoung, Claire1 aHuang, Chang-Jen1 aChou, Chih-Ming1 aKashir, Junaid1 aFissore, Rafael, A1 aParrington, John uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-and-functional-analysis-of-an-ovarian-form-of-the-egg-003274nas a2200613 4500008004100000245012100041210006900162260001300231300000900244490000700253520137700260653001001637653000901647653002101656653002701677653002501704653001101729653003801740653001101778653000901789653001601798653003601814653003001850653002801880653001701908100002401925700001901949700001701968700002301985700002902008700002402037700001902061700001902080700002102099700001902120700002202139700001702161700002202178700002202200700001802222700001802240700002502258700002402283700002002307700002802327700002702355700002402382700002102406700001902427700002202446700002102468700002302489856014802512 2010 eng d00aIdentification of a DMBT1 polymorphism associated with increased breast cancer risk and decreased promoter activity.0 aIdentification of a DMBT1 polymorphism associated with increased c2010 Jan a60-60 v313 aAccording to present estimations, the unfavorable combination of alleles with low penetrance but high prevalence in the population might account for the major part of hereditary breast cancer risk. Deleted in Malignant Brain Tumors 1 (DMBT1) has been proposed as a tumor suppressor for breast cancer and other cancer types. Genomewide mapping in mice further identified Dmbt1 as a potential modulator of breast cancer risk. Here, we report the association of two frequent and linked single-nucleotide polymorphisms (SNPs) with increased breast cancer risk in women above the age of 60 years: DMBT1 c.-93C>T, rs2981745, located in the DMBT1 promoter; and DMBT1 c.124A>C, p.Thr42Pro, rs11523871(odds ratio [OR]=1.66, 95% confidence interval [CI]=1.21-2.29, P=0.0017; and OR=1.66; 95% CI=1.21-2.28, P=0.0016, respectively), based on 1,195 BRCA1/2 mutation-negative German breast cancer families and 1,466 unrelated German controls. Promoter studies in breast cancer cells demonstrate that the risk-increasing DMBT1 -93T allele displays significantly decreased promoter activity compared to the DMBT1 -93C allele, resulting in a loss of promoter activity. The data suggest that DMBT1 polymorphisms in the 5'-region are associated with increased breast cancer risk. In accordance with previous results, these data link decreased DMBT1 levels to breast cancer risk.
10aAdult10aAged10aBreast Neoplasms10aBreast Neoplasms, Male10aCase-Control Studies10aFemale10aGenetic Predisposition to Disease10aHumans10aMale10aMiddle Aged10aPolymorphism, Single Nucleotide10aPromoter Regions, Genetic10aReceptors, Cell Surface10aRisk Factors1 aTchatchou, Sandrine1 aRiedel, Angela1 aLyer, Stefan1 aSchmutzhard, Julia1 aStrobel-Freidekind, Olga1 aGronert-Sum, Sabine1 aMietag, Carola1 aD'Amato, Mauro1 aSchlehe, Bettina1 aHemminki, Kari1 aSutter, Christian1 aDitsch, Nina1 aBlackburn, Anneke1 aHill, Linda, Zhai1 aJerry, Joseph1 aBugert, Peter1 aWeber, Bernhard, H F1 aNiederacher, Dieter1 aArnold, Norbert1 aVaron-Mateeva, Raymonda1 aWappenschmidt, Barbara1 aSchmutzler, Rita, K1 aEngel, Christoph1 aMeindl, Alfons1 aBartram, Claus, R1 aMollenhauer, Jan1 aBurwinkel, Barbara uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-a-dmbt1-polymorphism-associated-with-increased03201nas a2200169 4500008004100000245017000041210006900211260001600280300000900296490000800305520247800313100002002791700002402811700002202835700002102857856015302878 2010 eng d00aIn vivo priming and ex vivo activation of equine neutrophils in black walnut extract-induced equine laminitis is not attenuated by systemic lidocaine administration.0 aIn vivo priming and ex vivo activation of equine neutrophils in c2010 Nov 15 a60-90 v1383 aLaminitis is a crippling disease of horses characterized by an inflammatory response in the tissue that suspends the axial skeleton within the hoof. Pain is a common feature of laminitic pathology and its management is an important component of the treatment regime for this disease. Systemic lidocaine administration is commonly utilized to manage pain in equine laminitis; however, the potential anti-inflammatory effects of this drug during the treatment of equine laminitis have not been investigated. Here, we sought to determine if lidocaine concentrations achieved in the plasma (therapeutic concentrations) of horses systemically administered lidocaine are capable of attenuating neutrophil activation and associated inflammation. To identify markers of activation, purified neutrophils were stimulated in vitro with LPS or recombinant equine IL-8 (reqIL-8) and surface expression of CD13 and CD18 was ascertained by immunofluorescent staining. Activation with LPS or reqIL-8 in vitro induced an elevated expression of CD13 as well as a putative conformational change in CD18 detected by elevated staining with a sub-saturating concentration of anti-CD18 mAb. Lidocaine attenuated the activation-induced changes in CD13 and CD18 expression only when used at 30-70 times therapeutic concentrations. For in vivo analyses, horses were administered black walnut extract (BWE) to induce laminitis and either systemic lidocaine (n=6) or saline (n=6) as a control. Whole blood was collected and incubated with or without reqIL-8. Following which, leukocytes were stained for CD13 and CD18. Protein was extracted from laminar tissue and subjected to gelatin zymography to measure matrix metalloproteinase-9 (MMP-9) accumulation. Results obtained show that changes in neutrophil size, granularity/complexity, CD13 surface expression and CD18 staining intensity occurred over time post BWE administration irrespective of lidocaine treatment in response to incubation alone or with 100 ng/ml of reqIL-8. The mean fluorescence intensities of neutrophils stained for either CD13 or CD18 did not differ between lidocaine treated and saline controls, nor did lamellar MMP-9 content measured by gelatin zymography. Thus, using changes in surface expression of CD13 and CD18 as markers of neutrophil activation in the horse we have shown that BWE treatment activates neutrophils in vivo and this is not affected by systemic administration of lidocaine at levels used to manage pain.1 aLoftus, John, P1 aWilliams, Jarred, M1 aBelknap, James, K1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/in-vivo-priming-and-ex-vivo-activation-of-equine-neutrophils-in-black01471nas a2200181 4500008004100000245008400041210006900125260001300194300001100207490000800218520083300226100001601059700001701075700001501092700001401107700001601121856015201137 2010 eng d00aInduction and regulation of Trypanosoma brucei VSG-specific antibody responses.0 aInduction and regulation of Trypanosoma brucei VSGspecific antib c2010 Dec a2041-90 v1373 aThe review addresses how infection with Trypanosoma brucei affects the development, survival and functions of B lymphocytes in mice. It discusses (1) the contributions of antibodies to trypanosome clearance from the bloodstream, (2) how B lymphocytes, the precursors of antibody producing plasma cells, interact with membrane form variable surface glycoprotein (VSG), i.e. with monovalent antigen that is free to diffuse within the lipid bilayer of the trypanosome plasma membrane and consequently can cross-link B cell antigen specific receptors by indirect processes only and (3) the extent and underlying causes of dysregulation of humoral immune responses in infected mice, focusing on the impact of wild type and GPI-PLC⁻/⁻ trypanosomes on bone marrow and extramedullary B lymphopoiesis, B cell maturation and survival.1 aBlack, S, J1 aGuirnalda, P1 aFrenkel, D1 aHaynes, C1 aBockstal, V uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/induction-and-regulation-of-trypanosoma-brucei-vsg-specific-antibody03001nas a2200493 4500008004100000245012300041210006900164260001600233300001200249490000800261520148000269653002201749653001201771653001501783653004101798653002201839653001101861653001201872653000901884653000901893653002501902653002501927653001301952653001701965653001301982653002001995653003702015653001502052653002402067653002302091653002802114653001602142653002302158653001702181653001302198100001702211700001802228700002302246700002002269700002302289700002202312700002302334856015002357 2010 eng d00aInhibition of Ser/Thr phosphatases induces capacitation-associated signaling in the presence of Src kinase inhibitors.0 aInhibition of SerThr phosphatases induces capacitationassociated c2010 Mar 12 a7977-850 v2853 aSignaling events leading to mammalian sperm capacitation rely on activation/deactivation of proteins by phosphorylation. This cascade includes soluble adenylyl cyclase, an atypical bicarbonate-stimulated adenylyl cyclase, and is mediated by protein kinase A and the subsequent stimulation of protein tyrosine phosphorylation. Recently, it has been proposed that the capacitation-associated increase in tyrosine phosphorylation is governed by Src tyrosine kinase activity. This conclusion was based mostly on the observation that Src is present in sperm and that the Src kinase family inhibitor SU6656 blocked the capacitation-associated increase in tyrosine phosphorylation. Results in the present manuscript confirmed these observations and provided evidence that these inhibitors were also able to inhibit protein kinase A phosphorylation, sperm motility, and in vitro fertilization. However, the block of capacitation-associated parameters was overcome when sperm were incubated in the presence of Ser/Thr phosphatase inhibitors such as okadaic acid and calyculin-A at concentrations reported to affect only PP2A. Altogether, these data indicate that Src is not directly involved in the observed increase in tyrosine phosphorylation. More importantly, this work presents strong evidence that capacitation is regulated by two parallel pathways. One of them requiring activation of protein kinase A and the second one involving inactivation of Ser/Thr phosphatases.
10aAniline Compounds10aAnimals10aCyclic AMP10aCyclic AMP-Dependent Protein Kinases10aEnzyme Inhibitors10aFemale10aIndoles10aMale10aMice10aMice, Inbred Strains10aMice, Mutant Strains10aNitriles10aOkadaic Acid10aOxazoles10aPhosphorylation10aProtein-Serine-Threonine Kinases10aQuinolines10aSignal Transduction10aSperm Capacitation10aSperm-Ovum Interactions10aSpermatozoa10asrc-Family Kinases10aSulfonamides10aTyrosine1 aKrapf, Dario1 aArcelay, Enid1 aWertheimer, Eva, V1 aSanjay, Archana1 aPilder, Stephen, H1 aSalicioni, Ana, M1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/inhibition-of-ser-thr-phosphatases-induces-capacitation-associated02763nas a2200349 4500008004100000245010600041210006900147260001300216300001100229490000800240520164200248653001401890653001201904653002201916653002001938653001101958653001801969653001101987653001501998653003302013653004302046653000902089653000902098653004302107653001702150100001402167700002102181700001902202700001802221700002302239856015102262 2010 eng d00aInositol 1,4,5-trisphosphate receptor 1 degradation in mouse eggs and impact on [Ca2+]i oscillations.0 aInositol 145trisphosphate receptor 1 degradation in mouse eggs a c2010 Jan a238-470 v2223 aThe initiation of normal embryo development depends on the completion of all events of egg activation. In all species to date, egg activation requires an increase(s) in the intracellular concentration of calcium ([Ca(2+)](i)), which is almost entirely mediated by inositol 1,4,5-trisphosphate receptor 1 (IP(3)R1). In mammalian eggs, fertilization-induced [Ca(2+)](i) responses exhibit a periodic pattern that are called [Ca(2+)](i) oscillations. These [Ca(2+)](i) oscillations are robust at the beginning of fertilization, which occurs at the second metaphase of meiosis, but wane as zygotes approach the pronuclear stage, time after which in the mouse oscillations cease altogether. Underlying this change in frequency are cellular and biochemical changes associated with egg activation, including degradation of IP(3)R1, progression through the cell cycle, and reorganization of intracellular organelles. In this study, we investigated the system requirements for IP(3)R1 degradation and examined the impact of the IP(3)R1 levels on the pattern of [Ca(2+)](i) oscillations. Using microinjection of IP(3) and of its analogs and conditions that prevent the development of [Ca(2+)](i) oscillations, we show that IP(3)R1 degradation requires uniform and persistently elevated levels of IP(3). We also established that progressive degradation of the IP(3)R1 results in [Ca(2+)](i) oscillations with diminished periodicity while a near complete depletion of IP(3)R1s precludes the initiation of [Ca(2+)](i) oscillations. These results provide insights into the mechanism involved in the generation of [Ca(2+)](i) oscillations in mouse eggs.10aAdenosine10aAnimals10aCalcium Signaling10aDown-Regulation10aFemale10aFertilization10aHumans10aInjections10aInositol 1,4,5-Trisphosphate10aInositol 1,4,5-Trisphosphate Receptors10aMice10aOvum10aProtein Processing, Post-Translational10aTime Factors1 aLee, Bora1 aYoon, Sook-Young1 aMalcuit, Chris1 aParys, Jan, B1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/inositol-145-trisphosphate-receptor-1-degradation-in-mouse-eggs-and02777nas a2200205 4500008004100000245004500041210004400086260000900130520215400139653001202293653001502305653001102320653002702331653000902358653002702367100002102394700001502415700001802430856012302448 2010 eng d00aMammary epithelial transplant procedure.0 aMammary epithelial transplant procedure c20103 aThis article describes and compares the fat pad clearance procedure developed by DeOme KB et al. and the sparing procedure developed by Brill B et al., followed by the mammary epithelial transplant procedure. The mammary transplant procedure is widely used by mammary biologists because it takes advantage of the fact that significant development of the mammary epithelium doesn't occur until after puberty. At 3 weeks of age, growth of the mammary epithelial tree is confined to the vicinity of the nipple and the fat pad is largely devoid of mammary epithelium, but by 7 weeks of age the epithelial ductal tree extends throughout the entire fat pad. Therefore, if this small portion of the fat pad containing epithelium, the region between the nipple and the lymph node, is removed at 3 weeks of age, the endogenous epithelium will never populate the mammary fat pad and the fat pad is described as "cleared". At this time, mammary epithelium from another source can be transplanted in the cleared fat pad where it has the potential to extend mammary ductal trees through out the fat pad. This procedure has been utilized in many experimental models including the examination of tumor phenotype in transgenic mammary epithelial tissue without the confounding effects of genotype on the entire animal, in the identification of mammary stem cells by transplanting cells in limited dilution, determining if hyperplastic nodules proceed to mammary tumors, and to assess the effect of prior hormone exposure on the behavior of the mammary epithelium. Three week old host mice are anesthetized, cleaned and restrained on a surgical stage. A mid-sagittal incision is made through the skin, but not the peritoneum, extending from the pubis to the sternum. Oblique cuts are made through the skin from the mid-sagittal incision across the pelvis toward each leg. The skin is pulled away from the peritoneum to expose the 4th inguinal mammary gland. The fat pad is cleared by removing the fat pad tissue anterior to the lymph node. Epithelium fragments or epithelial cells are transplanted into the remaining cleared fat pad and the mouse is closed.
10aAnimals10aEpithelium10aFemale10aMammary Glands, Animal10aMice10aTissue Transplantation1 aDunphy, Karen, A1 aTao, Luwei1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mammary-epithelial-transplant-procedure01480nas a2200313 4500008004100000245006200041210006100103260001700164300001100181490000600192520046200198653001200660653001800672653001800690653002700708653004600735653000900781653005000790653002200840653002300862653001700885653002400902653001700926653001900943100002400962700002100986700001901007856014001026 2010 eng d00aMechanism of Xenopus cranial neural crest cell migration.0 aMechanism of Xenopus cranial neural crest cell migration c2010 Oct-Dec a553-600 v43 aThis review focuses on recent advances in the field of cranial neural crest cell migration in Xenopus laevis with specific emphasis on cell adhesion and the regulation of cell migration. Our goal is to combine the understanding of cell adhesion to the extracellular matrix with the regulation of cell-cell adhesion and the involvement of the planar cell polarity signaling-pathway in guiding the migration of cranial neural crest cells during embryogenesis.10aAnimals10aCell Adhesion10aCell Movement10aCentral Nervous System10aGene Expression Regulation, Developmental10aHead10aIntracellular Signaling Peptides and Proteins10aMembrane Proteins10aModels, Biological10aNeural Crest10aSignal Transduction10aWnt Proteins10aXenopus laevis1 aAlfandari, Dominque1 aCousin, Hélène1 aMarsden, Mungo uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mechanism-of-xenopus-cranial-neural-crest-cell-migration00885nas a2200145 4500008004100000245007500041210006900116260000900185300000900194490000600203520033200209100002300541700002300564856015200587 2010 eng d00aMechanisms of sperm-egg interactions: between sugars and broken bonds.0 aMechanisms of spermegg interactions between sugars and broken bo c2010 ape350 v33 aA model of the early events of mammalian fertilization has emerged during the past 30 years. However, studies during the past decade have used newly available mouse models to readdress these processes. Here, we will consider these new data in light of the existing model and point to areas of reconciliation and of controversy.1 aVisconti, Pablo, E1 aFlorman, Harvey, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mechanisms-of-sperm-egg-interactions-between-sugars-and-broken-bonds03267nas a2200517 4500008004100000245009600041210006900137260001300206300001200219490000800231520160400239653001201843653001101855653004301866653001301909653002701922653003001949653003601979653000902015653002402024653002902048653002802077653002102105653002402126653002102150653002802171653002402199653003302223100001702256700002502273700002202298700001502320700002002335700002502355700002402380700001902404700002302423700002502446700001702471700002302488700002402511700002502535700001902560700001802579856015202597 2010 eng d00aPathways contributing to development of spontaneous mammary tumors in BALB/c-Trp53+/- mice.0 aPathways contributing to development of spontaneous mammary tumo c2010 Mar a1421-320 v1763 aMutation and loss of function in p53 are common features among human breast cancers. Here we use BALB/c-Trp53+/- mice as a model to examine the sequence of events leading to mammary tumors. Mammary gland proliferation rates were similar in both BALB/c-Trp53+/- mice and wild-type controls. In addition, sporadic mammary hyperplasias were rare in BALB/c-Trp53+/- mice and not detectably different from those of wild-type controls. Among the 28 mammary tumors collected from BALB/c-Trp53+/- mice, loss of heterozygosity for Trp53 was detected in more than 90% of invasive mammary tumors. Transplantation of Trp53+/- ductal hyperplasias also indicated an association between loss of the wild-type allele of Trp53 and progression to invasive carcinomas. Therefore, loss of p53 function seems to be a rate-limiting step in progression. Moreover, expression of biomarkers such as estrogen receptor alpha, progesterone receptor, Her2/Neu, and activated Notch1 varied among mammary tumors, suggesting that multiple oncogenic lesions collaborate with loss of p53 function. Expression of biomarkers was retained when tumor fragments were transplanted to syngeneic hosts. Tumors expressing solely luminal or basal keratins were also observed (27 and 11%, respectively), but the largest class of tumors expressed both luminal and basal keratins (62%). Overall, this panel of transplantable tumors provides a resource for detailed evaluation of the cell lineages undergoing transformation and preclinical testing of therapeutic agents targeting a variety of oncogenic pathways including cancer stem cells.
10aAnimals10aFemale10aGene Expression Regulation, Neoplastic10aKeratins10aLoss of Heterozygosity10aMammary Neoplasms, Animal10aMammary Neoplasms, Experimental10aMice10aMice, Inbred BALB C10aNeoplasm Transplantation10aPrecancerous Conditions10aReceptor, erbB-210aReceptors, Estrogen10aReceptors, Notch10aReceptors, Progesterone10aSignal Transduction10aTumor Suppressor Protein p531 aYan, Haoheng1 aBlackburn, Anneke, C1 aMcLary, Christine1 aTao, Luwei1 aRoberts, Amy, L1 aXavier, Elizabeth, A1 aDickinson, Ellen, S1 aSeo, Jae, Hong1 aArenas, Richard, B1 aOtis, Christopher, N1 aCao, Qing, J1 aLawlor, Rebecca, G1 aOsborne, Barbara, A1 aKittrell, Frances, S1 aMedina, Daniel1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pathways-contributing-to-development-of-spontaneous-mammary-tumors-002644nas a2200313 4500008004100000245009600041210006900137260001300206300001200219490000800231520159700239100001701836700002501853700002201878700001501900700002001915700002501935700002401960700001901984700002302003700002502026700001702051700002302068700002402091700002502115700001902140700001802159856015302177 2010 eng d00aPathways contributing to development of spontaneous mammary tumors in BALB/c-Trp53+/- mice.0 aPathways contributing to development of spontaneous mammary tumo c2010 Mar a1421-320 v1763 aMutation and loss of function in p53 are common features among human breast cancers. Here we use BALB/c-Trp53+/- mice as a model to examine the sequence of events leading to mammary tumors. Mammary gland proliferation rates were similar in both BALB/c-Trp53+/- mice and wild-type controls. In addition, sporadic mammary hyperplasias were rare in BALB/c-Trp53+/- mice and not detectably different from those of wild-type controls. Among the 28 mammary tumors collected from BALB/c-Trp53+/- mice, loss of heterozygosity for Trp53 was detected in more than 90% of invasive mammary tumors. Transplantation of Trp53+/- ductal hyperplasias also indicated an association between loss of the wild-type allele of Trp53 and progression to invasive carcinomas. Therefore, loss of p53 function seems to be a rate-limiting step in progression. Moreover, expression of biomarkers such as estrogen receptor alpha, progesterone receptor, Her2/Neu, and activated Notch1 varied among mammary tumors, suggesting that multiple oncogenic lesions collaborate with loss of p53 function. Expression of biomarkers was retained when tumor fragments were transplanted to syngeneic hosts. Tumors expressing solely luminal or basal keratins were also observed (27 and 11%, respectively), but the largest class of tumors expressed both luminal and basal keratins (62%). Overall, this panel of transplantable tumors provides a resource for detailed evaluation of the cell lineages undergoing transformation and preclinical testing of therapeutic agents targeting a variety of oncogenic pathways including cancer stem cells.1 aYan, Haoheng1 aBlackburn, Anneke, C1 aMcLary, Christine1 aTao, Luwei1 aRoberts, Amy, L1 aXavier, Elizabeth, A1 aDickinson, Ellen, S1 aSeo, Jae, Hong1 aArenas, Richard, B1 aOtis, Christopher, N1 aCao, Qing, J1 aLawlor, Rebecca, G1 aOsborne, Barbara, A1 aKittrell, Frances, S1 aMedina, Daniel1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pathways-contributing-to-development-of-spontaneous-mammary-tumors-in02684nas a2200373 4500008004100000245010900041210006900150260001300219300001000232490000700242520150500249653001201754653002201766653002401788653002001812653001301832653001101845653004301856653001201899653000901911653003801920653001201958653002001970653002001990653001502010100001502025700002002040700001902060700001802079700001802097700002302115700002302138856014902161 2010 eng d00aPhosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes.0 aPhosphorylation of inositol 145triphosphate receptor 1 during in c2010 Feb a34-410 v813 aDuring fertilization in mammalian species, a sperm-induced intracellular Ca(2+) signal ([Ca(2+)](i)) mediates both exit of meiosis and oocyte activation. Recently, we demonstrated in mouse oocytes that the phosphorylation levels of inositol 1,4,5 trisphosphate receptor type1 (IP(3)R1), the channel responsible for Ca(2+) release and oscillations during fertilization, changed during maturation and fertilization. Therefore, we examined the expression and phosphorylation of IP(3)R1 during in vitro maturation of pig oocytes. Here, our present study shows that expression of IP(3)R1 protein did not change during maturation, although the phosphorylation status of the receptor, specifically at an MPM-2 epitope, did. We found that while at the beginning of maturation IP(3)R1 lacked MPM-2 immunoreactivity, it became MPM-2 reactive by 24 h and reached maximal reactivity by 36 h. Interestingly, the acquisition of MPM-2 reactivity coincided with the activation of p34(cdc2) kinase and mitogen-activated protein kinase (MAPK), which are involved in meiotic progression. Following completion of maturation, inactivation of MAPK by U0126 did not affect IP(3)R1 phosphorylation, although inactivation of p34(cdc2) kinase by roscovitine dramatically reduced IP(3)R1 phosphorylation. Neither inhibitor affected total expression of IP(3)R1. Altogether, our results show that IP(3)R1 undergoes dynamic phosphorylation during maturation and this might underlie the generation of oscillations at fertilization.10aAnimals10aCalcium Signaling10aCDC2 Protein Kinase10aCells, Cultured10aEpitopes10aFemale10aInositol 1,4,5-Trisphosphate Receptors10aMeiosis10aMice10aMitogen-Activated Protein Kinases10aOocytes10aPhosphoproteins10aPhosphorylation10aSus scrofa1 aIto, Junya1 aYoshida, Tomoko1 aKasai, Yasushi1 aWakai, Takuya1 aParys, Jan, B1 aFissore, Rafael, A1 aKashiwazaki, Naomi uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/phosphorylation-of-inositol-145-triphosphate-receptor-1-during-in02743nas a2200253 4500008004100000245010400041210006900145260001300214300001200227490000700239520183400246100002502080700003002105700002502135700002302160700002502183700001402208700002202222700002402244700002102268700002402289700002402313856015202337 2010 eng d00aPotent neutralization of staphylococcal enterotoxin B by synergistic action of chimeric antibodies.0 aPotent neutralization of staphylococcal enterotoxin B by synergi c2010 Jun a2801-110 v783 aStaphylococcal enterotoxin B (SEB), a shock-inducing exotoxin synthesized by Staphylococcus aureus, is an important cause of food poisoning and is a class B bioterrorism agent. SEB mediates antigen-independent activation of a major subset of the T-cell population by cross-linking T-cell receptors (TCRs) with class II major histocompatibility complex (MHC-II) molecules of antigen-presenting cells, resulting in the induction of antigen independent proliferation and cytokine secretion by a significant fraction of the T-cell population. Neutralizing antibodies inhibit SEB-mediated T-cell activation by blocking the toxin's interaction with the TCR or MHC-II and provide protection against the debilitating effects of this superantigen. We derived and searched a set of monoclonal mouse anti-SEB antibodies to identify neutralizing anti-SEB antibodies that bind to different sites on the toxin. A pair of non-cross-reactive, neutralizing anti-SEB monoclonal antibodies (MAbs) was found, and a combination of these antibodies inhibited SEB-induced T-cell proliferation in a synergistic rather than merely additive manner. In order to engineer antibodies more suitable than mouse MAbs for use in humans, the genes encoding the VL and VH gene segments of a synergistically acting pair of mouse MAbs were grafted, respectively, onto genes encoding the constant regions of human Igkappa and human IgG1, transfected into mammalian cells, and used to generate chimeric versions of these antibodies that had affinity and neutralization profiles essentially identical to their mouse counterparts. When tested in cultures of human peripheral blood mononuclear cells or splenocytes derived from HLA-DR3 transgenic mice, the chimeric human-mouse antibodies synergistically neutralized SEB-induced T-cell activation and cytokine production.1 aTilahun, Mulualem, E1 aRajagopalan, Govindarajan1 aShah-Mahoney, Nalini1 aLawlor, Rebecca, G1 aTilahun, Ashenafi, Y1 aXie, Chen1 aNatarajan, Kannan1 aMargulies, David, H1 aRatner, David, I1 aOsborne, Barbara, A1 aGoldsby, Richard, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/potent-neutralization-of-staphylococcal-enterotoxin-b-by-synergistic03141nas a2200469 4500008004100000245012500041210006900166260001500235300001100250490000600261520163300267653001001900653001601910653001101926653002101937653001401958653001601972653002601988653002002014653002102034653001102055653002802066653003302094653001102127653005002138653001402188653002202202653001602224653001602240653003002256653003902286653001702325653003002342653001602372100001902388700002502407700002802432700001902460700002302479700002402502856014502526 2010 eng d00aQuantitative analysis of promoter methylation in exfoliated epithelial cells isolated from breast milk of healthy women.0 aQuantitative analysis of promoter methylation in exfoliated epit c2010 Oct 1 a645-550 v53 aPromoter methylation analysis of genes frequently silenced in breast cancer is a promising indicator of breast cancer risk, as these methylation events are thought to occur long before presentation of disease. The numerous exfoliated epithelial cells present in breast milk may provide the breast epithelial DNA needed for detailed methylation analysis and assessment of breast cancer risk. Fresh breast milk samples and health, lifestyle, and reproductive history questionnaires were collected from 111 women. Pyrosequencing analysis was conducted on DNA isolated from the exfoliated epithelial cells immunomagnetically separated from the total cell population in the breast milk of 102 women. A total of 65 CpG sites were examined in six tumor suppressor genes: PYCARD (also known as ASC or TMS1), CDH1, GSTP1, RBP1 (also known as CRBP1), SFRP1, and RASSF1. A sufficient quantity of DNA was obtained for meaningful analysis of promoter methylation; women donated an average of 86 ml of milk with a mean yield of 32,700 epithelial cells per ml. Methylation scores were in general low as expected of benign tissue, but analysis of outlier methylation scores revealed a significant relationship between breast cancer risk, as indicated by previous biopsy, and methylation score for several CpG sites in CDH1, GSTP1, SFRP1, and RBP1. Methylation of RASSF1 was positively correlated with women's age irrespective of her reproductive history. Promoter methylation patterns in DNA from breast milk epithelial cells can likely be used to assess breast cancer risk. Additional studies of women at high breast cancer risk are warranted.10aAdult10aAge Factors10aBreast10aBreast Neoplasms10aCadherins10aCpG Islands10aCytoskeletal Proteins10aDNA Methylation10aEpithelial Cells10aFemale10aGenes, Tumor Suppressor10aGlutathione S-Transferase pi10aHumans10aIntercellular Signaling Peptides and Proteins10aLactation10aMembrane Proteins10aMiddle Aged10aMilk, Human10aPromoter Regions, Genetic10aRetinol-Binding Proteins, Cellular10aRisk Factors10aTumor Suppressor Proteins10aYoung Adult1 aWong, Chung, M1 aAnderton, Douglas, L1 aSmith-Schneider, Sallie1 aWing, Megan, A1 aGreven, Melissa, C1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/quantitative-analysis-of-promoter-methylation-in-exfoliated-002386nas a2200193 4500008004100000245012500041210006900166260001500235300001100250490000600261520163300267100001901900700002501919700002801944700001901972700002301991700002402014856015402038 2010 eng d00aQuantitative analysis of promoter methylation in exfoliated epithelial cells isolated from breast milk of healthy women.0 aQuantitative analysis of promoter methylation in exfoliated epit c2010 Oct 1 a645-550 v53 aPromoter methylation analysis of genes frequently silenced in breast cancer is a promising indicator of breast cancer risk, as these methylation events are thought to occur long before presentation of disease. The numerous exfoliated epithelial cells present in breast milk may provide the breast epithelial DNA needed for detailed methylation analysis and assessment of breast cancer risk. Fresh breast milk samples and health, lifestyle, and reproductive history questionnaires were collected from 111 women. Pyrosequencing analysis was conducted on DNA isolated from the exfoliated epithelial cells immunomagnetically separated from the total cell population in the breast milk of 102 women. A total of 65 CpG sites were examined in six tumor suppressor genes: PYCARD (also known as ASC or TMS1), CDH1, GSTP1, RBP1 (also known as CRBP1), SFRP1, and RASSF1. A sufficient quantity of DNA was obtained for meaningful analysis of promoter methylation; women donated an average of 86 ml of milk with a mean yield of 32,700 epithelial cells per ml. Methylation scores were in general low as expected of benign tissue, but analysis of outlier methylation scores revealed a significant relationship between breast cancer risk, as indicated by previous biopsy, and methylation score for several CpG sites in CDH1, GSTP1, SFRP1, and RBP1. Methylation of RASSF1 was positively correlated with women's age irrespective of her reproductive history. Promoter methylation patterns in DNA from breast milk epithelial cells can likely be used to assess breast cancer risk. Additional studies of women at high breast cancer risk are warranted.1 aWong, Chung, M1 aAnderton, Douglas, L1 aSmith-Schneider, Sallie1 aWing, Megan, A1 aGreven, Melissa, C1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/quantitative-analysis-of-promoter-methylation-in-exfoliated-epithelial02145nas a2200337 4500008004100000245006700041210006500108260000900173300001100182490000600193520105400199653001201253653002101265653002701286653001101313653002701324653000901351653002401360653003901384653003901423653002201462653003301484100002401517700002201541700001901563700002201582700001801604700002001622700002001642856014501662 2010 eng d00aRole of JNK in a Trp53-dependent mouse model of breast cancer.0 aRole of JNK in a Trp53dependent mouse model of breast cancer c2010 ae124690 v53 aThe cJun NH2-terminal kinase (JNK) signal transduction pathway has been implicated in mammary carcinogenesis. To test the role of JNK, we examined the effect of ablation of the Jnk1 and Jnk2 genes in a Trp53-dependent model of breast cancer using BALB/c mice. We detected no defects in mammary gland development in virgin mice or during lactation and involution in control studies of Jnk1(-/-) and Jnk2(-/-) mice. In a Trp53(-/+) genetic background, mammary carcinomas were detected in 43% of control mice, 70% of Jnk1(-/-) mice, and 53% of Jnk2(-/-) mice. These data indicate that JNK1 and JNK2 are not essential for mammary carcinoma development in the Trp53(-/+) BALB/c model of breast cancer. In contrast, this analysis suggests that JNK may partially contribute to tumor suppression. This conclusion is consistent with the finding that tumor-free survival of JNK-deficient Trp53(-/+) mice was significantly reduced compared with control Trp53(-/+) mice. We conclude that JNK1 and JNK2 can act as suppressors of mammary tumor development.
10aAnimals10aBreast Neoplasms10aDisease Models, Animal10aFemale10aMammary Glands, Animal10aMice10aMice, Inbred BALB C10aMitogen-Activated Protein Kinase 810aMitogen-Activated Protein Kinase 910aSurvival Analysis10aTumor Suppressor Protein p531 aCellurale, Cristina1 aWeston, Claire, R1 aReilly, Judith1 aGarlick, David, S1 aJerry, Joseph1 aSluss, Hayla, K1 aDavis, Roger, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/role-of-jnk-in-a-trp53-dependent-mouse-model-of-breast-cancer00805nas a2200205 4500008004100000245012300041210006900164260001600233300001100249490000600260100002300266700002100289700002100310700002300331700002200354700002100376700002400397700002400421856015400445 2010 eng d00aThe role of surface functionality on acute cytotoxicity, ROS generation and DNA damage by cationic gold nanoparticles.0 arole of surface functionality on acute cytotoxicity ROS generati c2010 Oct 18 a2246-90 v61 aChompoosor, Apiwat1 aSaha, Krishnendu1 aGhosh, Partha, S1 aMacarthy, Dylan, J1 aMiranda, Oscar, R1 aZhu, Zheng-Jiang1 aArcaro, Kathleen, F1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-role-of-surface-functionality-on-acute-cytotoxicity-ros-generation03106nas a2200361 4500008004100000245011900041210006900160260001300229300001100242490000700253520191300260653001202173653002202185653001102207653002002218653004302238653000902281653000902290653002002299653001202319653003702331653002502368653002302393653002402416653001602440100002302456700002502479700001902504700002402523700002302547700002102570856015302591 2010 eng d00aSpecies-specific differences in the activity and nuclear localization of murine and bovine phospholipase C zeta 1.0 aSpeciesspecific differences in the activity and nuclear localiza c2010 Jul a92-1010 v833 aInjection of mammalian sperm extracts or cRNA of the sperm-specific phospholipase C zeta 1 (PLCZ1) has been shown to trigger repetitive oscillations in the concentration of free calcium ([Ca(2+)](i)), leading to oocyte activation and embryo development in all mammals studied to date. While PLCZ1 has cross-species activity, it has also been observed that species-specific differences may exist in the frequency and pattern of the resulting [Ca(2+)](i) oscillations following PLCZ1 cRNA injection into oocytes of different species. Accordingly, we used a crossover design strategy to directly investigate the activity of murine and bovine PLCZ1 in both murine and bovine oocytes. In murine oocytes, injection of murine Plcz1 cRNA induced [Ca(2+)](i) oscillations at 10-fold lower concentrations than bovine PLCZ1, although in bovine oocytes bovine PLCZ1 was more effective than murine Plcz1 at inducing [Ca(2+)](i) oscillations. Investigation of ITPR1 (IP(3)R1) down-regulation in bovine oocytes by PLCZ1 cRNA also showed that bovine PLCZ1 was more active in homologous oocytes. To determine whether these PLCZs exhibited similar cellular distribution, Venus-tagged PLCZ1 cRNA was injected into oocytes, and PLCZ1 was overexpressed. Bovine PLCZ1 failed to accumulate in the pronucleus (PN) of bovine or murine zygotes, despite possessing a putative nuclear localization signal. Conversely, murine PLCZ1 accumulated in the PN of both murine and bovine zygotes. These results demonstrate that murine PLCZ1 and bovine PLCZ1 possess species-specific differences in activity and suggest potential differences in the mode of action of the protein between the two species. Variation in sperm PLCZ1 protein content among species, along with oocyte-specific differences in the localization and availability of PLCZ1 substrates, may further contribute to optimize the activation stimulus to enhance embryo development.10aAnimals10aCalcium Signaling10aCattle10aDown-Regulation10aInositol 1,4,5-Trisphosphate Receptors10aMale10aMice10aMicroinjections10aOocytes10aPhosphoinositide Phospholipase C10aRecombinant Proteins10aRNA, Complementary10aSpecies Specificity10aSpermatozoa1 aCooney, Melissa, A1 aMalcuit, Christopher1 aCheon, Banyoon1 aHolland, Michael, K1 aFissore, Rafael, A1 aD'Cruz, Nancy, T uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/species-specific-differences-in-the-activity-and-nuclear-localization00812nas a2200217 4500008004100000245010300041210006900144260001600213300001100229490000600240100002100246700002000267700002400287700001200311700002200323700002500345700002400370700002400394700002300418856015300441 2010 eng d00aSurface properties dictate uptake, distribution, excretion, and toxicity of nanoparticles in fish.0 aSurface properties dictate uptake distribution excretion and tox c2010 Oct 18 a2261-50 v61 aZhu, Zheng-Jiang1 aCarboni, Rachel1 aQuercio, Michael, J1 aYan, Bo1 aMiranda, Oscar, R1 aAnderton, Douglas, L1 aArcaro, Kathleen, F1 aRotello, Vincent, M1 aVachet, Richard, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/surface-properties-dictate-uptake-distribution-excretion-and-toxicity02436nas a2200325 4500008004100000245014700041210006900188260001300257300001100270490000800281520132900289653002401618653001201642653001801654653001701672653001101689653002501700653002801725653002101753653001201774653002201786653002701808653001701835653001901852100002101871700002201892700002301914700002201937856015101959 2010 eng d00aVitellogenesis in Bufo arenarum: identification, characterization and immunolocalization of high molecular mass lipovitellin during oogenesis.0 aVitellogenesis in Bufo arenarum identification characterization c2010 Mar a256-650 v1553 aVitellogenin (Vtg), a large lipoglycophosphoprotein, is the most important precursor of the yolk proteins, and the major source of nutrients for the developing embryo in oviparous species. After its uptake by the oocytes, Vtg is converted into lipovitellins (high and light) and phosvitin, which are deposited into crystalline yolk platelets. We describe here the presence of two high molecular mass lipovitellin isoforms in Bufo arenarum mature oocytes with masses of 113 and 100 kDa, respectively. The amino acid sequence analysis of p113 and p100 peptides showed a high sequence homology between both polypeptides and the complete reported sequences of Xenopus laevis vitellogenin. Using specific antibodies, we determined that the Vtg uptake begins early during oogenesis, at the previtellogenic stage, and continues until oocytes have reached their mature status. In addition, we found that large endocytic vesicles mediate Vtg uptake in stage I oocytes, and that the size of the endocytic vesicles declines with oogenesis progression. In terms of the Vtg protein trafficking, we detected the Vtg precursor (190 kDa) in the liver of estradiol-injected females. Finally, we propose a subclassification of B. arenarum stage II oocytes into three physiologically and morphologically distinct periods (early, mid and late).10aAmino Acid Sequence10aAnimals10aBufo arenarum10aEgg Proteins10aFemale10aImmunohistochemistry10aMolecular Sequence Data10aMolecular Weight10aOocytes10aProtein Transport10aSequence Analysis, DNA10aTime Factors10aVitellogenesis1 aO'Brien, Emma, D1 aSalicioni, Ana, M1 aCabada, Marcelo, O1 aArranz, Silvia, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/vitellogenesis-in-bufo-arenarum-identification-characterization-and03062nas a2200421 4500008004100000245007400041210006900115260001600184300001100200490000700211520171200218653001101930653002101941653002101962653002901983653002402012653001102036653003002047653004302077653001802120653001102138653002502149653001602174653003502190653003102225653001802256100002502274700002002299700002002319700001502339700002002354700002402374700002102398700002202419700001802441700002902459856015202488 2009 eng d00aActivation of host wound responses in breast cancer microenvironment.0 aActivation of host wound responses in breast cancer microenviron c2009 Nov 15 a7020-80 v153 aPURPOSE: Cancer progression is mediated by processes that are also important in wound repair. As a result, cancers have been conceptualized as overhealing wounds or "wounds that do not heal," and gene expression signatures reflective of wound repair have shown value as predictors of breast cancer survival. Despite the widespread acknowledgment of commonalities between host responses to wounds and host responses to cancer, the gene expression responses of normal tissue adjacent to cancers have not been well characterized. EXPERIMENTAL DESIGN: Using RNA extracted from histologically normal breast tissue from 107 patients, including 60 reduction mammoplasty patients and 47 cancer patients, we measured whole genome expression profiles and identified a gene expression signature that is induced in response to breast cancer. RESULTS: This signature represents an in vivo "wound response" signature that is differentially expressed in the normal tissue of breast cancer patients compared with those without disease and is highly accurate (at least 92% sensitivity and 98% specificity) in distinguishing diseased and nondiseased. The in vivo wound response signature is highly prognostic of breast cancer survival, and there is a strong association between the groups identified by this signature and those identified using serum-treated fibroblasts and other microenvironment-derived or microenvironment-related signatures. CONCLUSIONS: The prevalence of the wound response signature in histologically normal tissue adjacent to breast cancer suggests that microenvironment response is an important variable in breast cancer progression and may be an important target for clinical interventions.
10aBreast10aBreast Neoplasms10aCyclooxygenase 210aCysteine-Rich Protein 6110aDisease Progression10aFemale10aGene Expression Profiling10aGene Expression Regulation, Neoplastic10aGenome, Human10aHumans10aImmunohistochemistry10aMammaplasty10aNeovascularization, Pathologic10aReproducibility of Results10aWound Healing1 aTroester, Melissa, A1 aLee, Myung, Hee1 aCarter, Matthew1 aFan, Cheng1 aCowan, David, W1 aPerez, Erick, Roman1 aPirone, Jason, R1 aPerou, Charles, M1 aJerry, Joseph1 aSchneider, Sallie, Smith uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/activation-of-host-wound-responses-in-breast-cancer-microenvironment01091nas a2200157 4500008004100000245003600041210003500077260001300112300001100125490000700136520060600143100002500749700002400774700002100798856011400819 2009 eng d00aADAM function in embryogenesis.0 aADAM function in embryogenesis c2009 Apr a153-630 v203 aCleavage of proteins inserted into the plasma membrane (shedding) is an essential process controlling many biological functions including cell signaling, cell adhesion and migration as well as proliferation and differentiation. ADAM surface metalloproteases have been shown to play an essential role in these processes. Gene inactivation during embryonic development have provided evidence of the central role of ADAM proteins in nematodes, flies, frogs, birds and mammals. The relative contribution of four subfamilies of ADAM proteins to developmental processes is the focus of this review.
1 aAlfandari, Dominique1 aMcCusker, Catherine1 aCousin, Hélène uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/adam-function-in-embryogenesis02709nas a2200169 4500008004100000245008200041210006900123260001300192300001200205490000700217520207400224100001702298700002502315700002202340700002402362856015302386 2009 eng d00aAntigenic basis of diversity in the gammadelta T cell co-receptor WC1 family.0 aAntigenic basis of diversity in the gammadelta T cell coreceptor c2009 Aug a2565-750 v463 aWC1 co-receptors are transmembrane glycoproteins with 11 extracellular scavenger receptor cysteine rich (SRCR) domains. They are related to the CD163 family but are uniquely expressed by gammadelta T cells. We recently showed that at least 13 members comprise the WC1 gene family in cattle, a model animal species for studies of gammadelta T cell biology. Since WC1 co-receptors participate in directing functional responses by gammadelta T cells either through the ligands they bind or the signals they transduce, availability of reagents to identify the expression of individual WC1 molecules of this diverse family would be valuable in further elucidating mechanisms of gammadelta T cell responsiveness. Although monoclonal antibodies (mAbs) have been widely used to identify WC1 co-receptors on gammadelta T cells, the locations of the antigenic epitopes recognized are unknown. Here, we mapped the epitopes to particular SRCR domains and evaluated their distribution among WC1 molecules. To do this, cDNA representing the extracellular domains of seven different WC1 genes was expressed in mammalian cells and analyzed for reactivity with anti-WC1 mAbs using ELISA and Western blotting. The study included mAbs that are broadly reactive with WC1(+) gammadelta T cells and those that divide WC1(+) gammadelta T cells into functionally distinct subpopulations. We found that mAb CC15 is a pan-reactive anti-WC1 mAb recognizing an epitope in the closely related SRCR domains 2 and 7 and that this epitope is present in at least domain 2 or 7 of all seven WC1 molecules evaluated here. Five other anti-WC1 mAbs, typified by mAb IL-A29, were found to be broadly reactive, recognizing epitopes in the related SRCR domains 4 and 9 but each having a unique pattern of reactivity with the seven WC1 molecules. Finally, the subpopulation-specific anti-WC1 mAbs, including those that recognize either the archetypal WC1.1 or WC1.2 molecule, were found to react with epitopes in the most variable WC1 domain, i.e. domain 1, of a restricted number of WC1 co-receptors.
1 aChen, Chuang1 aHerzig, Carolyn, T A1 aTelfer, Janice, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/antigenic-basis-of-diversity-in-the-gammadelta-t-cell-co-receptor-wc102642nas a2200217 4500008004100000245008100041210006900122260001600191300001100207490000800218520187300226100002202099700002102121700002002142700002302162700002202185700002202207700002102229700002502250856014902275 2009 eng d00aCloning and expression of ADAM-related metalloproteases in equine laminitis.0 aCloning and expression of ADAMrelated metalloproteases in equine c2009 Jun 15 a231-410 v1293 aEquine laminitis is a debilitating disease affecting the digital laminae that suspend the distal phalanx within the hoof. While the clinical progression of the disease has been well documented, the molecular events associated with its pathogenesis remain largely unknown. Using real time quantitative PCR (RT-qPCR), we have investigated the expression of genes coding for proteins containing a Disintegrin and Metalloprotease domain (ADAM), as well as genes encoding the natural inhibitors of these enzymes (tissue inhibitor of metalloprotease; TIMP) in horses with naturally-acquired (acute, chronic and aggravated chronic clinical cases) or experimentally-induced (black walnut extract (BWE) and starch gruel models) laminitis. Changes in expression of these enzymes and regulators may underlie the pathologic remodeling of lamellar tissue in laminitis. Genes encoding ADAMs involved in inflammation (ADAM-10 and ADAM-17), as well as those implicated in arthritis (ADAMTS-1, ADAMTS-4 and ADAMTS-5) were cloned, and the sequences used to generate specific oligonucleotide primers for the RT-qPCR experiments. Our results show that genes encoding ADAM-10 and ADAM-17 were not induced in most laminitic animals, whereas ADAMTS-4 gene expression was strongly upregulated in nearly all horses with experimentally-induced and naturally-acquired laminitis. The expression of matrix metalloproteases (MMP)-9 and ADAMTS-5 was also increased in many of the laminitic horses. In addition, TIMP-2 gene expression was decreased in most laminitic horses, whereas expression of genes encoding other TIMPs, namely TIMP-1 and TIMP-3, was randomly increased or decreased in the various models. We conclude that increased expression of lamellar ADAMTS-4 is a common feature of laminitis consistent with a central role of the gene product in the pathophysiology of the disease.
1 aCoyne, Michael, J1 aCousin, Hélène1 aLoftus, John, P1 aJohnson, Philip, J1 aBelknap, James, K1 aGradil, Carlos, M1 aBlack, Samuel, J1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cloning-and-expression-of-adam-related-metalloproteases-in-equine01588nas a2200205 4500008004100000245011900041210006900160260001500229300001200244490000800256520082600264100001901090700002201109700001601131700002401147700001801171700001901189700002401208856015001232 2009 eng d00aDetection and differentiation of normal, cancerous, and metastatic cells using nanoparticle-polymer sensor arrays.0 aDetection and differentiation of normal cancerous and metastatic c2009 Jul 7 a10912-60 v1063 aRapid and effective differentiation between normal and cancer cells is an important challenge for the diagnosis and treatment of tumors. Here, we describe an array-based system for identification of normal and cancer cells based on a "chemical nose/tongue" approach that exploits subtle changes in the physicochemical nature of different cell surfaces. Their differential interactions with functionalized nanoparticles are transduced through displacement of a multivalent polymer fluorophore that is quenched when bound to the particle and fluorescent after release. Using this sensing strategy we can rapidly (minutes/seconds) and effectively distinguish (i) different cell types; (ii) normal, cancerous and metastatic human breast cells; and (iii) isogenic normal, cancerous and metastatic murine epithelial cell lines.1 aBajaj, Avinash1 aMiranda, Oscar, R1 aKim, Ik-Bum1 aPhillips, Ronnie, L1 aJerry, Joseph1 aBunz, Uwe, H F1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/detection-and-differentiation-of-normal-cancerous-and-metastatic-001855nas a2200313 4500008004100000245011900041210006900160260001500229300001200244490000800256520083300264653001201097653002601109653002101135653001101156653000901167653001801176653002401194653001401218653001301232100001901245700002201264700001601286700002401302700001801326700001901344700002401363856015401387 2009 eng d00aDetection and differentiation of normal, cancerous, and metastatic cells using nanoparticle-polymer sensor arrays.0 aDetection and differentiation of normal cancerous and metastatic c2009 Jul 7 a10912-60 v1063 aRapid and effective differentiation between normal and cancer cells is an important challenge for the diagnosis and treatment of tumors. Here, we describe an array-based system for identification of normal and cancer cells based on a "chemical nose/tongue" approach that exploits subtle changes in the physicochemical nature of different cell surfaces. Their differential interactions with functionalized nanoparticles are transduced through displacement of a multivalent polymer fluorophore that is quenched when bound to the particle and fluorescent after release. Using this sensing strategy we can rapidly (minutes/seconds) and effectively distinguish (i) different cell types; (ii) normal, cancerous and metastatic human breast cells; and (iii) isogenic normal, cancerous and metastatic murine epithelial cell lines.
10aAnimals10aBiosensing Techniques10aCell Line, Tumor10aHumans10aMice10aNanoparticles10aNeoplasm Metastasis10aNeoplasms10aPolymers1 aBajaj, Avinash1 aMiranda, Oscar, R1 aKim, Ik-Bum1 aPhillips, Ronnie, L1 aJerry, Joseph1 aBunz, Uwe, H F1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/detection-and-differentiation-of-normal-cancerous-and-metastatic-cells02419nas a2200133 4500008004100000245013600041210006900177260001500246300000800261490000700269520184000276100002002116856014902136 2009 eng d00aDetermination, mechanism and monitoring of knockdown resistance in permethrin-resistant human head lice, Pediculus humanus capitis.0 aDetermination mechanism and monitoring of knockdown resistance i c2009 Mar 1 a1-70 v123 aPermethrin resistance has been reported worldwide and clinical failures to commercial pediculicides containing permethrin have likewise occurred. Permethrin resistance in head lice populations from the U.S. is widespread but is not yet uniform and the level of resistance is relatively low (~4-8 fold). Permethrin-resistant lice are cross-resistant to pyrethrins, PBO-synergized pyrethrins and to DDT. Nix((R)), when applied to human hair tufts following manufacture's instructions, did not provide 100% control when assessed by the hair tuft bioassay in conjunction with the in vitro rearing system. Resistance to permethrin is due to knockdown resistance (kdr), which is the result of three point mutations within the alpha-subunit gene of the voltage-gated sodium channel that causes amino acid substitutions, leading to nerve insensitivity.A three-tiered resistance monitoring system has been established based on molecular resistance detection techniques. Quantitative sequencing (QS) has been developed to predict the kdr allele frequency in head lice at a population level. The speed, simplicity and accuracy of QS made it an ideal candidate for a routine primary resistance monitoring tool to screen a large number of louse populations as an alternative to conventional bioassay. As a secondary monitoring method, real-time PASA (rtPASA) has been devised for a more precise determination of low resistance allele frequencies. To obtain more detailed information on resistance allele zygosity, as well as allele frequency, serial invasive signal amplification reaction (SISAR) has been developed as an individual genotyping method. Our approach of using three tiers of molecular resistance detection should facilitate large-scale routine resistance monitoring of permethrin resistance in head lice using field-collected samples.1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/determination-mechanism-and-monitoring-of-knockdown-resistance-in01300nas a2200157 4500008004100000245005500041210005400096260001600150300001100166490000800177520075800185100001700943700002500960700002400985856013301009 2009 eng d00aExpressed gene sequence of bovine IL23A and IL23R.0 aExpressed gene sequence of bovine IL23A and IL23R c2009 Apr 15 a425-300 v1283 aThe cloning and characterization of bovine IL23A and IL23 receptor cDNA from total RNA of PBMC and the genomic organization of the coding sequences are reported. The IL23A partial coding region was found to be 578 nucleotides coded for in 4 exons and shared 84% and 76% identity with human and mouse sequences, respectively. The IL23R complete coding region had 1890 nucleotides coded for in 10 exons and shared 87% and 73% homology with the human and mouse sequences, respectively. Both bovine sequences were more closely related to the human sequences than were mouse sequence. This work was done as part of the U.S. Veterinary Immune Reagent Network whose goal is to develop reagents for investigating diseases in livestock species, poultry and fish.1 aChen, Chuang1 aHerzig, Carolyn, T A1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/expressed-gene-sequence-of-bovine-il23a-and-il23r01791nas a2200169 4500008004100000245012900041210006900170260001300239300001000252490000700262520111300269100002401382700002101406700002001427700002501447856014901472 2009 eng d00aExtracellular cleavage of cadherin-11 by ADAM metalloproteases is essential for Xenopus cranial neural crest cell migration.0 aExtracellular cleavage of cadherin11 by ADAM metalloproteases is c2009 Jan a78-890 v203 aCell adhesion molecules such as cadherins alternate their expression throughout cranial neural crest (CNC) development, yet our understanding of the role of these molecules during CNC migration remains incomplete. The "mesenchymal" cadherin-11 is expressed in the CNC during migration yet prevents migration when overexpressed in the embryo, suggesting that a defined level of cadherin-11-mediated cell adhesion is required for migration. Here we show that members of the meltrin subfamily of ADAM metalloproteases cleave the extracellular domain of cadherin-11 during CNC migration. We show that a fragment corresponding to the putative shed form of cadherin-11 retains biological activity by promoting CNC migration in vivo, in a non-cell-autonomous manner. Additionally, cleavage of cadherin-11 does not affect binding to beta-catenin and downstream signaling events. We propose that ADAM cleavage of cadherin-11 promotes migration by modifying its ability to support cell-cell adhesion while maintaining the membrane-bound pool of beta-catenin associated with the cadherin-11 cytoplasmic domain.
1 aMcCusker, Catherine1 aCousin, Hélène1 aNeuner, Russell1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/extracellular-cleavage-of-cadherin-11-by-adam-metalloproteases-is00645nas a2200133 4500008004100000245006100041210005900102260001600161300001000177490000800187520016200195100001600357856013800373 2009 eng d00aExtracellular matrix, leukocyte migration and laminitis.0 aExtracellular matrix leukocyte migration and laminitis c2009 Jun 15 a161-30 v1293 aThe structure and dynamic nature of extracellular matrix is discussed in the context of healthy and diseased tissues particularly the equine digital laminae.1 aBlack, S, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/extracellular-matrix-leukocyte-migration-and-laminitis01223nas a2200145 4500008004100000245012400041210006900165260000900234300000800243490000700251520061900258100002500877700002400902856015100926 2009 eng d00aGenomic organization and classification of the bovine WC1 genes and expression by peripheral blood gamma delta T cells.0 aGenomic organization and classification of the bovine WC1 genes c2009 a1910 v103 aWC1 co-receptors are group B scavenger receptor cysteine-rich molecules that are found exclusively on gammadeltaT cells and are thought to be encoded by a multi-gene family. Previous studies have shown gammadeltaT cells that respond to a particular stimulus have unique WC1 molecules expressed. Prior to the onset of the studies described here only one full-length WC1 nucleotide sequence was publicly available, though three WC1 molecules had been distinguished based on monoclonal antibody reactivity. Furthermore, the number of WC1 genes found in the bovine genome and their sequences had not yet been resolved.1 aHerzig, Carolyn, T A1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genomic-organization-and-classification-of-the-bovine-wc1-genes-and01997nas a2200193 4500008004100000245006300041210006200104260001300166300001100179490000600190520133600196100002501532700001901557700002301576700002001599700002601619700001701645856014101662 2009 eng d00aIdentification of novel oocyte and granulosa cell markers.0 aIdentification of novel oocyte and granulosa cell markers c2009 Sep a404-100 v93 aHere we present novel gene expression patterns in the ovary as part of an ongoing assessment of published micro-array data from mouse oocytes and embryos. We present the expression patterns of 13 genes that had been determined by micro-array to be expressed in the mature egg, but not during subsequent preimplantation development. In-situ hybridization of sectioned ovaries revealed that these genes were expressed in one of two distinct patterns: (1) oocyte-specific or (2) expressed in both the oocyte and surrounding granulosa cells. Despite the fact that micro-array data demonstrated expression in the egg, several of these genes are expressed at low levels in the oocyte, but strongly expressed in granulosa cells. Eleven of these genes have no reported function or expression during oogenesis, indicating that this approach is a necessary step towards functional annotation of the genome. Also of note is that while some of these gene products have been well characterized in other tissues and cell types, others are relatively unstudied in the literature. Our results provide novel gene expression information that may provide insights into the molecular mechanisms of follicular recruitment, oocyte maturation and ovulation and will direct further experimentation into the role these genes play during oogenesis.
1 aMalcuit, Christopher1 aTrask, Mary, C1 aSantiago, Laurelis1 aBeaudoin, Emily1 aTremblay, Kimberly, D1 aMager, Jesse uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-novel-oocyte-and-granulosa-cell-markers01219nas a2200181 4500008004100000245015100041210006900192260001600261300001100277490000800288520048400296100002000780700002300800700002200823700002200845700002100867856014900888 2009 eng d00aLeukocyte-derived and endogenous matrix metalloproteinases in the lamellae of horses with naturally acquired and experimentally induced laminitis.0 aLeukocytederived and endogenous matrix metalloproteinases in the c2009 Jun 15 a221-300 v1293 aInflammation and dysregulation of endogenous matrix metalloproteinase (MMP) production are implicated in the development of equine laminitis. In this study, we examine quantitative relationships among levels of leukocyte-derived proMMP-9 and MMP-9, lamellar proMMP-2 and MMP-2, and expression of proMMP-2 processing enzymes, MT1-MMP/PACE4, as steps towards determining whether inflammation and dysregulation of endogenous MMP production are independent or co-dependent processes.1 aLoftus, John, P1 aJohnson, Philip, J1 aBelknap, James, K1 aPettigrew, Amanda1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/leukocyte-derived-and-endogenous-matrix-metalloproteinases-in-the02172nas a2200145 4500008004100000245010700041210006900148260000900217300001000226490000600236520158600242100002701828700002501855856014601880 2009 eng d00aLife after proteolysis: Exploring the signaling capabilities of classical cadherin cleavage fragments.0 aLife after proteolysis Exploring the signaling capabilities of c c2009 a155-70 v23 aClassical cadherins are a group of Ca(++) dependent transmembrane cell adhesion molecules, mostly known for their ability to perform homophylic interactions with like-cadherin molecules on the surface of neighboring cells. Over the past decade, many studies have also established cadherins as key players of intracellular signaling events by modifying the activity of Rho GTPases, members of the Wnt signaling pathway, and receptor tyrosine kinases. Given the utility of these molecules, it is not surprising that they play multiple roles during different embryological and adult processes. Yet, these activities have been primarily tied to their full-length molecules. And, while the activity of full-length molecules is undoubtedly an essential part of how cadherins perform in vivo, it is becoming increasingly evident that the proteolytic fragments of these molecules may also play a role. This is an exciting development because proteolysis of cadherins was previously thought to be a simple clearing-mechanism meant to regulate the levels of cadherin molecules on the cell-surface.Here, we will further discuss our recent findings by McCusker and colleagues, showing that both N-terminal and C-terminal fragments of cadherin-11 retain biological activity in Xenopus embryos. We will also review the current literature demonstrating that both the extracellular and intracellular fragments of other classical cadherins are capable of activating certain signaling events tied to Epithelial to Mesenchymal Transitions (EMTs), cell survival, cell proliferation and cell migration.1 aMcCusker, Catherine, D1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/life-after-proteolysis-exploring-the-signaling-capabilities-of02651nas a2200169 4500008004100000245011400041210006900155260001300224300001200237490000700249520198600256100002502242700002002267700001902287700002302306856015202329 2009 eng d00aLocalization of low-density detergent-resistant membrane proteins in intact and acrosome-reacted mouse sperm.0 aLocalization of lowdensity detergentresistant membrane proteins c2009 May a897-9040 v803 aMammalian sperm become fertile after completing capacitation, a process associated with cholesterol loss and changes in the biophysical properties of the sperm membranes that prepares the sperm to undergo the acrosome reaction. Different laboratories have hypothesized that cholesterol efflux can influence the extent and/or movement of lipid raft microdomains. In a previous study, our laboratory investigated the identity of sperm proteins putatively associated with rafts. After extraction with Triton X-100 and ultracentrifugation in sucrose gradients, proteins distributing to the light buoyant-density fractions were cored from polyacrylamide gels and microsequenced. In this study, a subset of these proteins (TEX101, basigin, hexokinase 1, facilitated glucose transporter 3, IZUMO, and SPAM1) and other molecules known to be enriched in membrane rafts (caveolin 2, flotillin 1, flotillin 2, and the ganglioside GM3) were selected to investigate their localization in the sperm and their behavior during capacitation and the acrosome reaction. These molecules localize to multiple sperm domains, including the acrosomal cap (IZUMO, caveolin 2, and flotillin 2), equatorial segment (GM3), cytoplasmic droplet (TEX101), midpiece (basigin, facilitated glucose transporter 3, and flotillin 2), and principal piece (facilitated glucose transporter 3). Some of these markers modified their immunofluorescence pattern after sperm incubation under capacitating conditions, and these changes correlated with the occurrence of the acrosome reaction. While GM3 and caveolin 2 were not detected after the acrosome reaction, flotillin 2 was found in the equatorial segment of acrosome-reacted sperm, and IZUMO distributed along the sperm head, reaching the post- and para-acrosomal areas. Taking into consideration the requirement of the acrosome reaction for sperm to become fusogenic, these results suggest that membrane raft dynamics may have a role in sperm-egg membrane interaction.1 aMiranda, Patricia, V1 aAllaire, Alicia1 aSosnik, Julian1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/localization-of-low-density-detergent-resistant-membrane-proteins-in01531nas a2200205 4500008004100000245006500041210006300106260001600169300001100185490000800196520084600204100001801050700001901068700001701087700001901104700001601123700002001139700002401159856014201183 2009 eng d00aNotch regulates cytolytic effector function in CD8+ T cells.0 aNotch regulates cytolytic effector function in CD8 T cells c2009 Mar 15 a3380-90 v1823 aThe maturation of naive CD8(+) T cells into effector CTLs is a critical feature of a functional adaptive immune system. Development of CTLs depends, in part, upon the expression of the transcriptional regulator eomesodermin (EOMES), which is thought to regulate expression of two key effector molecules, perforin and granzyme B. Although EOMES is important for effector CTL development, the precise mechanisms regulating CD8(+) effector cell maturation remains poorly understood. In this study, we show that Notch1 regulates the expression of EOMES, perforin, and granzyme B through direct binding to the promoters of these crucial effector molecules. By abrogating Notch signaling, both biochemically as well as genetically, we conclude that Notch activity mediates CTL activity through direct regulation of EOMES, perforin, and granzyme B.1 aCho, Ok, Hyun1 aShin, Hyun, Mu1 aMiele, Lucio1 aGolde, Todd, E1 aFauq, Abdul1 aMinter, Lisa, M1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch-regulates-cytolytic-effector-function-in-cd8-t-cells02218nas a2200241 4500008004100000245010900041210006900150260001600219300001200235490000800247520136800255100001501623700002001638700001901658700002401677700002701701700001801728700002001746700001601766700001901782700002401801856015101825 2009 eng d00aNotch signaling mediates G1/S cell-cycle progression in T cells via cyclin D3 and its dependent kinases.0 aNotch signaling mediates G1S cellcycle progression in T cells vi c2009 Feb 19 a1689-980 v1133 aNotch signaling plays a role in normal lymphocyte development and function. Activating Notch1-mutations, leading to aberrant downstream signaling, have been identified in human T-cell acute lymphoblastic leukemia (T-ALL). While this highlights the contribution of Notch signaling to T-ALL pathogenesis, the mechanisms by which Notch regulates proliferation and survival in normal and leukemic T cells are not fully understood. Our findings identify a role for Notch signaling in G(1)-S progression of cell cycle in T cells. Here we show that expression of the G(1) proteins, cyclin D3, CDK4, and CDK6, is Notch-dependent both in vitro and in vivo, and we outline a possible mechanism for the regulated expression of cyclin D3 in activated T cells via CSL (CBF-1, mammals; suppressor of hairless, Drosophila melanogaster; Lag-1, Caenorhabditis elegans), as well as a noncanonical Notch signaling pathway. While cyclin D3 expression contributes to cell-cycle progression in Notch-dependent human T-ALL cell lines, ectopic expression of CDK4 or CDK6 together with cyclin D3 shows partial rescue from gamma-secretase inhibitor (GSI)-induced G(1) arrest in these cell lines. Importantly, cyclin D3 and CDK4 are highly overexpressed in Notch-dependent T-cell lymphomas, justifying the combined use of cell-cycle inhibitors and GSI in treating human T-cell malignancies.1 aJoshi, Ila1 aMinter, Lisa, M1 aTelfer, Janice1 aDemarest, Renée, M1 aCapobianco, Anthony, J1 aAster, Jon, C1 aSicinski, Piotr1 aFauq, Abdul1 aGolde, Todd, E1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch-signaling-mediates-g1-s-cell-cycle-progression-in-t-cells-via02399nas a2200397 4500008004100000245012300041210006900164260001300233300001000246490000700256520111000263653002401373653001201397653002701409653001201436653002401448653001401472653002001486653002401506653002301530653001301553653001101566653004301577653000901620653001201629653002001641653003701661653002801698100002501726700001801751700002001769700002201789700001801811700002301829856014901852 2009 eng d00aRegulation of inositol 1,4,5-trisphosphate receptor type 1 function during oocyte maturation by MPM-2 phosphorylation.0 aRegulation of inositol 145trisphosphate receptor type 1 function c2009 Jul a56-640 v463 aEgg activation and further embryo development require a sperm-induced intracellular Ca(2+) signal at the time of fertilization. Prior to fertilization, the egg's Ca(2+) machinery is therefore optimized. To this end, during oocyte maturation, the sensitivity, i.e. the Ca(2+) releasing ability, of the inositol 1,4,5-trisphosphate receptor type 1 (IP(3)R1), which is responsible for most of this Ca(2+) release, markedly increases. In this study, the recently discovered specific Polo-like kinase (Plk) inhibitor BI2536 was used to investigate the role of Plk1 in this process. BI2536 inactivates Plk1 in oocytes at the early stages of maturation and significantly decreases IP(3)R1 phosphorylation at an MPM-2 epitope at this stage. Moreover, this decrease in Plk1-dependent MPM-2 phosphorylation significantly lowers IP(3)R1 sensitivity. Finally, using in vitro phosphorylation techniques we identified T(2656) as a major Plk1 site on IP(3)R1. We therefore propose that the initial increase in IP(3)R1 sensitivity during oocyte maturation is underpinned by IP(3)R1 phosphorylation at an MPM-2 epitope(s).10aAmino Acid Sequence10aAnimals10aAntibodies, Monoclonal10aCalcium10aCell Cycle Proteins10aCell Line10aCells, Cultured10aComputer Simulation10aConsensus Sequence10aEpitopes10aFemale10aInositol 1,4,5-Trisphosphate Receptors10aMice10aOocytes10aPhosphorylation10aProtein-Serine-Threonine Kinases10aProto-Oncogene Proteins1 aVanderheyden, Veerle1 aWakai, Takuya1 aBultynck, Geert1 aDe Smedt, Humbert1 aParys, Jan, B1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-inositol-145-trisphosphate-receptor-type-1-function02477nas a2200409 4500008004100000245004500041210004400086260001600130300001200146490000800158520119600166653001201362653004001374653003901414653003701453653002801490653003501518653002001553653000901573653003201582653003001614100002001644700002101664700002401685700002101709700002101730700001901751700001901770700001801789700002001807700002201827700002301849700002101872700002201893700002901915856012301944 2009 eng d00aRunx proteins regulate Foxp3 expression.0 aRunx proteins regulate Foxp3 expression c2009 Oct 26 a2329-370 v2063 aRunx proteins are essential for hematopoiesis and play an important role in T cell development by regulating key target genes, such as CD4 and CD8 as well as lymphokine genes, during the specialization of naive CD4 T cells into distinct T helper subsets. In regulatory T (T reg) cells, the signature transcription factor Foxp3 interacts with and modulates the function of several other DNA binding proteins, including Runx family members, at the protein level. We show that Runx proteins also regulate the initiation and the maintenance of Foxp3 gene expression in CD4 T cells. Full-length Runx promoted the de novo expression of Foxp3 during inducible T reg cell differentiation, whereas the isolated dominant-negative Runt DNA binding domain antagonized de novo Foxp3 expression. Foxp3 expression in natural T reg cells remained dependent on Runx proteins and correlated with the binding of Runx/core-binding factor beta to regulatory elements within the Foxp3 locus. Our data show that Runx and Foxp3 are components of a feed-forward loop in which Runx proteins contribute to the expression of Foxp3 and cooperate with Foxp3 proteins to regulate the expression of downstream target genes.10aAnimals10aCore Binding Factor Alpha 3 Subunit10aCore Binding Factor alpha Subunits10aCore Binding Factor beta Subunit10aFeedback, Physiological10aForkhead Transcription Factors10aGenes, Dominant10aMice10aProtein Structure, Tertiary10aT-Lymphocytes, Regulatory1 aBruno, Ludovica1 aMazzarella, Luca1 aHoogenkamp, Maarten1 aHertweck, Arnulf1 aCobb, Bradley, S1 aSauer, Stephan1 aHadjur, Suzana1 aLeleu, Marion1 aNaoe, Yoshinori1 aTelfer, Janice, C1 aBonifer, Constanze1 aTaniuchi, Ichiro1 aFisher, Amanda, G1 aMerkenschlager, Matthias uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/runx-proteins-regulate-foxp3-expression02020nas a2200205 4500008004100000245007900041210006900120260001500189300001100204490000800215520127900223100001901502700002501521700002701546700002101573700002301594700002301617700002301640856015101663 2009 eng d00aTssk6 is required for Izumo relocalization and gamete fusion in the mouse.0 aTssk6 is required for Izumo relocalization and gamete fusion in c2009 Aug 1 a2741-90 v1223 aOne of the most important processes in fertilization is the fusion of egg and sperm; however, the molecular mechanisms involved in this process are not well understood. So far, using genetic approaches, only two proteins have been demonstrated to be necessary for this process: Izumo in sperm and CD9 in the egg. Here we demonstrate that sperm produced by Tssk6 (Sstk)-null mice present defects that prevent the successful fertilization of eggs in vitro and the fusion to zona-pellucida-free eggs. Tssk6 is a member of the testis-specific serine kinase family of proteins and is expressed postmeiotically in male germ cells. In order for fusion to occur, during the process known as acrosome reaction Izumo needs to relocate from the anterior head to other regions, including the postacrosomal compartment. Tssk6-null sperm fails to relocate Izumo during the acrosome reaction. Agents that interfere with actin dynamics blocked the acrosome-reaction-associated translocation of Izumo that is required for fusion in wild-type sperm. Additionally, actin polymerization was compromised in Tssk6-null sperm. Taken together, our results indicate that Tssk6 is involved in sperm-egg fusion through the regulation of actin polymerization and changes in Izumo localization.
1 aSosnik, Julian1 aMiranda, Patricia, V1 aSpiridonov, Nikolay, A1 aYoon, Sook-Young1 aFissore, Rafael, A1 aJohnson, Gibbes, R1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/tssk6-is-required-for-izumo-relocalization-and-gamete-fusion-in-the02081nas a2200181 4500008004100000245015200041210006900193260001300262300001100275490000700286520136100293100001401654700002001668700001401688700002401702700002201726856015101748 2009 eng d00aTyrosine phosphorylation of scavenger receptor cysteine-rich WC1 is required for the WC1-mediated potentiation of TCR-induced T-cell proliferation.0 aTyrosine phosphorylation of scavenger receptor cysteinerich WC1 c2009 Jan a254-660 v393 aWorkshop cluster 1 (WC1) molecules are transmembrane glycoproteins uniquely expressed by gammadelta T cells. They belong to the scavenger receptor cysteine-rich superfamily and are encoded by a multi-gene family, which is divided on the basis of antibody reactivity, into three groups, WC1.1, WC1.2, and WC1.3. The potential role of WC1 as a co-stimulatory molecule for the gammadelta TCR is suggested by the presence of several tyrosine-based motifs in their intracellular domains. In this study, we found that WC1 was constitutively phosphorylated in ex vivo bovine gammadelta T cells and associated with src family tyrosine kinases. Crosslinking of WC1 molecules resulted in an increase in WC1 phosphorylation and co-crosslinking of WC1 and gammadelta TCR together prolonged WC1 phosphorylation. We identified the second tyrosine residue as the primary phosphorylation target in WC1.1 and WC1.2 intracellular sequences in both in vitro and in vivo assays. The cytoplasmic tails of WC1.1 and WC1.2 were phosphorylated on serine and PKC activity was required for PMA-induced endocytosis of WC1.1 or WC1.2. We found that phosphorylation of the second tyrosine in the WC1 cytoplasmic domain was required for the WC1-mediated potentiation of TCR-induced T-cell proliferation, suggesting that WC1 acts as a co-stimulatory molecule for gammadelta TCR.
1 aWang, Fei1 aHerzig, Carolyn1 aOzer, Dar1 aBaldwin, Cynthia, L1 aTelfer, Janice, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/tyrosine-phosphorylation-of-scavenger-receptor-cysteine-rich-wc1-is00526nas a2200121 4500008004100000245008300041210006900124260001600193300001000209490000800219100002300227856015400250 2009 eng d00aUnderstanding the molecular basis of sperm capacitation through kinase design.0 aUnderstanding the molecular basis of sperm capacitation through c2009 Jan 20 a667-80 v1061 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/understanding-the-molecular-basis-of-sperm-capacitation-through-kinase02930nas a2200337 4500008004100000245016200041210006900203260001300272300001200285490000600297520178400303653002402087653001202111653002902123653002102152653002102173653000902194653002202203653000902225653002802234653002002262653002002282653001302302653002302315653001602338100001902354700002202373700002002395700002302415856015402438 2009 eng d00aUse of differential isotopic labeling and mass spectrometry to analyze capacitation-associated changes in the phosphorylation status of mouse sperm proteins.0 aUse of differential isotopic labeling and mass spectrometry to a c2009 Mar a1431-400 v83 aMammalian sperm need to reside in the female reproductive tract for a finite period of time before acquiring fertilizing competence. The biochemical changes associated with this process are collectively known as "capacitation". With the use of the mouse as an experimental model, we have previously demonstrated that capacitation is associated with a cAMP-dependent increase in protein tyrosine phosphorylation. However, little is known about the identity and function of the protein targets of this phosphorylation cascade. In the present work, we have used differential isotopic labeling coupled with immobilized metal affinity chromatography (IMAC)-based phosphopeptide enrichment and analysis on a hybrid linear ion trap/FT-ICR mass spectrometer to measure the changes in protein phosphorylation resulting from the capacitation process. As no kinase activators and/or phosphatase inhibitors were used in the preparation of the sperm samples, phosphorylated residues identified in this study represent in vivo sites of phosphorylation. Also, in contrast to other methods which rely on the incorporation of isotopically labeled amino acids at the protein level (e.g., SILAC), the present technique is based on the Fisher esterification of protein digests, allowing for the comparison of phosphorylation status in the absence of protein synthesis. This approach resulted in the identification of 55 unique, in vivo sites of phosphorylation and permitted the relative extent of phosphorylation, as a consequence of capacitation, to be calculated for 42 different phosphopeptides. This work represents the first effort to determine which specific protein phosphorylation sites change their phosphorylation status in vivo as a result of the mammalian capacitation process.
10aAmino Acid Sequence10aAnimals10aChromatography, Affinity10aFourier Analysis10aIsotope Labeling10aMale10aMass Spectrometry10aMice10aMolecular Sequence Data10aPhosphopeptides10aPhosphorylation10aProteome10aSperm Capacitation10aSpermatozoa1 aPlatt, Mark, D1 aSalicioni, Ana, M1 aHunt, Donald, F1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/use-of-differential-isotopic-labeling-and-mass-spectrometry-to-analyze01183nas a2200157 4500008004100000245006400041210006200105260001600167300001200183490000700195520062200202100001900824700001800843700002000861856014400881 2009 eng d00aVirus-inspired approach to nonviral gene delivery vehicles.0 aVirusinspired approach to nonviral gene delivery vehicles c2009 Aug 10 a2189-930 v103 aThe basic TAT peptide, responsible for translocation of the HIV-TAT protein, has been conjugated to a variety of artificial nanoscopic materials to transport them across the cellular membrane. However, attempts to translocate genes using the TAT-peptide had met with limited success. We hypothesized that the cationic nature of the peptide does not allow for displaying these peptides on the surface of the polyplex. To circumvent this potential issue, we have developed a new molecular design strategy where the TAT-peptide can be effectively displayed on the surface of the polyplex, thus enhancing gene expression.1 aRoy, Raghunath1 aJerry, Joseph1 aThayumanavan, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/virus-inspired-approach-to-nonviral-gene-delivery-vehicles-001518nas a2200289 4500008004100000245006400041210006200105260001600167300001200183490000700195520062900202653002100831653002000852653001100872653002300883653002900906653002000935653001100955653001100966653002200977653001300999653001701012100001901029700001801048700002001066856014201086 2009 eng d00aVirus-inspired approach to nonviral gene delivery vehicles.0 aVirusinspired approach to nonviral gene delivery vehicles c2009 Aug 10 a2189-930 v103 aThe basic TAT peptide, responsible for translocation of the HIV-TAT protein, has been conjugated to a variety of artificial nanoscopic materials to transport them across the cellular membrane. However, attempts to translocate genes using the TAT-peptide had met with limited success. We hypothesized that the cationic nature of the peptide does not allow for displaying these peptides on the surface of the polyplex. To circumvent this potential issue, we have developed a new molecular design strategy where the TAT-peptide can be effectively displayed on the surface of the polyplex, thus enhancing gene expression.
10aBreast Neoplasms10aCells, Cultured10aFemale10aGene Products, tat10aGene Transfer Techniques10aGenetic Vectors10aHumans10aKidney10aPeptide Fragments10aPolymers10aTransfection1 aRoy, Raghunath1 aJerry, Joseph1 aThayumanavan, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/virus-inspired-approach-to-nonviral-gene-delivery-vehicles02308nas a2200181 4500008004100000245007900041210006900120260001700189300001100206490000800217520164800225100002001873700002101893700002401914700001901938700002501957856014401982 2009 eng d00aXenopus ADAM19 is involved in neural, neural crest and muscle development.0 aXenopus ADAM19 is involved in neural neural crest and muscle dev c2009 Mar-Apr a240-550 v1263 aADAM19 is a member of the meltrin subfamily of ADAM metalloproteases. In Xenopus, ADAM19 is present as a maternal transcript. Zygotic expression starts during gastrulation and is apparent in the dorsal blastopore lip. ADAM19 expression through neurulation and tailbud formation becomes enriched in dorsal structures such as the neural tube, the notochord and the somites. Using morpholino knock-down, we show that a reduction of ADAM19 protein in gastrula stage embryos results in a decrease of Brachyury expression in the notochord concomitant with an increase in the dorsal markers, Goosecoid and Chordin. These changes in gene expression are accompanied by a decrease in phosphorylated AKT, a downstream target of the EGF signaling pathway, and occur while the blastopore closes at the same rate as the control embryos. During neurulation and tailbud formation, ADAM19 knock-down induces a reduction of the neural markers N-tubulin and NRP1 but not Sox2. In the somitic mesoderm, the expression of MLC is also decreased while MyoD is not. ADAM19 knockdown also reduces neural crest markers prior to cell migration. Neural crest induction is also decreased in embryos treated with an EGF receptor inhibitor suggesting that this pathway is necessary for neural crest cell induction. Using targeted knock-down of ADAM19 we show that the reduction of neural and neural crest markers is cell autonomous and that the migration if the cranial neural crest is perturbed. We further show that ADAM19 protein reduction affects somite organization, reduces 12-101 expression and perturbs fibronectin localization at the intersomitic boundary.
1 aNeuner, Russell1 aCousin, Hélène1 aMcCusker, Catherine1 aCoyne, Michael1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/xenopus-adam19-is-involved-in-neural-neural-crest-and-muscle02496nas a2200193 4500008004100000245011200041210006900153260001300222300001300235490000700248520176200255100002102017700001902038700002502057700002802082700002002110700002002130856015202150 2008 eng d00aBiochemical and molecular analysis of deltamethrin resistance in the common bed bug (Hemiptera: Cimicidae).0 aBiochemical and molecular analysis of deltamethrin resistance in c2008 Nov a1092-1010 v453 aThis study establishes deltamethrin resistance in a common bed bug, Cimex lectularius L., population collected from New York City (NY-BB). The NY-BB population was 264-fold more resistant to 1% deltamethrin in contact bioassay compared with an insecticide-susceptible population collected in Florida (FL-BB). General esterase, glutathione S-transferase, and 7-ethoxycoumarin O-deethylase activities of NY-BB were not statistically different from those of FL-BB. cDNA fragments that encoded the open reading frame of voltage-sensitive sodium channel alpha-subunit genes from the FL-BB and NY-BB populations, respectively, were obtained by homology probing polymerase chain reaction (PCR) and sequenced. Sequence alignment of the internal and 5' and 3' rapid amplification of cDNA ends (RACE) fragments generated a 6500-bp cDNA sequence contig, which was composed of a 6084-bp open reading frame (ORF) encoding 2027 amino acid residues and 186-bp 5' and 230-bp 3' untranslated regions (5' and 3' UTRs, respectively). Sequence comparisons of the open reading frames of the alpha-subunit genes identified two point mutations (V419L and L925I) that were presented only in the NY-BB population. L925I, located the intracellular loop between IIS4 and IIS5, has been previously found in a highly pyrethroid-resistant populations of whitefly (Bemisia tabaci). V419L, located in the IS6 transmembrane segment, is a novel mutation. A Val to Met mutation at the corresponding position of the bed bug V419, however, has been identified in the tobacco budworm as a kdr-type mutation. This evidence suggests that the two mutations are likely the major resistance-causing mutations in the deltamethrin-resistant NY-BB through a knockdown-type nerve insensitivity mechanism.1 aYoon, Kyong, Sup1 aKwon, Deok, Ho1 aStrycharz, Joseph, P1 aHollingsworth, Craig, S1 aLee, Si, Hyeock1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/biochemical-and-molecular-analysis-of-deltamethrin-resistance-in-the03230nas a2200457 4500008004100000245011700041210006900158260001600227300001300243490000800256520176900264653002202033653001202055653001502067653001802082653001402100653001502114653004102129653001102170653001502181653000902196653000902205653002002214653002402234653005102258653004102309653002302350653001902373653001602392653001302408653001902421100002302440700002202463700001602485700002502501700001802526700003702544700002102581700002302602856014702625 2008 eng d00aChloride Is essential for capacitation and for the capacitation-associated increase in tyrosine phosphorylation.0 aChloride Is essential for capacitation and for the capacitationa c2008 Dec 19 a35539-500 v2833 aAfter epididymal maturation, sperm capacitation, which encompasses a complex series of molecular events, endows the sperm with the ability to fertilize an egg. This process can be mimicked in vitro in defined media, the composition of which is based on the electrolyte concentration of the oviductal fluid. It is well established that capacitation requires Na(+), HCO(3)(-), Ca(2+), and a cholesterol acceptor; however, little is known about the function of Cl(-) during this important process. To determine whether Cl(-), in addition to maintaining osmolarity, actively participates in signaling pathways that regulate capacitation, Cl(-) was replaced by either methanesulfonate or gluconate two nonpermeable anions. The absence of Cl(-) did not affect sperm viability, but capacitation-associated processes such as the increase in tyrosine phosphorylation, the increase in cAMP levels, hyperactivation, the zona pellucidae-induced acrosome reaction, and most importantly, fertilization were abolished or significantly reduced. Interestingly, the addition of cyclic AMP agonists to sperm incubated in Cl(-)-free medium rescued the increase in tyrosine phosphorylation and hyperactivation suggesting that Cl(-) acts upstream of the cAMP/protein kinase A signaling pathway. To investigate Cl(-) transport, sperm incubated in complete capacitation medium were exposed to a battery of anion transport inhibitors. Among them, bumetanide and furosemide, two blockers of Na(+)/K(+)/Cl(-) cotransporters (NKCC), inhibited all capacitation-associated events, suggesting that these transporters may mediate Cl(-) movements in sperm. Consistent with these results, Western blots using anti-NKCC1 antibodies showed the presence of this cotransporter in mature sperm.
10aAcrosome Reaction10aAnimals10aBumetanide10aCell Survival10aChlorides10aCyclic AMP10aCyclic AMP-Dependent Protein Kinases10aFemale10aFurosemide10aMale10aMice10aPhosphorylation10aSignal Transduction10aSodium Potassium Chloride Symporter Inhibitors10aSodium-Potassium-Chloride Symporters10aSperm Capacitation10aSperm Motility10aSpermatozoa10aTyrosine10aZona Pellucida1 aWertheimer, Eva, V1 aSalicioni, Ana, M1 aLiu, Weimin1 aTreviño, Claudia, L1 aChavez, Julio1 aHernández-González, Enrique, O1 aDarszon, Alberto1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/chloride-is-essential-for-capacitation-and-for-the-capacitation00482nas a2200157 4500008004100000245006800041210006700109260002300176300001200199490000600211100001400217700001400231700001800245700001500263856004600278 2008 eng d00aElite control of HIV infection: implications for vaccine design0 aElite control of HIV infection implications for vaccine design bInforma Healthcare a55–690 v91 aBaker, BM1 aBlock, BL1 aRothchild, AC1 aWalker, BD uhttps://doi.org/10.1517/1471259080257192801087nas a2200181 4500008004100000245013800041210006900179260000900248300000800257490000700265520037300272100002100645700002500666700001700691700002500708700001800733856015400751 2008 eng d00aEstrogen and progesterone induce persistent increases in p53-dependent apoptosis and suppress mammary tumors in BALB/c-Trp53+/- mice.0 aEstrogen and progesterone induce persistent increases in p53depe c2008 aR430 v103 aTreatment with estrogen and progesterone (E+P) mimics the protective effect of parity on mammary tumors in rodents and depends upon the activity of p53. The following experiments tested whether exogenous E+P primes p53 to be more responsive to DNA damage and whether these pathways confer resistance to mammary tumors in a mouse model of Li-Fraumeni syndrome.
1 aDunphy, Karen, A1 aBlackburn, Anneke, C1 aYan, Haoheng1 aO'Connell, Lauren, R1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/estrogen-and-progesterone-induce-persistent-increases-in-p53-dependent02153nas a2200337 4500008004100000245012000041210006900161260001300230300001200243490000600255520096500261653001201226653001601238653001601254653000901270653003801279653002401317653004901341653003701390653002401427653006801451653001401519100002801533700001701561700001801578700002301596700001201619700001701631700001901648856014801667 2008 eng d00aThe function of TRADD in signaling through tumor necrosis factor receptor 1 and TRIF-dependent Toll-like receptors.0 afunction of TRADD in signaling through tumor necrosis factor rec c2008 Sep a1047-540 v93 aThe physiological function of the adaptor protein TRADD remains unclear because of the unavailability of a TRADD-deficient animal model. By generating TRADD-deficient mice, we found here that TRADD serves an important function in tumor necrosis factor receptor 1 (TNFR1) signaling by orchestrating the formation of TNFR1 signaling complexes. TRADD was essential for TNFR1 signaling in mouse embryonic fibroblasts but was partially dispensable in macrophages; abundant expression of the adaptor RIP in macrophages may have allowed some transmission of TNFR1 signals in the absence of TRADD. Although morphologically normal, TRADD-deficient mice were resistant to toxicity induced by TNF, lipopolysaccharide and polyinosinic-polycytidylic acid. TRADD was also required for TRIF-dependent Toll-like receptor signaling in mouse embryonic fibroblasts but not macrophages. Our findings definitively establish the biological function of TRADD in TNF signaling.
10aAnimals10aFibroblasts10aMacrophages10aMice10aMitogen-Activated Protein Kinases10aSignal Transduction10aTNF Receptor-Associated Death Domain Protein10aTNF Receptor-Associated Factor 110aToll-Like Receptors10aTumor Necrosis Factor Receptor-Associated Peptides and Proteins10aUbiquitin1 aPobezinskaya, Yelena, L1 aKim, You-Sun1 aChoksi, Swati1 aMorgan, Michael, J1 aLi, Tao1 aLiu, Chengyu1 aLiu, Zhenggang uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-function-of-tradd-in-signaling-through-tumor-necrosis-factor01865nas a2200157 4500008004100000245008500041210006900126260001600195300001200211490000700223520125700230100002301487700002401510700002001534856015301554 2008 eng d00aGolfer exposure to chlorpyrifos and carbaryl following application to turfgrass.0 aGolfer exposure to chlorpyrifos and carbaryl following applicati c2008 Aug 13 a6616-220 v563 aExposure of golfers to pesticides following their application to turfgrass is of concern to regulators, turfgrass professionals, and consumers. Multipathway exposures were evaluated for golfers on turfgrass treated with chlorpyrifos and carbaryl. Air concentrations and transferable foliar residues (TFRs) were measured to assess potential respiratory and dermal exposures, respectively. At the same time, exposure to individuals simulating the play of golf was determined by dosimetry and urinary biomonitoring. Individual golfer exposure was determined in 76 rounds of golf following eight applications of chlorpyrifos and two applications of carbaryl. Estimated exposures to golfers following full course and full rate applications of chlorpyrifos and carbaryl were 19-68 times below current U.S. EPA acute reference dose (Rfd) values, indicating safe exposures under U.S. EPA hazard quotient criteria. Dermal exposure was determined to be the dominant exposure pathway to golfers, accounting for approximately 60% of the chlorpyrifos absorbed dose and 100% of the carbaryl absorbed dose. This study also provides a set of transfer factors (TFs) that may be used to determine dermal exposure of golfers to pesticides using transferable residue data.1 aPutnam, Raymond, A1 aDoherty, Jeffery, J1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/golfer-exposure-to-chlorpyrifos-and-carbaryl-following-application-to02592nas a2200337 4500008004100000245013300041210006900174260001300243300001200256490000800268520146900276653001201745653002601757653001101783653000901794653003701803653001601840100002101856700002101877700002601898700002001924700001901944700001801963700002101981700001702002700002102019700002302040700001702063700002302080856015102103 2008 eng d00aHuman sperm devoid of PLC, zeta 1 fail to induce Ca(2+) release and are unable to initiate the first step of embryo development.0 aHuman sperm devoid of PLC zeta 1 fail to induce Ca2 release and c2008 Nov a3671-810 v1183 aEgg activation, which is the first step in the initiation of embryo development, involves both completion of meiosis and progression into mitotic cycles. In mammals, the fertilizing sperm delivers the activating signal, which consists of oscillations in free cytosolic Ca(2+) concentration ([Ca(2+)](i)). Intracytoplasmic sperm injection (ICSI) is a technique that in vitro fertilization clinics use to treat a myriad of male factor infertility cases. Importantly, some patients who repeatedly fail ICSI also fail to induce egg activation and are, therefore, sterile. Here, we have found that sperm from patients who repeatedly failed ICSI were unable to induce [Ca(2+)](i) oscillations in mouse eggs. We have also shown that PLC, zeta 1 (PLCZ1), the sperm protein thought to induce [Ca(2+)](i) oscillations, was localized to the equatorial region of wild-type sperm heads but was undetectable in sperm from patients who had failed ICSI. The absence of PLCZ1 in these patients was further confirmed by Western blot, although genomic sequencing failed to reveal conclusive PLCZ1 mutations. Using mouse eggs, we reproduced the failure of sperm from these patients to induce egg activation and rescued it by injection of mouse Plcz1 mRNA. Together, our results indicate that the inability of human sperm to initiate [Ca(2+)](i) oscillations leads to failure of egg activation and sterility and that abnormal PLCZ1 expression underlies this functional defect.
10aCalcium10aEmbryonic Development10aHumans10aMale10aPhosphoinositide Phospholipase C10aSpermatozoa1 aYoon, Sook-Young1 aJellerette, Teru1 aSalicioni, Ana, Maria1 aLee, Hoi, Chang1 aYoo, Myung-Sik1 aCoward, Kevin1 aParrington, John1 aGrow, Daniel1 aCibelli, Jose, B1 aVisconti, Pablo, E1 aMager, Jesse1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/human-sperm-devoid-of-plc-zeta-1-fail-to-induce-ca2-release-and-are02662nas a2200337 4500008004100000245010000041210006900141260000900210300001100219490000700230520159000237653001201827653002801839653003501867653000901902653000901911653002301920653002001943653001301963653002301976653001601999653001102015653001202026653001302038653003702051100001802088700002202106700002002128700002302148856015302171 2008 eng d00aIdentification of proteins undergoing tyrosine phosphorylation during mouse sperm capacitation.0 aIdentification of proteins undergoing tyrosine phosphorylation d c2008 a463-720 v523 aMammalian sperm are not able to fertilize immediately upon ejaculation; they become fertilization-competent after undergoing changes in the female reproductive tract collectively termed capacitation. Although it has been established that capacitation is associated with an increase in tyrosine phosphorylation, little is known about the role of this event in sperm function. In this work we used a combination of two dimensional gel electrophoresis and mass spectrometry to identify proteins that undergo tyrosine phosphorylation during capacitation. Some of the identified proteins are the mouse orthologues of human sperm proteins known to undergo tyrosine phosphorylation. Among them we identified VDAC, tubulin, PDH E1 beta chain, glutathione S-transferase, NADH dehydrogenase (ubiquinone) Fe-S protein 6, acrosin binding protein precursor (sp32), proteasome subunit alpha type 6b and cytochrome b-c1 complex. In addition to previously described proteins, we identified two testis-specific aldolases as substrates for tyrosine phosphorylation. Genomic and EST analyses suggest that these aldolases are retroposons expressed exclusively in the testis, as has been reported elsewhere. Because of the importance of glycolysis for sperm function, we hypothesize that tyrosine phosphorylation of these proteins can play a role in the regulation of glycolysis during capacitation. However, neither the Km nor the Vmax of aldolase changed as a function of capacitation when its enzymatic activity was assayed in vitro, suggesting other levels of regulation for aldolase function.
10aAnimals10aExpressed Sequence Tags10aFructose-Bisphosphate Aldolase10aMale10aMice10aModels, Biological10aPhosphorylation10aProteins10aSperm Capacitation10aSpermatozoa10aTestis10aTubulin10aTyrosine10aVoltage-Dependent Anion Channels1 aArcelay, Enid1 aSalicioni, Ana, M1 aWertheimer, Eva1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-proteins-undergoing-tyrosine-phosphorylation-during02070nas a2200241 4500008004100000245012400041210006900165260001600234300001100250490000800261520119900269100001501468700002101483700001401504700002501518700002001543700002601563700002501589700002201614700001801636700002301654856015101677 2008 eng d00aInositol 1,4,5-trisphosphate receptor 1, a widespread Ca2+ channel, is a novel substrate of polo-like kinase 1 in eggs.0 aInositol 145trisphosphate receptor 1 a widespread Ca2 channel is c2008 Aug 15 a402-130 v3203 aTo initiate embryo development, the sperm induces in the egg release of intracellular calcium ([Ca2+](i)). During oocyte maturation, the inositol 1,4,5-trisphosphate receptor (IP(3)R1), the channel implicated, undergoes modifications that enhance its function. We found that IP(3)R1 becomes phosphorylated during maturation at an MPM-2 epitope and that this persists until the fertilization-associated [Ca2+](i) responses cease. We also reported that maturation without ERK activity diminishes IP(3)R1 MPM-2 reactivity and [Ca2+](i) responses. Here, we show that IP(3)R1 is a novel target for Polo-like kinase1 (Plk1), a conserved M-phase kinase, which phosphorylates it at an MPM-2 epitope. Plk1 and IP(3)R1 interact in an M-phase preferential manner, and they exhibit close co-localization in the spindle/spindle poles area. This co-localization is reduced in the absence of ERK activity, as the ERK pathway regulates spindle organization and IP(3)R1 cortical re-distribution. We propose that IP(3)R1 phosphorylation by Plk1, and possibly by other M-phase kinases, underlies the delivery of spatially and temporally regulated [Ca2+](i) signals during meiosis/mitosis and cytokinesis.
1 aIto, Junya1 aYoon, Sook-Young1 aLee, Bora1 aVanderheyden, Veerle1 aVermassen, Elke1 aWojcikiewicz, Richard1 aAlfandari, Dominique1 aDe Smedt, Humbert1 aParys, Jan, B1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/inositol-145-trisphosphate-receptor-1-a-widespread-ca2-channel-is-a03310nas a2200481 4500008004100000245013300041210006900174260001500243300000900258490000700267520174400274653001202018653001202030653004902042653004202091653002002133653003002153653002002183653002602203653001102229653004602240653002302286653002002309653001102329653003402340653000902374653000902383653002302392653002102415653001302436653001402449653003802463653001902501653002202520653001302542100002202555700001702577700002102594700001702615700002402632700002602656856014602682 2008 eng d00aManipulations of mouse embryos prior to implantation result in aberrant expression of imprinted genes on day 9.5 of development.0 aManipulations of mouse embryos prior to implantation result in a c2008 Jan 1 a1-140 v173 aIn vitro culture of mouse embryos results in loss of imprinting. The aim of the present study was to examine how two of the techniques commonly used during assisted reproduction, namely embryo culture and embryo transfer, affect genomic imprinting after implantation in the mouse. F1 hybrid mouse embryos were subjected to three experimental conditions: control (unmanipulated), embryo transfer and in-vitro-culture followed by embryo transfer. Concepti were collected on d9.5 of development and allelic expression determination of ten imprinted genes (H19, Snrpn, Igf2, Kcnq1ot1, Cdkn1c, Kcnq1, Mknr3, Ascl2, Zim1, Peg3) was performed. Although control concepti had monoallelic imprinted gene expression in all tissues, both manipulated groups had aberrant expression of one or more imprinted genes in the yolk sac and placenta. Culture further exacerbated the effects of transfer by increasing the number of genes with aberrant allelic expression in extraembryonic, as well as embryonic tissues. Additionally, placentae of both groups of manipulated concepti exhibited reduced levels of Igf2 mRNA and increased levels of Ascl2 mRNA when compared with their unmanipulated counterparts. Furthermore, we show that biallelic expression of Kcnq1ot1 coincided with loss of methylation on the maternal allele of the KvDMR1 locus, a phenotype often associated with the human syndrome Beckwith-Wiedemann. In conclusion, our results show that even the most basic manipulation used during human-assisted reproduction, namely, embryo transfer, can lead to misexpression of several imprinted genes during post-implantation development. Additionally, our results serve as a cautionary tale for gene expression studies in which embryo transfer is used.10aAlleles10aAnimals10aBasic Helix-Loop-Helix Transcription Factors10aCyclin-Dependent Kinase Inhibitor p5710aDNA Methylation10aEmbryo Culture Techniques10aEmbryo Transfer10aEmbryonic Development10aFemale10aGene Expression Regulation, Developmental10aGenomic Imprinting10aGestational Age10aHumans10aInsulin-Like Growth Factor II10aMale10aMice10aMice, Inbred C57BL10aMice, Transgenic10aPlacenta10aPregnancy10aReproductive Techniques, Assisted10aRNA, Messenger10aRNA, Untranslated10aYolk Sac1 aRivera, Rocío, M1 aStein, Paula1 aWeaver, Jamie, R1 aMager, Jesse1 aSchultz, Richard, M1 aBartolomei, Marisa, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/manipulations-of-mouse-embryos-prior-to-implantation-result-in01150nas a2200145 4500008004100000245011900041210006900160260001300229300001100242490000700253520055300260100002000813700002500833856014600858 2008 eng d00aNeurotoxic implications of the agonistic action of CS-syndrome pyrethroids on the N-type Ca(v)2.2 calcium channel.0 aNeurotoxic implications of the agonistic action of CSsyndrome py c2008 Jun a628-380 v643 aCismethrin (T-syndrome) and deltamethrin (CS-syndrome) pyrethroids have been previously shown to increase membrane depolarization and calcium influx, but only deltamethrin increased Ca(2+)-dependent neurotransmitter release from rat brain synaptosomes. Deltamethrin's action was blocked by omega-conotoxin GVIA, delineating a separate action at N-type Ca(v)2.2 channels that is consistent with the in vivo release of neurotransmitter. It is hypothesized that other CS-syndrome pyrethroids will elicit similar actions at presynaptic nerve terminals.1 aClark, Marshall1 aSymington, Steven, B uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/neurotoxic-implications-of-the-agonistic-action-of-cs-syndrome01668nas a2200157 4500008004100000245011100041210006900152260001300221300001000234490000700244520104400251100002501295700002101320700002001341856014901361 2008 eng d00aA new ivermectin formulation topically kills permethrin-resistant human head lice (Anoplura: Pediculidae).0 anew ivermectin formulation topically kills permethrinresistant h c2008 Jan a75-810 v453 aThis study examines the effectiveness of a new ivermectin formulation for the topical treatment of the human head louse, Pediculus humanus capitis De Geer (Anoplura: Pediculidae). Permethrin-resistant lice originally obtained from south Florida and maintained on an in vitro rearing system were 100% susceptible to ivermectin formulations by using a semiclinical hair tuft bioassay. The formulation was 100% effective at killing lice using 1, 0.5, and 0.25% ivermectin concentrations after 10-min exposures. As judged by the lethal time (LT)50 and LT95 values, 0.5% formulated ivermectin was 3.8 and 3.2 times faster at killing lice, respectively, than 0.5% nonformulated ivermectin, indicating that the formulation may facilitate the penetration of ivermectin into the louse. The hair tuft-based bioassay in conjunction with the in vitro rearing system provides a standardized method to assess the comparative efficacy of pediculicide formulations in a reproducible format that mimics the exposure scenario that occurs on the human scalp.1 aStrycharz, Joseph, P1 aYoon, Kyong, Sup1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/a-new-ivermectin-formulation-topically-kills-permethrin-resistant02110nas a2200229 4500008004100000245011800041210006900159260001500228300001200243490000800255520128900263100002101552700002201573700002001595700002201615700001701637700001601654700001601670700001901686700002401705856015101729 2008 eng d00aNotch1 and TGFbeta1 cooperatively regulate Foxp3 expression and the maintenance of peripheral regulatory T cells.0 aNotch1 and TGFbeta1 cooperatively regulate Foxp3 expression and c2008 Sep 1 a1813-210 v1123 aNotch and its ligands have been implicated in the regulation and differentiation of various CD4(+) T-helper cells. Regulatory T cells (T(regs)), which express the transcription factor Foxp3, suppress aberrant immune responses that are typically associated with autoimmunity or excessive inflammation. Previous studies have shown that transforming growth factor beta (TGFbeta1) induces Foxp3 expression and a regulatory phenotype in peripheral T cells. Here, we show that pharmacologic inhibition of Notch signaling using gamma-secretase inhibitor (GSI) treatment blocks (1) TGFbeta1-induced Foxp3 expression, (2) the up-regulation of Foxp3-target genes, and (3) the ability to suppress naive T-cell proliferation. In addition, the binding of Notch1, CSL, and Smad to conserved binding sites in the foxp3 promoter can be inhibited by treatment with GSI. Finally, in vivo administration of GSI results in reduced Foxp3 expression and development of symptoms consistent with autoimmune hepatitis, a disease previously found to result from dysregulation of TGFbeta signaling and regulatory T cells. Together, these findings indicate that the Notch and TGFbeta signaling pathways cooperatively regulate Foxp3 expression and regulatory T-cell maintenance both in vitro and in vivo.
1 aSamon, Jeremy, B1 aChamphekar, Ameya1 aMinter, Lisa, M1 aTelfer, Janice, C1 aMiele, Lucio1 aFauq, Abdul1 aDas, Pritam1 aGolde, Todd, E1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch1-and-tgfbeta1-cooperatively-regulate-foxp3-expression-and-the02012nas a2200157 4500008004100000245007500041210006900116260001500185300001000200490000800210520141200218100002101630700002501651700002501676856015301701 2008 eng d00aPACSIN2 regulates cell adhesion during gastrulation in Xenopus laevis.0 aPACSIN2 regulates cell adhesion during gastrulation in Xenopus l c2008 Jul 1 a86-990 v3193 aWe previously identified the adaptor protein PACSIN2 as a negative regulator of ADAM13 proteolytic function. In Xenopus embryos, PACSIN2 is ubiquitously expressed, suggesting that PACSIN2 may control other proteins during development. To investigate this possibility, we studied PACSIN2 function during Xenopus gastrulation and in XTC cells. Our results show that PACSIN2 is localized to the plasma membrane via its coiled-coil domain. We also show that increased levels of PACSIN2 in embryos inhibit gastrulation, fibronectin (FN) fibrillogenesis and the ability of ectodermal cells to spread on a FN substrate. These effects require PACSIN2 coiled-coil domain and are not due to a reduction of FN or integrin expression and/or trafficking. The expression of a Mitochondria Anchored PACSIN2 (PACSIN2-MA) sequesters wild type PACSIN2 to mitochondria, and blocks gastrulation without interfering with cell spreading or FN fibrillogenesis but perturbs both epiboly and convergence/extension. In XTC cells, the over-expression of PACSIN2 but not PACSIN2-MA prevents the localization of integrin beta1 to focal adhesions (FA) and filamin to stress fiber. PACSIN2-MA prevents filamin localization to membrane ruffles but not to stress fiber. We propose that PACSIN2 may regulate gastrulation by controlling the population of activated alpha5beta1 integrin and cytoskeleton strength during cell movement.
1 aCousin, Hélène1 aDesimone, Douglas, W1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pacsin2-regulates-cell-adhesion-during-gastrulation-in-xenopus-laevis02290nas a2200289 4500008004100000245007000041210006600111260001600177300001300193490000700206520137200213653001001585653002901595653002901624653001101653653001801664653001101682653001801693653002201711653001601733100001301749700002701762700001901789700002401808700002301832856014501855 2008 eng d00aPerfluorinated compounds in human milk from Massachusetts, U.S.A.0 aPerfluorinated compounds in human milk from Massachusetts USA c2008 Apr 15 a3096-1010 v423 aPerfluorinated compounds (PFCs), notably perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA), have been reported in human blood. Furthermore, the occurrence of PFCs in the blood of newborn babies, coupled with the need to study the potential association of PFC exposure with birth outcomes in neonates, suggests the need for determining the sources and magnitude of exposure in infants. In this study, nine PFCs were measured in 45 human breast milk samples collected in 2004 from Massachusetts, U.S.A. PFOS and PFOA were the predominant PFCs found at mean concentrations of 131 and 43.8 pg/mL, respectively. Comparison of the ratio of PFOS to PFOA in human milk with the ratios published for human serum from the U.S. female population suggested preferential partitioning of PFOA to milk. Concentrations of PFOA were significantly higher in the milk of mothers nursing for the first time (n = 34) than in the milk of mothers who have previously nursed (n = 8). Based on the estimated body weight and milk intake, the average and highest daily intakes of total PFCs by infants were 23.5 and 87.1 ng/kg bw, respectively. We found that the daily ingestion rates of PFOS and PFOA did not exceed the tolerable daily intake recommended by the U.K. Food Standards Agency. This is the first study to measure the occurrence of PFCs in human milk from the U.S.A.10aAdult10aEnvironmental Monitoring10aEnvironmental Pollutants10aFemale10aFluorocarbons10aHumans10aMassachusetts10aMaternal Exposure10aMilk, Human1 aTao, Lin1 aKannan, Kurunthachalam1 aWong, Chung, M1 aArcaro, Kathleen, F1 aButenhoff, John, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/perfluorinated-compounds-in-human-milk-from-massachusetts-usa01239nas a2200193 4500008004100000245010000041210006900141260000900210300001400219490000700233520053000240100002400770700001700794700002500811700001400836700001800850700002400868856015300892 2008 ENG d00aProtein-passivated Fe(3)O(4) nanoparticles: low toxicity and rapid heating for thermal therapy.0 aProteinpassivated Fe3O4 nanoparticles low toxicity and rapid hea c2008 a1204-12080 v183 aThermotherapy is a promising technique for the minimally invasive elimination of solid tumors. Here we report the fabrication of protein-coated iron oxide NPs (12 nm core) for use as thermal therapeutic agents. These albumin-passivated NPs are stable under physiological conditions, with rapid heating and cell killing capacity upon alternating magnetic field (AMF) exposure. The mode of action is specific: no measurable cytotoxicity was observed for the particle without AMF or for AMF exposure without the particle.
1 aSamanta, Bappaditya1 aYan, Haoheng1 aFischer, Nicholas, O1 aShi, Jing1 aJerry, Joseph1 aRotello, Vincent, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/protein-passivated-fe3o4-nanoparticles-low-toxicity-and-rapid-heating01054nas a2200157 4500008004100000245004400041210004300085260000900128300000800137490000700145520057000152100001800722700001500740700001700755856012400772 2008 eng d00aRegulation of cancer stem cells by p53.0 aRegulation of cancer stem cells by p53 c2008 a3040 v103 aThe hypothesis that cancer stem cells are responsible for the chemoresistant and metastatic phenotypes of many breast cancers has gained support using cell-sorting strategies to enrich the tumor-initiating population of cells. The mechanisms regulating the cancer stem cell pool, however, are less clear. Two recent publications suggest that loss of p53 permits expansion of presumptive cancer stem cells in mouse mammary tumors and in human breast cell lines. These results add restriction of cancer stem cells as a new tumor suppressor activity attributed to p53.1 aJerry, Joseph1 aTao, Luwei1 aYan, Haoheng uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-cancer-stem-cells-by-p53-001414nas a2200289 4500008004100000245004400041210004300085260000900128300000800137490000700145520057700152653001200729653004300741653001500784653001100799653000900810653002400819653002300843653001300866653002600879653001400905653003300919100001800952700001500970700001700985856012201002 2008 eng d00aRegulation of cancer stem cells by p53.0 aRegulation of cancer stem cells by p53 c2008 a3040 v103 aThe hypothesis that cancer stem cells are responsible for the chemoresistant and metastatic phenotypes of many breast cancers has gained support using cell-sorting strategies to enrich the tumor-initiating population of cells. The mechanisms regulating the cancer stem cell pool, however, are less clear. Two recent publications suggest that loss of p53 permits expansion of presumptive cancer stem cells in mouse mammary tumors and in human breast cell lines. These results add restriction of cancer stem cells as a new tumor suppressor activity attributed to p53.
10aAnimals10aGene Expression Regulation, Neoplastic10aGenes, p5310aHumans10aMice10aMice, Inbred BALB C10aModels, Biological10aMutation10aNeoplastic Stem Cells10aPhenotype10aTumor Suppressor Protein p531 aJerry, Joseph1 aTao, Luwei1 aYan, Haoheng uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-cancer-stem-cells-by-p5302621nas a2200265 4500008004100000245011900041210006900160260001300229300001200242490000800254520168800262100001601950700002101966700001901987700001702006700002002023700002302043700002502066700002402091700002102115700002402136700002602160700001802186856015102204 2008 eng d00aTranscriptional responses to estrogen and progesterone in mammary gland identify networks regulating p53 activity.0 aTranscriptional responses to estrogen and progesterone in mammar c2008 Oct a4809-200 v1493 aEstrogen and progestins are essential for mammary growth and differentiation but also enhance the activity of the p53 tumor suppressor protein in the mammary epithelium. However, the pathways by which these hormones regulate p53 activity are unknown. Microarrays were used to profile the transcriptional changes within the mammary gland after administration of either vehicle, 17beta-estradiol (E), or progesterone (P) individually and combined (EP). Treatment with EP yielded 1182 unique genes that were differentially expressed compared to the vehicle-treated group. Although 30% of genes were responsive to either E or P individually, combined treatment with both EP had a synergistic effect accounting for 60% of the differentially regulated genes. Analysis of protein-protein interactions identified p53, RelA, Snw1, and Igfals as common targets of genes regulated by EP. RelA and p53 form hubs within a network connected by genes that are regulated by EP and that may coordinate the competing functions of RelA and p53 in proliferation and survival of cells. Induction of early growth response 1 (Egr1) and Stratifin (Sfn) (also known as 14-3-3sigma) by EP was confirmed by reverse transcription-quantitative PCR and shown to be p53 independent. In luciferase reporter assays, Egr1 was shown to enhance transcriptional activation by p53 and inhibit nuclear factor kappaB activity. These results identify a gene expression network that provides redundant activation of RelA to support proliferation as well as sensitize p53 to ensure proper surveillance and integration of their competing functions through factors such as Egr1, which both enhance p53 and inhibit RelA.
1 aLu, Shaolei1 aBecker, Klaus, A1 aHagen, Mary, J1 aYan, Haoheng1 aRoberts, Amy, L1 aMathews, Lesley, A1 aSchneider, Sallie, S1 aSiegelmann, Hava, T1 aMacBeth, Kyle, J1 aTirrell, Stephen, M1 aBlanchard, Jeffrey, L1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transcriptional-responses-to-estrogen-and-progesterone-in-mammary-003359nas a2200529 4500008004100000245011900041210006900160260001300229300001200242490000800254520168800262653002001950653001201970653002101982653002702003653002102030653003602051653002102087653001502108653001402123653001102137653003002148653001102178653002702189653000902216653002402225653002502249653001502274653004402289653001602333653001702349653002702366653003302393100001602426700002102442700001902463700001702482700002002499700002302519700002502542700002402567700002102591700002402612700002602636700001802662856014902680 2008 eng d00aTranscriptional responses to estrogen and progesterone in mammary gland identify networks regulating p53 activity.0 aTranscriptional responses to estrogen and progesterone in mammar c2008 Oct a4809-200 v1493 aEstrogen and progestins are essential for mammary growth and differentiation but also enhance the activity of the p53 tumor suppressor protein in the mammary epithelium. However, the pathways by which these hormones regulate p53 activity are unknown. Microarrays were used to profile the transcriptional changes within the mammary gland after administration of either vehicle, 17beta-estradiol (E), or progesterone (P) individually and combined (EP). Treatment with EP yielded 1182 unique genes that were differentially expressed compared to the vehicle-treated group. Although 30% of genes were responsive to either E or P individually, combined treatment with both EP had a synergistic effect accounting for 60% of the differentially regulated genes. Analysis of protein-protein interactions identified p53, RelA, Snw1, and Igfals as common targets of genes regulated by EP. RelA and p53 form hubs within a network connected by genes that are regulated by EP and that may coordinate the competing functions of RelA and p53 in proliferation and survival of cells. Induction of early growth response 1 (Egr1) and Stratifin (Sfn) (also known as 14-3-3sigma) by EP was confirmed by reverse transcription-quantitative PCR and shown to be p53 independent. In luciferase reporter assays, Egr1 was shown to enhance transcriptional activation by p53 and inhibit nuclear factor kappaB activity. These results identify a gene expression network that provides redundant activation of RelA to support proliferation as well as sensitize p53 to ensure proper surveillance and integration of their competing functions through factors such as Egr1, which both enhance p53 and inhibit RelA.
10a14-3-3 Proteins10aAnimals10aBreast Neoplasms10aCell Line, Transformed10aCell Line, Tumor10aEarly Growth Response Protein 110aEpithelial Cells10aEpithelium10aEstradiol10aFemale10aGene Expression Profiling10aHumans10aMammary Glands, Animal10aMice10aMice, Inbred BALB C10aMice, Mutant Strains10aNF-kappa B10aOligonucleotide Array Sequence Analysis10aOvariectomy10aProgesterone10aTranscription, Genetic10aTumor Suppressor Protein p531 aLu, Shaolei1 aBecker, Klaus, A1 aHagen, Mary, J1 aYan, Haoheng1 aRoberts, Amy, L1 aMathews, Lesley, A1 aSchneider, Sallie, S1 aSiegelmann, Hava, T1 aMacBeth, Kyle, J1 aTirrell, Stephen, M1 aBlanchard, Jeffrey, L1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transcriptional-responses-to-estrogen-and-progesterone-in-mammary02399nas a2200169 4500008004100000245012600041210006900167260001300236300001100249490000700260520173200267100002301999700002002022700001702042700002102059856014902080 2007 eng d00aAnti-Trypanosoma brucei activity in Cape buffalo serum during the cryptic phase of parasitemia is mediated by antibodies.0 aAntiTrypanosoma brucei activity in Cape buffalo serum during the c2007 Oct a1391-90 v373 aCape buffalo are reservoir hosts of African trypanosomes. They rapidly suppress population growth of the highly antigenically variable extracellular haemoprotozoa and subsequently maintain a cryptic infection. Here we use in vitro cultures of trypanosomes cloned from Cape buffalo blood during cryptic infection, as well as related and unrelated trypanosomes, to identify anti-trypanosome components present in cryptic-phase infection serum. Trypanosome clone-specific complement-dependent trypanolytic IgM and IgG arose after appearance of target trypanosomes during cryptic infection. Serum collected late in the cryptic phase of infection contained complement-independent growth-inhibitory IgG which varied in activity among target trypanosomes. Removal of protein A/G-binding IgG from the serum restored its capacity to support trypanosome growth in vitro. Recovered growth-inhibitory IgG reacted with the variable surface glycoprotein (VSG) of parasites most affected by it, and reacted with trypanosome common antigens, notably the endosome-restricted tomato lectin-binding glycoproteins (TL-antigens). The inclusion of purified TL-antigens in culture medium did not affect the trypanosome growth-inhibitory activity of immune Cape buffalo serum. In addition, hyperimmune rabbit IgG against TL-antigens showed little or no binding to intact trypanosomes and did not affect trypanosome growth in vitro although it did react strongly with TL-antigens and trypanosome endosomes. We conclude that antibodies, particularly clone-specific (putatively VSG-specific) antibodies are responsible for the anti-trypanosome activity of cryptic phase infection serum consistent with a dominant role in parasite control in Cape buffalo.1 aGuirnalda, Patrick1 aMurphy, Noel, B1 aNolan, Derek1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/anti-trypanosoma-brucei-activity-in-cape-buffalo-serum-during-the02318nas a2200241 4500008004100000245011000041210006900151260001300220300001000233490000600243520148100249100002401730700002001754700001701774700002001791700001901811700001901830700001901849700001701868700002201885700002401907856014501931 2007 eng d00aBrucella abortus bacA mutant induces greater pro-inflammatory cytokines than the wild-type parent strain.0 aBrucella abortus bacA mutant induces greater proinflammatory cyt c2007 Jan a55-620 v93 aThe inner-membrane protein BacA affects Brucella LPS structure. A bacA deletion mutant of Brucella abortus, known as KL7 (bacA(mut)-KL7), is attenuated in BALB/c mice and protects against challenge. Thus, bacA mutation was a candidate for incorporation into live attenuated vaccines. We assessed bacA(mut)-KL7 in 2 additional mouse strains: the more resistant C57BL/6 that produces interferon-gamma throughout the infection and the highly susceptible interferon-gamma-deficient C57BL/6 in which brucellae exhibit continual exponential growth. While it was hypothesized that bacA(mut)-KL7 would exhibit even greater attenuation relative to its parent strain B. abortus 2308 in C57BL/6 mice than it did in BALB/c mice, this was not the case. Moreover, it was more pathogenic in C57BL/6 interferon-gamma-deficient mice than 2308 causing abscesses and wasting even though the splenic loads of bacA(mut)-KL7 were significantly lower. These 2 observations were correlated, respectively, with an ability of IFNgamma-activated macrophages to equivalently control strains 2308 and bacA(mut)-KL7 and the ability of bacA(mut)-KL7 organism and its LPS to induce greater amounts of pro-inflammatory cytokines than 2308. We conclude that attenuation properties of bacA mutation are dependent upon the nature of the host but more importantly that bacterial gene deletion can result in increased host pathology without an increase in bacterial load, crucial considerations for vaccine design.1 aParent, Michelle, A1 aGoenka, Radhika1 aMurphy, Erin1 aLevier, Kristen1 aCarreiro, Nuno1 aGolding, Basil1 aFerguson, Gail1 aRoop, Martin1 aWalker, Graham, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/brucella-abortus-baca-mutant-induces-greater-pro-inflammatory02479nas a2200181 4500008004100000245014300041210006900184260001300253300001200266490000700278520175300285100002302038700002502061700001402086700002202100700002402122856015102146 2007 eng d00aComparison of gene expression by co-cultured WC1+ gammadelta and CD4+ alphabeta T cells exhibiting a recall response to bacterial antigen.0 aComparison of gene expression by cocultured WC1 gammadelta and C c2007 Mar a2023-350 v443 aImmunization of cattle with a Leptospira borgpetersenii serovar hardjo-bovis vaccine results in the development of a recall response by WC1(+) gammadelta T cells and CD4(+) alphabeta T cells characterized by proliferation and interferon-gamma production. It was hypothesized that these two T cell subpopulations had largely redundant effector functions, principally differing in their requirements for activation. To test this, gene expression in cells proliferating to antigen were compared utilizing RT-PCR and bovine microarrays. Both T cell populations had similar transcript profiles for effector molecules, including IFN-gamma, FasL and granzyme B. In contrast, transcripts for costimulatory receptors and ligands were notably different following activation, as WC1(+) T cells expressed no or lower levels of transcripts for CD28 and CD40L, while CD4(+) T cells expressed substantial levels of both. However, both cell types had high levels of CTLA-4 transcript suggesting the cells may be regulated similarly following activation but differ in their need for and ability to provide costimulation. Microarray analyses to extend the number of genes examined revealed that while both subpopulations upregulated anti-apoptotic genes as well as those involved in cell activation and protein biosynthesis, overall there were limited differences between the two antigen-activated cell populations. Those genes that did differ were involved in cell signaling, protein production and intracellular protein trafficking. These results strengthen the hypothesis that these particular activated WC1(+) and CD4(+) T cells have overlapping effector functions and therefore may differ principally with regard to how they are recruited into immune responses.1 aBlumerman, Seth, L1 aHerzig, Carolyn, T A1 aWang, Fei1 aCoussens, Paul, M1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/comparison-of-gene-expression-by-co-cultured-wc1-gammadelta-and-cd402609nas a2200433 4500008004100000245012900041210006900170260001300239300001100252490000700263520114300270653001201413653001801425653001801443653002301461653002501484653001901509653003501528653002401563653005701587653003701644653000901681653002101690653004301711653001801754653001701772100002601789700002401815700002001839700001701859700002301876700002201899700001901921700001901940700002201959700002301981700001902004856015202023 2007 eng d00aDeletion of CD4 and CD8 coreceptors permits generation of alphabetaT cells that recognize antigens independently of the MHC.0 aDeletion of CD4 and CD8 coreceptors permits generation of alphab c2007 Nov a735-500 v273 aThe thymus generates major histocompatibility complex (MHC)-restricted alphabetaT cells that only recognize antigenic ligands in association with MHC or MHC-like molecules. We hypothesized that MHC specificity might be imposed on a broader alphabetaTCR repertoire during thymic selection by CD4 and CD8 coreceptors that bind and effectively sequester the tyrosine kinase Lck, thereby preventing T cell receptor (TCR) signaling by non-MHC ligands that do not engage either coreceptor. This hypothesis predicts that, in coreceptor-deficient mice, alphabeta thymocytes would be signaled by non-MHC ligands to differentiate into alphabetaT cells lacking MHC specificity. We now report that MHC-independent alphabetaT cells were indeed generated in mice deficient in both coreceptors as well as MHC ("quad-deficient" mice) and that such mice contained a diverse alphabetaT cell repertoire whose MHC independence was confirmed at the clonal level. We conclude that CD4 and CD8 coreceptors impose MHC specificity on a broader alphabetaTCR repertoire during thymic selection by preventing thymocytes from being signaled by non-MHC ligands.
10aAnimals10aAntigens, CD410aAntigens, CD810aBlotting, Northern10aCell Differentiation10aFlow Cytometry10aFluorescent Antibody Technique10aImmunoprecipitation10aLymphocyte Specific Protein Tyrosine Kinase p56(lck)10aMajor Histocompatibility Complex10aMice10aMice, Transgenic10aReceptors, Antigen, T-Cell, alpha-beta10aT-Lymphocytes10aThymus Gland1 aVan Laethem, Francois1 aSarafova, Sophia, D1 aPark, Jung-Hyun1 aTai, Xuguang1 aPobezinsky, Leonid1 aGuinter, Terry, I1 aAdoro, Stanley1 aAdams, Anthony1 aSharrow, Susan, O1 aFeigenbaum, Lionel1 aSinger, Alfred uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/deletion-of-cd4-and-cd8-coreceptors-permits-generation-of-alphabetat00975nas a2200181 4500008004100000245010600041210006900147260001300216300001200229490000700241520028400248100002000532700002100552700002200573700002400595700002200619856015200641 2007 eng d00aEarly laminar events involving endothelial activation in horses with black walnut- induced laminitis.0 aEarly laminar events involving endothelial activation in horses c2007 Nov a1205-110 v683 aTo determine proinflammatory gene expression, endothelial adhesion molecule gene expression, and matrix metalloproteinase (MMP) concentrations in laminar specimens at 1.5 hours after administration of black walnut extract (BWE) and to compare these values with later time points.1 aLoftus, John, P1 aBlack, Samuel, J1 aPettigrew, Amanda1 aAbrahamsen, Eric, J1 aBelknap, James, K uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/early-laminar-events-involving-endothelial-activation-in-horses-with03205nas a2200553 4500008004100000245011100041210006900152260001300221300001200234490000800246520150100254653001201755653002101767653002301788653001101811653003001822653003801852653001301890653001101903653002101914653002701935653002601962653000901988653002401997653002302021653001102044653004402055653001702099653001802116653003302134100002502167700001902192700002002211700001402231700001302245700001602258700002202274700001702296700001602313700002502329700001702354700002502371700002202396700002102418700002302439700002002462700001802482856015102500 2007 eng d00aGenetic mapping in mice identifies DMBT1 as a candidate modifier of mammary tumors and breast cancer risk.0 aGenetic mapping in mice identifies DMBT1 as a candidate modifier c2007 Jun a2030-410 v1703 aLow-penetrance breast cancer susceptibility alleles seem to play a significant role in breast cancer risk but are difficult to identify in human cohorts. A genetic screen of 176 N2 backcross progeny of two Trp53(+/-) strains, BALB/c and C57BL/6, which differ in their susceptibility to mammary tumors, identified a modifier of mammary tumor susceptibility in an approximately 25-Mb interval on mouse chromosome 7 (designated SuprMam1). Relative to heterozygotes, homozygosity for BALB/c alleles of SuprMam1 significantly decreased mammary tumor latency from 70.7 to 61.1 weeks and increased risk twofold (P = 0.002). Dmbt1 (deleted in malignant brain tumors 1) was identified as a candidate modifier gene within the SuprMam1 interval because it was differentially expressed in mammary tissues from BALB/c-Trp53(+/-) and C57BL/6-Trp53(+/-) mice. Dmbt1 mRNA and protein was reduced in mammary glands of the susceptible BALB/c mice. Immunohistochemical staining demonstrated that DMBT1 protein expression was also significantly reduced in normal breast tissue from women with breast cancer (staining score, 1.8; n = 46) compared with cancer-free controls (staining score, 3.9; n = 53; P < 0.0001). These experiments demonstrate the use of Trp53(+/-) mice as a sensitized background to screen for low-penetrance modifiers of cancer. The results identify a novel mammary tumor susceptibility locus in mice and support a role for DMBT1 in suppression of mammary tumors in both mice and women.
10aAnimals10aBreast Neoplasms10aChromosome Mapping10aFemale10aGene Expression Profiling10aGenetic Predisposition to Disease10aGenotype10aHumans10aIntestine, Small10aMammary Glands, Animal10aMammary Glands, Human10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aMucins10aOligonucleotide Array Sequence Analysis10aRisk Factors10aSurvival Rate10aTumor Suppressor Protein p531 aBlackburn, Anneke, C1 aHill, Linda, Z1 aRoberts, Amy, L1 aWang, Jun1 aAud, Dee1 aJung, Jimmy1 aNikolcheva, Tania1 aAllard, John1 aPeltz, Gary1 aOtis, Christopher, N1 aCao, Qing, J1 aRicketts, Reva, St J1 aNaber, Stephen, P1 aMollenhauer, Jan1 aPoustka, Annemarie1 aMalamud, Daniel1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genetic-mapping-in-mice-identifies-dmbt1-as-a-candidate-modifier-of02557nas a2200325 4500008004100000245011100041210006900152260001300221300001200234490000800246520149100254100002501745700001901770700002001789700001401809700001301823700001601836700002201852700001701874700001601891700002501907700001701932700002501949700002201974700002101996700002302017700002002040700001802060856015302078 2007 eng d00aGenetic mapping in mice identifies DMBT1 as a candidate modifier of mammary tumors and breast cancer risk.0 aGenetic mapping in mice identifies DMBT1 as a candidate modifier c2007 Jun a2030-410 v1703 aLow-penetrance breast cancer susceptibility alleles seem to play a significant role in breast cancer risk but are difficult to identify in human cohorts. A genetic screen of 176 N2 backcross progeny of two Trp53(+/-) strains, BALB/c and C57BL/6, which differ in their susceptibility to mammary tumors, identified a modifier of mammary tumor susceptibility in an approximately 25-Mb interval on mouse chromosome 7 (designated SuprMam1). Relative to heterozygotes, homozygosity for BALB/c alleles of SuprMam1 significantly decreased mammary tumor latency from 70.7 to 61.1 weeks and increased risk twofold (P = 0.002). Dmbt1 (deleted in malignant brain tumors 1) was identified as a candidate modifier gene within the SuprMam1 interval because it was differentially expressed in mammary tissues from BALB/c-Trp53(+/-) and C57BL/6-Trp53(+/-) mice. Dmbt1 mRNA and protein was reduced in mammary glands of the susceptible BALB/c mice. Immunohistochemical staining demonstrated that DMBT1 protein expression was also significantly reduced in normal breast tissue from women with breast cancer (staining score, 1.8; n = 46) compared with cancer-free controls (staining score, 3.9; n = 53; P < 0.0001). These experiments demonstrate the use of Trp53(+/-) mice as a sensitized background to screen for low-penetrance modifiers of cancer. The results identify a novel mammary tumor susceptibility locus in mice and support a role for DMBT1 in suppression of mammary tumors in both mice and women.1 aBlackburn, Anneke, C1 aHill, Linda, Z1 aRoberts, Amy, L1 aWang, Jun1 aAud, Dee1 aJung, Jimmy1 aNikolcheva, Tania1 aAllard, John1 aPeltz, Gary1 aOtis, Christopher, N1 aCao, Qing, J1 aRicketts, Reva, St J1 aNaber, Stephen, P1 aMollenhauer, Jan1 aPoustka, Annemarie1 aMalamud, Daniel1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genetic-mapping-in-mice-identifies-dmbt1-as-a-candidate-modifier-of-000890nas a2200169 4500008004100000245014000041210006900181260001300250300001000263490000700273520021400280100001700494700001800511700002200529700001600551856015300567 2007 eng d00aLaminar xanthine oxidase, superoxide dismutase and catalase activities in the prodromal stage of black-walnut induced equine laminitis.0 aLaminar xanthine oxidase superoxide dismutase and catalase activ c2007 Jan a48-530 v393 aREASONS FOR STUDY: Xanthine oxidase (XO)-dependent production of superoxide anion and hydrogen peroxide, a characteristic of ischaemia-reperfusion injury, may contribute to the development of equine laminitis.1 aLoftus, J, P1 aBelknap, J, K1 aStankiewicz, K, M1 aBlack, S, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/laminar-xanthine-oxidase-superoxide-dismutase-and-catalase-activities02536nas a2200409 4500008004100000245006600041210006400107260001300171300001000184490000700194520129900201653001201500653002101512653001101533653001501544653003801559653001701597653000901614653003001623653000901653653002401662653002301686653000901709653003701718100002001755700001701775700001801792700002301810700002201833700001901855700001801874700002501892700001601917700002501933700002401958856014401982 2007 eng d00aMammary tumor modifiers in BALB/cJ mice heterozygous for p53.0 aMammary tumor modifiers in BALBcJ mice heterozygous for p53 c2007 May a300-90 v183 aBALB/c mice are predisposed to developing spontaneous mammary tumors, which are further increased in a p53 heterozygous state. C57BL/6J mice are resistant to induced mammary tumors and develop less than 1% mammary tumors in both wild-type and p53+/- states. To map modifiers of mammary tumorigenesis, we have established F1 and F2 crosses and backcrosses to BALB/cJ (N2-BALB/cJ) and C57BL/6J (N2-C57BL/6J) strains. All cohorts developed mammary carcinomas in p53+/- females, suggesting that multiple loci dominantly and recessively contributed to mammary tumorigenesis. We mapped two modifiers of mammary tumorigenesis in the BALB/cJ strain. Mtsm1 (mammary tumor susceptibility modifier), a dominant-acting modifier, is located on chromosome 7. Mtsm1 is suggestive for linkage to mammary tumorigenesis (p = 0.001). We have analyzed the Mtsm1 region to locate candidate genes by comparing it to a rat modifier region, Mcs3, which shares syntenic conservation with Mtsm1. Expression data and SNPs were also taken into account. Five potential candidate genes within Mtsm1 are Aldh1a3, Chd2, Nipa2, Pcsk6, and Tubgcp5. The second modifier mapped is Mtsm2, a recessive-acting modifier. Mtsm2 is located on chromosome X and is significantly linked to mammary tumorigenesis (p = 1.03 x 10(-7)).
10aAnimals10aCrosses, Genetic10aFemale10aGenes, p5310aGenetic Predisposition to Disease10aHeterozygote10aMale10aMammary Neoplasms, Animal10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aRats10aSpecific Pathogen-Free Organisms1 aKoch, Joanna, G1 aGu, Xiangjun1 aHan, Younghun1 aEl-Naggar, Adel, K1 aOlson, Melissa, V1 aMedina, Daniel1 aJerry, Joseph1 aBlackburn, Anneke, C1 aPeltz, Gary1 aAmos, Christopher, I1 aLozano, Guillermina uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mammary-tumor-modifiers-in-balb-cj-mice-heterozygous-for-p5301766nas a2200169 4500008004100000245009500041210006900136260000900205300001100214490000700225520112500232100002301357700001401380700002501394700002401419856015301443 2007 eng d00aMolecular cloning of bovine chemokine receptors and expression by WC1+ gammadelta T cells.0 aMolecular cloning of bovine chemokine receptors and expression b c2007 a87-1020 v313 aChemokine receptors mediate leukocyte migration into secondary lymphoid tissues and localization to peripheral inflammation sites. We describe full-length cDNA sequences of bovine chemokine receptors CCR5, CCR7, CXCR3 and CXCR5 and transcript expression by WC1(+)gammadelta T cells, a unique cell population with proinflammatory characteristics that comprises a large proportion of mononuclear cells in young ruminants. Bovine chemokine sequences were more similar to those of humans than were murine sequences to humans', ranging from 84% to 91%. Transcript analysis showed that antigen stimulation of WC1(+)gammadelta T cells induced IFN-gamma production and substantially increased CCR5 and CXCR3 expression when compared with freshly isolated (ex vivo) cells. CCR7 transcripts were minimally expressed in ex vivo and proliferating WC1(+)gammadelta T cells and CXCR5 expression was negligible. These results confirm the proinflammatory nature of WC1(+)gammadelta T cells is reflected by its chemokine receptor expression and suggest WC1(+)gammadelta T cells are unlikely to transit through secondary lymphoid tissues.1 aBlumerman, Seth, L1 aWang, Fei1 aHerzig, Carolyn, T A1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/molecular-cloning-of-bovine-chemokine-receptors-and-expression-by-wc101005nas a2200145 4500008004100000245007800041210006900119260001300188300001000201490000600211520045800217100002400675700002000699856014000719 2007 eng d00aNotch signalling during peripheral T-cell activation and differentiation.0 aNotch signalling during peripheral Tcell activation and differen c2007 Jan a64-750 v73 aFor many years, researchers have focused on the contribution of Notch signalling to lymphoid development. Only recently have investigators begun to ask what role, if any, Notch has during the activation and differentiation of naive CD4(+) T cells in the periphery. As interest in this issue grows, it is becoming increasingly clear that the main role of Notch signalling, to regulate cell-fate decisions, might also be influential in peripheral T cells.1 aOsborne, Barbara, A1 aMinter, Lisa, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch-signalling-during-peripheral-t-cell-activation-and02778nas a2200349 4500008004100000245011100041210006900152260001300221300001200234490000600246520159800252653001001850653002701860653001101887653004101898653001101939653003001950653002001980653001602000653001602016653001502032653002902047653002002076653001802096653005002114100002802164700001902192700001602211700002102227700002702248856015302275 2007 eng d00aPolybrominated diphenyl ethers and organochlorine pesticides in human breast milk from Massachusetts, USA.0 aPolybrominated diphenyl ethers and organochlorine pesticides in c2007 Nov a1205-120 v93 aConcentrations of polybrominated diphenyl ethers (PBDEs), and organochlorine pesticides (OCPs; DDTs, HCHs, CHLs, and HCB) were measured in human breast milk samples collected across Massachusetts, USA, in 2004. Seventeen PBDE congeners were found in the samples, ranging in concentration from 0.06 to 1910 ng g(-1) lipid wt. BDE-47 (2,2',4,4'-tetraBDE), BDE-99 (2,2',4,4',5-pentaBDE), and BDE-100 (2,2',4,4',6-pentaBDE) were the major congeners detected in breast milk samples. Overall mean (+/-SD) concentrations of DDTs, HCHs, CHLs, and HCB were 64.5 +/- 75, 18.9 +/- 19, 32.4 +/- 36, and 2.3 +/- 2.2 ng g(-1) lipid wt, respectively. Concentrations of PBDEs were strongly correlated with concentrations of OCPs in the samples. Based on the concentrations of organohalogens and the intake rates of breast milk by infants in the United States, daily ingestion rates of contaminants were calculated. The median ingestion rates for PBDEs, HCHs, DDTs, CHLs, and HCB were 4.0, 212, 141, 44, and 5.79 ng kg(-1) body wt day(-1), respectively. The estimated daily intake of organohalogens by infants was compared with threshold reference values suggested by the United States Environmental Protection Agency (USEPA) and the Agency for Toxic Substances and Disease Registry (ATSDR), for calculation of hazard quotients (HQs). HQs for individual organohalogens and the sum of HQ for all organohalogens were calculated as HQ indices (HQI). The results suggest that one or more of the contaminants analyzed in this study exceeded the threshold reference values in at least 26% of the breast milk samples.10aAdult10aEnvironmental Exposure10aFemale10aGas Chromatography-Mass Spectrometry10aHumans10aHydrocarbons, Chlorinated10aInfant, Newborn10aMiddle Aged10aMilk, Human10aPesticides10aPolybrominated Biphenyls10aQuality Control10aUnited States10aUnited States Environmental Protection Agency1 aJohnson-Restrepo, Boris1 aAddink, Rudolf1 aWong, Chung1 aArcaro, Kathleen1 aKannan, Kurunthachalam uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/polybrominated-diphenyl-ethers-and-organochlorine-pesticides-in-human02231nas a2200193 4500008004100000245010700041210006900148260001500217300001100232490000800243520150600251100002101757700002201778700002501800700001901825700001901844700002301863856015101886 2007 eng d00aProteolytic processing of phospholipase Czeta and [Ca2+]i oscillations during mammalian fertilization.0 aProteolytic processing of phospholipase Czeta and Ca2i oscillati c2007 Dec 1 a407-180 v3123 aPhospholipase Czeta (PLCzeta) is a sperm-specific PLC capable of causing repetitive intracellular Ca2+ ([Ca2+]i) release ([Ca2+]i oscillations) in mammalian eggs. Accumulating evidence suggests that PLCzeta is the sperm factor responsible for inducing egg activation. Nevertheless, some sperm fractions devoid of 72-kDa PLCzeta showed [Ca2+]i oscillation-inducing and PLCzeta-like PLC activity (Kurokawa et al., (2005) Dev. Biol. 285, 376-392). Here, we report that PLCzeta remains functional after proteolytic cleavage at the X-Y linker region. We found that N-terminal (33 and 37 kDa) and C-terminal fragments (27 kDa), presumably the result of PLCzeta cleavage at the X-Y linker region, were present in fresh sperm as well as in sperm extracts and remained associated as functional complexes. Protease V8 cleaved 72-kDa PLCzeta into 33/37 and 27 kDa fragments, while PLC activity and [Ca2+]i oscillation-inducing activity persisted until degradation of the fragments. Immunodepletion or affinity depletion of these fragments abolished PLC activity and [Ca2+]i oscillation-inducing activity from sperm extracts. Lastly, co-expression of cRNAs encoding residues 1-361 and 362-647 of mouse PLCzeta, mimicking cleavage at the X-Y linker region, induced [Ca2+]i oscillations and embryo development in mouse eggs. Our results support the hypothesis that PLCzeta is the sole mammalian sperm factor and that its linker region may have important regulatory functions during mammalian fertilization.
1 aKurokawa, Manabu1 aYoon, Sook, Young1 aAlfandari, Dominique1 aFukami, Kiyoko1 aSato, Ken-ichi1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/proteolytic-processing-of-phospholipase-czeta-and-ca2i-oscillations01898nas a2200145 4500008004100000245007600041210006900117260001300186300000900199490000600208520134000214100002001554700002501574856015301599 2007 eng d00aPyrethroid action on calcium channels: neurotoxicological implications.0 aPyrethroid action on calcium channels neurotoxicological implica c2007 Mar a3-160 v73 aActions of cismethrin versus deltamethrin were compared using two functional attributes of rat brain synaptosomes. Both pyrethroids increased calcium influx but only deltamethrin increased Ca(2+)-dependent neurotransmitter release following K(+)-stimulated depolarization. The action of deltamethrin was stereospecific, concentration-dependent, and blocked by omega-conotoxin GVIA. These findings delineate a separate action for deltamethrin and implicate N-type rat brain Ca(v)2.2 voltage-sensitive calcium channels (VSCC) as target sites that are consistent with the in vivo release of neurotransmitter caused by deltamethrin. Deltamethrin (10(-7) M) reduced the peak current (approx. -47%) of heterologously expressed wild type Ca(v)2.2 in a stereospecific manner. Mutation of threonine 422 to glutamic acid (T422E) in the alpha(1)-subunit results in a channel that functions as if it were permanently phosphorylated. Deltamethrin now increased peak current (approx. +49%) of T422E Ca(v)2.2 in a stereospecific manner. Collectively, these results substantiate that Ca(v)2.2 is directly modified by deltamethrin but the resulting perturbation is dependent upon the phosphorylation state of Ca(v)2.2. Our findings may provide a partial explanation for the different toxic syndromes produced by these structurally-distinct pyrethroids.1 aClark, Marshall1 aSymington, Steven, B uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pyrethroid-action-on-calcium-channels-neurotoxicological-implications01788nas a2200289 4500008004100000245006900041210006800110260000900178300000800187490000600195520086600201653001201067653002601079653002101105653003601126653002701162653001401189653001101203653004301214653001101257653003001268653000901298653001701307653000901324100001801333856014701351 2007 eng d00aRoles for estrogen and progesterone in breast cancer prevention.0 aRoles for estrogen and progesterone in breast cancer prevention c2007 a1020 v93 aPrevention has long been the holy grail of breast cancer research. The significant reduction in breast cancer risk afforded by a full-term pregnancy early in life suggests the great potential of preventive strategies. In contrast to the risks associated with prolonged exposures, exogenous estrogen and progesterone for short durations can mimic the protective effects of pregnancy in carcinogen-induced mammary tumor models. Rajkumar and coworkers have now demonstrated that these hormones protect mice from mammary tumors initiated by a spectrum of oncogenic alterations that are common in breast cancers. Although differences between rodent models and humans remain, the results reveal that exogenous estrogen and progesterone potently inhibit tumorigenesis through multiple pathways and establish a foundation for strategies to prevent breast cancer.
10aAnimals10aAntineoplastic Agents10aBreast Neoplasms10aCell Transformation, Neoplastic10aDisease Models, Animal10aEstrogens10aFemale10aGene Expression Regulation, Neoplastic10aHumans10aMammary Neoplasms, Animal10aMice10aProgesterone10aRats1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/roles-for-estrogen-and-progesterone-in-breast-cancer-prevention01950nas a2200337 4500008004100000245005600041210005500097260000900152300001100161490000700172520096000179653002201139653001201161653001801173653001101191653000901202653001201211653002001223653002801243653002401271653002301295653001601318100002201334700001901356700002301375700001801398700002001416700001901436700002301455856013401478 2007 eng d00aSignalling pathways involved in sperm capacitation.0 aSignalling pathways involved in sperm capacitation c2007 a245-590 v653 aAfter ejaculation, mammalian sperm have not yet acquired full fertilising capacity. They will require a finite period of residence in the female reproductive tract before they become fertilisation competent. The molecular, biochemical, and physiological changes that occur to sperm while in the female tract are collectively referred to as capacitation. During capacitation, changes in membrane properties, enzyme activities, and motility render spermatozoa responsive to stimuli that induce the acrosome reaction and prepare spermatozoa for penetration of the egg investments prior to fertilisation. These changes are facilitated by the activation of cell signalling cascades in the female reproductive tract in vivo or in defined media in vitro. The purposes of this review are to consider some recent contributions towards our understanding of capacitation, to summarise open questions in this field, and to discuss future avenues of research.
10aAcrosome Reaction10aAnimals10aCell Membrane10aFemale10aMale10aMammals10aPhosphorylation10aSeminal Plasma Proteins10aSignal Transduction10aSperm Capacitation10aSpermatozoa1 aSalicioni, Ana, M1 aPlatt, Mark, D1 aWertheimer, Eva, V1 aArcelay, Enid1 aAllaire, Alicia1 aSosnik, Julian1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/signalling-pathways-involved-in-sperm-capacitation03369nas a2200325 4500008004100000245006800041210006700109260001500176300001200191490000700203520238000210653001002590653001602600653002702616653001102643653001102654653001102665653001802676653001602694653001202710653002502722653002702747653001802774653001202792100002302804700001902827700002402846700002702870856014602897 2007 eng d00aSynthetic musk fragrances in human milk from the United States.0 aSynthetic musk fragrances in human milk from the United States c2007 Jun 1 a3815-200 v413 aSynthetic musk compounds are used as additives in many consumer products, including perfumes, deodorants, and detergents. Earlier studies have reported the occurrence of synthetic musks in environmental and wildlife samples collected in the United States. In this study, human breast milk samples collected from Massachusetts, were analyzed for the determination of concentrations of synthetic musks such as musk xylene (1-tert-butyl-3,5-dimethyl-2,4,6-trinitrobenzene), musk ketone (4-tert-butyl-2,6-dimethyl-3,5-dinitroacetophenone), HHCB (1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta[gamma]-2-benzopyran), AHTN (7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydronaphthalene), and HHCB-lactone, the oxidation product of HHCB. In addition, we estimated the daily intake of synthetic musks by infants based on the ingestion rate of breast milk. Synthetic musks were found in most of the samples analyzed, and the concentrations ranged from < 2 to 150 ng musk xylene/g, < 2 to 238 ng musk ketone/ g, < 5 to 917 ng HHCB/g, < 5 to 144 ng AHTN/g, and < 10 to 88.0 ng HHCB-lactone/g, on a lipid weight basis. The concentrations of HHCB were higher than the concentrations of other synthetic musks in breast milk samples. The mean concentration of HHCB (220 ng/g, lipid weight) was 5 times greater than the concentrations reported 10 years ago for breast milk samples collected in Germany and Denmark. Maternal age was not correlated with the concentrations of musk xylene, musk ketone, HHCB, or AHTN. There was a trend of decreasing concentrations of musk xylene, musk ketone, HHCB, and AHTN, with the number of children previously breast-fed, although the correlation was not significant. Based on average daily ingestion rate of breast milk, an infant is estimated to ingest 297 +/- 229 ng musk xylene, 780 +/- 805 ng musk ketone, 1830 +/- 1170 ng HHCB, 565 +/- 614 ng AHTN, and 649 +/- 598 ng HHCB-lactone per day. The ingestion rate of synthetic musks by infants in the United States is lower than that estimated for persistent organic pollutants (POPs) such as polychlorinated biphenyls (PCBs). Based on the residue patterns and accumulation features, it can be concluded that the exposure characteristics for synthetic musks are different from those of POPs, and that the major source of exposure to synthetic musks is probably via dermal absorption or inhalation.10aAdult10aBenzopyrans10aEnvironmental Exposure10aFemale10aHumans10aInfant10aMassachusetts10aMilk, Human10aPerfume10aPolycyclic Compounds10aTetrahydronaphthalenes10aUnited States10aXylenes1 aReiner, Jessica, L1 aWong, Chung, M1 aArcaro, Kathleen, F1 aKannan, Kurunthachalam uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/synthetic-musk-fragrances-in-human-milk-from-the-united-states02022nas a2200157 4500008004100000245008500041210006900126260001300195300001200208490000700220520141100227100002301638700002501661700002401686856015401710 2007 eng d00aWC1+ gammadelta T cell memory population is induced by killed bacterial vaccine.0 aWC1 gammadelta T cell memory population is induced by killed bac c2007 May a1204-160 v373 aLimited studies have addressed the ability of gammadelta T cells to become memory populations. We previously demonstrated that WC1.1(+) gammadelta T cells from ruminants vaccinated with killed Leptospira borgpetersenii proliferate and produce IFN-gamma in recall responses. Here we show that this response is dependent upon antigen-responsive CD4 T cells, at least across transwell membranes; this requirement cannot be replaced by IL-2. The response was also dependent upon in vivo priming, since gammadelta T cells from leptospira vaccine-naive animals did not respond to antigen even when co-cultured across membranes from antigen-responsive PBMC. Gammadelta T cells were the major antigen-responding T cell population for the first 4 wks following vaccination and replicated more rapidly than CD4 T cells. Primed WC1(+) gammadelta T cells circulated as CD62L(hi)/CD45RO(int)/CD44(lo), characteristics of T(CM) cells. When stimulated with antigen, they decreased CD62L, increased CD44 and CD25, and had no change in CD45RO expression. These changes paralleled those of the leptospira antigen-responsive CD4 T cells but differed from those of gammadelta T cells proliferating to mitogen stimulation. This system for in vivo gammadelta T cell priming is unique, since it relies on a killed antigen to induce memory and may be pertinent to designing vaccines that require type 1 pro-inflammatory cytokines.1 aBlumerman, Seth, L1 aHerzig, Carolyn, T A1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/wc1-gammadelta-t-cell-memory-population-is-induced-by-killed-bacterial01886nas a2200217 4500008004100000245011400041210006900155260001300224300001100237490000800248520109100256100002001347700002501367700001801392700002201410700002501432700002001457700002201477700002401499856014501523 2006 eng d00aCharacterization of WC1 co-receptors on functionally distinct subpopulations of ruminant gamma delta T cells.0 aCharacterization of WC1 coreceptors on functionally distinct sub c2006 Feb a151-610 v2393 aWC1 molecules are implicated in augmenting cellular activation as well as inducing cell cycle arrest of gammadelta T cells. Since WC1 is a large multigene family differences in outcome could result from modulation of different WC1 molecules. To further investigate this family of molecules, peripheral blood WC1(+) gammadelta T cell subpopulations were evaluated by 2-D Western blotting and RT-PCR. We found 13 cDNA intracytoplasmic tail sequences with differences in signaling motifs among them and at least 20 biochemically distinguishable WC1 spots associated with cell membranes, with some in lipid rafts. An understanding of the diversity of 2-D spots could not be resolved by evaluating T cell clones, removing sialyated carbohydrates or blotting with anti-WC1.1 or anti-WC1.2-specific antibodies. Nevertheless, while the major gammadelta T cell subpopulations in blood (WC1.1(+)/WC1.2(-) and WC1.2(+)/WC1.1(-)) both had complex 2-D patterns, virtually all spots associated with WC1.2(+)/WC1.1(-) cells bore the WC1.2 epitope, distinguishing them from the WC1.1(+) cells.
1 aRogers, Aric, N1 aVanburen, Denille, G1 aZou, Baixiang1 aLahmers, Kevin, K1 aHerzig, Carolyn, T A1 aBrown, Wendy, C1 aTelfer, Janice, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/characterization-of-wc1-co-receptors-on-functionally-distinct02535nas a2200181 4500008004100000245014500041210006900186260001300255300001100268490000700279520180000286100002302086700002502109700002002134700002202154700002402176856015302200 2006 eng d00aDifferential TCR gene usage between WC1- and WC1+ ruminant gammadelta T cell subpopulations including those responding to bacterial antigen.0 aDifferential TCR gene usage between WC1 and WC1 ruminant gammade c2006 Aug a680-920 v583 aRuminant gammadelta T cells are divided into subpopulations based on the presence or absence of WC1 co-receptors (scavenger-receptor-cysteine-rich family members uniquely expressed on gammadelta T cells). Evidence suggests WC1+ are inflammatory while WC1- are regulatory and that they also differ in their tissue distribution. Recently, this paradigm was refined further as cells that produce interferon-gamma and proliferate to autologous antigens, leptospira antigens, or IL-12 were largely found within the WC1+ subpopulation that bears the WC1.1 antigenic epitope but not that bearing the WC1.2 epitope. Here, the T cell receptor gene expression by these different subpopulations (WC1-, WC1.1+, and WC1.2+) was compared using flow cytometrically-purified cells and reverse transcriptase-polymerase chain reaction (RT-PCR). The WC1- gammadelta T cells had transcripts for all 11 possible combinations of the TRG subgroup V and C genes while those in both WC1+ subpopulations were restricted to TRGV3-TRGC5 and TRGV7-TRGC5. In contrast, all three subpopulations expressed transcripts from all four known bovine TRDV genes. Further analysis of the WC1+ gammadelta T cells that proliferated in leptospira antigen-stimulated cultures indicated that they do not represent a unique subpopulation within the larger WC1+ population based on their TCR gene usage. Moreover, sequencing of 65 transcripts showed that their junctional regions were diverse as TRGJ5-1, TRGJ5-2, TRDJ1, and TRDJ3 were used, and CDR3s ranged from 9 to 24 amino acids. The restricted but shared gammadelta TCR gene usage for WC1.1+, WC1.2+, and WC1(+)-antigen-responsive cells leaves open the possibility that the WC1 co-receptor is an important determining element in the activation process and subsequent response.
1 aBlumerman, Seth, L1 aHerzig, Carolyn, T A1 aRogers, Aric, N1 aTelfer, Janice, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/differential-tcr-gene-usage-between-wc1-and-wc1-ruminant-gammadelta-t03280nas a2200541 4500008004100000245010900041210006900150260001300219300001000232490000700242520164800249653003201897653002201929653001201951653002701963653002301990653002202013653001602035653001502051653001102066653001602077653001502093653001502108653001102123653000902134653000902143653003902152653003902191653001202230653001302242653002002255653002002275653001202295653004302307653003702350653002602387653002302413653002302436653001602459100001402475700002002489700001502509700001702524700001402541700001502555700001902570856014902589 2006 eng d00aEvidence for the involvement of proline-directed serine/threonine phosphorylation in sperm capacitation.0 aEvidence for the involvement of prolinedirected serinethreonine c2006 Dec a781-90 v123 aTo become fertilization competent, mammalian sperm undergo changes in the female reproductive tract termed capacitation. Capacitation correlates with an increase in tyrosine phosphorylation; however, less is known about the role of serine/threonine phosphorylation in this process. Proline-directed phosphorylation is one of the major regulatory phosphorylation events in many cellular processes such as cell proliferation and differentiation. Using mitotic phosphoprotein monoclonal-2 (MPM-2) antibody in this study, we observed that several mouse sperm proteins in the range of 70-250 kDa underwent increased serine/threonine-proline phosphorylation during capacitation. In contrast to the time course of tyrosine phosphorylation, proline-directed phosphorylation could be observed at shorter time points of sperm incubation, and it was found to be independent of NaHCO(3) and adenosine 3'5'-cyclic monophosphate (cAMP). Similar to the regulation of the increase in tyrosine phosphorylation, cholesterol acceptors such as bovine serum albumin (BSA) or 2-hydroxypropyl-beta-cyclodextrin (2-OH-propyl-beta-CD) were essential for the regulation of proline-directed phosphorylation in mouse sperm. Furthermore, it was also found to be BSA dependent in human sperm. Among proline-directed kinases, extracellular signal-regulated kinase 1/2 (ERK1/2) is present in mammalian sperm; nevertheless, U0126 and PD098059, two inhibitors of the ERK pathway, did not block this phosphorylation in mouse sperm. In conclusion, capacitation is associated with an increase in proline-directed phosphorylation linked to cholesterol efflux in the sperm.
10a1-Methyl-3-isobutylxanthine10aAmino Acid Motifs10aAnimals10aAntibodies, Monoclonal10abeta-Cyclodextrins10aBlotting, Western10aBucladesine10aButadienes10aCattle10aCholesterol10aCyclic AMP10aFlavonoids10aHumans10aMale10aMice10aMitogen-Activated Protein Kinase 110aMitogen-Activated Protein Kinase 310aMitosis10aNitriles10aPhosphoproteins10aPhosphorylation10aProline10aProtein Processing, Post-Translational10aProtein-Serine-Threonine Kinases10aSerum Albumin, Bovine10aSodium Bicarbonate10aSperm Capacitation10aSpermatozoa1 aJha, K, N1 aSalicioni, A, M1 aArcelay, E1 aChertihin, O1 aKumari, S1 aHerr, J, C1 aVisconti, P, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evidence-for-the-involvement-of-proline-directed-serine-threonine02442nas a2200445 4500008004100000245009400041210006900135260001300204300001100217490000700228520105500235653001201290653002001302653002301322653003001345653002401375653001301399653003001412653001501442653001001457653003201467653002201499653001301521653000901534653002301543653002101566653001801587653003501605653002001640653001301660653002401673100002101697700001901718700002601737700002101763700002301784700002001807700002201827856014701849 2006 eng d00aAn FGF response pathway that mediates hepatic gene induction in embryonic endoderm cells.0 aFGF response pathway that mediates hepatic gene induction in emb c2006 Sep a339-480 v113 aWhile particular combinations of mesodermal signals are known to induce distinct tissue-specific programs in the endoderm, there is little information about the response pathways within endoderm cells that control their specification. We have used signaling inhibitors on embryo tissue explants and whole-embryo cultures as well as genetic approaches to reveal part of an intracellular network by which FGF signaling helps induce hepatic genes and stabilize nascent hepatic cells within the endodermal epithelium. Specifically, we found that hepatic gene induction is elicited by an FGF/MAPK pathway. Although the PI3K pathway is activated in foregut endoderm cells, its inhibition does not block hepatic gene induction in explants; however, it does block tissue growth. We also found that at the onset of hepatogenesis, the FGF/MAPK and PI3K pathways do not crossregulate in the endoderm. The finding of separate pathways for endoderm tissue specification and growth provides insights for guiding cellular regeneration and stem cell differentiation.10aAnimals10aBody Patterning10aCell Proliferation10aEmbryo Culture Techniques10aEmbryonic Induction10aEndoderm10aFibroblast Growth Factors10aIntegrases10aLiver10aMAP Kinase Signaling System10aMembrane Proteins10aMesoderm10aMice10aMice, Inbred C57BL10aMice, Transgenic10aOrganogenesis10aPhosphatidylinositol 3-Kinases10aPhosphorylation10aProteins10aSignal Transduction1 aCalmont, Amélie1 aWandzioch, Ewa1 aTremblay, Kimberly, D1 aMinowada, George1 aKaestner, Klaus, H1 aMartin, Gail, R1 aZaret, Kenneth, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/an-fgf-response-pathway-that-mediates-hepatic-gene-induction-in02601nas a2200421 4500008004100000245013700041210006900178260001500247300001000262490000800272520125700280653001201537653002001549653002501569653002301594653001301617653001501630653001101645653004601656653002201702653002501724653001001749653000901759653000901768653002301777653002501800653001301825653001801838653001801856653002101874653002601895100001601921700002101937700002301958700003001981700002202011856014602033 2006 eng d00aHex homeobox gene controls the transition of the endoderm to a pseudostratified, cell emergent epithelium for liver bud development.0 aHex homeobox gene controls the transition of the endoderm to a p c2006 Feb 1 a44-560 v2903 aLittle is known about the mechanism by which embryonic liver, lung, and pancreas progenitor cells emerge from the endodermal epithelium to initiate organogenesis. Understanding this process and its genetic control provides insight into ontogeny, developmental abnormalities, and tissue regeneration. We find that shortly after hepatic endoderm cells are specified, they undergo a transition from a columnar, gut morphology to a pseudostratified morphology, with concomitant "interkinetic nuclear migration" (INM) during cell division. INM is a hallmark of pseudostratified epithelia and the process used by neural progenitors to emerge from the neural epithelium. We find that the transition of the hepatic endoderm, but not the neural epithelium, to a pseudostratified epithelium is dependent upon the cell-autonomous activity of the homeobox gene Hex. In the absence of Hex, hepatic endoderm cells survive but maintain a columnar, simple epithelial phenotype and ectopically express Shh and other genes characteristic of the midgut epithelium. Thus, Hex promotes endoderm organogenesis by promoting the transition to a pseudostratified epithelium, which in turn allows hepatoblasts to emerge into the stromal environment and continue differentiating.10aAnimals10aBody Patterning10aCell Differentiation10aCell Proliferation10aEndoderm10aEpithelium10aFemale10aGene Expression Regulation, Developmental10aHedgehog Proteins10aHomeodomain Proteins10aLiver10aMale10aMice10aMice, Inbred C57BL10aMice, Mutant Strains10aMutation10aOrganogenesis10aStromal Cells10aTrans-Activators10aTranscription Factors1 aBort, Roque1 aSignore, Massimo1 aTremblay, Kimberly1 aBarbera, Juan, Pedro Mart1 aZaret, Kenneth, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/hex-homeobox-gene-controls-the-transition-of-the-endoderm-to-a02071nas a2200145 4500008004100000245013500041210006900176260000900245300001100254490000700265520145600272100002401728700002001752856015301772 2006 eng d00aHost immune responses to the intracellular bacteria Brucella: does the bacteria instruct the host to facilitate chronic infection?0 aHost immune responses to the intracellular bacteria Brucella doe c2006 a407-420 v263 aBrucella spp. are intracellular gram-negative bacteria that include a number of virulent species that cause chronic infections in a variety of mammalian hosts. Human infections are proportional to the level of disease in domestic animals because humans are infected zoonotically after contact with infected animals or their products. The chronicity of infection results from the ability of some brucellae to survive reactive oxygen intermediate and nitric oxide killing in host phagocytes, following which they activate bacterial genes in response to the acidic phagosome environment, prevent phagolysosomal fusion by remodeling the intracellular compartment, and subsequently replicate intracellularly. The crucial component of immunity that results in survival of the host and thus maintenance of this chronic infective state is interferon-gamma (IFN-gamma). Production of IFN-gamma results from the ability of brucella components, including lipid A, to interact with Toll-like receptors for the production of IL-12 and TNF-alpha, although the regulatory cytokine IL-10 is also produced and decreases control of the infection. Although CD4 and CD8 T cells are clearly involved in the production of IFN-gamma, and CD8 T cells may be cytotoxic, a role for NK cells and cytotoxicity in protective immunity to brucellosis has not been substantiated experimentally. Moreover, antibodies have been shown to have a limited role in passive transfer studies.1 aBaldwin, Cynthia, L1 aGoenka, Radhika uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/host-immune-responses-to-the-intracellular-bacteria-brucella-does-the02763nas a2200277 4500008004100000245010700041210006900148260001300217300001000230490000700240520177600247653001202023653001502035653001102050653003002061653000902091653002302100653004402123653005202167653002702219100001702246700002402263700002002287700002602307856015202333 2006 eng d00aIdentification of candidate maternal-effect genes through comparison of multiple microarray data sets.0 aIdentification of candidate maternaleffect genes through compari c2006 Sep a941-90 v173 aTranscriptional profiling by microarray hybridization has become a standard method to analyze global gene expression and has resulted in the availability of enormous amounts of experimental data. Given the number of different microarray platforms currently in use, it is critical to determine how reproducible results are from one platform to another. Additional variability may also arise from tissue collection and protocol differences among laboratories. In an effort to identify genes whose maternal mRNA pools are critical during preimplantation development, we compared published results of three independent studies of the mouse preimplantation embryo transcriptome, each performed in a different laboratory using different microarray platforms. We searched the combined data set for genes whose expression patterns were consistent among the three experiments. Querying for presence or absence at single developmental windows indicates that between 52% and 60% of genes are in agreement among the three experiments. Searching for expression patterns across three developmental windows (oocyte + 1-cell, 2- through 8-cell, and blastocyst stage) revealed approximately 33% agreement among the three experiments, although the majority of these genes were either always present or always absent. Using this approach, we identified 51 genes with a predicted expression pattern of maternal RNA only (not present during 2-cell through 8-cell or at the blastocyst stage). RT-PCR validation indicates 37 (72%) of these candidates have the microarray-predicted expression pattern and represent candidate maternal-effect genes. Based on our analysis, we conclude that data mining microarray experiments in this way greatly enhances candidate gene expression pattern accuracy.10aAnimals10aBlastocyst10aFemale10aGene Expression Profiling10aMice10aModels, Biological10aOligonucleotide Array Sequence Analysis10aReverse Transcriptase Polymerase Chain Reaction10aRNA, Messenger, Stored1 aMager, Jesse1 aSchultz, Richard, M1 aBrunk, Brian, P1 aBartolomei, Marisa, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-candidate-maternal-effect-genes-through-comparison02001nas a2200157 4500008004100000245012400041210006900165260001300234300001100247490000700258520135200265100002501617700002301642700002401665856015401689 2006 eng d00aIdentification of three new bovine T-cell receptor delta variable gene subgroups expressed by peripheral blood T cells.0 aIdentification of three new bovine Tcell receptor delta variable c2006 Sep a746-570 v583 aTo understand the biology of gammadelta T cells in ruminants, it is necessary to have a comprehensive picture of gammadelta T-cell receptor gene diversity and expression. In this study, three new subgroups of bovine T-cell receptor delta (TRD) variable genes were identified by RT-PCR and sequencing and homology with TRDV genes from other mammals determined. Previously unidentified TRDV subgroup genes described in this study include the bovine homologues of ovine TRDV2, TRDV3, and TRDV4 which were named accordingly. TRDV2 subgroup has two genes (TRDV2-1 and TRDV2-2) while we found the previously identified TRDV1 has at least eight genes corresponding to separate genomic sequences. Nucleotide and amino acid sequences for particular gene subgroups between cattle and sheep were more than 87% identical but identities among TRDV subgroups within a species were much less, with bovine TRDV4 having <45% identity to the other three bovine TRDV gene subgroups. Analysis of circulating bovine gammadelta T cells revealed that genes from all four TRDV subgroups were expressed in combination with TRDJ1, TRDJ3, and TRDC, although TRDV4 was the least represented, and all displayed a variety of CDR3 junctional lengths. Finally, some genes within the TRDV1, TRDV2, and TRDV3 subgroups recombined with TRAV incorporating TRAJs, suggesting dual use.1 aHerzig, Carolyn, T A1 aBlumerman, Seth, L1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-three-new-bovine-t-cell-receptor-delta-variable-gene01958nas a2200193 4500008004100000245005900041210005800100260001600158300001000174490000800184520132100192100002101513700001501534700001701549700001801566700002101584700002201605856013701627 2006 eng d00aLeukocyte emigration in the early stages of laminitis.0 aLeukocyte emigration in the early stages of laminitis c2006 Jan 15 a161-60 v1093 aThe mechanisms that initiate the pathophysiologic changes in the digital laminae in equine laminitis are poorly understood. Due to the fact that (1) the horse at risk of laminitis has many similarities clinically to the human sepsis patient and (2) our recent finding of marked laminar proinflammatory cytokine expression at the developmental time point of the black walnut extract (BWE) model of laminitis, we tested the possibility that, similar to organ damage in human sepsis, leukocyte emigration is an early event in laminitis. Using immunoperoxidase methods with an anti-equine CD13 monoclonal antibody that recognizes neutrophils and monocytes, we discovered that, whereas the dermal microvasculature of the skin commonly has a marginal pool of leukocytes, the normal laminar dermal microvasculature has minimal to no perivascular leukocytes. However, increases in leukocyte numbers occurred around the dermal vasculature of both the laminae and the skin in the majority of BWE-treated horses in the developmental stage and at the onset of clinical signs of lameness in the BWE model. These findings indicate that, similar to organ failure in human sepsis, leukocyte emigration is likely to play a significant role in initiating numerous pathophysiologic mechanisms that lead to the development of laminitis.1 aBlack, Samuel, J1 aLunn, Paul1 aYin, Cailing1 aHwang, Misako1 aLenz, Stephen, D1 aBelknap, James, K uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/leukocyte-emigration-in-the-early-stages-of-laminitis02305nas a2200157 4500008004100000245011900041210006900160260001600229300001100245490000800256520166700264100002001931700002201951700002101973856015301994 2006 eng d00aMatrix metalloproteinase-9 in laminae of black walnut extract treated horses correlates with neutrophil abundance.0 aMatrix metalloproteinase9 in laminae of black walnut extract tre c2006 Oct 15 a267-760 v1133 aWe sought to determine whether a correlation exists between neutrophil infiltration and tissue matrix metalloproteinase-9 (MMP-9) content in digital laminae collected during the prodromal and acute phases of laminitis in horses treated with an aqueous black walnut heartwood extract (BWE). Hoof laminar tissue was obtained at the onset of leukopenia and at the onset of clinical signs of lameness from BWE-treated horses and at equivalent times from control horses. Thin sections of laminae were screened for neutrophils by immunohistochemistry with an anti-CD13 monoclonal antibody and extracts of the same tissues were screened for SDS-renaturable and native MMP-9 activities by denaturing and non-denaturing gelatin zymography. Samples were also screened for MMP-2 and MMP-9 gene expression by RT-qPCR. Control laminae were devoid of both MMP-9 and neutrophils, whereas neutrophils and SDS-renaturable MMP-9 activity were detected in laminae from BWE-treated horses and were strongly correlated at the acute stage of the disease at which time laminar MMP-9 gene expression was significantly (15-fold) elevated. In contrast, BWE-treatment did not significantly elevate MMP-2 gene or protein expression in the laminae. Interestingly, MMP-9 that was present in extracts of laminae from BWE-treated horses at both the prodromal and acute stages of the disease was mainly in the zymogen form, suggesting that the accumulation of the MMP did not contribute to pathology during these stages. However, elevated presence of the MMP-9 zymogen in the tissue would predispose it to catastrophic damage should conditions arise that cleave the regulatory propeptide domain.1 aLoftus, John, P1 aBelknap, James, K1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/matrix-metalloproteinase-9-in-laminae-of-black-walnut-extract-treated02030nas a2200217 4500008004100000245007800041210006900119260001600188300001100204490000700215520127800222100001901500700002001519700001801539700002301557700001901580700001901599700002401618700002401642856014601666 2006 eng d00aNotch1 augments NF-kappaB activity by facilitating its nuclear retention.0 aNotch1 augments NFkappaB activity by facilitating its nuclear re c2006 Jan 11 a129-380 v253 aNotch1 specifically upregulates expression of the cytokine interferon-gamma in peripheral T cells through activation of NF-kappaB. However, how Notch mediates NF-kappaB activation remains unclear. Here, we examined the temporal relationship between Notch signaling and NF-kappaB induction during T-cell activation. NF-kappaB activation occurs within minutes of T-cell receptor (TCR) engagement and this activation is sustained for at least 48 h following TCR signaling. We used gamma-secretase inhibitor (GSI) to prevent the cleavage and subsequent activation of Notch family members. We demonstrate that GSI blocked the later, sustained NF-kappaB activation, but did not affect the initial activation of NF-kappaB. Using biochemical approaches, as well as confocal microscopy, we show that the intracellular domain of Notch1 (N1IC) directly interacts with NF-kappaB and competes with IkappaBalpha, leading to retention of NF-kappaB in the nucleus. Additionally, we show that N1IC can directly regulate IFN-gamma expression through complexes formed on the IFN-gamma promoter. Taken together, these data suggest that there are two 'waves' of NF-kappaB activation: an initial, Notch-independent phase, and a later, sustained activation of NF-kappaB, which is Notch dependent.1 aShin, Hyun, Mu1 aMinter, Lisa, M1 aCho, Ok, Hyun1 aGottipati, Sridevi1 aFauq, Abdul, H1 aGolde, Todd, E1 aSonenshein, Gail, E1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch1-augments-nf-kappab-activity-by-facilitating-its-nuclear03106nas a2200457 4500008004100000245012200041210006900163260001300232300001200245490000800257520165900265653002401924653001201948653003301960653001201993653001502005653002202020653001102042653001802053653001102071653004302082653003202125653000902157653003802166653002802204653001202232653002002244653002302264653001902287653001102306100001402317700002002331700002102351700002502372700001502397700002502412700002202437700001802459700002302477856014802500 2006 eng d00aPhosphorylation of IP3R1 and the regulation of [Ca2+]i responses at fertilization: a role for the MAP kinase pathway.0 aPhosphorylation of IP3R1 and the regulation of Ca2i responses at c2006 Nov a4355-650 v1333 aA sperm-induced intracellular Ca2+ signal ([Ca2+]i) underlies the initiation of embryo development in most species studied to date. The inositol 1,4,5 trisphosphate receptor type 1 (IP3R1) in mammals, or its homologue in other species, is thought to mediate the majority of this Ca2+ release. IP3R1-mediated Ca2+ release is regulated during oocyte maturation such that it reaches maximal effectiveness at the time of fertilization, which, in mammalian eggs, occurs at the metaphase stage of the second meiosis (MII). Consistent with this, the [Ca2+]i oscillations associated with fertilization in these species occur most prominently during the MII stage. In this study, we have examined the molecular underpinnings of IP3R1 function in eggs. Using mouse and Xenopus eggs, we show that IP3R1 is phosphorylated during both maturation and the first cell cycle at a MPM2-detectable epitope(s), which is known to be a target of kinases controlling the cell cycle. In vitro phosphorylation studies reveal that MAPK/ERK2, one of the M-phase kinases, phosphorylates IP3R1 at at least one highly conserved site, and that its mutation abrogates IP3R1 phosphorylation in this domain. Our studies also found that activation of the MAPK/ERK pathway is required for the IP3R1 MPM2 reactivity observed in mouse eggs, and that eggs deprived of the MAPK/ERK pathway during maturation fail to mount normal [Ca2+]i oscillations in response to agonists and show compromised IP3R1 function. These findings identify IP3R1 phosphorylation by M-phase kinases as a regulatory mechanism of IP3R1 function in eggs that serves to optimize [Ca2+]i release at fertilization.
10aAmino Acid Sequence10aAnimals10aAntibodies, Phospho-Specific10aCalcium10aCell Cycle10aEnzyme Activation10aFemale10aFertilization10aHumans10aInositol 1,4,5-Trisphosphate Receptors10aMAP Kinase Signaling System10aMice10aMitogen-Activated Protein Kinases10aMolecular Sequence Data10aOocytes10aPhosphorylation10aSequence Alignment10aXenopus laevis10aZygote1 aLee, Bora1 aVermassen, Elke1 aYoon, Sook-Young1 aVanderheyden, Veerle1 aIto, Junya1 aAlfandari, Dominique1 aDe Smedt, Humbert1 aParys, Jan, B1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/phosphorylation-of-ip3r1-and-the-regulation-of-ca2i-responses-at02478nas a2200229 4500008004100000245012200041210006900163260001300232300001200245490000800257520165000265100001401915700002001929700002101949700002501970700001501995700002502010700002202035700001802057700002302075856015002098 2006 eng d00aPhosphorylation of IP3R1 and the regulation of [Ca2+]i responses at fertilization: a role for the MAP kinase pathway.0 aPhosphorylation of IP3R1 and the regulation of Ca2i responses at c2006 Nov a4355-650 v1333 aA sperm-induced intracellular Ca2+ signal ([Ca2+]i) underlies the initiation of embryo development in most species studied to date. The inositol 1,4,5 trisphosphate receptor type 1 (IP3R1) in mammals, or its homologue in other species, is thought to mediate the majority of this Ca2+ release. IP3R1-mediated Ca2+ release is regulated during oocyte maturation such that it reaches maximal effectiveness at the time of fertilization, which, in mammalian eggs, occurs at the metaphase stage of the second meiosis (MII). Consistent with this, the [Ca2+]i oscillations associated with fertilization in these species occur most prominently during the MII stage. In this study, we have examined the molecular underpinnings of IP3R1 function in eggs. Using mouse and Xenopus eggs, we show that IP3R1 is phosphorylated during both maturation and the first cell cycle at a MPM2-detectable epitope(s), which is known to be a target of kinases controlling the cell cycle. In vitro phosphorylation studies reveal that MAPK/ERK2, one of the M-phase kinases, phosphorylates IP3R1 at at least one highly conserved site, and that its mutation abrogates IP3R1 phosphorylation in this domain. Our studies also found that activation of the MAPK/ERK pathway is required for the IP3R1 MPM2 reactivity observed in mouse eggs, and that eggs deprived of the MAPK/ERK pathway during maturation fail to mount normal [Ca2+]i oscillations in response to agonists and show compromised IP3R1 function. These findings identify IP3R1 phosphorylation by M-phase kinases as a regulatory mechanism of IP3R1 function in eggs that serves to optimize [Ca2+]i release at fertilization.1 aLee, Bora1 aVermassen, Elke1 aYoon, Sook-Young1 aVanderheyden, Veerle1 aIto, Junya1 aAlfandari, Dominique1 aDe Smedt, Humbert1 aParys, Jan, B1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/phosphorylation-of-ip3r1-and-the-regulation-of-ca2i-responses-at-002962nas a2200505 4500008004100000245016800041210006900209260001600278300001200294490000800306520121400314653001201528653001101540653001401551653001801565653002301583653001301606653003101619653004201650653004001692653001601732653002501748653001501773653003101788653001101819653003401830653005501864653002201919653000901941653002901950653003501979653002002014653001502034653001902049653005202068653001902120653002402139653002302163653003002186100002002216700002602236700002402262700002202286856014802308 2006 eng d00aRegulation of the composition of the extracellular matrix by low density lipoprotein receptor-related protein-1: activities based on regulation of mRNA expression.0 aRegulation of the composition of the extracellular matrix by low c2006 Mar 17 a7332-400 v2813 aLow density lipoprotein receptor-related protein-1 (LRP-1) is a catabolic receptor for extracellular matrix (ECM) structural proteins and for proteins that bind to ECM. LRP-1 also is implicated in integrin maturation. In this study, we applied a proteomics strategy to identify novel proteins involved in ECM modeling that are regulated by LRP-1. We show that LRP-1 deficiency in murine embryonic fibroblasts (MEFs) is associated with increased levels of type III collagen and pigment epithelium-derived factor, which accumulate in the substratum surrounding cells. The collagen receptor, uPAR-AP/Endo-180, is also increased in LRP-1-deficient MEFs. Human LRP-1 reversed the changes in protein expression associated with LRP-1 deficiency; however, the endocytic activity of LRP-1 was not involved. Instead, regulation occurred at the mRNA level. Inhibition of c-Jun amino-terminal kinase (JNK) blocked type III collagen expression in LRP-1-deficient MEFs, suggesting regulation of JNK activity as a mechanism by which LRP-1 controls mRNA expression. The ability of LRP-1 to regulate expression of the factors identified here suggests a role for LRP-1 in determining blood vessel structure and in angiogenesis.10aAnimals10aBiotin10aCell Line10aCell Membrane10aCloning, Molecular10aCollagen10aCulture Media, Conditioned10aElectrophoresis, Gel, Two-Dimensional10aElectrophoresis, Polyacrylamide Gel10aEndocytosis10aExtracellular Matrix10aFibrinogen10aGene Expression Regulation10aHumans10aLDL-Receptor Related Proteins10aLow Density Lipoprotein Receptor-Related Protein-110aMass Spectrometry10aMice10aMicroscopy, Fluorescence10aNeovascularization, Pathologic10aPhosphorylation10aProteomics10aReceptors, LDL10aReverse Transcriptase Polymerase Chain Reaction10aRNA, Messenger10aSignal Transduction10aSurface Properties10aTumor Suppressor Proteins1 aGaultier, Alban1 aSalicioni, Ana, Maria1 aArandjelovic, Sanja1 aGonias, Steven, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-the-composition-of-the-extracellular-matrix-by-low02588nas a2200241 4500008004100000245013700041210006900178260001500247300001200262490000800274520166400282100003701946700001901983700002202002700002502024700002802049700003002077700002102107700002002128700002102148700002302169856015402192 2006 eng d00aSodium and epithelial sodium channels participate in the regulation of the capacitation-associated hyperpolarization in mouse sperm.0 aSodium and epithelial sodium channels participate in the regulat c2006 Mar 3 a5623-330 v2813 aIn a process called capacitation, mammalian sperm gain the ability to fertilize after residing in the female tract. During capacitation the mouse sperm plasma membrane potential (E(m)) hyperpolarizes. However, the mechanisms that regulate sperm E(m) are not well understood. Here we show that sperm hyperpolarize when external Na(+) is replaced by N-methyl-glucamine. Readdition of external Na(+) restores a more depolarized E(m) by a process that is inhibited by amiloride or by its more potent derivative 5-(N-ethyl-N-isopropyl)-amiloride hydrochloride. These findings indicate that under resting conditions an electrogenic Na(+) transporter, possibly involving an amiloride sensitive Na(+) channel, may contribute to the sperm resting E(m). Consistent with this proposal, patch clamp recordings from spermatogenic cells reveal an amiloride-sensitive inward Na(+) current whose characteristics match those of the epithelial Na(+) channel (ENaC) family of epithelial Na(+) channels. Indeed, ENaC-alpha and -delta mRNAs were detected by reverse transcription-PCR in extracts of isolated elongated spermatids, and ENaC-alpha and -delta proteins were found on immunoblots of sperm membrane preparations. Immunostaining indicated localization of ENaC-alpha to the flagellar midpiece and of ENaC-delta to the acrosome. Incubations known to produce capacitation in vitro or induction of capacitation by cell-permeant cAMP analogs decreased the depolarizing response to the addition of external Na(+). These results suggest that increases in cAMP content occurring during capacitation may inhibit ENaCs to produce a required hyperpolarization of the sperm membrane.1 aHernández-González, Enrique, O1 aSosnik, Julian1 aEdwards, Jennifer1 aAcevedo, Juan, José1 aMendoza-Lujambio, Irene1 aLópez-González, Ignacio1 aDemarco, Ignacio1 aWertheimer, Eva1 aDarszon, Alberto1 aVisconti, Pablo, E uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sodium-and-epithelial-sodium-channels-participate-in-the-regulation-of01880nas a2200217 4500008004100000245009600041210006900137260001300206300000800219490000800227520109500235100002601330700002001356700002101376700002001397700002001417700002701437700002301464700002401487856015101511 2006 eng d00aTreatment of Brucella-susceptible mice with IL-12 increases primary and secondary immunity.0 aTreatment of Brucellasusceptible mice with IL12 increases primar c2006 Sep a1-90 v2433 aBrucella spp. cause disease in humans and livestock and are potential biowarfare agents. Defining the protective immune response is necessary to design vaccines. This has largely been done with mice, brucella-susceptible BALB/c and resistant C57BL strains. Since interferon-gamma is key to brucella resistance, contrary to expectations, we found that ex vivo splenocytes from naïve BALB/c mice produced IL-12 and interferon-gamma in cultures with brucellae at levels comparable to those of splenocytes from the more resistant C57BL/10 mice. Moreover, both IL-12 and interferon-gamma were produced in the first week following infection of BALB/c mice. However, by the third week of infection we found decreased IL-12Rbeta2 expression by BABL/c splenocytes, corresponding to their inability to produce interferon-gamma in Brucella recall responses at this time as reported previously. Administering recombinant IL-12 to these mice ameliorated the interferon-gamma hiatus, resulted in a 1000-fold reduction in CFU during primary infection and increased survival following secondary challenge.1 aSathiyaseelan, Janaki1 aGoenka, Radhika1 aParent, Michelle1 aBenson, Rita, M1 aMurphy, Erin, A1 aFernandes, Dancella, M1 aFoulkes, Andrea, S1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/treatment-of-brucella-susceptible-mice-with-il-12-increases-primary02604nas a2200193 4500008004100000245010000041210006900141260001300210300001100223490000600234520188300240100002202123700002402145700002302169700002502192700001702217700002402234856015202258 2006 eng d00aUse of an aggressive MCF-7 cell line variant, TMX2-28, to study cell invasion in breast cancer.0 aUse of an aggressive MCF7 cell line variant TMX228 to study cell c2006 Dec a905-130 v43 aAn estrogen receptor-negative variant of the MCF-7 breast cancer cell line, TMX2-28, was used as a model in which to study breast cancer cell invasion. Using a reconstituted basement membrane (Matrigel) assay to evaluate cell invasion, we determined that TMX2-28 cells are more invasive than MCF-7 cells and that the invasiveness of TMX2-28 is similar to that of the aggressive MDA-MB-231 breast cancer cell line. TMX2-28 cells displayed a rounded, epithelial cell-like morphology, suggesting an amoeboid mode of cell invasion, in contrast to the mesenchymal mode of invasion characteristic of spindle-shaped, fibroblast-like MDA-MB-231 cells. Using real-time reverse transcription-PCR, we found that mitogen-inducible gene 2 (MIG2) is expressed at a 17-fold higher level in TMX2-28 cells than in nonaggressive MCF-7 cells and that MIG2 mRNA levels are low in the nontumorigenic human mammary epithelial cell line, 184. We determined that MIG2 plays a role in cell invasion by using small interfering RNA (siRNA) to suppress the expression of MIG2 mRNA levels in TMX2-28 cells. TMX2-28 cell invasion was reduced by 48% when the cells were transfected with siRNAs targeting MIG2, relative to cells transfected with siRNAs against glyceraldehyde-3-phosphate dehydrogenase. Finally, MIG2 expression was evaluated in reductive mammoplasty and breast tumor tissue. Although all 21 normal tissues from reduction mammoplasty showed immunoreactivity for MIG2, ranging from weak (62%) to strong (24%), only half of the 34 formalin-fixed breast tumors showed immunoreactivity for MIG2. Of these 17 positive cases, 10 were considered to overexpress MIG2 (moderate to strong staining). Examination of 30 frozen breast tumors supported the finding that MIG2 is overexpressed in a subset of breast cancers. We suggest that MIG2's normal regulation and function are disrupted in breast cancer.1 aGozgit, Joseph, M1 aPentecost, Brian, T1 aMarconi, Sharon, A1 aOtis, Christopher, N1 aWu, Chuanyue1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/use-of-an-aggressive-mcf-7-cell-line-variant-tmx2-28-to-study-cell-002908nas a2200301 4500008004100000245010000041210006900141260001300210300001100223490000600234520188300240653002102123653002802144653002102172653002602193653001102219653001102230653002602241653003002267653002402297100002202321700002402343700002302367700002502390700001702415700002402432856015002456 2006 eng d00aUse of an aggressive MCF-7 cell line variant, TMX2-28, to study cell invasion in breast cancer.0 aUse of an aggressive MCF7 cell line variant TMX228 to study cell c2006 Dec a905-130 v43 aAn estrogen receptor-negative variant of the MCF-7 breast cancer cell line, TMX2-28, was used as a model in which to study breast cancer cell invasion. Using a reconstituted basement membrane (Matrigel) assay to evaluate cell invasion, we determined that TMX2-28 cells are more invasive than MCF-7 cells and that the invasiveness of TMX2-28 is similar to that of the aggressive MDA-MB-231 breast cancer cell line. TMX2-28 cells displayed a rounded, epithelial cell-like morphology, suggesting an amoeboid mode of cell invasion, in contrast to the mesenchymal mode of invasion characteristic of spindle-shaped, fibroblast-like MDA-MB-231 cells. Using real-time reverse transcription-PCR, we found that mitogen-inducible gene 2 (MIG2) is expressed at a 17-fold higher level in TMX2-28 cells than in nonaggressive MCF-7 cells and that MIG2 mRNA levels are low in the nontumorigenic human mammary epithelial cell line, 184. We determined that MIG2 plays a role in cell invasion by using small interfering RNA (siRNA) to suppress the expression of MIG2 mRNA levels in TMX2-28 cells. TMX2-28 cell invasion was reduced by 48% when the cells were transfected with siRNAs targeting MIG2, relative to cells transfected with siRNAs against glyceraldehyde-3-phosphate dehydrogenase. Finally, MIG2 expression was evaluated in reductive mammoplasty and breast tumor tissue. Although all 21 normal tissues from reduction mammoplasty showed immunoreactivity for MIG2, ranging from weak (62%) to strong (24%), only half of the 34 formalin-fixed breast tumors showed immunoreactivity for MIG2. Of these 17 positive cases, 10 were considered to overexpress MIG2 (moderate to strong staining). Examination of 30 frozen breast tumors supported the finding that MIG2 is overexpressed in a subset of breast cancers. We suggest that MIG2's normal regulation and function are disrupted in breast cancer.10aBreast Neoplasms10aCell Adhesion Molecules10aCell Line, Tumor10aCytoskeletal Proteins10aFemale10aHumans10aNeoplasm Invasiveness10aPolymerase Chain Reaction10aReceptors, Estrogen1 aGozgit, Joseph, M1 aPentecost, Brian, T1 aMarconi, Sharon, A1 aOtis, Christopher, N1 aWu, Chuanyue1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/use-of-an-aggressive-mcf-7-cell-line-variant-tmx2-28-to-study-cell02551nas a2200313 4500008004100000245011700041210006900158260001500227300001000242490000800252520153800260653001201798653002001810653001701830653001801847653002301865653002101888653002201909653002501931653001301956653001001969653000901979653002101988653001802009653001502027100002602042700002202068856014702090 2005 eng d00aDistinct populations of endoderm cells converge to generate the embryonic liver bud and ventral foregut tissues.0 aDistinct populations of endoderm cells converge to generate the c2005 Apr 1 a87-990 v2803 aThe location and movement of mammalian gut tissue progenitors, prior to the expression of tissue-specific genes, has been unknown, but this knowledge is essential to identify transitions that lead to cell type specification. To address this, we used vital dyes to label exposed anterior endoderm cells of early somite stage mouse embryos, cultured the embryos into the tissue bud phase of development, and determined the tissue fate of the dye labeled cells. This approach was performed at three embryonic stages that are prior to, or coincident with, foregut tissue patterning (1-3 somites, 4-6 somites, and 7-10 somites). Short-term labeling experiments tracked the movement of tissue progenitor cells during foregut closure. Surprisingly, we found that two distinct types of endoderm-progenitor cells, lateral and medial, arising from three spatially separated embryonic domains, converge to generate the epithelial cells of the liver bud. Whereas the lateral endoderm-progenitors give rise to descendants that are constrained in tissue fate and position along the anterior-posterior axis of the gut, the medial gut endoderm-progenitors give rise to descendants that stream along the anterior-posterior axis at the ventral midline and contribute to multiple gut tissues. The fate map reveals extensive morphogenetic movement of progenitors prior to tissue specification, it permits a detailed analysis of endoderm tissue patterning, and it illustrates that diverse progenitor domains can give rise to individual tissue cell types.10aAnimals10aBody Patterning10aCell Lineage10aCell Movement10aCell Proliferation10aDigestive System10aEmbryo, Mammalian10aEmbryonic Structures10aEndoderm10aLiver10aMice10aMice, Inbred C3H10aMorphogenesis10aStem Cells1 aTremblay, Kimberly, D1 aZaret, Kenneth, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/distinct-populations-of-endoderm-cells-converge-to-generate-the02665nas a2200373 4500008004100000245012900041210006900170260001600239300001200255490000700267520141100274653001201685653002301697653002201720653001501742653001401757653002501771653002701796653000901823653002401832653001901856653002901875653001701904653003601921653003301957100002101990700001602011700002402027700002102051700002002072700002902092700001802121856015202139 2005 eng d00aEstrogen and progesterone regulate radiation-induced p53 activity in mammary epithelium through TGF-beta-dependent pathways.0 aEstrogen and progesterone regulate radiationinduced p53 activity c2005 Sep 22 a6345-530 v243 aDNA damage normally induces p53 activity, but responses to ionizing radiation in the mammary epithelium vary among developmental stages. The following studies examined the hormones and growth factors that regulate radiation-responsiveness of p53 in mouse mammary epithelium. Immunoreactive p21/WAF1 and TUNEL staining were used as indicators of p53 activity following exposure to ionizing radiation. In ovariectomized mice, radiation-induced accumulation of p21/WAF1 was minimal in the mammary epithelial cells (<1%). Systemic injections of estrogen and progesterone (E+P) for 72 h were necessary to recover maximal expression of p21/WAF1 following ionizing radiation (55%). The effects of E+P on radiation-induced p21/WAF1 were p53-dependent as responses were absent in Trp53-/- mice. Though hormonal treatments stimulated increases in the proportion of cycling cells (PCNA-positive), this was not directly correlated with p53 activity. Whole organ cultures were used to determine whether E+P act directly upon the mammary gland. Treatment with E+P was sufficient to render p53 responsive to radiation, but TGF-beta-neutralizing antibodies blocked responsiveness. In the absence of E+P, TGF-beta1 alone did not alter p53 activity. These results demonstrate that estrogen and progesterone together with TGF-beta signaling are necessary for maintenance of p53 activity in the mammary epithelium.
10aAnimals10aBlotting, Northern10aBlotting, Western10aEpithelium10aEstrogens10aImmunohistochemistry10aMammary Glands, Animal10aMice10aMice, Inbred BALB C10aMice, Knockout10aOrgan Culture Techniques10aProgesterone10aTransforming Growth Factor beta10aTumor Suppressor Protein p531 aBecker, Klaus, A1 aLu, Shaolei1 aDickinson, Ellen, S1 aDunphy, Karen, A1 aMathews, Lesley1 aSchneider, Sallie, Smith1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/estrogen-and-progesterone-regulate-radiation-induced-p53-activity-in02194nas a2200205 4500008004100000245012900041210006900170260001600239300001200255490000700267520141100274100002101685700001601706700002401722700002101746700002001767700002901787700001801816856015401834 2005 eng d00aEstrogen and progesterone regulate radiation-induced p53 activity in mammary epithelium through TGF-beta-dependent pathways.0 aEstrogen and progesterone regulate radiationinduced p53 activity c2005 Sep 22 a6345-530 v243 aDNA damage normally induces p53 activity, but responses to ionizing radiation in the mammary epithelium vary among developmental stages. The following studies examined the hormones and growth factors that regulate radiation-responsiveness of p53 in mouse mammary epithelium. Immunoreactive p21/WAF1 and TUNEL staining were used as indicators of p53 activity following exposure to ionizing radiation. In ovariectomized mice, radiation-induced accumulation of p21/WAF1 was minimal in the mammary epithelial cells (<1%). Systemic injections of estrogen and progesterone (E+P) for 72 h were necessary to recover maximal expression of p21/WAF1 following ionizing radiation (55%). The effects of E+P on radiation-induced p21/WAF1 were p53-dependent as responses were absent in Trp53-/- mice. Though hormonal treatments stimulated increases in the proportion of cycling cells (PCNA-positive), this was not directly correlated with p53 activity. Whole organ cultures were used to determine whether E+P act directly upon the mammary gland. Treatment with E+P was sufficient to render p53 responsive to radiation, but TGF-beta-neutralizing antibodies blocked responsiveness. In the absence of E+P, TGF-beta1 alone did not alter p53 activity. These results demonstrate that estrogen and progesterone together with TGF-beta signaling are necessary for maintenance of p53 activity in the mammary epithelium.
1 aBecker, Klaus, A1 aLu, Shaolei1 aDickinson, Ellen, S1 aDunphy, Karen, A1 aMathews, Lesley1 aSchneider, Sallie, Smith1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/estrogen-and-progesterone-regulate-radiation-induced-p53-activity-in-003007nas a2200193 4500008004100000245009300041210006900134260001600203300001000219490000800229520229000237100002002527700002502547700002002572700002502592700002202617700002402639856015002663 2005 eng d00aFunction of ruminant gammadelta T cells is defined by WC1.1 or WC1.2 isoform expression.0 aFunction of ruminant gammadelta T cells is defined by WC11 or WC c2005 Oct 18 a211-70 v1083 aWC1 is a transmembrane glycoprotein and member of the scavenger receptor cysteine rich (SRCR) family that is uniquely expressed on gammadelta T cells. The WC1 isoforms referred to as WC1.1, WC1.2, and WC1.3 are expressed on discrete subpopulations of gammadelta T cells with WC1.1 and WC1.2 expressed on mostly nonoverlapping gammadelta T cell populations. Studies have demonstrated a potential role for WC1 in modulating the response of gammadelta T cells but have not converged into a single accepted paradigm. Recent investigations that examined changing representation among mononuclear cells with age and patterns of proliferation and cytokine production by subsets bearing one or more of the previously identified variants of the WC1 molecule are summarized here. While the decrease in percentages within blood in the first year of life was found to be precipitous for WC1.1+ gammadelta T cells it was not for WC1.2+ cells. While both populations proliferated to mitogen stimulation there was a bias towards responses by WC1.2+ cells. In leptospira antigen-stimulated cultures and autologous mixed lymphocyte reaction (AMLR) cultures WC1.1+ cells proliferated and produced interferon-gamma (IFN-gamma) while WC1.2+ cells did to a much lower extent. This suggested functional differences related to the isoform of WC1 expressed. Under Th1-polarizing conditions, the WC1.1+ cells also made IFN-gamma whereas the vast majority of cells expressing WC1.2 did not. Despite the difference in IFN-gamma production, cells bearing either WC1 isoform had similar transcription levels of the high affinity IL-12 receptor subunit (IL-12Rbeta2) as well as of the transcription factors T-bet and GATA-3 when cultured with IL-12. Both populations transcribed low levels of IL-10 mRNA under Th1-polarizing conditions and TGF-beta transcripts were ubiquitously expressed by each of these cell types. Cloning and sequencing of the cytoplasmic tails of the WC1 isoforms revealed a consensus ITAM in all three isoforms but a DENY sequence adjacent to one of the SH-2 binding sites of WC1.1 only. The results suggest that WC1+ gammadelta T cells differentiated on the basis of WC1 isoform expression play distinct roles in immune responses that may be dictated by WC1 intracellular signaling.
1 aRogers, Aric, N1 aVanburen, Denille, G1 aHedblom, Emmett1 aTilahun, Mulualem, E1 aTelfer, Janice, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/function-of-ruminant-gammadelta-t-cells-is-defined-by-wc11-or-wc1202386nas a2200217 4500008004100000245013800041210006900179260001600248300001100264490000800275520157900283100002101862700001901883700001201902700001601914700002501930700002101955700001901976700002301995856015002018 2005 eng d00aFunctional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm.0 aFunctional biochemical and chromatographic characterization of t c2005 Sep 15 a376-920 v2853 aA cytosolic sperm protein(s), referred to as sperm factor (SF), is delivered into eggs by the sperm during mammalian fertilization to induce repetitive increases in the intracellular concentration of free Ca2+ ([Ca2+]i) that are referred to as [Ca2+]i oscillations. [Ca2+]i oscillations are essential for egg activation and early embryonic development. Recent evidence shows that the novel sperm-specific phospholipase C (PLC), PLCzeta, may be the long sought after [Ca2+]i oscillation-inducing SF. Here, we demonstrate the complete extraction of SF from porcine sperm and show that regardless of the method of extraction a single molecule/complex appears to be responsible for the [Ca2+]i oscillation-inducing activity of these extracts. Consistent with this notion, all sperm fractions that induced [Ca2+]i oscillations, including FPLC-purified fractions, exhibited high in vitro PLC activity at basal Ca2+ levels (0.1-5 microM), a hallmark of PLCzeta. Notably, we detected immunoreactive 72-kDa PLCzeta in an inactive fraction, and several fractions capable of inducing oscillations were devoid of 72-kDa PLCzeta. Nonetheless, in the latter fractions, proteolytic fragments, presumably corresponding to cleaved forms of PLCzeta, were detected by immunoblotting. Therefore, our findings corroborate the hypothesis that a sperm-specific PLC is the main component of the [Ca2+]i oscillation-inducing activity of sperm but provide evidence that the presence of 72-kDa PLCzeta does not precisely correspond with the Ca2+ releasing activity of porcine sperm fractions.
1 aKurokawa, Manabu1 aSato, Ken-ichi1 aWu, Hua1 aHe, Changli1 aMalcuit, Christopher1 aBlack, Samuel, J1 aFukami, Kiyoko1 aFissore, Rafael, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/functional-biochemical-and-chromatographic-characterization-of-the02483nas a2200193 4500008004100000245007300041210006900114260001600183300001200199490000800211520178000219100002001999700002502019700002302044700002502067700002202092700002402114856015102138 2005 eng d00aGammadelta T cell function varies with the expressed WC1 coreceptor.0 aGammadelta T cell function varies with the expressed WC1 corecep c2005 Mar 15 a3386-930 v1743 aWC1 molecules are transmembrane glycoproteins belonging to the scavenger receptor cysteine-rich family and uniquely expressed on gammadelta T cells. Although participation of WC1+ gammadelta T cells in immune responses is well established, very little is understood regarding the significance of expressing different forms of the WC1 molecule. Two forms previously identified by mAbs, i.e., WC1.1 and WC1.2, are expressed by largely nonoverlapping subpopulations of gammadelta T cells. In this study it was shown that expression of the WC1.1 coreceptor was the main indicator of proliferation and IFN-gamma production in response to autologous and bacterial Ags as well as for IFN-gamma production without proliferation in Th1-polarizing, IL-12-containing cultures. Nevertheless, after culture in either Th1-polarizing or neutral conditions, mRNA was present for both T-bet and GATA-3 as well as for IL-12Rbeta2 in WC1.1+ and WC1.2+ subpopulations, and neither produced IL-4 under any conditions. Although the steady decrease in the proportion of WC1.1+ cells, but not WC1.2+ cells, within PBMC with animal aging suggested that the two subpopulations may have different roles in immune regulation, cells bearing either WC1.1 or WC1.2 expressed mRNA for regulatory cytokines IL-10 and TGF-beta, with TGF-beta being constitutively expressed by ex vivo cells. Overall, the results demonstrate that the form of the WC1 coreceptor expressed on gammadelta T cells divides them into functional subsets according to IFN-gamma production and proliferative capacity to specific stimuli as well as with regard to representation within PBMC. Finally, evidence is provided for minor differences in the intracytoplasmic tail sequences of WC1.1 and WC1.2 that may affect signaling.
1 aRogers, Aric, N1 aVanburen, Denille, G1 aHedblom, Emmett, E1 aTilahun, Mulualem, E1 aTelfer, Janice, C1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/gammadelta-t-cell-function-varies-with-the-expressed-wc1-coreceptor02181nas a2200337 4500008004100000245013300041210006900174260001300243300001000256490000600266520105200272100002001324700002401344700001601368700001901384700001501403700002301418700001801441700002001459700002301479700002201502700001801524700001901542700002301561700002201584700001701606700001901623700002301642700002401665856015401689 2005 eng d00aInhibitors of gamma-secretase block in vivo and in vitro T helper type 1 polarization by preventing Notch upregulation of Tbx21.0 aInhibitors of gammasecretase block in vivo and in vitro T helper c2005 Jul a680-80 v63 aNotch receptors are processed by gamma-secretase acting in synergy with T cell receptor signaling to sustain peripheral T cell activation. Activated CD4+ T cells differentiate into T helper type 1 (TH1) or TH2 subsets. Molecular cues directing TH1 differentiation include expression of the TH1-specific transcription factor T-bet, encoded by Tbx21. However, the regulation of Tbx21 remains incompletely defined. Here we report that Notch1 can directly regulate Tbx21 through complexes formed on the Tbx21 promoter. In vitro, gamma-secretase inhibitors extinguished expression of Notch, interferon-gamma and Tbx21 in TH1-polarized CD4+ cells, whereas ectopic expression of activated Notch1 restored Tbx21 transcription. In vivo, administration of gamma-secretase inhibitors substantially impeded TH1-mediated disease progression in the mouse experimental autoimmune encephalomyelitis model of multiple sclerosis. Thus, using gamma-secretase inhibitors to modulate Notch signaling may prove beneficial in treating TH1-mediated autoimmunity.
1 aMinter, Lisa, M1 aTurley, Danielle, M1 aDas, Pritam1 aShin, Hyun, Mu1 aJoshi, Ila1 aLawlor, Rebecca, G1 aCho, Ok, Hyun1 aPalaga, Tanapat1 aGottipati, Sridevi1 aTelfer, Janice, C1 aKostura, Lisa1 aFauq, Abdul, H1 aSimpson, Katherine1 aSuch, Kimberly, A1 aMiele, Lucio1 aGolde, Todd, E1 aMiller, Stephen, D1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/inhibitors-of-gamma-secretase-block-in-vivo-and-in-vitro-t-helper-type02550nas a2200397 4500008004100000245006700041210006500108260001500173300001000188490000800198520126100206653001201467653002401479653001701503653003101520653003101551653001501582653004001597653004101637653000901678653002401687653002101711653002301732653001901755653002101774653001301795653003101808100002501839700002201864700002601886700002001912700002401932700002201956700002901978856014502007 2005 eng d00aRestricted MHC-peptide repertoire predisposes to autoimmunity.0 aRestricted MHCpeptide repertoire predisposes to autoimmunity c2005 Jul 4 a73-840 v2023 aMHC molecules associated with autoimmunity possess known structural features that limit the repertoire of peptides that they can present. Such limitation gives a selective advantage to TCRs that rely on interaction with the MHC itself, rather than with the peptide residues. At the same time, negative selection is impaired because of the lack of negatively selecting peptide ligands. The combination of these factors may predispose to autoimmunity. We found that mice with an MHC class II-peptide repertoire reduced to a single complex demonstrated various autoimmune reactions. Transgenic mice bearing a TCR (MM14.4) cloned from such a mouse developed severe autoimmune dermatitis. Although MM14.4 originated from a CD4+ T cell, dermatitis was mediated by CD8+ T cells. It was established that MM14.4+ is a highly promiscuous TCR with dual MHC class I/MHC class II restriction. Furthermore, mice with a limited MHC-peptide repertoire selected elevated numbers of TCRs with dual MHC class I/MHC class II restriction, a likely source of autoreactivity. Our findings may help to explain the link between MHC class I responses that are involved in major autoimmune diseases and the well-established genetic linkage of these diseases with MHC class II.
10aAnimals10aAutoimmune Diseases10aAutoimmunity10aCD4-Positive T-Lymphocytes10aCD8-Positive T-Lymphocytes10aDermatitis10aHistocompatibility Antigens Class I10aHistocompatibility Antigens Class II10aMice10aMice, Inbred BALB C10aMice, Inbred C3H10aMice, Inbred C57BL10aMice, Knockout10aMice, Transgenic10aPeptides10aReceptors, Antigen, T-Cell1 aLogunova, Nadezda, N1 aViret, Christophe1 aPobezinsky, Leonid, A1 aMiller, Sara, A1 aKazansky, Dmitri, B1 aSundberg, John, P1 aChervonsky, Alexander, V uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/restricted-mhc-peptide-repertoire-predisposes-to-autoimmunity02121nas a2200169 4500008004100000245011700041210006900158260001300227300001100240490000600251520146500257100001401722700001801736700001801754700002901772856015001801 2005 eng d00aSensitivity to DNA damage is a common component of hormone-based strategies for protection of the mammary gland.0 aSensitivity to DNA damage is a common component of hormonebased c2005 Aug a435-420 v33 aAn early full-term pregnancy significantly reduces the risk of getting breast cancer in women. In animals, this protection can be mimicked by a short-term exposure to physiologic doses of estrogen plus progesterone. Sensitization of p53 and up-regulation of transforming growth factor beta are believed to be important aspects of the mechanism by which protection is imparted. Little is known, however, about the use of this pathway in response to other chemopreventive agents. In this article, we investigated the ability of retinoids, such as 9-cis retinoic acid, all-trans retinoic acid, and N-4-hydroxyphenylretinamide (4-HPR), to sensitize the ductal epithelial cells of virgin mammary glands to DNA damage responses. Using a whole-organ culture system, we observed enhanced cell death in response to gamma-irradiation in the virgin tissues treated with retinoids for 72 hours. These retinoids were partially dependent on p53 and transforming growth factor beta to exert their radiosensitizing effects. However, 4-HPR seemed to sensitize other cells or activate these pathways in a different manner as costimulation with ovarian hormones and 4-HPR was additive, whereas coculture of ovarian hormones and the natural retinoids did not increase amount of death. Taken together, these data suggest that sensitization of the mammary epithelium to p53-dependent apoptosis is a common pathway, which is engaged by retinoids as well as ovarian hormones.
1 aTu, Yifan1 aJerry, Joseph1 aPazik, Brooke1 aSchneider, Sallie, Smith uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sensitivity-to-dna-damage-is-a-common-component-of-hormone-based-002888nas a2200457 4500008004100000245011700041210006900158260001300227300001100240490000600251520146500257653001201722653001401734653001501748653001501763653003701778653001401815653001101829653001601840653001501856653001501871653002501886653003001911653002701941653000901968653002401977653001002001653001702011653002502028653001702053653003202070653003602102653001402138653003302152653001802185100001402203700001802217700001802235700002902253856014802282 2005 eng d00aSensitivity to DNA damage is a common component of hormone-based strategies for protection of the mammary gland.0 aSensitivity to DNA damage is a common component of hormonebased c2005 Aug a435-420 v33 aAn early full-term pregnancy significantly reduces the risk of getting breast cancer in women. In animals, this protection can be mimicked by a short-term exposure to physiologic doses of estrogen plus progesterone. Sensitization of p53 and up-regulation of transforming growth factor beta are believed to be important aspects of the mechanism by which protection is imparted. Little is known, however, about the use of this pathway in response to other chemopreventive agents. In this article, we investigated the ability of retinoids, such as 9-cis retinoic acid, all-trans retinoic acid, and N-4-hydroxyphenylretinamide (4-HPR), to sensitize the ductal epithelial cells of virgin mammary glands to DNA damage responses. Using a whole-organ culture system, we observed enhanced cell death in response to gamma-irradiation in the virgin tissues treated with retinoids for 72 hours. These retinoids were partially dependent on p53 and transforming growth factor beta to exert their radiosensitizing effects. However, 4-HPR seemed to sensitize other cells or activate these pathways in a different manner as costimulation with ovarian hormones and 4-HPR was additive, whereas coculture of ovarian hormones and the natural retinoids did not increase amount of death. Taken together, these data suggest that sensitization of the mammary epithelium to p53-dependent apoptosis is a common pathway, which is engaged by retinoids as well as ovarian hormones.
10aAnimals10aApoptosis10aCell Death10aDNA Damage10aDose-Response Relationship, Drug10aEstrogens10aFemale10aFenretinide10aGamma Rays10aGenes, p5310aImmunohistochemistry10aIn Situ Nick-End Labeling10aMammary Glands, Animal10aMice10aMice, Inbred BALB C10aOvary10aProgesterone10aRetinoid X Receptors10aRisk Factors10aSensitivity and Specificity10aTransforming Growth Factor beta10aTretinoin10aTumor Suppressor Protein p5310aUp-Regulation1 aTu, Yifan1 aJerry, Joseph1 aPazik, Brooke1 aSchneider, Sallie, Smith uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sensitivity-to-dna-damage-is-a-common-component-of-hormone-based02019nas a2200157 4500008004100000245012800041210006900169260001300238300001100251490000700262520138400269100001601653700001901669700002001688856015301708 2005 eng d00aSequencing and characterization of a cDNA encoding a ferritin subunit of Colorado potato beetle, Leptinotarsa decemlineata.0 aSequencing and characterization of a cDNA encoding a ferritin su c2005 Nov a140-500 v603 aA differentially expressed cDNA fragment (P311) from Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), was identified by restriction fragment differential display-polymerase chain reaction (RFDD-PCR) technique, and showed a strong similarity to ferritin heavy chain subunits of other organisms. Based on P311, we constructed specific primers and obtained a 840-bp cDNA fragment spanning the open reading frame of CPB ferritin subunit using the rapid amplification of cDNA ends (RACE) technique. The sequence encodes 213 amino acid residues, including a 19 amino acid signal peptide. The sequence has a conserved cysteine in the N-terminus and has the seven conserved residues that comprise the ferroxidase center, which is the feature of heavy chain ferritins of vertebrates. The CPB ferritin subunit has high amino acid sequence identity with the Apriona germari (69.3%), Galleria mellonela (54.5%), Manduca sexta (54.0%), Drosophila melanogaster (53.2%), Calpodes ethlius (51.4%), and Nilaparvata lugens (47.6%) but lower identity with the Anopheles gambiae (38.7%) and Aedes aegypti (37.8%). Using Northern blot analysis, the subunit mRNA was identified from fat body and midgut of 4th instars with much higher mRNA levels found in midgut than that in fat body (2.5-fold). Nevertheless, only the levels of mRNA in fat body was induced by dexamethasone (1.5-fold).1 aQiu, Lihong1 aGao, Jian-Rong1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/sequencing-and-characterization-of-a-cdna-encoding-a-ferritin-subunit01671nas a2200289 4500008004100000245006600041210006500107260001300172300001300185490000700198520077600205653001200981653002000993653002401013653003101037653002801068653001101096653001301107653000901120653002001129653001601149653001401165653001501179100001701194700002601211856014401237 2005 eng d00aStrategies for dissecting epigenetic mechanisms in the mouse.0 aStrategies for dissecting epigenetic mechanisms in the mouse c2005 Nov a1194-2000 v373 aEpigenetics generally refers to heritable changes in gene expression that are independent of nucleotide sequence. With complete genome sequences in hand, understanding the epigenetic control of genomes is the next step towards comprehending how the same DNA sequence gives rise to different cells, lineages and organs. Epigenetics also contributes to individual variation in normal biology and in disease states. The mouse provides a unique opportunity to understand how epigenetic differences contribute to both development and disease in a tractable mammalian system. Here we discuss current approaches and protocols used to study epigenetics in the mouse, including loss-of-function studies, mutagenesis screens, somatic cell nuclear transfer, genomics and proteomics.10aAnimals10aDNA Methylation10aEpigenesis, Genetic10aGene Expression Regulation10aGenes, Tumor Suppressor10aGenome10aGenomics10aMice10aModels, Genetic10aMutagenesis10aPhenotype10aProteomics1 aMager, Jesse1 aBartolomei, Marisa, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/strategies-for-dissecting-epigenetic-mechanisms-in-the-mouse01866nas a2200169 4500008004100000245012000041210006900161260001700230300001100247490000700258520119800265100002401463700001601487700002501503700002301528856014501551 2004 eng d00aBeta-galactosidase and alpha-mannosidase inhibit formation of multicellular nodules in breast cancer cell cultures.0 aBetagalactosidase and alphamannosidase inhibit formation of mult c2004 Jan-Feb a139-440 v243 aIn response to an estrogen, confluent monolayers of MCF-7 cell cultures develop multi-cellular nodules, termed foci. Post-confluent development of foci occurs with physiologic levels of 17beta-estradiol and are inhibited by various anti-estrogens acting through either the estrogen or aryl hydrocarbon receptors. In the present paper we report that disruption of the terminal sugars on membrane receptors results in inhibition of foci. Treatment with 0.013-0.05 units/ml of beta-galactosidase completely inhibited the development of foci while leaving the monolayer of cells intact. Trials with alpha-mannosidase resulted in a similar but less potent inhibition of foci. Lectin-fluorescent conjugates, RCA (Ricinus communis agglutinin), and ConA (Canavalia ensiformis agglutinin) were used to identify membrane surface carbohydrates on MCF-7 cells. Binding of the RCA-fluorescent conjugate was inhibited by co-treatment with galactose or lactose. Binding of ConA-fluorescent conjugate was significantly inhibited by mannose and n-acetyl-glucosamine. This is the first report of inhibition of foci development in MCF-7 cell cultures by disruption of surface carbohydrates on membrane receptors.1 aArcaro, Kathleen, F1 aWang, Joann1 aOtis, Christopher, N1 aZuckerman, Bert, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/beta-galactosidase-and-alpha-mannosidase-inhibit-formation-of02527nas a2200181 4500008004100000245016600041210006900207260001500276300001000291490000800301520176700309100002202076700002402098700002202122700002402144700002402168856015302192 2004 eng d00aDifferential action of polycyclic aromatic hydrocarbons on endogenous estrogen-responsive genes and on a transfected estrogen-responsive reporter in MCF-7 cells.0 aDifferential action of polycyclic aromatic hydrocarbons on endog c2004 Apr 1 a58-670 v1963 aPolycyclic aromatic hydrocarbons (PAHs) are common environmental pollutants that have been extensively studied for multiple toxicological endpoints in both laboratory animals and humans. The purpose of this study was to investigate the estrogenicity of PAHs in the human breast cancer cell line MCF-7. We investigated 14 PAHs for their ability to bind either the estrogen receptor (ER) or the aryl hydrocarbon receptor (AhR) and to activate target gene expression. PAHs were tested in a human recombinant estrogen receptor (hrER) competitive binding assay, and in both an estrogen response element (ERE)- and xenobiotic response element (XRE)-mediated reporter gene assay. We used quantitative RT-PCR to examine selected PAHs that showed activity in the ERE reporter gene assay for their ability to upregulate estrogen-responsive genes HEM45, progesterone receptor, and pS2, and the aryl hydrocarbon-responsive CYP1A1 gene. None of the 14 PAHs bound the hrER, but five of the PAHs (anthracene, B[a]A, chrysene, B[b]F, and B[a]P) induced ER-reporter activity. This activity was dependent on the metabolism of PAHs in MCF-7 cells via the AhR pathway, which resulted in the formation of metabolites that bound the ER. None of the five PAHs that induced the ER-reporter were found to upregulate estrogen-responsive genes, yet four of the five PAHs induced AhR-dependent CYP1A1 gene expression. In contrast, a metabolite of B[a]P, 3'OH-B[a]P, and a PCB metabolite, 4'OH-2,4,6-BP, did weakly upregulate all three estrogen-responsive genes. Data from these studies indicate that induction of ER-reporter activity alone does not necessarily parallel endogenous gene transcription, and that the reporter gene assay may detect interactions that are not functional in vivo.1 aGozgit, Joseph, M1 aNestor, Kathleen, M1 aFasco, Michael, J1 aPentecost, Brian, T1 aArcaro, Kathleen, F uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/differential-action-of-polycyclic-aromatic-hydrocarbons-on-endogenous02065nas a2200217 4500008004100000245011300041210006900154260001300223300001000236490000700246520130800253100001701561700001401578700001601592700001601608700002101624700001701645700001601662700001601678856015301694 2004 eng d00aField validation of Aedes aegypti (Diptera: Culicidae) age estimation by analysis of cuticular hydrocarbons.0 aField validation of Aedes aegypti Diptera Culicidae age estimati c2004 Mar a231-80 v413 aIn previous studies, we developed linear regression models to age-grade female Aedes aegypti L. reared and maintained under controlled laboratory conditions. The models were based on temporal differences between two cuticular hydrocarbons, pentacosane (C25H52) and nonacosane (C29H60), which were extracted from Ae. aegypti legs and analyzed by gas-liquid chromatography. These initial models predicted adult female age up to 165 DD (12-15 calendar d at 28 degrees C). The age of older mosquitoes, however, could not be accurately predicted. In this study, our original regression models were tested using age data obtained from mosquitoes maintained in a field laboratory and those that were marked, released, and recaptured in northwestern Thailand. Our field data led to the development of two new regression models: one for the cool-dry season (February-March) and one for the rainy season (July-August). Both models resulted in better estimates of age than the original model and thus improved our ability to predict the age of Ae. aegypti to 15 calendar d. Females older than 15 d can be identified as such, but their exact age cannot yet be estimated. The new models will be useful for epidemiological studies and evaluating the impact of Ae. aegypti control interventions for disease prevention.1 aGerade, B, B1 aLee, S, H1 aScott, T, W1 aEdman, J, D1 aHarrington, L, C1 aKitthawee, S1 aJones, J, W1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/field-validation-of-aedes-aegypti-diptera-culicidae-age-estimation-by02861nas a2200517 4500008004100000245011200041210006900153260001500222300001200237490000800249520116100257653002401418653001201442653001801454653003101472653003101503653002501534653004001559653004001599653002501639653002001664653001001684653002201694653000901716653002301725653002101748653002101769653002801790653001901818653002901837653002201866653002101888653003201909653002801941653002301969653002201992653001102014653001702025653002602042100002202068700002302090700002502113700002602138700002502164856015402189 2004 eng d00aLocalization of the domains in Runx transcription factors required for the repression of CD4 in thymocytes.0 aLocalization of the domains in Runx transcription factors requir c2004 Apr 1 a4359-700 v1723 aThe runt family transcription factors Runx1 and Runx3 are expressed in developing murine thymocytes. We show that enforced expression of full-length Runx1 in CD4(-)CD8(-) thymocytes results in a profound suppression of immature CD4/CD8 double-positive thymocytes and mature CD4 single-positive thymocytes compared with controls. This effect arises from Runx1- or Runx3-mediated repression of CD4 expression, and is independent of positively selecting signals. Runx1 is able to repress CD4 in CD4/CD8 double-positive thymocytes, but not in mature splenic T cells. Runx-mediated CD4 repression is independent of association with the corepressors Groucho/TLE or Sin3. Two domains are required for complete Runx-mediated CD4 repression. These are contained within Runx1 aa 212-262 and 263-360. The latter region contains the nuclear matrix targeting sequence, which is highly conserved among runt family transcription factors across species. The presence of the nuclear matrix targeting sequence is required for Runx-mediated CD4 repression, suggesting that Runx transcription factors are stabilized on the CD4 silencer via association with the nuclear matrix.10aAmino Acid Sequence10aAnimals10aAntigens, CD410aCD4-Positive T-Lymphocytes10aCD8-Positive T-Lymphocytes10aCell Differentiation10aCore Binding Factor Alpha 2 Subunit10aCore Binding Factor Alpha 3 Subunit10aDNA-Binding Proteins10aDown-Regulation10aFetus10aGrowth Inhibitors10aMice10aMice, Inbred C57BL10aMice, Inbred DBA10aMice, Transgenic10aMolecular Sequence Data10aNuclear Matrix10aOrgan Culture Techniques10aPeptide Fragments10aProtein Isoforms10aProtein Structure, Tertiary10aProto-Oncogene Proteins10aRepressor Proteins10aSequence Deletion10aSpleen10aThymus Gland10aTranscription Factors1 aTelfer, Janice, C1 aHedblom, Emmett, E1 aAnderson, Michele, K1 aLaurent, Micheline, N1 aRothenberg, Ellen, V uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/localization-of-the-domains-in-runx-transcription-factors-required-for03923nas a2200505 4500008004100000245014800041210006900189260001500258300001100273490000700284520232200291653001202613653002302625653001602648653002102664653002702685653001502712653001502727653001102742653001702753653001602770653002702786653000902813653003602822653000902858653002402867653002302891653001902914653001202933653002702945653003302972100002503005700002203030700001803052700001803070700002303088700002703111700002203138700002103160700001803181700002203199700002503221700001803246856015303264 2004 eng d00aLoss of heterozygosity occurs via mitotic recombination in Trp53+/- mice and associates with mammary tumor susceptibility of the BALB/c strain.0 aLoss of heterozygosity occurs via mitotic recombination in Trp53 c2004 Aug 1 a5140-70 v643 aLoss of heterozygosity (LOH) occurs commonly in cancers causing disruption of tumor suppressor genes and promoting tumor progression. BALB/c-Trp53(+/-) mice are a model of Li-Fraumeni syndrome, exhibiting a high frequency of mammary tumors and other tumor types seen in patients. However, the frequency of mammary tumors and LOH differs among strains of Trp53(+/-) mice, with mammary tumors occurring only on a BALB/c genetic background and showing a high frequency of LOH, whereas Trp53(+/-) mice on a 129/Sv or (C57BL/6 x 129/Sv) mixed background have a very low frequency of mammary tumors and show LOH for Trp53 in only approximately 50% of tumors. We have performed studies on tumors from Trp53(+/-) mice of several genetic backgrounds to examine the mechanism of LOH in BALB/c-Trp53(+/-) mammary tumors. By Southern blotting, 96% (24 of 25) of BALB/c-Trp53(+/-) mammary tumors displayed LOH for Trp53. Karyotype analysis indicated that cells lacking one copy of chromosome 11 were present in all five mammary tumors analyzed but were not always the dominant population. Comparative genomic hybridization analysis of these five tumors indicated either loss or retention of the entire chromosome 11. Thus chromosome loss or deletions within chromosome 11 do not account for the LOH observed by Southern blotting. Simple sequence length polymorphism analysis of (C57BL/6 x BALB/c) F1-Trp53(+/-) mammary tumors showed that LOH occurred over multiple loci and that a combination of maternal and paternal alleles were retained, indicating that mitotic recombination is the most likely mechanism of LOH. Nonmammary tumors of BALB/c mice also showed a high frequency of LOH (22 of 26, 85%) indicating it was not a mammary tumor specific phenomenon but rather a feature of the BALB/c strain. In (C57BL/6 x BALB/c) F1-Trp53(+/-) mice LOH was observed in 93% (13 of 14) of tumors, indicating that the high frequency of LOH was a dominant genetic trait. Thus the high frequency of LOH for Trp53 in BALB/c-Trp53(+/-) mammary tumors occurs via mitotic recombination and is a dominant genetic trait that associates with the occurrence of mammary tumors in (C57BL/6 x BALB/c) F1-Trp53(+/-) mice. These results further implicate double-strand DNA break repair machinery as important contributors to mammary tumorigenesis.
10aAnimals10aBlotting, Southern10aChromosomes10aCrosses, Genetic10aDisease Susceptibility10aDNA Damage10aDNA Repair10aFemale10aHeterozygote10aKaryotyping10aLoss of Heterozygosity10aMale10aMammary Neoplasms, Experimental10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aMice, Knockout10aMitosis10aRecombination, Genetic10aTumor Suppressor Protein p531 aBlackburn, Anneke, C1 aMcLary, Christine1 aNaeem, Rizwan1 aLuszcz, Jason1 aStockton, David, W1 aDonehower, Lawrence, A1 aMohammed, Mansoor1 aMailhes, John, B1 aSoferr, Tamar1 aNaber, Stephen, P1 aOtis, Christopher, N1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/loss-of-heterozygosity-occurs-via-mitotic-recombination-in-trp53-mice03281nas a2200265 4500008004100000245014800041210006900189260001500258300001100273490000700284520231500291100002502606700002202631700001802653700001802671700002302689700002702712700002202739700002102761700001802782700002202800700002502822700001802847856015002865 2004 eng d00aLoss of heterozygosity occurs via mitotic recombination in Trp53+/- mice and associates with mammary tumor susceptibility of the BALB/c strain.0 aLoss of heterozygosity occurs via mitotic recombination in Trp53 c2004 Aug 1 a5140-70 v643 aLoss of heterozygosity (LOH) occurs commonly in cancers causing disruption of tumor suppressor genes and promoting tumor progression. BALB/c-Trp53(+/-) mice are a model of Li-Fraumeni syndrome, exhibiting a high frequency of mammary tumors and other tumor types seen in patients. However, the frequency of mammary tumors and LOH differs among strains of Trp53(+/-) mice, with mammary tumors occurring only on a BALB/c genetic background and showing a high frequency of LOH, whereas Trp53(+/-) mice on a 129/Sv or (C57BL/6 x 129/Sv) mixed background have a very low frequency of mammary tumors and show LOH for Trp53 in only approximately 50% of tumors. We have performed studies on tumors from Trp53(+/-) mice of several genetic backgrounds to examine the mechanism of LOH in BALB/c-Trp53(+/-) mammary tumors. By Southern blotting, 96% (24 of 25) of BALB/c-Trp53(+/-) mammary tumors displayed LOH for Trp53. Karyotype analysis indicated that cells lacking one copy of chromosome 11 were present in all five mammary tumors analyzed but were not always the dominant population. Comparative genomic hybridization analysis of these five tumors indicated either loss or retention of the entire chromosome 11. Thus chromosome loss or deletions within chromosome 11 do not account for the LOH observed by Southern blotting. Simple sequence length polymorphism analysis of (C57BL/6 x BALB/c) F1-Trp53(+/-) mammary tumors showed that LOH occurred over multiple loci and that a combination of maternal and paternal alleles were retained, indicating that mitotic recombination is the most likely mechanism of LOH. Nonmammary tumors of BALB/c mice also showed a high frequency of LOH (22 of 26, 85%) indicating it was not a mammary tumor specific phenomenon but rather a feature of the BALB/c strain. In (C57BL/6 x BALB/c) F1-Trp53(+/-) mice LOH was observed in 93% (13 of 14) of tumors, indicating that the high frequency of LOH was a dominant genetic trait. Thus the high frequency of LOH for Trp53 in BALB/c-Trp53(+/-) mammary tumors occurs via mitotic recombination and is a dominant genetic trait that associates with the occurrence of mammary tumors in (C57BL/6 x BALB/c) F1-Trp53(+/-) mice. These results further implicate double-strand DNA break repair machinery as important contributors to mammary tumorigenesis.1 aBlackburn, Anneke, C1 aMcLary, Christine1 aNaeem, Rizwan1 aLuszcz, Jason1 aStockton, David, W1 aDonehower, Lawrence, A1 aMohammed, Mansoor1 aMailhes, John, B1 aSoferr, Tamar1 aNaber, Stephen, P1 aOtis, Christopher, N1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/loss-of-heterozygosity-occurs-via-mitotic-recombination-in-trp53-003291nas a2200241 4500008004100000245009800041210006900139260001300208300001200221490000700233520243200240653001202672653001802684653003402702653001102736653005502747653002202802653001302824100002202837700001402859700002602873856015002899 2004 eng d00aLow density lipoprotein receptor-related protein: regulation of the plasma membrane proteome.0 aLow density lipoprotein receptorrelated protein regulation of th c2004 Jun a1056-640 v913 aProteins in the plasma membrane anchor the cell within its microenvironment and sense changes occurring outside the cell. The anchoring interactions are cell type-specific and may involve adjacent cells or extracellular matrix proteins (ECMPs). In development, wound healing, and in various forms of pathology, including thrombosis and atherosclerosis, the microenvironment of the cell may change rapidly and dramatically. How the cell responds is strongly dependent on the protein composition of its plasma membrane, which we refer to as the plasma membrane proteome. Processes that regulate the plasma membrane proteome may alter cellular response. Low density lipoprotein receptor-related protein-1 (LRP-1) is a member of the LDL receptor family; however, LRP-1 and other less well studied members of this gene family demonstrate multiple activities unrelated to lipid homeostasis. LRP-1 binds and internalizes numerous, structurally diverse ligands, delivering most but not all these ligands to lysosomes for degradation. The intracellular tail of LRP-1 binds signaling adaptor proteins and thus may function in cell signaling. Biological activities of LRP-1 include antigen presentation, phagocytosis, removal of apoptotic cells, and regulation of vascular permeability. This review focuses on an emerging view of LRP-1 activity, in which LRP-1 regulates the protein composition of the plasma membrane and thereby "models" or "landscapes" the cell surface. In some cases, plasma membrane modeling results from the binding to bifunctional ligands or intracellular adaptor proteins, so that LRP-1 is bridged to another plasma membrane protein and the entire complex undergoes endocytosis. Membrane proteins already known to be subject to this form of regulation include urokinase-type plasminogen activator receptor, amyloid precursor protein, tissue factor, and alpha(V)-containing integrins. LRP-1 also controls the plasma membrane proteome by regulating maturation and transport of proteins in the secretory pathway. At the same time, LRP-1 serves as a receptor for specific ECMPs, including fibronectin and thrombospondin. Although ECMP-binding to LRP-1 results in endocytosis and catabolism, these receptor-ligation events also may be coupled, directly or indirectly, to cell-signaling. Based on these novel activities, LRP-1 emerges as a protein capable of modeling the interface of the cell with its microenvironment.10aAnimals10aCell Membrane10aExtracellular Matrix Proteins10aHumans10aLow Density Lipoprotein Receptor-Related Protein-110aMembrane Proteins10aProteome1 aGonias, Steven, L1 aWu, Lihua1 aSalicioni, Ana, Maria uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/low-density-lipoprotein-receptor-related-protein-regulation-of-the03340nas a2200457 4500008004100000245012500041210006900166260001600235300001300251490000800264520181900272653001202091653001902103653002502122653001802147653001402165653001802179653002002197653001402217653001502231653001702246653003702263653002002300653004002320653001602360653002602376653001702402653002502419653001102444653001902455653005502474653000902529653006302538653001702601100002602618700002002644700002302664700002402687700002202711856014902733 2004 eng d00aLow density lipoprotein receptor-related protein-1 promotes beta1 integrin maturation and transport to the cell surface.0 aLow density lipoprotein receptorrelated protein1 promotes beta1 c2004 Mar 12 a10005-120 v2793 aLow density lipoprotein receptor-related protein-1 (LRP-1) mediates the endocytosis of multiple plasma membrane proteins and thereby models the composition of the cell surface. LRP-1 also functions as a catabolic receptor for fibronectin, limiting fibronectin accumulation in association with cells. The goal of the present study was to determine whether LRP-1 regulates cell surface levels of the beta(1) integrin subunit. We hypothesized that LRP-1 may down-regulate cell surface beta(1) by promoting its internalization; however, unexpectedly, LRP-1 expression was associated with a substantial increase in cell surface beta(1) integrin in two separate cell lines, murine embryonic fibroblasts (MEFs) and CHO cells. The total amount of beta(1) integrin was unchanged because LRP-1-deficient cells retained increased amounts of beta(1) in the endoplasmic reticulum (ER). Expression of human LRP-1 in LRP-1-deficient MEFs reversed the shift in subcellular beta(1) integrin distribution. Metabolic labeling experiments demonstrated that the precursor form of newly synthesized beta(1) integrin (p105) is converted into mature beta(1) (p125) more slowly in LRP-1-deficient cells. Although low levels of cell surface beta(1) integrin, in LRP-1-deficient MEFs, were associated with decreased adhesion to fibronectin, the subcellular distribution of beta(1) integrin was most profoundly dependent on LRP-1 only after the cell cultures became confluent. A mutagen-treated CHO cell line, in which LRP-1 is expressed but retained in the secretory pathway, also demonstrated nearly complete ER retention of beta(1) integrin. These studies support a model in which LRP-1 either directly or indirectly promotes maturation of beta(1) integrin precursor and thereby increases the level of beta(1) integrin at the cell surface.10aAnimals10aAntigens, CD2910aBiological Transport10aCell Adhesion10aCell Line10aCell Membrane10aCells, Cultured10aCHO Cells10aCricetinae10aDensitometry10aDose-Response Relationship, Drug10aDown-Regulation10aElectrophoresis, Polyacrylamide Gel10aEndocytosis10aEndoplasmic Reticulum10aFibronectins10aGlycoside Hydrolases10aHumans10aImmunoblotting10aLow Density Lipoprotein Receptor-Related Protein-110aMice10aPeptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase10aTime Factors1 aSalicioni, Ana, Maria1 aGaultier, Alban1 aBrownlee, Cristina1 aCheezum, Michael, K1 aGonias, Steven, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/low-density-lipoprotein-receptor-related-protein-1-promotes-beta101849nas a2200145 4500008004100000245013400041210006900175260001600244300001100260490000800271520122500279100002101504700002501525856015301550 2004 eng d00aA PTP-PEST-like protein affects alpha5beta1-integrin-dependent matrix assembly, cell adhesion, and migration in Xenopus gastrula.0 aPTPPESTlike protein affects alpha5beta1integrindependent matrix c2004 Jan 15 a416-320 v2653 aDuring amphibian gastrulation, mesodermal cell movements depend on both cell-cell and cell-matrix interactions. Ectodermal cells from the blastocoel roof use alpha5beta1 integrins to assemble a fibronectin-rich extracellular matrix on which mesodermal cells migrate using the same alpha5beta1 integrin. In this report, we show that the tyrosine phosphatase xPTP-PESTr can prevent fibronectin fibril formation when overexpressed in ectodermal cells resulting in delayed gastrulation. In addition, isolated ectodermal cells overexpressing xPTP-PESTr are able to spread on fibronectin using the alpha5beta1 integrin in the absence of activin-A induction and before the onset of gastrulation. We further show that while the inhibition of fibrillogenesis depends on the phosphatase activity of xPTP-PESTr, induction of cell spreading does not. Finally, while cell spreading is usually associated with cell migration, xPTP-PESTr promotes ectodermal cell spreading on fibronectin but also reduces cell migration in response to activin-A, suggesting an adverse effect on cell translocation. We propose that xPTP-PESTr overexpression adversely affect cell migration by preventing de-adhesion of cells from the substrate.
1 aCousin, Hélène1 aAlfandari, Dominique uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/a-ptp-pest-like-protein-affects-alpha5beta1-integrin-dependent-matrix02708nas a2200409 4500008004100000245009800041210006900139260001600208300001100224490000700235520137600242653001201618653001201630653004201642653003301684653002501717653001101742653001501753653001501768653003801783653001501821653000901836653003601845653000901881653002401890653002301914653002101937653002601958653003701984100002502021700002302046700002202069700002502091700001802116700001802134856014602152 2003 eng d00aBALB/c alleles for Prkdc and Cdkn2a interact to modify tumor susceptibility in Trp53+/- mice.0 aBALBc alleles for Prkdc and Cdkn2a interact to modify tumor susc c2003 May 15 a2364-80 v633 aIn mice heterozygous for p53 (Trp53(+/-)), the incidence of mammary tumors varies among strains, with C57BL/6 being resistant and BALB/c being susceptible. Mammary tumor phenotypes were examined in female Trp53(+/-) F1 mice (C57BL/6 x BALB/c;n = 19) and N2 backcross mice [(C57BL/6 x BALB/c) x BALB/c] (n = 224). Susceptibility to mammary tumors segregated as a dominant phenotype in F1 females, but a higher frequency and shorter latency in N2 mice indicated a contribution from recessive-acting modifiers. Segregation of the hypomorphic BALB/c alleles for DNA-dependent protein kinase catalytic subunit (Prkdc) and p16(INK4A) (Cdkn2a) was analyzed in the N2 mice. The time to first tumor (considering all tumor types) was significantly different among the four genotype combinations (P = 0.01). Cdkn2a had a strong effect (P = 0.008) but was restricted to Prkdc(B/B) mice (P = 0.001), indicating a strong interaction between the loci. Differences in mammary tumor occurrence among genotypes for Prkdc and Cdkn2a in N2 mice were not statistically significant. This study indicates that BALB/c Prkdc and Cdkn2a alleles do modify tumor incidence in Trp53(+/-) mice and highlights the complexity of gene interaction effects in determining cancer phenotypes but discounts these alleles as major recessive loci contributing to spontaneous mammary tumor susceptibility.
10aAlleles10aAnimals10aCyclin-Dependent Kinase Inhibitor p1610aDNA-Activated Protein Kinase10aDNA-Binding Proteins10aFemale10aGenes, p1610aGenes, p5310aGenetic Predisposition to Disease10aInbreeding10aMale10aMammary Neoplasms, Experimental10aMice10aMice, Inbred BALB C10aMice, Inbred C57BL10aNuclear Proteins10aPolymorphism, Genetic10aProtein-Serine-Threonine Kinases1 aBlackburn, Anneke, C1 aBrown, Jennifer, S1 aNaber, Stephen, P1 aOtis, Christopher, N1 aWood, Jeff, T1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/balb-c-alleles-for-prkdc-and-cdkn2a-interact-to-modify-tumor-002080nas a2200193 4500008004100000245009800041210006900139260001600208300001100224490000700235520136900242100002501611700002301636700002201659700002501681700001801706700001801724856014401742 2003 eng d00aBALB/c alleles for Prkdc and Cdkn2a interact to modify tumor susceptibility in Trp53+/- mice.0 aBALBc alleles for Prkdc and Cdkn2a interact to modify tumor susc c2003 May 15 a2364-80 v633 aIn mice heterozygous for p53 (Trp53(+/-)), the incidence of mammary tumors varies among strains, with C57BL/6 being resistant and BALB/c being susceptible. Mammary tumor phenotypes were examined in female Trp53(+/-) F1 mice (C57BL/6 x BALB/c;n = 19) and N2 backcross mice [(C57BL/6 x BALB/c) x BALB/c] (n = 224). Susceptibility to mammary tumors segregated as a dominant phenotype in F1 females, but a higher frequency and shorter latency in N2 mice indicated a contribution from recessive-acting modifiers. Segregation of the hypomorphic BALB/c alleles for DNA-dependent protein kinase catalytic subunit (Prkdc) and p16(INK4A) (Cdkn2a) was analyzed in the N2 mice. The time to first tumor (considering all tumor types) was significantly different among the four genotype combinations (P = 0.01). Cdkn2a had a strong effect (P = 0.008) but was restricted to Prkdc(B/B) mice (P = 0.001), indicating a strong interaction between the loci. Differences in mammary tumor occurrence among genotypes for Prkdc and Cdkn2a in N2 mice were not statistically significant. This study indicates that BALB/c Prkdc and Cdkn2a alleles do modify tumor incidence in Trp53(+/-) mice and highlights the complexity of gene interaction effects in determining cancer phenotypes but discounts these alleles as major recessive loci contributing to spontaneous mammary tumor susceptibility.1 aBlackburn, Anneke, C1 aBrown, Jennifer, S1 aNaber, Stephen, P1 aOtis, Christopher, N1 aWood, Jeff, T1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/balb-c-alleles-for-prkdc-and-cdkn2a-interact-to-modify-tumor01232nas a2200145 4500008004100000245006200041210005800103260001300161300001100174490000700185520071200192100002000904700002400924856013800948 2003 eng d00aCell death in the thymus--it' s all a matter of contacts.0 aCell death in the thymusit s all a matter of contacts c2003 Jun a135-440 v153 aApoptosis, or programmed cell death, plays a critical role in shaping the T cell repertoire, deleting unproductive as well as potentially autoreactive T cells. Our understanding of how thymocyte apoptosis is regulated is continually evolving, as new essential modulators of this process are discovered. A conundrum that remains, however, is how signaling through essentially the same receptors and cascades evokes distinct biological responses: death by neglect, positive or negative selection. We hypothesize that the immunological synapse (IS) may be critical to transducing survival signals during thymocyte development, and suggest that factors affecting IS assembly may also influence T cell selection.1 aMinter, Lisa, M1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cell-death-in-the-thymus-it-s-all-a-matter-of-contacts01590nas a2200193 4500008004100000245014200041210006900183260001500252300001200267490000700279520084200286100002101128700002001149700001501169700001801184700001701202700002401219856015301243 2003 eng d00aComparison of three different leptospiral vaccines for induction of a type 1 immune response to Leptospira borgpetersenii serovar Hardjo.0 aComparison of three different leptospiral vaccines for induction c2003 Oct 1 a4448-580 v213 aLeptospira serovar Hardjo are bacterial pathogens of cattle that cause zoonotic infections of humans. Monovalent serovar Hardjo vaccines protect cattle from serovar Hardjo while pentavalent vaccines do not even though they contain serovar Hardjo organisms. Here, cattle vaccinated with either of two monovalent vaccines had lymphocytes that made interferon-gamma (IFN-gamma) and IgG(1) and IgG(2) antibodies to Hardjo antigen while those from cattle vaccinated with a pentavalent Leptospira vaccine did not. IFN-gamma-producing cells were mainly CD4(+), but included CD8(+) and gamma delta TCR(+) cells. Despite their monovalent composition, those vaccines also induced IFN-gamma responses to serovar Grippotyphosa antigens. Thus, induction of a type 1 immune response is consistent with protective immunity to serovar Hardjo infections.1 aBrown, Rachel, A1 aBlumerman, Seth1 aGay, Cyril1 aBolin, Carole1 aDuby, Robert1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/comparison-of-three-different-leptospiral-vaccines-for-induction-of-a02635nas a2200241 4500008004100000245007500041210006900116260001600185300001100201490000800212520184200220653001202062653001502074653001302089653001302102653002502115653000902140653002602149100002802175700001702203700002002220856015302240 2003 eng d00aDynamic morphogenetic events characterize the mouse visceral endoderm.0 aDynamic morphogenetic events characterize the mouse visceral end c2003 Sep 15 a470-870 v2613 aSeveral lines of evidence suggest that the extraembryonic endoderm of vertebrate embryos plays an important role in the development of rostral neural structures. In mice, neural inductive signals are thought to reside in an area of visceral endoderm that expresses the Hex gene. Here, we have conducted a morphological and lineage analysis of visceral endoderm cells spanning pre- and postprimitive streak stages. Our results show that Hex-expressing cells have a tall, columnar epithelial morphology, which distinguishes them from other visceral endoderm cells. This region of visceral endoderm thickening (VET) is found overlying first the distal and then one side of the epiblast at stages between 5.5 and 5.75 days post coitum (d.p.c.). In addition, we show that the epiblast has an anteroposterior-compressed appearance that is aligned with the position of the VET. Intracellular labeling of VET/Hex-expressing cells reveals an anterior and anterolateral shift from their distal epiblast position. VET/Hex-expressing cells are first localized to the anterior side of the epiblast by 5.75 d.p.c. and form a crescent on the anterior half of the embryo at the onset of gastrulation. Subsequently, VET descendants are distributed along the embryonic/extraembryonic boundary by headfold stages at 7.5 d.p.c. The morphological characteristics and position of VET/Hex-expressing cells distinguishes the future anteroposterior axis of the embryo and provide landmarks to stage mouse embryos at preprimitive streak stages. Moreover, the morphological characteristics of pregastrulation mouse embryos together with the stereotyped shift in the position of visceral endoderm cells reveal similarities among amniote embryos that suggest an evolutionary conservation of the mechanisms that pattern the rostral neurectoderm at pregastrula stages.10aAnimals10aBlastocyst10aEndoderm10aGastrula10aHomeodomain Proteins10aMice10aTranscription Factors1 aRivera-Pérez, Jaime, A1 aMager, Jesse1 aMagnuson, Terry uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/dynamic-morphogenetic-events-characterize-the-mouse-visceral-endoderm02392nas a2200181 4500008004100000245011200041210006900153260001500222300001200237490000800249520170100257100002101958700002201979700001602001700002402017700002402041856014502065 2003 eng d00aGlucocorticoid-induced apoptosis of thymocytes: requirement of proteasome-dependent mitochondrial activity.0 aGlucocorticoidinduced apoptosis of thymocytes requirement of pro c2003 Mar 1 a2469-780 v1703 aThymocytes undergo negative and positive selection during development in the thymus. During this selection process, the majority of thymocytes are eliminated by apoptosis through signaling via TCR or die by neglect, possibly mediated through glucocorticoids. In this study, we report that thymocytes require molecular oxygen to undergo apoptosis induced by dexamethasone (DEX), a synthetic glucocorticoid, and treatment with N-acetyl-L-cysteine (NAC), a thiol antioxidant, inhibits thymocyte apoptosis in vivo as well as ex vivo. We detected elevated intracellular levels of hydrogen peroxide (H(2)O(2)) during DEX-induced apoptosis, which is reduced by NAC treatment, indicating that the elevated levels of intracellular H(2)O(2) are proapoptotic. We also show that loss of mitochondrial membrane potential, cytochrome c release, as well as caspase-3 activation induced by DEX are attenuated by NAC treatment. We identified the production site for H(2)O(2) as the ubiquinone cycle at complex III of mitochondria by using various inhibitors of the mitochondrial electron transport chain, and we show that the cell death events mediated by mitochondria are also significantly reduced when the inhibitors were used. Through inhibition of the proteasome, we also show that the production of H(2)O(2) and the cell death events mediated by mitochondria are regulated by proteosomal activities in DEX-induced thymocyte apoptosis. We conclude that in DEX-treated thymocytes, the increased production of H(2)O(2) originates from mitochondria and is proapoptotic for cell death mediated by mitochondria. We also conclude that all the apoptotic events mediated by mitochondria are regulated by proteasomes.1 aTonomura, Noriko1 aMcLaughlin, Kelly1 aGrimm, Lisa1 aGoldsby, Richard, A1 aOsborne, Barbara, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/glucocorticoid-induced-apoptosis-of-thymocytes-requirement-of02531nas a2200145 4500008004100000245019200041210006900233260001300302300001200315490000700327520186500334100002402199700001902223856014302242 2003 eng d00aOestradiol-dependent and -independent modulation of tyrosine hydroxylase mRNA levels in subpopulations of A1 and A2 neurones with oestrogen receptor (ER)alpha and ER beta gene expression.0 aOestradioldependent and independent modulation of tyrosine hydro c2003 Mar a296-3030 v153 aOestradiol (E2) induces luteinizing hormone-releasing hormone (LHRH) hypersecretion, thereby triggering LH surge release in ovariectomized (OVX) rats. Neural signals responsible for the surge are marked by a morning increase in LHRH gene expression and an afternoon increase in LHRH release. Evidence suggests that subpopulations of noradrenergic neurones may be responsible for one or both of these signals. To further investigate this issue, we examined effects of E2 on the activity of A1 and A2 noradrenergic neurones, as reflected in changes in tyrosine hydroxylase (TH) mRNA expression, on the day of LH surge release. We then used dual-label in situ hybridization to determine whether E2-induced changes occurred primarily in A1 and A2 subdivisions wherein most noradrenergic neurones expressed oestrogen receptor (ER)alpha and/or ER beta mRNA. We found that in all subdivisions, levels of TH mRNA were higher in E2- than oil-treated rats at 12.00 h. These differences resulted from a decline in TH mRNA expression in oil-treated rats, as well as a rise in levels in E2-treated rats between 10.00 h and 12.00 h. During the afternoon, TH mRNA expression in most A1 and A2 subdivisions peaked at 14.00 h when LH surge release began. However, in all but the middle and caudal A2 subdivisons, levels were similar in E2-treated and control rats at this time. This was attributable to a widespread increase in TH mRNA expression between 12.00 h and 14.00 h in OVX rats. There was no evidence that E2 induced changes in TH mRNA expression preferentially in regions wherein most neurones contained ER alpha or ER beta mRNA. Our findings suggest that E2 activation of middle and caudal A2 neurones, in conjunction with the widespread E2-independent activation of noradrenergic neurones in other subdivisions, may play a role in the induction of LH surge release.1 aCurran-Rauhut, M, A1 aPetersen, S, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/oestradiol-dependent-and-independent-modulation-of-tyrosine00869nas a2200193 4500008004100000245008200041210006900123260001300192300001300205490000800218520018700226100002100413700001900434700002000453700002000473700001600493700001800509856014800527 2003 eng d00aPermethrin-resistant human head lice, Pediculus capitis, and their treatment.0 aPermethrinresistant human head lice Pediculus capitis and their c2003 Aug a994-10000 v1393 aTo compare the pediculicidal activity of Ovide lotion and its active ingredient, 0.5% malathion, with Nix and its active ingredient, 1% permethrin, in permethrin-resistant head lice.1 aYoon, Kyong, Sup1 aGao, Jian-Rong1 aLee, Si, Hyeock1 aClark, Marshall1 aBrown, Leon1 aTaplin, David uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/permethrin-resistant-human-head-lice-pediculus-capitis-and-their02387nas a2200157 4500008004100000245009100041210006900132260001500201300001000216490000700226520178000233100002302013700002002036700002002056856015302076 2003 eng d00aThe persistence and degradation of chlorothalonil and chlorpyrifos in a cranberry bog.0 apersistence and degradation of chlorothalonil and chlorpyrifos i c2003 Jan 1 a170-60 v513 aThe effect of a spray-tank adjuvant on the persistence, distribution, and degradation of two pesticides, chlorothalonil and chlorpyrifos, was studied in a commercial cranberry bog. Pesticides were applied according to label instructions to cranberry plants in paired plot studies. Dislodgeable foliar and whole fruit residues of both pesticides and several degradation products were assessed over a growing season. Residues were also assessed in soil samples collected at fruit harvest. Adjuvant increased both fruit and foliar residues but did not significantly alter the dissipation rate or metabolism of either pesticide. The dissipation of dislodgeable foliar chlorothalonil and chlorpyrifos residues followed first-order kinetics, with estimated half-lives of 12.7 and 3.5 d, respectively. All residue levels on harvested fruit were well below the current U.S. EPA tolerances for fresh cranberries. Chlorothalonil (58%) was the major residue in fruit at harvest (76 d post-chlorothalonil application), with 4-hydroxy-2,5,6-trichloroisophthalonitrile and 1,3-dicarbamoyl-2,4,5,6-tetrachlorobenzene accounting for 26% and 6% of the total residues, respectively. Degradation products accounted for 88% of the total chlorothalonil residues in soil at fruit harvest. The products 1,3-dicarbamoyl-2,4,5,6-tetrachlorobenzene, 1-carbamoyl-3-cyano-4-hydroxy-2,5,6-trichlorobenzene, 2,5,6-trichloro-4-methylthioisophthalonitrile, and 2,4,5-trichloroisophthalonitrile have not been previously identified in cranberry bog environments. Chlorpyrifos was detected in fruit at harvest (62 d post-chlorpyrifos application), but no metabolites were found. Chlorpyrifos-oxon and 3,5,6-trichloro-2-pyridinol, however, were detected in earlier fruit samples and in foliage and soil samples.1 aPutnam, Raymond, A1 aNelson, Judd, O1 aClark, Marshall uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-persistence-and-degradation-of-chlorothalonil-and-chlorpyrifos-in02064nas a2200217 4500008004100000245010500041210006900146260001600215300001000231490000700241520132000248100001801568700002101586700001401607700001601621700001301637700001701650700001401667700001601681856014901697 2002 eng d00aActivation of bovine peripheral blood gammadelta T cells for cell division and IFN-gamma production.0 aActivation of bovine peripheral blood gammadelta T cells for cel c2002 Sep 10 a251-90 v873 aBovine peripheral blood gammadelta T cells have been evaluated for effector function (IFN-gamma production) and clonal expansion in a variety of systems including following activation by mitogens, IL-12, and stimulation, through the T cell receptor (TCR) with anti-CD3 monoclonal antibody (mAb), a cell-bound molecule and a soluble antigenic extract. To evaluate cell division, carboxyfluorescein succinimidyl ester (CFSE) loading of cells and flow cytometric analysis were used, while IFN-gamma production was evaluated by intracytoplasmic staining. It was found that bovine gammadelta T cells produced IFN-gamma and clonally expanded when stimulated through the TCR/CD3 complex by a cell-associated autologous molecule on monocyte, by bacterial components following in vivo sensitization of gammadelta T cells with a leptospira vaccine or by anti-CD3 mAb. In addition, gammadelta T cells were activated efficiently for effector function but not clonal expansion by culturing with IL-12. In contrast, stimulation by Con A or PMA/ionomycin induced efficient replication but only low level IFN-gamma production which was not enhanced by the presence of IL-12. In several systems the amount of IFN-gamma produced per cell by gammadelta T cells was less than that produced by CD4 T cells in the same cultures.
1 aBaldwin, C, L1 aSathiyaseelan, T1 aNaiman, B1 aWhite, A, M1 aBrown, R1 aBlumerman, S1 aRogers, A1 aBlack, S, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/activation-of-bovine-peripheral-blood-gammadelta-t-cells-for-cell02111nas a2200193 4500008004100000245011800041210006900159260001300228300001100241490000700252520137400259100002401633700002301657700002001680700001701700700002201717700002401739856015401763 2002 eng d00aBrucella abortus siderophore 2,3-dihydroxybenzoic acid (DHBA) facilitates intracellular survival of the bacteria.0 aBrucella abortus siderophore 23dihydroxybenzoic acid DHBA facili c2002 May a239-480 v323 aSiderophores are low molecular weight molecules that allow bacteria to acquire iron from host cell proteins. 2,3-dihydroxybenzoic acid (DHBA) is the only known siderophore produced by the intracellular pathogen Brucella abortus. Here its role in virulence was assessed by evaluating the ability of a mutant with a disruption of the entC gene to survive and replicate in vitro in murine and bovine cells and in vivo in resistant and susceptible murine hosts. It was hypothesized that DHBA is vital for bacterial virulence by its ability to chelate intracellular iron thereby preventing generation of anti-bacterial hydroxyl radicals via the Haber-Weiss reaction, to scavenge reactive oxygen intermediates and for acquisition of iron needed for nutritional purposes. The data showed DHBA played a significant role for bacterial survival in host cells after infection including in murine macrophages cultured in the presence and absence of exogenous interferon-gamma (IFN-gamma) and in bovine trophoblasts supplemented with erythritol. In severely iron-depleted conditions, DHBA was also found to be essential for growth in murine macrophages. Despite these deficiencies, the absence of DHBA had no long-term significant effect on the number of CFU recovered in vivo from either the Brucella-resistant C57BL/6 mice or Brucella-susceptible IFN-gamma knock-out C57BL/6 mice.1 aParent, Michelle, A1 aBellaire, Bryan, H1 aMurphy, Erin, A1 aRoop, Martin1 aElzer, Phillip, H1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/brucella-abortus-siderophore-23-dihydroxybenzoic-acid-dhba-facilitates03056nas a2200397 4500008004100000245012200041210006900163260001300232300001400245490000800259520177400267653001202041653002102053653002502074653001802099653001702117653004202134653001202176653001502188653001402203653001102217653001502228653003102243653002702274653000902301653002502310653002902335653001402364653001702378653003302395100002002428700002402448700002202472700001802494856014602512 2002 eng d00aEpithelial cell cycling predicts p53 responsiveness to gamma-irradiation during post-natal mammary gland development.0 aEpithelial cell cycling predicts p53 responsiveness to gammairra c2002 Jun a2997-30080 v1293 aThe tumor suppressor gene, TP53, plays a major role in surveillance and repair of radiation-induced DNA damage. In multiple cell types, including mammary epithelial cells, abrogation of p53 (encoded by Trp53) function is associated with increased tumorigenesis. We examined gamma-irradiated BALB/c-Trp53(+/+) and -Trp53(-/-) female mice at five stages of post-natal mammary gland development to determine whether radiation-induced p53 activity is developmentally regulated. Our results show that p53-mediated responses are attenuated in glands from irradiated virgin and lactating mice, as measured by induction of p21/WAF1 (encoded by Cdkn1a) and apoptosis, while irradiated early- and mid-pregnancy glands exhibit robust p53 activity. There is a strong correlation between p53-mediated apoptosis and the degree of cellular proliferation, independent of the level of differentiation. In vivo, proliferation is intimately influenced by steroid hormones. To determine whether steroid hormones directly modulate p53 activity, whole organ cultures of mammary glands were induced to proliferate using estrogen plus progesterone or epidermal growth factor plus transforming growth factor-alpha and p53 responses to gamma-irradiation were measured. Regardless of mitogens used, proliferating mammary epithelial cells show comparable p53 responses to gamma-irradiation, including expression of nuclear p53 and p21/WAF1 and increased levels of apoptosis, compared to non-proliferating irradiated control cultures. Our study suggests that differences in radiation-induced p53 activity during post-natal mammary gland development are influenced by the proliferative state of the gland, and may be mediated indirectly by the mitogenic actions of steroid hormones in vivo.
10aAnimals10aAnimals, Newborn10aCell Differentiation10aCell Division10aCell Nucleus10aCyclin-Dependent Kinase Inhibitor p2110aCyclins10aEpithelium10aEstrogens10aFemale10aGamma Rays10aGene Expression Regulation10aMammary Glands, Animal10aMice10aMice, Mutant Strains10aOrgan Culture Techniques10aPregnancy10aProgesterone10aTumor Suppressor Protein p531 aMinter, Lisa, M1 aDickinson, Ellen, S1 aNaber, Stephen, P1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/epithelial-cell-cycling-predicts-p53-responsiveness-to-gamma-002439nas a2200169 4500008004100000245012200041210006900163260001300232300001400245490000800259520177400267100002002041700002402061700002202085700001802107856014402125 2002 eng d00aEpithelial cell cycling predicts p53 responsiveness to gamma-irradiation during post-natal mammary gland development.0 aEpithelial cell cycling predicts p53 responsiveness to gammairra c2002 Jun a2997-30080 v1293 aThe tumor suppressor gene, TP53, plays a major role in surveillance and repair of radiation-induced DNA damage. In multiple cell types, including mammary epithelial cells, abrogation of p53 (encoded by Trp53) function is associated with increased tumorigenesis. We examined gamma-irradiated BALB/c-Trp53(+/+) and -Trp53(-/-) female mice at five stages of post-natal mammary gland development to determine whether radiation-induced p53 activity is developmentally regulated. Our results show that p53-mediated responses are attenuated in glands from irradiated virgin and lactating mice, as measured by induction of p21/WAF1 (encoded by Cdkn1a) and apoptosis, while irradiated early- and mid-pregnancy glands exhibit robust p53 activity. There is a strong correlation between p53-mediated apoptosis and the degree of cellular proliferation, independent of the level of differentiation. In vivo, proliferation is intimately influenced by steroid hormones. To determine whether steroid hormones directly modulate p53 activity, whole organ cultures of mammary glands were induced to proliferate using estrogen plus progesterone or epidermal growth factor plus transforming growth factor-alpha and p53 responses to gamma-irradiation were measured. Regardless of mitogens used, proliferating mammary epithelial cells show comparable p53 responses to gamma-irradiation, including expression of nuclear p53 and p21/WAF1 and increased levels of apoptosis, compared to non-proliferating irradiated control cultures. Our study suggests that differences in radiation-induced p53 activity during post-natal mammary gland development are influenced by the proliferative state of the gland, and may be mediated indirectly by the mitogenic actions of steroid hormones in vivo.
1 aMinter, Lisa, M1 aDickinson, Ellen, S1 aNaber, Stephen, P1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/epithelial-cell-cycling-predicts-p53-responsiveness-to-gamma02692nas a2200205 4500008004100000245019100041210007000232260001300302300001200315490000700327520186000334100002102194700002002215700001502235700002102250700001802271700002102289700002402310856015202334 2002 eng d00aEvaluation of type 1 immune response in naïve and vaccinated animals following challenge with Leptospira borgpetersenii serovar Hardjo: involvement of WC1(+) gammadelta and CD4 T cells.0 aEvaluation of type 1 immune response in naïve and vaccinated ani c2002 Nov a6147-570 v703 aOrganisms within the Hardjo serovar of Leptospira species are harbored in cattle throughout the world, causing abortion in pregnant animals as well as being shed in the urine, thereby providing sources of zoonotic infection for humans. We recently showed that sterile immunity in vaccinated cattle is associated with induction of a type 1 (Th1) cell-mediated immune response. Here naïve and previously vaccinated pregnant cattle were challenged with a virulent strain of serovar Hardjo and subsequently evaluated for expression of a type 1 immune response. Lymphocytes that responded in a recall response to antigen by undergoing blast transformation were evident in cultures of peripheral blood mononuclear cells (PBMC) from vaccinated cattle throughout the postchallenge test period while those from naïve cattle were evident at one time point only. Nevertheless, beginning at 2 weeks after challenge, gamma interferon (IFN-gamma) was measured in supernatants of antigen-stimulated PBMC cultures from nonvaccinated animals although the amount produced was always less than that in cultures of PBMC from vaccinated animals. IFN-gamma(+) cells were also evident in antigen-stimulated cultures of PBMC from vaccinated but not from nonvaccinated animals throughout the postchallenge period. The IFN-gamma(+) cells included CD4(+) and WC1(+) gammadelta T cells, and a similar proportion of these two subpopulations were found among the dividing cells in antigen-stimulated cultures as ascertained by carboxyfluorescein succinimidyl ester loading. Finally, while naïve and vaccinated animals had similar levels of antigen-specific immunoglobulin G1 (IgG1) following challenge, vaccinated animals had twofold-more IgG2. In conclusion, while infection may induce a type 1 response we suggest that it is too weak to prevent establishment of chronic infection.1 aNaiman, Brian, M1 aBlumerman, Seth1 aAlt, David1 aBolin, Carole, A1 aBrown, Rachel1 aZuerner, Richard1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evaluation-of-type-1-immune-response-in-naive-and-vaccinated-animals01834nas a2200169 4500008004100000245006500041210006300106260001600169300001100185490000700196520123400203100002201437700002001459700001801479700002401497856014301521 2002 eng d00aExpression of the bovine high affinity IL-12 receptor beta2.0 aExpression of the bovine high affinity IL12 receptor beta2 c2002 Jan 15 a127-420 v843 aFour fragments of the bovine IL-12 receptor beta2 were sequenced following generation by reverse transcriptase polymerase chain reaction (RT-PCR) amplification of RNA from mitogen-activated bovine peripheral blood mononuclear cells (PBMC). Primers were based on sequences within regions of the human IL-12Rbeta2 gene that displayed high levels of similarity with the mouse IL-12Rbeta2 gene sequence. The amplified bovine IL-12Rbeta2 fragments had 82-87% similarity at the nucleotide level with human IL-12Rbeta2 and 70-88% similarity at the predicted amino acid level. Bovine IL-12Rbeta2 gene expression was induced following culture of PBMC with Concanavalin A (Con A), with immobilized monoclonal antibody to CD3 or with human recombinant IL-12 p70 and correlated with interferon-gamma (IFN-gamma) production. Expression of bovine IL-12Rbeta2 by PBMC was detected by 2h of culture with Con A and sustained for at least 5 days when cultured with rHuIL-12. Expression, however, did not require cellular proliferation since IL-12 did not induce proliferation, although both Con A and anti-CD3 monoclonal antibody did do so. Addition of rHuIL-10 inhibited IFN-gamma production without abrogating bovine IL-12Rbeta2 gene expression.1 aWhite, Ann, Marie1 aBlumerman, Seth1 aNaiman, Brian1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/expression-of-the-bovine-high-affinity-il-12-receptor-beta203026nas a2200145 4500008004100000245012900041210006900170260001600239300001100255490000700266520240800273100002402681700002102705856015402726 2002 eng d00aFundamentals of host immune response against Brucella abortus: what the mouse model has revealed about control of infection.0 aFundamentals of host immune response against Brucella abortus wh c2002 Dec 20 a367-820 v903 aThe studies reviewed here evaluated the role cellular immune system components play in control of brucellosis by conducting comparative studies with brucella-resistant C57BL/10 or C57BL/6 mice and susceptible BALB/c mice. We have shown by both in vitro and in vivo studies that activation of macrophages with interferon-gamma (IFN-gamma) is an important factor for control of infection with B. abortus in the mouse model and that the mechanism of anti-brucella activity largely involved reactive oxygen intermediates. Differences in control of the organism by resistant and susceptible mice was not related to inherent differences in the ability of their macrophages to control infection either with or without IFN-gamma activation nor was it attributable to NK cells since we found no role for them in control of brucellosis in either mouse strain. However, relative resistance to brucellosis did correlate with increased production of IFN-gamma by CD4 T cells during the first weeks after infection while IL-10 contributed to susceptibility in BALB/c mice. Moreover, by 3 weeks post-infection splenocytes from the susceptible BALB/c mice failed to produce IFN-gamma and relied on TNF-alpha as well as CD8 T cells to control infection until the end of the plateau phase around 6 weeks post-infection when IFN-gamma production resumed and clearance began. In contrast, IFN-gamma was crucial for control throughout the infection in the more resistant C57BL/6 mice and the mice died in its absence by 6 weeks post-infection compared to 12 weeks for the more susceptible mice that relied on additional mechanisms of control. In contrast to the IFN-gamma knock-out mice, both beta2 microglobulin knock-out C57BL/6 mice, which do not express conventional MHC class I molecules and thus cannot present antigen to CD8 T cells, or perforin knock-out C57BL/6 mice, which have no T cell cytotoxic activity, controlled and cleared the infection as well as normal C57BL/6 mice. The hiatus of IFN-gamma production in BALB/c mice correlated with very high levels of total IL-12 and it was postulated that the lack of IFN-gamma was a consequence of p40 homodimer blocking activity. However, reduction of p40 IL-12 in vivo through administration of indomethacin reduced the infection without a concomitant measurable increase in IFN-gamma. Current studies are aimed at elucidating the mechanism of the IFN-gamma hiatus.1 aBaldwin, Cynthia, L1 aParent, Michelle uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/fundamentals-of-host-immune-response-against-brucella-abortus-what-the01849nas a2200265 4500008004100000245004900041210004800090260000900138300000900147490000600156520103300162653002101195653002001216653001101236653002801247653003801275653001301313653001101326653003301337100001801370700002001388700002101408700002501429856012901454 2002 eng d00aHormonal control of p53 and chemoprevention.0 aHormonal control of p53 and chemoprevention c2002 a91-40 v43 aImprovements in the detection and treatment of breast cancer have dramatically altered its clinical course and outcome. However, prevention of breast cancer remains an elusive goal. Parity, age of menarche, and age at menopause are major risk factors drawing attention to the important role of the endocrine system in determining the risk of breast cancer, while heritable breast cancer susceptibility syndromes have implicated tumor suppressor genes as important targets. Recent work demonstrating hormonal modulation of the p53 tumor suppressor pathway draws together these established determinants of risk to provide a model of developmental susceptibility to breast cancer. In this model, the mammary epithelium is rendered susceptible due to impaired p53 activity during specific periods of mammary gland development, but specific endocrine stimuli serve to activate p53 function and to mitigate this risk. The results focus attention on p53 as a molecular target for therapies to reduce the risk of breast cancer.
10aBreast Neoplasms10aChemoprevention10aFemale10aGenes, Tumor Suppressor10aGenetic Predisposition to Disease10aHormones10aHumans10aTumor Suppressor Protein p531 aJerry, Joseph1 aMinter, Lisa, M1 aBecker, Klaus, A1 aBlackburn, Anneke, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/hormonal-control-of-p53-and-chemoprevention-001576nas a2200169 4500008004100000245004900041210004800090260000900138300000900147490000600156520103300162100001801195700002001213700002101233700002501254856012701279 2002 eng d00aHormonal control of p53 and chemoprevention.0 aHormonal control of p53 and chemoprevention c2002 a91-40 v43 aImprovements in the detection and treatment of breast cancer have dramatically altered its clinical course and outcome. However, prevention of breast cancer remains an elusive goal. Parity, age of menarche, and age at menopause are major risk factors drawing attention to the important role of the endocrine system in determining the risk of breast cancer, while heritable breast cancer susceptibility syndromes have implicated tumor suppressor genes as important targets. Recent work demonstrating hormonal modulation of the p53 tumor suppressor pathway draws together these established determinants of risk to provide a model of developmental susceptibility to breast cancer. In this model, the mammary epithelium is rendered susceptible due to impaired p53 activity during specific periods of mammary gland development, but specific endocrine stimuli serve to activate p53 function and to mitigate this risk. The results focus attention on p53 as a molecular target for therapies to reduce the risk of breast cancer.
1 aJerry, Joseph1 aMinter, Lisa, M1 aBecker, Klaus, A1 aBlackburn, Anneke, C uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/hormonal-control-of-p53-and-chemoprevention02130nas a2200181 4500008004100000245006500041210006400106260001300170300001200183490000600195520151500201100001801716700001601734700001701750700001801767700002001785856014301805 2002 eng d00aHyperbaric oxygen enhances apoptosis in hematopoietic cells.0 aHyperbaric oxygen enhances apoptosis in hematopoietic cells c2002 Dec a499-5100 v73 aHyperbaric oxygen (HBO) is 100% oxygen administered at elevated atmospheric pressure. In this study, we examined the effect of HBO on hematopoietic cell apoptosis. Cells exposed to HBO were incubated in a chamber containing 97.9% O(2) and 2.1% CO(2) at 2.4 atmospheres absolute (ATA). HBO enhanced spontaneous HL-60 cell apoptosis in a time-dependent manner; a 12 h exposure increased apoptosis by 42%. Exposing these cells to hyperoxia at standard atmospheric pressure (95% O(2), 5% CO(2) at 1 ATA) or increased pressure alone (8.75% O(2), 2.1% CO(2) at 2.4 ATA) had minimal effect on apoptosis. HBO also enhanced stimulus-induced apoptosis. HL-60 cells stimulated to die using gamma radiation underwent 33% more apoptosis than cells exposed to radiation alone. HBO enhanced melphalan, camptothecin, and chlorambucil-induced apoptosis by 22%, 13%, and 8%, respectively. Jurkat cells stimulated to die with anti-Fas antibody underwent 44% more apoptosis when exposed to HBO. Spontaneous apoptosis was increased by 15% in HBO-exposed murine thymocytes. HBO's effect on apoptosis did not require new protein synthesis. As expected, HBO exposure increased the intracellular concentration of H(2)O(2). Incubating HL-60 cells in the presence of dehydroascorbic acid partially abrogated HBO-induced increases in intracellular H(2)O(2) and apoptosis. In summary, HBO enhances spontaneous and stimulus-induced apoptosis in hematopoietic cells, at least in part, by enhancing the intracellular accumulation of H(2)O(2).1 aGanguly, B, J1 aTonomura, N1 aBenson, R, M1 aOsborne, B, A1 aGranowitz, E, V uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/hyperbaric-oxygen-enhances-apoptosis-in-hematopoietic-cells00657nas a2200133 4500008004100000245003000041210002900071260001600100300001000116490000700126520025800133100002400391856010800415 2002 eng d00aImmune response overview.0 aImmune response overview c2002 Dec 20 a365-60 v903 aA short synopsis of the history of identification of the protective cellular immune response to Brucella is given along with indication of the current major research focuses in this area. Finally, critical areas of research for the future are suggested.1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/immune-response-overview02664nas a2200193 4500008004100000245010200041210006900143260001300212300001000225490000800235520194400243100003402187700001802221700001802239700001902257700002102276700002402297856014902321 2002 eng d00aImmunological characterization of a gammadelta T-cell stimulatory ligand on autologous monocytes.0 aImmunological characterization of a gammadelta Tcell stimulatory c2002 Feb a181-90 v1053 aBovine gammadelta T cells are stimulated to proliferate by autologous monocytes. This is referred to as the autologous mixed leucocyte reaction (AMLR). It has been shown previously that the stimulatory component is constitutively expressed on the monocyte plasma membrane and is a protein or has a protein moiety. Here we showed that gammadelta T-cell responses to the monocytes requires interaction with the T-cell receptor because Fab1 fragments of a monoclonal antibody (mAb) that reacts with the delta chain of the T-cell receptor blocked proliferation in the AMLR. Monocyte molecules involved in stimulation were also characterized further by biochemical and immunological methods. A mAb, named M5, was generated by immunizing mice with bovine monocytes and shown to block the ability of monocytes to stimulate in the AMLR. Treatment of monocytes or monocyte membranes with high salt, chelating agents or phospholipase C did not affect their ability to stimulate gammadelta T-cell proliferation or reactivity with mAb M5 indicating the ability of monocytes to stimulate does not involve peripheral membrane components or a glycosyl-phosphatidylinsositol (GPI)-anchored components. Hence it was concluded that the stimulation occurred as a result of intergral membrane proteins including that recognized by mAb M5. The ligand for mAb M5 was on all bovine monocytes and to a lower level on granulocytes but not on lymphocytes. MAb M5 also reacted with sheep monocytes but not with human monocytes or murine macrophages, in agreement with a previous reports that sheep monocytes but not human or mouse mononuclear phagocytes have the capacity to stimulate bovine gammadelta T cells in in vitro cultures. The level of expression of the M5 ligand was not altered by gamma-irradiation or culture of monocytes with lipopolysaccharide but it was decreased following culture with interferon-gamma-containing cell culture supernatants.
1 aSathiyaseelan, Thillainayagam1 aNaiman, Brian1 aWelte, Stefan1 aMachugh, Niall1 aBlack, Samuel, J1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/immunological-characterization-of-a-gammadelta-t-cell-stimulatory01847nas a2200145 4500008004100000245009200041210006900133260000900202300001100211490000600222520127800228100002501506700001801531856015201549 2002 eng d00aKnockout and transgenic mice of Trp53: what have we learned about p53 in breast cancer?0 aKnockout and transgenic mice of Trp53 what have we learned about c2002 a101-110 v43 aThe human p53 tumor suppressor gene TP53 is mutated at a high frequency in sporadic breast cancer, and Li-Fraumeni syndrome patients who carry germline mutations in one TP53 allele have a high incidence of breast cancer. In the 10 years since the first knockout of the mouse p53 tumor suppressor gene (designated Trp53) was published, much has been learned about the contribution of p53 to biology and tumor suppression in the breast through the use of p53 transgenic and knockout mice. The original mice deficient in p53 showed no mammary gland phenotype. However, studies using BALB/c-Trp53-deficient mice have demonstrated a delayed involution phenotype and a mammary tumor phenotype. Together with other studies of mutant p53 transgenes and p53 bitransgenics, a greater understanding has been gained of the role of p53 in involution, of the regulation of p53 activity by hormones, of the effect of mouse strain and modifier genes on tumor phenotype, and of the cooperation between p53 and other oncogenic pathways, chemical carcinogens and hormonal stimulation in mammary tumorigenesis. Both p53 transgenic and knockout mice are important in vivo tools for understanding breast cancer, and are yet to be exploited for developing therapeutic strategies in breast cancer.1 aBlackburn, Anneke, C1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/knockout-and-transgenic-mice-of-trp53-what-have-we-learned-about-p5302087nas a2200241 4500008004100000245009200041210006900133260000900202300001100211490000600222520128500228653001201513653002101525653002701546653001101573653001501584653000901599653001901608653002101627100002501648700001801673856015401691 2002 eng d00aKnockout and transgenic mice of Trp53: what have we learned about p53 in breast cancer?0 aKnockout and transgenic mice of Trp53 what have we learned about c2002 a101-110 v43 aThe human p53 tumor suppressor gene TP53 is mutated at a high frequency in sporadic breast cancer, and Li-Fraumeni syndrome patients who carry germline mutations in one TP53 allele have a high incidence of breast cancer. In the 10 years since the first knockout of the mouse p53 tumor suppressor gene (designated Trp53) was published, much has been learned about the contribution of p53 to biology and tumor suppression in the breast through the use of p53 transgenic and knockout mice. The original mice deficient in p53 showed no mammary gland phenotype. However, studies using BALB/c-Trp53-deficient mice have demonstrated a delayed involution phenotype and a mammary tumor phenotype. Together with other studies of mutant p53 transgenes and p53 bitransgenics, a greater understanding has been gained of the role of p53 in involution, of the regulation of p53 activity by hormones, of the effect of mouse strain and modifier genes on tumor phenotype, and of the cooperation between p53 and other oncogenic pathways, chemical carcinogens and hormonal stimulation in mammary tumorigenesis. Both p53 transgenic and knockout mice are important in vivo tools for understanding breast cancer, and are yet to be exploited for developing therapeutic strategies in breast cancer.
10aAnimals10aBreast Neoplasms10aDisease Models, Animal10aFemale10aGenes, p5310aMice10aMice, Knockout10aMice, Transgenic1 aBlackburn, Anneke, C1 aJerry, Joseph uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/knockout-and-transgenic-mice-of-trp53-what-have-we-learned-about-p53-002879nas a2200385 4500008004100000245014500041210006900186260001500255300001200270490000800282520162300290653001201913653001401925653001801939653001401957653001501971653002201986653001602008653001702024653001102041653001302052653005502065653001502120653000902135653002502144653002502169653001702194100002202211700002302233700002102256700002302277700002602300700002202326856014502348 2002 eng d00aThe low density lipoprotein receptor-related protein mediates fibronectin catabolism and inhibits fibronectin accumulation on cell surfaces.0 alow density lipoprotein receptorrelated protein mediates fibrone c2002 May 3 a16160-60 v2773 aLow density lipoprotein receptor-related protein (LRP) is a member of the low density lipoprotein receptor family, which functions as an endocytic receptor for diverse ligands. In this study, we demonstrate that murine embryonic fibroblasts (MEF-2 cells) and 13-5-1 Chinese hamster ovary cells, which are LRP-deficient, accumulate greatly increased levels of cell-surface fibronectin (Fn), compared with LRP-expressing MEF-1 and CHO-K1 cells. Increased Fn was also detected in conditioned medium from LRP-deficient MEF-2 cells; however, biosynthesis of Fn by MEF-1 and MEF-2 cells was not significantly different. When LRP-deficient cells were dissociated from monolayer culture, increased levels of Fn remained with the cells, as determined by cell-surface protein biotinylation, suggesting an intimate relationship with cell surface-binding sites. The LRP antagonist, receptor-associated protein (RAP), promoted Fn accumulation in association with MEF-1 cells, whereas expression of full-length LRP in MEF-2 cells substantially decreased Fn accumulation, confirming the role of LRP in this process. Purified LRP bound directly to immobilized Fn, and this interaction was inhibited by RAP. Furthermore, MEF-1 cells degraded (125)I-Fn at an increased rate, compared with MEF-2 cells. (125)I-Fn degradation by MEF-1 cells was inhibited by RAP. These results demonstrate that LRP functions as a catabolic receptor for Fn. The function of LRP in Fn degradation and the ability of LRP to regulate levels of other plasma membrane proteins represent possible mechanisms whereby LRP prevents Fn accumulation on cell surfaces.10aAnimals10aCell Line10aCell Membrane10aCHO Cells10aCricetinae10aEmbryo, Mammalian10aFibroblasts10aFibronectins10aHumans10aKinetics10aLow Density Lipoprotein Receptor-Related Protein-110aMethionine10aMice10aRecombinant Proteins10aSulfur Radioisotopes10aTransfection1 aSalicioni, Ana, M1 aMizelle, Kellie, S1 aLoukinova, Elena1 aMikhailenko, Irina1 aStrickland, Dudley, K1 aGonias, Steven, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-low-density-lipoprotein-receptor-related-protein-mediates02660nas a2200205 4500008004100000245020900041210006900250260001600319300001200335490000700347520184100354100001402195700002002209700001402229700001702243700001502260700001602275700001802291856014502309 2002 eng d00aMajor histocompatibility complex class I and II expression on macrophages containing a virulent strain of Brucella abortus measured using green fluorescent protein-expressing brucellae and flow cytometry.0 aMajor histocompatibility complex class I and II expression on ma c2002 Jul 12 a191-2000 v333 aImmune responses appropriate for control of an intracellular pathogen are generated in mice infected with Brucella abortus, shown by the ability of T cells to adoptively transfer resistance to naive mice. The infection nevertheless persists for months. It was hypothesized that one factor in maintaining the infection despite the presence of immune T cells was suboptimal expression of major histocompatibility complex (MHC) molecules on macrophages containing brucellae. This would allow B. abortus to elude detection by the host's immune system. To test this, B. abortus organisms expressing green fluorescent protein (GFP-Brucella) were constructed and three-color flow cytometry used to evaluate MHC expression on macrophages following in vitro or in vivo infection. When infected in vitro, the levels of MHC class I and class II expression on J774 macrophages containing GFP-Brucella were the same or higher than on macrophages without GFP-Brucella in the same cultures. Similarly, the MHC expression was higher on GFP(+) peritoneal exudate cells following infection or phagocytosis of heat-killed GFP-Brucella than it was on uninfected peritoneal exudate cells. Following in vivo infection of mice the level of MHC class I and II expression on GFP(+) cells in their spleens (the main site of infection) also tended to be as high as or higher than that on the GFP-negative cells. The only in vivo GFP(+) cells that showed a decreased MHC expression was a population of splenic Mac1(+) cells recovered from interferon-gamma gene-disrupted mice at the time of their death due to an overwhelming number of bacteria per spleen. Overall, it was concluded that decreased MHC expression is not a general principle associated with brucella infection of macrophages and thus not likely to contribute to maintenance of the chronic infection.1 aMurphy, E1 aRobertson, G, T1 aParent, M1 aHagius, S, D1 aRoop, R, M1 aElzer, P, H1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/major-histocompatibility-complex-class-i-and-ii-expression-on01542nas a2200169 4500008004100000245011600041210006900157260001300226300001000239490000700249520087900256100002001135700001801155700002101173700002401194856015401218 2002 eng d00aPartial cDNA sequences of bovine CD72 and CD166/ALCAM, ligands for SRCR-family accessory molecules CD5 and CD6.0 aPartial cDNA sequences of bovine CD72 and CD166ALCAM ligands for c2002 Mar a233-90 v853 aAccessory/co-stimulatory molecules on the surface of T cells are capable of regulating activation signals. Two of these, CD5 and CD6, are molecules from the scavenger receptor cysteine rich (SRCR) superfamily. Partial sequences for the ligands of these molecules, known as CD72 and CD166 (or ALCAM), respectively, are provided for Bos taurus in this communication. Using highly conserved regions between the corresponding human and mouse genes, primers were designed and reverse transcription polymerase chain reaction was used to generate cDNA from bovine PBMC RNA. cDNA clones of several hundred base pairs in length were created and sequenced. The results showed 81% homology between bovine and human CD72 nucleotide sequences and 93% homology for the CD166 sequences. Similar levels of homology are seen between the corresponding human and mouse cDNA sequences.
1 aRogers, Aric, N1 aWelte, Stefan1 aBlack, Samuel, J1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/partial-cdna-sequences-of-bovine-cd72-and-cd166-alcam-ligands-for-srcr02431nas a2200145 4500008004100000245018800041210006900229260001300298300001000311490000700321520176000328100002402088700001902112856015402131 2002 eng d00aRegulation of glutamic acid decarboxylase 65 and 67 gene expression by ovarian steroids: identification of two functionally distinct populations of GABA neurones in the preoptic area.0 aRegulation of glutamic acid decarboxylase 65 and 67 gene express c2002 Apr a310-70 v143 aGABA neurones in the preoptic area (POA) are critical for oestradiol (E2)-dependent surge release of luteinizing hormone (LH); however, it is not clear which population(s) of POA GABA neurones is involved. The goals of the present studies were: (i) to determine whether E2 regulates GABA neurones similarly in two subdivisions of the POA that play a role in LH surge release, the rostral POA region that contains the organum vasculosum of the lamina terminalis (rPOA/OVLT), and the region containing the anteroventral periventricular nucleus (AVPV) and medial preoptic nucleus (MPN) and (ii) to determine whether GABA neurones in either or both regions exhibit temporal changes consistent with a role in the regulation of LH surge release. To accomplish these goals, we measured glutamic acid decarboxylase (GAD) 65 and 67 mRNA levels at several time points in ovariectomized (OVX), E2-treated OVX rats exhibiting LH surge release, and in E2-treated OVX rats in which LH surge release was blocked by prior administration of progesterone (P4). Our findings demonstrate that, despite their close proximity, GABA neurones in the AVPV/MPN region are regulated differently from those in the rPOA/OVLT. Only neurones in the AVPV/MPN region show temporal changes in GAD 67 mRNA expression that appear to be linked to positive-feedback effects of E2 on luteinizing hormone-releasing hormone (LHRH) and LH release. Our findings also indicate that a morning rise and an afternoon fall in GAD 67 mRNA levels marks two E2-dependent signals required for LHRH and LH surge release. Finally, our results suggest that there are distinct E2-induced signals to the rPOA/OVLT and AVPV/MPN regions and that these signals differentially regulate GAD 65 and 67 gene expression.1 aCurran-Rauhut, M, A1 aPetersen, S, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-glutamic-acid-decarboxylase-65-and-67-gene-expression-by03070nas a2200445 4500008004100000245009100041210006900132260001600201300001200217490000800229520160200237653001201839653001401851653001801865653003001883653002201913653001601935653001701951653004401968653001202012653005502024653000902079653001902088653002902107653003802136653002002174653002902194653002802223653004702251653001702298653001602315100001402331700002102345700001902366700001902385700002602404700001902430700002202449856015302471 2002 eng d00aRegulation of Rac1 activation by the low density lipoprotein receptor-related protein.0 aRegulation of Rac1 activation by the low density lipoprotein rec c2002 Dec 23 a1061-700 v1593 aThe low density lipoprotein receptor-related protein (LRP-1) binds and mediates the endocytosis of multiple ligands, transports the urokinase-type plasminogen activator receptor (uPAR) and other membrane proteins into endosomes, and binds intracellular adaptor proteins involved in cell signaling. In this paper, we show that in murine embryonic fibroblasts (MEFs) and L929 cells, LRP-1 functions as a major regulator of Rac1 activation, and that this activity depends on uPAR. LRP-1-deficient MEFs demonstrated increased Rac1 activation compared with LRP-1-expressing MEFs, and this property was reversed by expressing the VLDL receptor, a member of the same gene family as LRP-1, with overlapping ligand-binding specificity. Neutralizing the activity of LRP-1 with receptor-associated protein (RAP) increased Rac1 activation and cell migration in MEFs and L929 cells. The same parameters were unaffected by RAP in uPAR-/- MEFs, prepared from uPAR gene knockout embryos, and in uPAR-deficient LM-TK- cells. Untreated uPAR+/+ MEFs demonstrated substantially increased Rac1 activation compared with uPAR-/- MEFs. In addition to Rac1, LRP-1 suppressed activation of extracellular signal-regulated kinase (ERK) in MEFs; however, it was Rac1 (and not ERK) that was responsible for the effects of LRP-1 on MEF migration. Thus, LRP-1 regulates two signaling proteins in the same cell (Rac1 and ERK), both of which may impact on cell migration. In uPAR-negative cells, LRP-1 neutralization does not affect Rac1 activation, and other mechanisms by which LRP-1 may regulate cell migration are not unmasked.10aAnimals10aCell Line10aCell Movement10aCulture Media, Serum-Free10aEnzyme Activation10aFibroblasts10aFibronectins10aGene Expression Regulation, Enzymologic10aLigands10aLow Density Lipoprotein Receptor-Related Protein-110aMice10aMice, Knockout10aMicroscopy, Fluorescence10aMitogen-Activated Protein Kinases10aProtein Binding10arac1 GTP-Binding Protein10aReceptors, Cell Surface10aReceptors, Urokinase Plasminogen Activator10aTransfection10aVitronectin1 aMa, Zhong1 aThomas, Keena, S1 aWebb, Donna, J1 aMoravec, Radim1 aSalicioni, Ana, Maria1 aMars, Wendy, M1 aGonias, Steven, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-rac1-activation-by-the-low-density-lipoprotein-receptor02565nas a2200157 4500008004100000245006900041210006800110260001300178300001100191490000700202520198900209100002102198700001702219700002402236856014702260 2002 eng d00aResponse of bovine gammadelta T cells to activation through CD3.0 aResponse of bovine gammadelta T cells to activation through CD3 c2002 Dec a155-680 v903 aSince the T cell receptor of gammadelta T cells is associated with CD3 molecules, it is a reasonable postulate that signal transduction through CD3 would occur in gammadelta T cells as it does in alphabeta T cells. However, while a small percentage of bovine gammadelta T cells divided in cultures of peripheral blood mononuclear cells (PBMCs) in response to stimulation by anti-CD3 monoclonal antibody (mAb) the majority of viable gammadelta T cells at the end of the culture period had not. This was assessed by carboxyfluorescein succinimidyl ester (CFSE) loading of cells and flow cytometric analysis here and previously [Res. Vet. Sci. 69 (2000) 275]. When intracytoplasmic staining for interferon-gamma (IFN-gamma) was also used here to assess activation through CD3, a small proportion of gammadelta T cells (approximately 14%) produced IFN-gamma during the first 4 h of culture and by 72 h of culture that number had doubled. By comparison, a much larger proportion of CD4 and CD8 T cells stimulated with anti-CD3 mAb divided and although the percentage of CD4 and CD8 T cells that produced IFN-gamma at 4 h was similar to that of gammadelta T cells, by 72 h the majority of CD4 and CD8 T cells were IFN-gamma(+). Addition of IL-2 did not increase the proportion of gammadelta T cells that responded to anti-CD3 stimulation by cell division. To test the hypothesis that gammadelta T cells were inhibited from responding by other mononuclear cell populations within PBMC, monocytes were removed from the PBMC or gammadelta T cells were purified by magnetic-bead sorting. Only a small distinct population of the sorted cells underwent multiple cell divisions in response to anti-CD3 mAb and removal of monocytes resulted in only a moderate increase in gammadelta T cell replication. The anti-CD3 mAb stimulation system may provide a useful system to evaluate the difference in the requirements for activation and clonal expansion for gammadelta T cells versus alphabeta T cells.1 aSathiyaseelan, T1 aRogers, Aric1 aBaldwin, Cynthia, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/response-of-bovine-gammadelta-t-cells-to-activation-through-cd302092nas a2200217 4500008004100000245010400041210006900145260001300214300001100227490000600238520132600244100001401570700002301584700002001607700001401627700001801641700002201659700002001681700002101701856015201722 2002 eng d00aSerum xanthine oxidase: origin, regulation, and contribution to control of trypanosome parasitemia.0 aSerum xanthine oxidase origin regulation and contribution to con c2002 Feb a161-780 v43 aAfrican trypanosomiasis is caused by Salivarian trypanosomes, tsetse fly-transmitted protozoa that inhabit the blood plasma, lymph and interstitial fluids, and, in the case of Trypanosoma brucei species, also the cerebrospinal fluid of mammal hosts. Trypanosomiasis in people and domestic animals manifests as recurring waves of parasites in the blood and is typically fatal. In contrast, trypanosomiasis in Cape buffaloes, which are naturally selected to resist the disease, is characterized by the development of only one or a few waves of parasitemia, after which the infection becomes cryptic, being maintained by the presence of 1-20 mammal-infective organisms/ml of blood. The control of the acute phase of parasitemia in Cape buffaloes correlates with a decline in blood catalase activity and the generation of trypanocidal H(2)O(2) in serum during the catabolism of endogenous purine by xanthine oxidase. Here we review features of this response, and of trypanosome metabolism, that facilitate H(2)O(2)-mediated killing of the parasites with minimal damage to the host. We also discuss the origin and regulation of serum xanthine oxidase and catalase, and show how recovery of serum catalase in infected Cape buffaloes precludes a role for H(2)O(2) in the long-term, stable suppression of trypanosome parasitemia.1 aWang, Jun1 aVan Praagh, Andrew1 aHamilton, Erika1 aWang, Qin1 aZou, Baixiang1 aMuranjan, Madhavi1 aMurphy, Noel, B1 aBlack, Samuel, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/serum-xanthine-oxidase-origin-regulation-and-contribution-to-control02753nas a2200181 4500008004100000245016500041210006900206260001300275300001100288490000700299520204100306100001602347700001402363700001402377700001502391700001302406856015202419 2001 eng d00aDNA-based genotyping techniques for the detection of point mutations associated with insecticide resistance in Colorado potato beetle Leptinotarsa decemlineata.0 aDNAbased genotyping techniques for the detection of point mutati c2001 Oct a968-740 v573 aThree DNA-based genotyping techniques, bi-directional PCR amplification of specific allele (bi-PASA), single-stranded conformational polymorphism (SSCP) and minisequencing, have been developed and compared for the detection of the S291G (insensitive acetylcholinesterase) and L1014F (insensitive sodium channel) mutations associated with azinphos-methyl and permethrin resistance, respectively, in the Colorado potato beetle (Leptinotarsa decemlineata). Extraction of genomic DNA from individual neonates that were hatched from previously collected egg masses is the most efficient and reliable means to obtain suitable templates in terms of convenience, economy, speed and DNA quality. Bi-PASA, employing two allele-specific primers, appears to be the most efficient and rapid genotyping method for the simultaneous detection of both resistant/susceptible homozygous (SS, RR) and heterozygous (SR) alleles. Its resolution, however, is strongly dependent on the quality of template genomic DNA. SSCP also allows unambiguous genotyping, including the detection of heterozygous alleles, and is less dependent on template DNA quality, but requires a longer processing time. Minisequencing is amenable to a 96-well microtiter plate format for the processing of a large number of samples and allows direct detection of resistant/susceptible homozygous alleles but is not as efficient as the PASA and SSCP in detecting heterozygous alleles. In considering the advantages and disadvantages of each technique, DNA-based genotyping is best employed in combinations, with the bi-PASA as the primary method and the SSCP and minisequencing as the secondary validating methods. These methods are rugged, rapid, cost-effective and capable of resolving SS, RR and SR individuals. The availability of such DNA-based genotyping techniques, using neonate genomic DNA as templates, will enable the precise monitoring of the resistant and susceptible allele frequencies, including those of heterozygote individuals, in field populations of L. decemlineata.1 aClark, J, M1 aLee, S, H1 aKim, H, J1 aYoon, K, S1 aZhang, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/dna-based-genotyping-techniques-for-the-detection-of-point-mutations02128nas a2200157 4500008004100000245004400041210004300085260001300128300001000141490000800151520164000159100001301799700001801812700001801830856012201848 2001 eng d00aEarly Vlambda diversification in sheep.0 aEarly Vlambda diversification in sheep c2001 May a26-340 v1033 aThis study examined a number of tissues during early gestation in foetal sheep to determine the earliest site of Vlambda expression and time of generation of the Vlambda repertoire. Tissues, including spleen, liver, gut, blood and bone marrow, were obtained from 48, 55, 60 and 63 gestational day (g.d.) ovine foetuses and cDNA libraries were prepared from them by reverse transcription-polymerase chain reaction. Clones were randomly selected from cDNA libraries and subjected to sequencing. Analysis of these sequences and comparison with a pool of germline genes led to the following conclusions. The expression of Vlambda occurs earlier in spleen (48 g.d.) than in all of the other tissues examined. Also, diversity is seen earlier and at higher levels in early foetal spleen than in all of the other tissues examined. In this regard, it is notable that splenic Vlambda expression is readily apparent even before such gut-associated lymphoid tissue as the ileal Peyer's patch (IPP) has developed. Two germline Vlambda genes, 5.1 and 5.3 predominate in early immunoglobulin lambda light-chain gene rearrangement. Examination of Jlambda usage revealed the existence of a new Jlambda gene and its utilization during the early phases of the development of the ovine antibody repertoire. This study indicates that sites other than the IPP contribute to the diversification of the Vlambda repertoire in sheep. We suggest that it is likely that foetal spleen may provide a partially diversified B-cell repertoire before the IPP becomes active as a major site for massive clonal expansion and extensive diversification of B cells.
1 aJeong, Y1 aOsborne, B, A1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/early-vlambda-diversification-in-sheep01982nas a2200181 4500008004100000245007100041210006900112260001300181300000900194490000700203520135800210100001701568700001401585700002101599700001301620700001801633856014901651 2001 eng d00aImmune control of Brucella abortus 2308 infections in BALB/c mice.0 aImmune control of Brucella abortus 2308 infections in BALBc mice c2001 Dec a85-80 v323 aBALB/c mice infected with Brucella abortus strain 2308 have 10-fold higher levels of bacteria during the plateau phase of infection (the time period when the number of colony-forming units in vivo remains consistent) than the more resistant C57BL/10 mice. This is due to a cessation of interferon-gamma (IFN-gamma) production that begins after the first week of infection and continues until the end of the plateau phase at least 6 weeks post infection. Despite the lack of IFN-gamma production during this time BALB/c mice are able to prevent an increase in bacterial colony-forming units. Here it was shown that both tumor necrosis factor (TNF)-alpha and CD8 T cells were involved in controlling bacterial numbers in BALB/c mice during this time. That is, neutralization of TNF-alpha or depletion of CD8 T cells with monoclonal antibodies resulted in a significant increase in the number of splenic colony-forming units recovered at 3 weeks post infection. In the absence of CD8 T cells there was also a significant increase in splenic macrophages. The role of TNF-alpha may depend upon the presence of interferon-gamma early in the infection since when TNF-alpha was neutralized in interferon-gamma gene knockout mice there was a marked increase in splenic macrophages, NK cells and neutrophils but not a significant increase in colony-forming units.1 aMurphy, E, A1 aParent, M1 aSathiyaseelan, J1 aJiang, X1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/immune-control-of-brucella-abortus-2308-infections-in-balb-c-mice00797nas a2200169 4500008004100000245007700041210006900118260001500187300001000202490000700212520020100219100001600420700001700436700001400453700001300467856014700480 2001 eng d00aInnate and acquired control of trypanosome parasitaemia in Cape buffalo.0 aInnate and acquired control of trypanosome parasitaemia in Cape c2001 May 1 a562-50 v313 aThe review discusses the roles of serum xanthine oxidase, serum catalase and trypanosome-specific immune responses in the regulation of the level of trypanosome parasitaemic waves in Cape buffalo.1 aBlack, S, J1 aSicard, E, L1 aMurphy, N1 aNolan, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/innate-and-acquired-control-of-trypanosome-parasitaemia-in-cape00755nas a2200157 4500008004100000245006300041210006200104260001300166300000800179490000700187520021400194100001600408700001500424700001700439856014100456 2001 eng d00aInnate and acquired resistance to African trypanosomiasis.0 aInnate and acquired resistance to African trypanosomiasis c2001 Feb a1-90 v873 aThe review discusses the current field status of human and bovine trypanosomiases, and focuses on the molecular basis of innate and acquired control of African trypanosomes in people, cattle, and Cape buffalo.1 aBlack, S, J1 aSeed, J, R1 aMurphy, N, B uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/innate-and-acquired-resistance-to-african-trypanosomiasis02301nas a2200181 4500008004100000245013000041210006900171260001300240300001000253490000800263520161000271100001701881700002101898700001701919700001101936700001801947856015401965 2001 eng d00aInterferon-gamma is crucial for surviving a Brucella abortus infection in both resistant C57BL/6 and susceptible BALB/c mice.0 aInterferongamma is crucial for surviving a Brucella abortus infe c2001 Aug a511-80 v1033 aBrucella abortus is an intracellular bacterial pathogen that causes chronic infections in humans and a number of agriculturally important species of animals. It has been shown that BALB/c mice are more susceptible to infections with virulent strains of Brucella abortus than C57BL/6 or C57BL/10 strains. In experiments described here, gene knock-out mice were utilized to elucidate some of the salient components of resistance. Resistant C57BL/6 mice with gene deletions or disruptions in the interferon-gamma (IFN-gamma), perforin or beta(2)-microglobulin genes had decreased abilities to control intracellular infections with B. abortus strain 2308 during the first week after infection. However, only the IFN-gamma knock-out mice had a sustained inability to control infections and this resulted in death of the mice at approximately 6 weeks post-infection. These mice had a continual increase in the number of bacterial colony-forming units (CFU) in their spleens until death. When BALB/c mice with the disrupted IFN-gamma gene were infected they had more splenic CFU at one week post-infection than control mice but the increase was not statistically significant and by 3 weeks they did not have more CFU than control mice. Moreover, the number of splenic bacteria did not increase in the BALB/c IFN-gamma knock-out mice between 6 and 10.5 weeks, although they died at 10.5 weeks, the time by which normal BALB/c mice were clearing the infection. Death in both strains of IFN-gamma gene disrupted mice coincided with symptoms of cachexia and macrophages comprised > or= 75% of the splenic leucocytes.1 aMurphy, E, A1 aSathiyaseelan, J1 aParent, M, A1 aZou, B1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/interferon-gamma-is-crucial-for-surviving-a-brucella-abortus-infection00648nas a2200205 4500008004100000245005800041210005700099260000900156300001100165490000700176100001700183700001000200700001200210700001800222700001500240700001400255700001800269700001900287856013600306 2001 eng d00aModel cell lines for the study of apoptosis in vitro.0 aModel cell lines for the study of apoptosis in vitro c2001 a417-360 v661 aValavanis, C1 aHu, Y1 aYang, Y1 aOsborne, B, A1 aChouaib, S1 aGreene, L1 aAshwell, J, D1 aSchwartz, L, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/model-cell-lines-for-the-study-of-apoptosis-in-vitro03369nas a2200469 4500008004100000245010800041210006900149260001300218300001200231490000800243520194100251653001202192653003202204653002502236653001702261653002102278653002502299653001302324653003602337653002902373653001302402653001802415653001302433653001502446653001702461653001302478653000902491653002502500653001402525653002002539653001802559653001802577653001802595653001802613653001502631653002402646653002102670100001902691700001502710700002002725856015402745 2001 eng d00aMouse embryos lacking Smad1 signals display defects in extra-embryonic tissues and germ cell formation.0 aMouse embryos lacking Smad1 signals display defects in extraembr c2001 Sep a3609-210 v1283 aThe Smad proteins are important intracellular mediators of the transforming growth factor beta (TGFbeta) family of secreted growth factors. Smad1 is an effector of signals provided by the bone morphogenetic protein (BMP) sub-group of TGFbeta molecules. To understand the role of Smad1 in mouse development, we have generated a Smad1 loss-of-function allele using homologous recombination in ES cells. Smad1-/- embryos die by 10.5 dpc because they fail to connect to the placenta. Mutant embryos are first recognizable by 7.0 dpc, owing to a characteristic localized outpocketing of the visceral endoderm at the posterior embryonic/extra-embryonic junction, accompanied by a dramatic twisting of the epiblast and nascent mesoderm. Chimera analysis reveals that these two defects are attributable to a requirement for Smad1 in the extra-embryonic tissues. By 7.5 dpc, Smad1-deficient embryos show a marked impairment in allantois formation. By contrast, the chorion overproliferates, is erratically folded within the extra-embryonic space and is impeded in proximal migration. BMP signals are known to be essential for the specification and proliferation of primordial germ cells. We find a drastic reduction of primordial germ cells in Smad1-deficient embryos, suggesting an essential role for Smad1-dependent signals in primordial germ cell specification. Surprisingly, despite the key involvement of BMP signaling in tissues of the embryo proper, Smad1-deficient embryos develop remarkably normally. An examination of the expression domains of Smad1, Smad5 and Smad8 in early mouse embryos show that, while Smad1 is uniquely expressed in the visceral endoderm at 6.5 dpc, in other tissues Smad1 is co-expressed with Smad5 and/or Smad8. Collectively, these data have uncovered a unique function for Smad1 signaling in coordinating the growth of extra-embryonic structures necessary to support development within the uterine environment.10aAnimals10aBone Morphogenetic Proteins10aCell Differentiation10aCell Lineage10aCrosses, Genetic10aDNA-Binding Proteins10aEctoderm10aEmbryonic and Fetal Development10aExtraembryonic Membranes10aGastrula10aGenes, Lethal10aGenotype10aGerm Cells10aHeterozygote10aMesoderm10aMice10aMice, Mutant Strains10aPhenotype10aPhosphoproteins10aSmad Proteins10aSmad1 Protein10aSmad5 Protein10aSmad8 Protein10aStem Cells10aTissue Distribution10aTrans-Activators1 aTremblay, K, D1 aDunn, N, R1 aRobertson, E, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mouse-embryos-lacking-smad1-signals-display-defects-in-extra-embryonic02397nas a2200181 4500008004100000245014200041210006900183260001300252300001100265490000700276520170100283100001701984700001102001700001602012700001502028700001802043856015402061 2001 eng d00aProtective killed Leptospira borgpetersenii vaccine induces potent Th1 immunity comprising responses by CD4 and gammadelta T lymphocytes.0 aProtective killed Leptospira borgpetersenii vaccine induces pote c2001 Dec a7550-80 v693 aLeptospira borgpetersenii serovar hardjo is the most common cause of bovine leptospirosis and also causes zoonotic infections of humans. A protective killed vaccine against serovar hardjo was shown to induce strong antigen-specific proliferative responses by peripheral blood mononuclear cells (PBMC) from vaccinated cattle by 2 months after the first dose of vaccine. This response was absent from nonvaccinated control cattle. The mean response peaked by 2 months after completion of the two-dose vaccination regimen, and substantial proliferation was measured in in vitro cultures throughout the 7 months of the study period. Variations in magnitude of the response occurred among the vaccinated animals, but by 7 months postvaccination there was a substantial antigen-specific response with PBMC from all vaccinated animals. Up to one-third of the PBMC from vaccinated animals produced gamma interferon (IFN-gamma) after 7 days in culture with antigen, as ascertained by flow cytometric analysis, and significant levels of IFN-gamma were measured in culture supernatants by enzyme-linked immunosorbent assay. Two-color immunofluorescence revealed that one-third of the IFN-gamma-producing cells were gammadelta T cells, with the remaining cells being CD4(+) T cells. The significance of this study is the very potent Th1-type immune response induced and sustained following vaccination with a killed bacterial vaccine adjuvanted with aluminum hydroxide and the involvement of gammadelta T cells in the response. Moreover, induction of this Th1-type cellular immune response is associated with the protection afforded by the bovine leptospiral vaccine against L. borgpetersenii serovar hardjo.1 aNaiman, B, M1 aAlt, D1 aBolin, C, A1 aZuerner, R1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/protective-killed-leptospira-borgpetersenii-vaccine-induces-potent-th102397nas a2200205 4500008004100000245016600041210006900207260001300276300001100289490000800300520160400308100001801912700001901930700002101949700001601970700002001986700001602006700001902022856015002041 2000 eng d00aDetection of estrogen receptor-beta messenger ribonucleic acid and 125I-estrogen binding sites in luteinizing hormone-releasing hormone neurons of the rat brain.0 aDetection of estrogen receptorbeta messenger ribonucleic acid an c2000 Sep a3506-90 v1413 aLuteinizing hormone-releasing hormone (LHRH) neurons of the forebrain play a pivotal role in the neuroendocrine control of reproduction. Although serum estrogen levels influence many aspects of LHRH neuronal activity in the female, earlier studies were unable to detect estrogen receptors (ERs) within LHRH neurons, thus shaping a consensus view that the effects of estradiol on the LHRH neuronal system are mediated by interneurons and/or the glial matrix. The present studies used dual-label in situ hybridization histochemistry (ISHH) and combined LHRH-immunocytochemistry/125I-estrogen binding to readdress the estrogen-receptivity of LHRH neurons in the female rat. In ISHH experiments we found that the majority of LHRH neurons exhibited hybridization signal for the "beta" form of ER (ER-beta). The degree of colocalization was similar in topographically distinct populations of LHRH neurons and was not significantly altered by estradiol (67.2+/-1.8% in ovariectomized and 73.8+/-4.2% in ovariectomized and estradiol-treated rats). In contrast, the mRNA encoding the classical ER-alpha could not be detected within LHRH neurons. In addition, in vivo binding studies using 125I-estrogen revealed a subset of LHRH-immunoreactive neurons (8.8%) which accumulated the radioligand thus providing evidence for the translation of ER protein(s) within these cells. The findings that most LHRH neurons in the female rat express ER-beta mRNA and at least some are capable of binding 125I-estrogen challenge the current opinion that estrogen does not exert direct effects upon the LHRH neuronal system.1 aHrabovszky, E1 aShughrue, P, J1 aMerchenthaler, I1 aHajszán, T1 aCarpenter, C, D1 aLiposits, Z1 aPetersen, S, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/detection-of-estrogen-receptor-beta-messenger-ribonucleic-acid-and02445nas a2200193 4500008004100000245019600041210006900237260001600306300001100322490000800333520165800341100001901999700001702018700001802035700002002053700001802073700001702091856014302108 2000 eng d00aDistribution of mRNAs encoding the arylhydrocarbon receptor, arylhydrocarbon receptor nuclear translocator, and arylhydrocarbon receptor nuclear translocator-2 in the rat brain and brainstem.0 aDistribution of mRNAs encoding the arylhydrocarbon receptor aryl c2000 Nov 20 a428-390 v4273 aDioxin exposure alters a variety of neural functions, most likely through activation of the arylhydrocarbon receptor (AhR) pathway. Many of the adverse effects, including disruption of circadian changes in hormone release and depressed appetite, seem to be mediated by hypothalamic and/or brainstem neurons. However, it is unclear whether these effects are direct or indirect, because there have been no comprehensive studies mapping the expression of components of the AhR pathway in the brain. Therefore, we used a sensitive in situ hybridization histochemical (ISHH) method to map the neural expression of AhR mRNA, as well as those of the mRNAs encoding the AhR dimerization partners, arylhydrocarbon receptor nuclear translocator (ARNT) and ARNT2. We found that AhR, ARNT, and ARNT2 mRNAs were widely distributed throughout the brain and brainstem. There was no neuroanatomic evidence that AhR is preferentially colocalized with ARNT or ARNT2. However, ARNT2, unlike ARNT expression, was relatively high in most regions. The most noteworthy regions in which we found AhR, ARNT, and ARNT2 mRNA were several hypothalamic and brainstem regions involved in the regulation of appetite and circadian rhythms, functions that are disrupted by dioxin exposure. These regions included the arcuate nucleus (Arc), ventromedial hypothalamus (VMH), paraventricular nucleus (PVN), suprachiasmatic nucleus (SCN), nucleus of the solitary tract (NTS), and the dorsal and median raphe nuclei. This neuroanatomic information provides important clues as to the sites and mechanisms underlying the previously unexplained effects of dioxins in the central nervous system.1 aPetersen, S, L1 aCurran, M, A1 aMarconi, S, A1 aCarpenter, C, D1 aLubbers, L, S1 aMcAbee, M, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/distribution-of-mrnas-encoding-the-arylhydrocarbon-receptor01745nas a2200145 4500008004100000245017700041210006900218260001300287300001100300490000700311520109400318100002101412700001801433856014801451 2000 eng d00aEvaluation of cell replication by bovine T cells in polyclonally activated cultures using carboxyfluorescein succinimidyl ester (CFSE) loading and flow cytometric analysis.0 aEvaluation of cell replication by bovine T cells in polyclonally c2000 Dec a275-810 v693 aStudies reported here demonstrated that carboxyfluorescein succinimidyl ester (CFSE) loading of lymphocytes and flow cytometric analysis is a powerful assay to assess the kinetics and extent of cellular replication by bovine T-cell subpopulations in heterogeneous cultures of peripheral blood mononuclear cells (PBMC) where subpopulation interactions can occur. As CFSE analysis allows determination of the proportion of lymphocytes that divided, as well as the number of cell divisions each cell underwent, distinctions in responses among mitogen-stimulated cultures could be made even when(3)H-thymidine incorporation was equivalent. When combined with surface staining for detection of differentiation antigens, differences among T-cell subpopulations with regard to the number of divisions their members had undergone, were found. Anti-CD3 mAb stimulated both CD8(+)and CD4(+)T cells to undergo several cell divisions in 72 hours, while there was essentially no division by gamma delta T cells. In contrast, in concanavalin A-stimulated cultures, all T-cell subpopulations had divided.1 aSathiyaseelan, T1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evaluation-of-cell-replication-by-bovine-t-cells-in-polyclonally02871nas a2200433 4500008004100000245008000041210006900121260001300190300001200203490000800215520154100223653001201764653002301776653002501799653001701824653002501841653002201866653001301888653001101901653004601912653001501958653000901973653001301982653000901995653002302004653002502027653002502052653001802077653002402095653001802119653001802137653002102155653003602176100001902212700001902231700001702250700002002267856015002287 2000 eng d00aFormation of the definitive endoderm in mouse is a Smad2-dependent process.0 aFormation of the definitive endoderm in mouse is a Smad2dependen c2000 Jul a3079-900 v1273 aTGFbeta growth factors specify cell fate and establish the body plan during early vertebrate development. Diverse cellular responses are elicited via interactions with specific cell surface receptor kinases that in turn activate Smad effector proteins. Smad2-dependent signals arising in the extraembryonic tissues of early mouse embryos serve to restrict the site of primitive streak formation and establish anteroposterior identity in the epiblast. Here we have generated chimeric embryos using lacZ-marked Smad2-deficient ES cells. Smad2 mutant cells extensively colonize ectodermal and mesodermal populations without disturbing normal development, but are not recruited into the definitive endoderm lineage during gastrulation. These experiments provide the first evidence that TGFbeta signaling pathways are required for specification of the definitive endoderm lineage in mammals and identify Smad2 as a key mediator that directs epiblast derivatives towards an endodermal as opposed to a mesodermal fate. In largely Smad2-deficient chimeras, asymmetric nodal gene expression is maintained and expression of pitx2, a nodal target, is also unaffected. These results strongly suggest that other Smad(s) act downstream of Nodal signals in mesodermal populations. We found Smad2 and Smad3 transcripts both broadly expressed in derivatives of the epiblast. However, Smad2 and not Smad3 mRNA is expressed in the visceral endoderm, potentially explaining why the primary defect in Smad2 mutant embryos originates in this cell population.10aAnimals10abeta-Galactosidase10aCell Differentiation10aCell Lineage10aDNA-Binding Proteins10aEmbryo, Mammalian10aEndoderm10aFemale10aGene Expression Regulation, Developmental10aHomozygote10aMale10aMesoderm10aMice10aMice, Inbred C57BL10aMice, Inbred Strains10aMice, Mutant Strains10aNodal Protein10aSignal Transduction10aSmad2 Protein10aSmad3 Protein10aTrans-Activators10aTransforming Growth Factor beta1 aTremblay, K, D1 aHoodless, P, A1 aBikoff, E, K1 aRobertson, E, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/formation-of-the-definitive-endoderm-in-mouse-is-a-smad2-dependent02034nas a2200157 4500008004100000245009200041210006900133260001300202300001100215490000800226520144600234100002101680700001301701700001801714856014401732 2000 eng d00aGrowth of Brucella abortus in macrophages from resistant and susceptible mouse strains.0 aGrowth of Brucella abortus in macrophages from resistant and sus c2000 Aug a289-940 v1213 aC57Bl/10 mice have a superior ability to control chronic infections with virulent strains of the intracellular bacteria Brucella abortus compared with BALB/c mice. While a number of differences in the cytokines produced by lymphocytes following infection of these two strains of mice have been shown, macrophages have not been evaluated for their role in conveying relative resistance. The importance of macrophages in control of brucella infections is demonstrated by the observations that intracellular survival of various strains of B. abortus directly correlates with their virulence in vivo, and the ability of macrophages to control brucellae in vitro has been shown to correlate with a brucella-resistant phenotype in ruminants. While both BALB/c and C57Bl are Nramp-susceptible mouse strains, additional differences in macrophage function outside of the Nramp1 gene effects could influence susceptibility to brucellosis. The studies conducted here comparing the ability of macrophages from C57Bl/10 and BALB/c mice indicate that the macrophages from resistant mice did not control intracellular growth of B. abortus strain 2308 more efficiently than those from the susceptible mice, either in the absence of, or following, interferon-gamma activation or iron supplementation. A number of different conditions for culturing macrophages were evaluated to rule out the influence of antibiotics on the conclusions drawn from the results.1 aSathiyaseelan, J1 aJiang, X1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/growth-of-brucella-abortus-in-macrophages-from-resistant-and03606nas a2200565 4500008004100000245017300041210006900214260001500283300001000298490000700308520176300315653002202078653002402100653001202124653002302136653003202159653003102191653002302222653002302245653001402268653002302282653001102305653002002316653002802336653001102364653004002375653000902415653002902424653002202453653002802475653002002503653002102523653003202544653001302576653001902589653002502608653002302633653002702656653003402683653002402717653003002741100002002771700001002791700002102801700001502822700001602837700001902853700001702872856015102889 2000 eng d00aIdentification and structural analysis of human RBM8A and RBM8B: two highly conserved RNA-binding motif proteins that interact with OVCA1, a candidate tumor suppressor.0 aIdentification and structural analysis of human RBM8A and RBM8B c2000 Oct 1 a54-620 v693 aThe OVCA1 gene is a candidate for the breast and ovarian tumor suppressor gene at chromosome 17p13.3. To help determine the function(s) of OVCA1, we used a yeast two-hybrid screening approach to identify OVCA1-associating proteins. One such protein, which we initially referred to as BOV-1 (binder of OVCA1-1) is 173 or 174 amino acids in length and appears to be a new member of a highly conserved RNA-binding motif (RBM) protein family that is highly conserved evolutionarily. Northern blot analysis revealed that BOV-1 is ubiquitously expressed and that three distinct messenger RNA species are expressed, 1-, 3.2-, and 5.8-kb transcripts. The 1-kb transcript is the most abundant and is expressed at high levels in the testis, heart, placenta, spleen, thymus, and lymphocytes. Using fluorescence in situ hybridization and the 5.8-kb complementary DNA probe, we determined that BOV-1 maps to both chromosome 5q13-q14 and chromosome 14q22-q23. Further sequence analysis determined that the gene coding the 1- and the 3.2-kb transcripts (HGMW-approved gene symbol RBM8A) maps to 14q22-q23, whereas a second highly related gene coding for the 5.8-kb transcript resides at chromosome 5q13-q14 (HGMW-approved gene symbol RBM8B). The predicted proteins encoded by RBM8A and RBM8B are identical except that RBM8B is 16 amino acids shorter at its N-terminus. Molecular modeling of the RNA-binding domain of RBM8A and RBM8B, based on homology to the sex-lethal protein of Drosophila, identifies conserved residues in the RBM8 protein family that are likely to contact RNA in a protein-RNA complex. The conservation of sequence and structure through such an evolutionarily divergent group of organisms suggests an important function for the RBM8 family of proteins.10aAmino Acid Motifs10aAmino Acid Sequence10aAnimals10aChromosome Mapping10aChromosomes, Human, Pair 1410aChromosomes, Human, Pair 510aCloning, Molecular10aConserved Sequence10aCOS Cells10aDNA, Complementary10aFemale10aGene Expression10aGenes, Tumor Suppressor10aHumans10aIn Situ Hybridization, Fluorescence10aMale10aMicroscopy, Fluorescence10aModels, Molecular10aMolecular Sequence Data10aProtein Binding10aProtein Isoforms10aProtein Structure, Tertiary10aProteins10aRNA, Messenger10aRNA-Binding Proteins10aSequence Alignment10aSequence Analysis, DNA10aSequence Homology, Amino Acid10aTissue Distribution10aTumor Suppressor Proteins1 aSalicioni, A, M1 aXi, M1 aVanderveer, L, A1 aBalsara, B1 aTesta, J, R1 aDunbrack, R, L1 aGodwin, A, K uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-and-structural-analysis-of-human-rbm8a-and-rbm8b-two02940nas a2200481 4500008004100000245007000041210006800111260001500179300001200194490000800206520144700214653001801661653002401679653001201703653001701715653004601732653002501778653002601803653002201829653002601851653002001877653002301897653002801920653001701948653001401965653002101979653002002000653001302020653003202033653001902065653002302084653003402107653002502141653002602166653001902192653002102211100001402232700001602246700001302262700001802275700001702293856014802310 2000 eng d00aPACSIN2 is a regulator of the metalloprotease/disintegrin ADAM13.0 aPACSIN2 is a regulator of the metalloproteasedisintegrin ADAM13 c2000 Nov 1 a197-2100 v2273 aADAM13 is a cell surface metalloprotease expressed in cephalic neural crest cells during early Xenopus development. The cytoplasmic domain of ADAM13 contains three potential SH3 (Src homology type 3) binding sites, suggesting that this region may support interactions with intracellular proteins. In this report we describe the identification, by a new strategy, of three proteins that bind the ADAM13 cytoplasmic domain in vitro: X-Src1, X-An4, and X-PACSIN2. We focused our study on X-PACSIN2 protein because it colocalizes with ADAM13 in migrating neural crest cells during embryonic development. Using pull-down experiments we show that X-PACSIN2 binds to ADAM13 in vitro. Using Xenopus XTC cells, we demonstrate that ADAM13 and X-PACSIN2 colocalize to membrane ruffles and cytoplasmic vesicles. We also show that X-PACSIN2 overexpression can rescue developmental alterations induced by overexpression of ADAM13, suggesting that both proteins interact in vivo. Finally, our results suggest that X-PACSIN2 overexpression reduces endogenous ADAM13 function while a truncated X-PACSIN2 (DeltaSH3) increases this activity in cephalic neural crest cells. We propose that X-PACSIN2 may regulate ADAM13 activity by influencing either its subcellular localization or its catalytic activity. In agreement with this model, elimination of the ADAM13 cytoplasmic domain increased developmental alterations attributable to ADAM13 proteolytic activity.10aADAM Proteins10aAmino Acid Sequence10aAnimals10aDisintegrins10aGene Expression Regulation, Developmental10aImmunohistochemistry10aIn Situ Hybridization10aMembrane Proteins10aMetalloendopeptidases10aMicroinjections10aModels, Biological10aMolecular Sequence Data10aNeural Crest10aPhenotype10aPrecipitin Tests10aProtein Binding10aProteins10aRecombinant Fusion Proteins10aRNA, Messenger10aSequence Alignment10aSequence Homology, Amino Acid10asrc Homology Domains10aSubstrate Specificity10aXenopus laevis10aXenopus Proteins1 aCousin, H1 aGaultier, A1 aBleux, C1 aDarribère, T1 aAlfandari, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/pacsin2-is-a-regulator-of-the-metalloprotease-disintegrin-adam1301848nas a2200157 4500008004100000245008800041210006900129260001300198300001000211490000800221520127300229100001201502700001601514700001601530856014401546 2000 eng d00aPurine requirements for the expression of Cape buffalo serum trypanocidal activity.0 aPurine requirements for the expression of Cape buffalo serum try c2000 Jan a25-320 v1253 aCape buffalo serum contains xanthine oxidase which generates trypanocidal H(2)O(2) during the catabolism of hypoxanthine and xanthine. The present studies show that xanthine oxidase-dependent trypanocidal activity in Cape buffalo serum was also elicited by purine nucleotides, nucleosides, and bases even though xanthine oxidase did not catabolize those purines. The paradox was explained in part, by the presence in serum of purine nucleoside phosphorylase and adenosine deaminase, that, together with xanthine oxidase, catabolized adenosine, inosine, hypoxanthine, and xanthine to uric acid yielding trypanocidal H(2)O(2). In addition, purine catabolism by trypanosomes provided substrates for serum xanthine oxidase and was implicated in the triggering of xanthine oxidase-dependent trypanocidal activity by purines that were not directly catabolized to uric acid in Cape buffalo serum, namely guanosine, guanine, adenine monophosphate, guanosine diphosphate, adenosine 3':5-cyclic monophosphate, and 1-methylinosine. The concentrations of guanosine and guanine that elicited xanthine oxidase-dependent trypanocidal activity were 30-270-fold lower than those of other purines requiring trypanosome-processing which suggests differential processing by the parasites.1 aWang, Q1 aHamilton, E1 aBlack, S, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/purine-requirements-for-the-expression-of-cape-buffalo-serum02199nas a2200169 4500008004100000245009300041210006900134260001300203300001100216490000700227520157400234100001801808700002101826700001401847700001601861856015201877 2000 eng d00aRapid changes occur in the percentage of circulating bovine WC1(+)gamma delta Th1 cells.0 aRapid changes occur in the percentage of circulating bovine WC1g c2000 Oct a175-800 v693 agamma delta T cells found in the peripheral blood of cattle include a major subpopulation distinguished by expression of WC1. These cells are distinct from the WC1(-)gamma delta T cell population based on T cell receptor gene usage. We documented that a group of 6-month-old calves allowed free-range grazing and access to their mothers had a significantly greater proportion of total gamma delta T cells in their blood, attributable to the WC1(+)gamma delta T cell subpopulation, compared to age and breed-matched calves held in conventional housing. When the animals with the greater proportion of gamma delta T cells were transferred to conventional housing there was a decrease in the WC1(+)population so that by 3 weeks after transfer there was no longer a significant difference between the two groups. To investigate the biological activities of WC1(+)gamma delta T cells, the cells were purified by flow cytometric sorting. In vitro, they responded to stimulation by irradiated monocytes in autologous mixed leukocyte reaction (AMLR) cultures but not to direct stimulation through the T cell receptor (T c R) by anti-delta monoclonal antibody. After stimulation in the AMLR, WC1(+)gamma delta T cells had a Th1 cytokine profile characterised by production of IFN -gamma and lack of IL -4. Thus we propose that higher levels of the WC1(+)gamma delta T cells may provide calves with a mechanism to produce Th1 cytokines and that the level of these cells may be modulated according to environment or stress since both groups of calves were apparently disease-free.1 aBaldwin, C, L1 aSathiyaseelan, T1 aRocchi, M1 aMcKeever, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/rapid-changes-occur-in-the-percentage-of-circulating-bovine-wc1gamma02729nas a2200373 4500008004100000245018100041210006900222260001300291300001100304490000700315520148000322653001801802653001101820653002101831653002901852653002201881653003001903653003601933653002101969653001101990653001102001653002802012653002202040653000802062653002602070100002802096700001302124700001602137700001002153700002002163700001202183700001302195856014702208 2000 eng d00aS100P calcium-binding protein overexpression is associated with immortalization of human breast epithelial cells in vitro and early stages of breast cancer development in vivo.0 aS100P calciumbinding protein overexpression is associated with i c2000 Feb a231-400 v163 aThe mechanism of cell immortalization of human breast epithelial cells leading to neoplastic transformation is not clear. The isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10F, have provided a valuable tool to identify genes involved in this process. Using the technique of differential display, we have identified seven cDNA bands differentially displayed in the MCF-10F cells when compared with the mortal S130 cells from which MCF-10F was originated. One of these bands was isolated and cloned. Sequence analysis revealed 99% homology to the EF-hand calcium-binding protein S100P (Placental). The clone was overexpressed in the immortal cell line MCF-10F when compared to the mortal counterpart S130 or other primary cultures of human breast epithelial cells. In addition, it was highly expressed in chemically transformed breast epithelial cell lines (BP1E and D3. 1), breast cancer cell line T47D, as well as in three invasive ductal carcinomas when compared to their normal adjacent tissue. The S100P protein was localized by immunohistochemistry, using a monoclonal antibody against the same amino acid sequence of the gene cloned, in ductal hyperplasias, in situ and invasive ductal carcinoma, but not in the normal tissues. We concluded that S100P overexpression is an early event that might play an important role in the immortalization of human breast epithelial cells in vitro and tumor progression in vivo.10aBase Sequence10aBreast10aBreast Neoplasms10aCalcium-Binding Proteins10aCarcinoma in Situ10aCarcinoma, Ductal, Breast10aCell Transformation, Neoplastic10aEpithelial Cells10aFemale10aHumans10aMolecular Sequence Data10aNeoplasm Proteins10aRNA10aTumor Cells, Cultured1 aDa Silva, I, D Guerreir1 aHu, Y, F1 aRusso, I, H1 aAo, X1 aSalicioni, A, M1 aYang, X1 aRusso, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/s100p-calcium-binding-protein-overexpression-is-associated-with02456nas a2200409 4500008004100000245015200041210006900193260001300262300001100275490000700286520114700293653004301440653001501483653001201498653001401510653001101524653002401535653001601559653001601575653001701591653000901608653001701617653002501634653003201659653001501691653001401706653002601720653003201746653001601778100001601794700001501810700002001825700001301845700001701858700001701875856015401892 2000 eng d00aSerotonin inhibits luteinizing hormone release via 5-HT1A receptors in the zona incerta of ovariectomised, anaesthetised rats primed with steroids.0 aSerotonin inhibits luteinizing hormone release via 5HT1A recepto c2000 Nov a272-830 v723 aThe zona incerta (ZI), an area in the dorsal hypothalamus, contains neuronal systems that appear to control gonadotropin release. Previous findings show that there is an inverse relationship between serotonin (5-HT) activity in the ZI and plasma luteinizing hormone (LH) levels, indicating that the 5-HT system in this area has an inhibitory effect on LH release. Employing anaesthetised, ovariectomised rats primed with 5 microg oestradiol benzoate followed at 48 h by 0.5 mg progesterone, we have shown that 2 microg/side 5-HT in the ZI inhibits the LH surge that normally occurs 4 h after the progesterone treatment. This effect was mimicked by 2 microg/side 8-OH-DPAT, a 5-HT1A agonist, but not by DOI, a 5-HT2 agonist, BMY7378, a presynaptic 5-HT1A agonist or MCPP, a 2B & 2C agonist. The inhibitory effect of 5-HT and 8-OH-DPAT was prevented by pretreatment, 1 h before, with either 2 mg/kg i.p. WAY100135, a 5-HT1A antagonist or 0.25 mg/kg i.p. ritanserin, a 5-HT2 antagonist. These results indicate that 5-HT in the ZI exerts its inhibitory effect on LH release via 5-HT1A receptors but that another 5-HT subtype may also be involved.10a8-Hydroxy-2-(di-n-propylamino)tetralin10aAnesthesia10aAnimals10aEstradiol10aFemale10aLuteinizing Hormone10aOvariectomy10aPiperazines10aProgesterone10aRats10aRats, Wistar10aReceptors, Serotonin10aReceptors, Serotonin, 5-HT110aRitanserin10aSerotonin10aSerotonin Antagonists10aSerotonin Receptor Agonists10aSubthalamus1 aSiddiqui, A1 aKotecha, K1 aSalicioni, A, M1 aKalia, V1 aMurray, J, F1 aWilson, C, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/serotonin-inhibits-luteinizing-hormone-release-via-5-ht1a-receptors-in02687nas a2200157 4500008004100000245015900041210006900200260001300269300001100282490000700293520203200300100001302332700001802345700001602363856015002379 2000 eng d00aToxicity and residual effectiveness of insecticides on insecticide-treated spheres for controlling females of Rhagoletis pomonella (Diptera: Tephritidae).0 aToxicity and residual effectiveness of insecticides on insectici c2000 Apr a403-110 v933 aThis study evaluated the toxicity of five technical-grade insecticides of four different classes to apple maggot females, Rhagoletis pomonella (Walsh), following a 10-min exposure period in insecticide-coated glass jars, with or without a feeding stimulant (sucrose) present. According to LC90 values for toxicity by ingestion and tarsal contact, imidacloprid was 1.5 times more toxic than dimethoate or abamectin, diazinon was less toxic, and phloxine B (a phototoxic dye) least toxic. Based on LC90 values for tarsal contact alone, dimethoate was 2.3, 4.0, and 18.4 times more toxic than imidacloprid, abamectin, and diazinon, respectively. Contact alone with phloxine B caused no mortality. When exposure was assessed using spheres coated with a latex paint mixture containing sucrose and formulated dimethoate (Digon 400 EC) or imidacloprid (Provado 1.6 F) at concentrations ranging from 5 to 70 g (AI)/cm2, both insecticides showed reduced effectiveness compared with toxicities from glass jar tests, with Digon two times more toxic than Provado. After exposure to artificial rainfall and retreatment with sucrose, Digon- and Provado-treated spheres exhibited greatest residual effectiveness, with diazinon-treated spheres less effective. Spheres treated with formulated abamectin (Agri-Mek 0.15 EC) at 1.0% (AI) performed only slightly better than phloxine B-treated spheres, which completely lost effectiveness after exposure to rainfall. Spheres treated with formulated imidacloprid (Merit 75 WP) at 1.5% (AI) showed equal or better residual efficacy in killing apple maggot flies (> 80% mortality, shorter lethal duration of feeding) over a 12-wk exposure period to outdoor weather than spheres treated with Digon at 1.0% (AI) after both types were retreated with sucrose. Our results indicate that imidacloprid is a promising safe substitute for dimethoate as a fly killing agent on lure-kill spheres. Imidacloprid formulated as Merit 75 WP had greater residual efficacy than imidacloprid formulated as Provado 1.6 F.1 aHu, X, P1 aProkopy, R, J1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/toxicity-and-residual-effectiveness-of-insecticides-on-insecticide00935nas a2200133 4500008004100000245005100041210005000092260001300142300001000155490000700165520048200172100001800654856012900672 2000 eng d00aTranscriptional control of T cell development.0 aTranscriptional control of T cell development c2000 Jun a301-60 v123 aTranscriptional control of T cell development is a complex and rapidly moving area of investigation. Recent advances reveal critical roles for several transcription factors in T cell commitment, differentiation and selection. In particular, new roles for E proteins as well as members of the Notch signaling pathway have been described. Additionally, a unique function of Ikaros in chromatin remodeling reveals a novel mechanism by which transcriptional control may be exerted.1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transcriptional-control-of-t-cell-development01989nas a2200181 4500008004100000245010700041210006900148260001300217300001100230490000700241520132800248100001701576700001601593700001601609700002001625700001601645856014601661 1999 eng d00aAedes aegypti (Diptera: Culicidae) age determination by cuticular hydrocarbon analysis of female legs.0 aAedes aegypti Diptera Culicidae age determination by cuticular h c1999 Nov a824-300 v363 aWe previously described methods for age-grading female Aedes aegypti (L.) by gas chromatographic (GC) analysis of whole-body cuticular hydrocarbon patterns. Two regression models were developed that were based on the age-dependent, relative abundance of 2 cuticular hydrocarbons, pentacosane (GC peak 1) and nonacosane (GC peak 5). We have refined this method so that only the legs are required to age individual females. Two new regression models were developed that also use the relative abundance of a 3rd cuticular hydrocarbon, octacosane (GC peak 4). These models improve the overall accuracy of the cuticular hydrocarbon method for aging female mosquitoes, especially for older females from 132 to 165 degree-days (DD) of age (12-15 calendar days at 28 degrees C). The correlation coefficients (R2) for the best-fitted linear regression models for aging females from 0 to 132 and 0-165 DD were 0.80 and 0.81, respectively (P < 0.001 in all cases). The use of leg cuticular hydrocarbons for estimating the age of female Ae. aegypti has a significant advantage over whole-body extracts as indicated by the decreased variability associated with the relative abundance of pentacosane and the expanded range over which the models were able to predict age accurately by the addition of the relative abundance of octacosane.1 aDesena, M, L1 aEdman, J, D1 aClark, J, M1 aSymington, S, B1 aScott, T, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/aedes-aegypti-diptera-culicidae-age-determination-by-cuticular02306nas a2200205 4500008004100000245006100041210005900102260001300161300001000174490000700184520166400191100001601855700001201871700001801883700001301901700001801914700001401932700001501946856013901961 1999 eng d00aAnti-Trypanosoma brucei activity of nonprimate zoo sera.0 aAntiTrypanosoma brucei activity of nonprimate zoo sera c1999 Feb a48-530 v853 aConstitutive anti-Trypanosoma brucei subsp. brucei S 427 clone 1 and 22 activities were evaluated in sera from 22 species of nonprimate mammals. The sera fell into 5 categories. Sera from Cape buffalo, giraffe, and greater kudu showed a concentration-dependent inhibition of replication of the 2 clones of organisms, which was dependent on the presence of xanthine oxidase. Sera from warthog and springbok also severely limited trypanosome replication but lacked xanthine oxidase. Their antitrypanosome activity was inactivated by heating at 56 C for 30 min but not affected by absorbing with trypanosomes at 4 C. Sera from lion and leopard showed a concentration-dependent inhibition of the growth of T. brucei S427 clone 1 organisms, but not clone 22 organisms. These sera lacked xanthine oxidase. Their anti-T. brucei S 427 clone 1 activity was inactivated by heating at 56 C for 30 min but not removed by absorbing with trypanosomes. Serum from Grant's gazelle prevented replication of both T. brucei clones, lacked xanthine oxidase, and was not affected by heating at 56 C. Sera from waterbuck, Thompson's gazelle, sitatunga, Cape hartebeeste, gerenuk, Grant's zebra, cow, several cat, cougar, bobcat, and domestic cat were fully supportive of trypanosome replication irrespective of concentration tested up to a maximum of 48% v/v in culture medium. Sera from different individuals of the same mammal species had similar effects on trypanosomes, and samples collected from the same individual at different times also had similar activities indicating species-specific stable expression, or lack thereof, of constitutive serum antitrypanosome components.1 aBlack, S, J1 aWang, Q1 aMakadzange, T1 aLi, Y, L1 aVan Praagh, A1 aLoomis, M1 aSeed, J, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/anti-trypanosoma-brucei-activity-of-nonprimate-zoo-sera03140nas a2200145 4500008004100000245003400041210003300075260000900108300001100117490000700128520271300135100001602848700001802864856011202882 1999 eng d00aApoptosis and the proteasome.0 aApoptosis and the proteasome c1999 a209-280 v233 aThe ubiquitin-proteasome pathway is responsible for the regular turnover of a wide variety of proteins and is a critical regulator of many cellular processes. Although this pathway is abundant and ubiquitous, it is also discriminating. This specificity is achieved because there are multiple levels of regulation at work in the pathway. X-ray crystallographic data on the eukaryotic 20S proteasome suggest that substantial rearrangement of the alpha rings, probably mediated by the association of additional regulatory complexes, is required to allow access of substrates into the inner core of the complex. The associated complexes also confer a ubiquitin-dependence on the proteasome, requiring that potential substrates be tagged with chains of ubiquitin proteins. The presence of multiple ubiquitinating enzymes that favor distinct substrates provides a way for a cell to regulate what proteins are to be ubiquitinated. In some cases ubiquitination is not required, but we now know that other modifications, such as phosphorylation and protein-protein interactions, are also important for targeting proteins for degradation. Even with the existence of so many regulatory controls, it is difficult to imagine how one complex can perform so many tasks. As more information is gathered about the proteasome, we begin to understand that all proteasomes are not exactly the same. For example, there is strong evidence that proteasomes involved in antigen presentation differ in both composition and function from proteasomes involved in other processes. The past image of the proteasome as a static structure is being shed, and a new image is emerging that portrays the complex as dynamic and flexible, able to tailor its composition and function to meet a particular need. With this new image of the proteasome in mind, investigators are looking at the potential involvement of the proteasome in cell death. Inhibitor studies have demonstrated a requirement for proteasomes during apoptosis in noncycling and differentiated cells. Similar studies in cycling cells suggest that the proteasome may regulate a cell's decision to proliferate, differentiate, or die. It will be necessary in the future to supplement the peptide and lactacystin studies with work that is not inhibitor-driven since the specificity of an inhibitor for a particular protease is always in question. In addition, a real understanding of how proteasomes may regulate this process awaits the identification of its substrates. With cell death investigators showing increased interest in proteasomes, it may be possible in the next few years to determine the precise role of the proteasome in the pathways that lead to the death of a cell.1 aGrimm, L, M1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/apoptosis-and-the-proteasome01355nas a2200169 4500008004100000245007400041210006900115260001600184300001000200490000800210520075400218100001500972700001400987700001501001700001801016856015101034 1999 eng d00aCutting edge: protective effects of notch-1 on TCR-induced apoptosis.0 aCutting edge protective effects of notch1 on TCRinduced apoptosi c1999 Jan 15 a635-80 v1623 aThe Notch receptor protein was originally identified in Drosophila and is known to mediate cell to cell communication and influence cell fate decisions. Members of this family have been isolated from invertebrates as well as vertebrates. We isolated mouse Notch-1 in a yeast two-hybrid screen with Nur77, which is a protein that has been shown previously to be required for apoptosis in T cell lines. The data presented below indicate that Notch-1 expression provides significant protection to T cell lines from TCR-mediated apoptosis. These data demonstrate a new antiapoptotic role for Notch-1, providing evidence that, in addition to regulating cell fate decisions, Notch-1 can play a critical role in controlling levels of cell death in T cells.1 aJehn, B, M1 aBielke, W1 aPear, W, S1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cutting-edge-protective-effects-of-notch-1-on-tcr-induced-apoptosis01705nas a2200145 4500008004100000245012500041210006900166260001300235300001200248490000700260520111400267100001801381700001601399856014401415 1999 eng d00aHapten design in the development of competitive enyme-linked immunosorbent assays for genotoxic metabolites of alachlor.0 aHapten design in the development of competitive enymelinked immu c1999 Sep a3925-330 v473 aThe acetanilide compounds 2-chloro-2',6'-diethylacetanilide (CDA) and 2-hydroxy-2',6'-diethylacetanilide (HDA) are environmental degradative products of the chloroacetanilide herbicide alachlor. CDA, HDA, and alachlor are ground and surface water contaminants. CDA and HDA are genotoxic in bacterial and mammalian test systems. This paper reports hapten design in the development of two competitive enzyme-linked immunosorbent assays (cELISA) for the detection of CDA and HDA. Chloroacetanilide herbicides and other alachlor metabolites that may be present in environmental samples do not cross-react with the detection of CDA and HDA. Solid-phase extraction of CDA and HDA residues from aqueous samples results in a 1000-fold concentration factor, resulting in an effective detection limit of 15 pg/mL for both assays. The specificity of the cELISAs required preservation of the degree of substitution of the acetanilide moiety in the hapten design. The hapten synthesis strategies are suitable for metabolites of other chloroacetanilide herbicides (i.e., acetochlor, butachlor, metolachlor, and propachlor).1 aTessier, D, M1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/hapten-design-in-the-development-of-competitive-enyme-linked02136nas a2200157 4500008004100000245010200041210006900143260001300212300001300225490000700238520153900245100001201784700001401796700001601810856015201826 1999 eng d00aInfection-associated decline of cape buffalo blood catalase augments serum trypanocidal activity.0 aInfectionassociated decline of cape buffalo blood catalase augme c1999 Jun a2797-8030 v673 aClearance of trypanosomes from the blood of infected Cape buffalo was associated with the development of two responses: (i) complement-dependent and clone-specific lytic activity and (ii) complement-independent trypanocidal activity that was not restricted by trypanosome clone or species. This latter activity was mediated by H2O2 and required the presence of xanthine oxidase in serum but not the addition of purine substrates. Expression of the xanthine oxidase-dependent trypanocidal activity in Cape buffalo serum was coincident with, and required, a decline in its H2O2 catabolic activity. The H2O2 catabolic activity of Cape buffalo serum was due solely to catalase and declined by eightfold around the time that trypanosomes were cleared from the blood, accompanied by a fivefold drop in erythrocyte-associated catalase activity. The Cape buffalo did not develop subsequent parasitemic waves. Clearance of parasitemia in similarly infected cattle was also associated with development of trypanosome clone-specific lytic activity, but not with the acquisition of H2O2-dependent trypanocidal activity in serum, and the cattle supported recurring parasitemia. The lack of trypanocidal activity in pre- and postinfection cattle sera was due to their low content of xanthine oxidase and sustained catalase activity. These data strongly suggest that an infection-induced serum oxidative response, the efficacy of which is amplified by a decline in blood catalase, contributes to suppression of recurring parasitemia in Cape buffalo.1 aWang, Q1 aMurphy, N1 aBlack, S, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/infection-associated-decline-of-cape-buffalo-blood-catalase-augments02759nas a2200205 4500008004100000245015500041210006900196260001300265300001100278490000700289520199000296100001702286700001602303700001602319700002002335700001602355700001602371700001702387856014902404 1999 eng d00aPotential for aging female Aedes aegypti (Diptera: Culicidae) by gas chromatographic analysis of cuticular hydrocarbons, including a field evaluation.0 aPotential for aging female Aedes aegypti Diptera Culicidae by ga c1999 Nov a811-230 v363 aGas chromatography with flame-ionization detection was used to measure the time-associated, quantitative changes in the cuticular hydrocarbons of female Aedes aegypti (L.). Cohorts of unstressed Ae. aegypti, Rockefeller strain, were reared and held at 3 constant temperatures (24, 28, and 30 degrees C). Five females from each cohort were taken at 33 degree-day (DD) intervals from 0 to 231 DD (using 17 degrees C as the threshold temperature). Quantitative changes over time of cuticular hydrocarbons associated with gas chromatographic peaks 1 and 5 were identified as having promise for age grading. The relative abundance of peak 1 (pentacosane) decreased linearly from 0 to 132 DD, whereas peak 5 (nonacosane) increased linearly over the same period. Suboptimal larval conditions (crowded and starved), which resulted in physiological stress (decreased size), had negligible effect on the relative abundance of pentacosane and nonacosane. Additionally, the rate of change in the relative abundance of pentacosane and nonacosane were the same for both a recently colonized Chachoengsao (Thailand) strain of Ae. aegypti compared with the long-colonized Rockefeller (Caribbean) strain over a 0-99 DD interval. Two linear regression models, one based on the relative abundance of pentacosane and the other on the logit transformation of these values, were developed for aging female Ae. aegypti. A blind study using laboratory-reared mosquitoes and a mark-release-recapture experiment using field mosquitoes validated these age-grading models and produced promising results for aging females up to 132 DD (19, 12, and 10 calendar days at 24, 28 and 30 degrees C, respectively). Therefore the regression models, based on the relative abundance of these 2 cuticular hydrocarbons, appeared to be a useful approach for age-grading Ae. aegypti up to at least 12 d of age regardless of environmental conditions (temperature and stress) and population history (origin and colonization time).1 aDesena, M, L1 aClark, J, M1 aEdman, J, D1 aSymington, S, B1 aScott, T, W1 aClark, G, G1 aPeters, T, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/potential-for-aging-female-aedes-aegypti-diptera-culicidae-by-gas02093nas a2200457 4500008004100000245007500041210006900116260001600185300001100201490000800212520079000220653001201010653003401022653001501056653001101071653001501082653001801097653001701115653002001132653001601152653002201168653002001190653001101210653001001221653001601231653001301247653000901260653003201269653001201301653001701313653001501330100001801345700001601363700001801379700001501397700001301412700001701425700002501442700001501467856015301482 1998 eng d00aCloned transgenic calves produced from nonquiescent fetal fibroblasts.0 aCloned transgenic calves produced from nonquiescent fetal fibrob c1998 May 22 a1256-80 v2803 aAn efficient system for genetic modification and large-scale cloning of cattle is of importance for agriculture, biotechnology, and human medicine. Here, actively dividing fetal fibroblasts were genetically modified with a marker gene, a clonal line was selected, and the cells were fused to enucleated mature oocytes. Out of 28 embryos transferred to 11 recipient cows, three healthy, identical, transgenic calves were generated. Furthermore, the life-span of near senescent fibroblasts could be extended by nuclear transfer, as indicated by population doublings in fibroblast lines derived from a 40-day-old fetal clone. With the ability to extend the life-span of these primary cultured cells, this system would be useful for inducing complex genetic modifications in cattle.
10aAnimals10aAnimals, Genetically Modified10aBlastocyst10aCattle10aCell Aging10aCell Division10aCell Nucleus10aCells, Cultured10aClone Cells10aCloning, Organism10aEmbryo Transfer10aFemale10aFetus10aFibroblasts10aG1 Phase10aMale10aNuclear Transfer Techniques10aOocytes10aTransfection10aTransgenes1 aCibelli, J, B1 aStice, S, L1 aGolueke, P, J1 aKane, J, J1 aJerry, J1 aBlackwell, C1 ade León, F, A Ponce1 aRobl, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cloned-transgenic-calves-produced-from-nonquiescent-fetal-fibroblasts01736nas a2200289 4500008004100000245012800041210006900169260001300238300001000251490000700261520076300268653001101031653002101042653002401063653001801087653003601105653002001141653002101161653001101182653001101193653001401204653002601218100002001244700001601264700001301280856015301293 1998 eng d00aCorrelation between cell cycle regulators and the immortalization and transformation of human breast epithelial cell lines.0 aCorrelation between cell cycle regulators and the immortalizatio c1998 Jul a65-710 v133 aCellular proliferation, essential for normal development, may result in neoplastic growth when the cell cycle clock is disrupted. In order to determine whether the protein expression of cell cycle regulators differs among normal, immortalized non-tumorigenic and malignant human breast epithelial cells (HBECs), we analyzed the protein expression of cyclins D1, D3, A and E, cyclin-dependent kinase (cdk) 4 and c-fos in exponentially growing MCF-10M, MCF-10F, and MCF-7 cells. The tumorigenicity of HBECs in vivo correlated with both cell cycle regulators and early-gene protein expression in vitro. The differential expression of cyclin E- and cyclin A-related proteins and their putative relevance in the tumorigenic properties of HBECs are also discussed.10aBreast10aBreast Neoplasms10aCell Cycle Proteins10aCell Division10aCell Transformation, Neoplastic10aCells, Cultured10aEpithelial Cells10aFemale10aHumans10aPhenotype10aTumor Cells, Cultured1 aSalicioni, A, M1 aRusso, I, H1 aRusso, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/correlation-between-cell-cycle-regulators-and-the-immortalization-and02191nas a2200193 4500008004100000245005800041210005700099260001600156300001200172490000800184520157000192100001701762700001901779700001301798700001401811700001801825700001801843856013601861 1998 eng d00aMultiple sites of V lambda diversification in cattle.0 aMultiple sites of V lambda diversification in cattle c1998 Nov 15 a5438-440 v1613 aIg repertoire diversification in cattle was studied in the ileal Peyer's patch (IPP) follicles of young calves and in the spleens of late first-trimester bovine fetuses. To investigate follicular diversification, individual IPP follicles were isolated by microdissection; VA diversity was examined by RT-PCR and subsequent cloning and sequencing. When 52 intrafollicular sequences from a 4-wk-old calf were determined and compared, two major groups, one of 23 members and the other of 25, could be delineated. An examination of these groups revealed clear genealogic relationships that implicated in situ diversification of V lambda sequences within the confines of an IPP follicle. V lambda expression was also examined in early (95 and 110 gestational day) fetal bovine spleens. Although earlier studies in cattle and sheep implicated the IPP as a likely site of Ab diversification, a close investigation of V lambda sequences in late first-trimester fetal calves revealed that diversity appears in the early fetal spleen before the establishment of a diverse repertoire in the ileum. When the sequences for the fetal spleen were compared with an existing pool of germline sequences, we found evidence of possible gene conversion events and possible untemplated point mutations occurring in sequences recovered from fetal spleens. We conclude that IPP is not the sole site of VA diversification in cattle. Also, as suggested for rabbits, cattle may use both gene conversion and untemplated somatic point mutation to diversify their primary VA repertoire.
1 aLucier, M, R1 aThompson, R, E1 aWaire, J1 aLin, A, W1 aOsborne, B, A1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/multiple-sites-of-v-lambda-diversification-in-cattle02099nas a2200181 4500008004100000245010000041210006900141260001600210300001100226490000800237520145000245100001001695700001301705700001201718700001601730700001801746856015301764 1998 eng d00aPartial characterization of the calcium-releasing activity of porcine sperm cytosolic extracts.0 aPartial characterization of the calciumreleasing activity of por c1998 Nov 15 a369-810 v2033 aInjection of sperm cytosolic extracts into mammalian eggs has been shown to elicit intracellular calcium ([Ca2+]i) oscillations that are similar in amplitude, duration, and frequency to those observed following fertilization. Thus, to characterize the Ca2+-release component(s) in porcine sperm cytosolic extracts, a combination of fractionation techniques was used. The fraction with Ca2+ releasing activity was precipitated by 50% saturating solutions of ammonium sulfate and Western blot analysis showed that the pellets contained glucosamine-6-phosphate deaminase (gpd)/oscillin, a protein which has been suggested to be the sperm's active component. Single and double isoelectrofocusing (IEF) of porcine sperm extracts generated fractions with different Ca2+-releasing activities. Fractions with maximal Ca2+-releasing activity did not contain material that was immunoreactive with antibodies against gpd/oscillin; adjacent fractions containing gpd/oscillin had no Ca2+-releasing activity. These findings were confirmed by IEF coupled with size exclusion chromatography on Superose 12 and with hydroxyapatite chromatography. These procedures predict an isoelectric point of our active component of 6.5-7.0 and a relative molecular weight ranging from 29 to 68 kDa. In summary, the data show that the Ca2+ release-inducing component(s) of porcine sperm extracts can be fractionated and that gpd/oscillin is not the pig sperm Ca2+ oscillogen.1 aWu, H1 aHe, C, L1 aJehn, B1 aBlack, S, J1 aFissore, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/partial-characterization-of-the-calcium-releasing-activity-of-porcine01848nas a2200397 4500008004100000245009300041210006900134260001300203300001000216490000700226520066200233653001200895653003400907653002300941653001500964653001100979653002500990653001601015653001201031653001301043653002001056653001301076653002901089653002001118653001301138653001501151100001801166700001601184700001801200700001501218700001301233700001701246700002501263700001501288856014701303 1998 eng d00aTransgenic bovine chimeric offspring produced from somatic cell-derived stem-like cells.0 aTransgenic bovine chimeric offspring produced from somatic celld c1998 Jul a642-60 v163 aWe have developed a method, using nuclear transplantation, to produce transgenic embryonic stem (ES)-like cells from fetal bovine fibroblasts. These cells, when reintroduced into preimplantation embryos, differentiated into derivatives from the three embryonic germ layers, ectoderm, mesoderm, and endoderm, in 5-month-old animals. Six out of seven (86%) calves born were found to be chimeric for at least one tissue. These experiments demonstrate that somatic cells can be genetically modified and then de-differentiated by nuclear transfer into ES-like cells, opening the possibility of using them in differentiation studies and human cell therapy.
10aAnimals10aAnimals, Genetically Modified10abeta-Galactosidase10aBlastocyst10aCattle10aCell Differentiation10aCell Fusion10aChimera10aEctoderm10aEmbryo Transfer10aEndoderm10aGene Transfer Techniques10aGenetic Therapy10aMesoderm10aStem Cells1 aCibelli, J, B1 aStice, S, L1 aGolueke, P, J1 aKane, J, J1 aJerry, J1 aBlackwell, C1 ade León, F, A Ponce1 aRobl, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transgenic-bovine-chimeric-offspring-produced-from-somatic-cell03124nas a2200421 4500008004100000245011000041210006900151260001300220300001000233490000700243520177400250653001802024653001102042653002102053653003002074653002302104653002002127653002302147653002302170653002102193653001102214653001402225653003102239653004302270653001702313653001102330653002302341653002802364653002602392653002702418653002602445100001602471700002002487700001602507700001602523700001302539856015002552 1997 eng d00aDifferential expression of human ferritin H chain gene in immortal human breast epithelial MCF-10F cells.0 aDifferential expression of human ferritin H chain gene in immort c1997 Dec a332-90 v203 aSubtractive hybridization was used to isolate genes expressed uniquely in the immortalized human breast epithelial cell (HBEC) line MCF-10F and not in the mortal HBEC line S-130, from which MCF-10F cells were derived. We identified a 233-bp cDNA that was expressed in MCF-10F cells and not in their mortal counterpart S-130 cells. Sequence comparison with the GenBank database revealed that the cDNA was identical to the gene encoding human ferritin heavy H chain. Northern blot analysis using the isolated cDNA as a probe showed a differentially expressed 1.1-kb transcript of ferritin H in total RNA from the immortal MCF-10F cells, MCF-10F cells treated with the chemical carcinogens 7,12-dimethylbenz[a]anthracene and benzo[a]pyrene, and the breast cancer cell lines MCF-7, HBL-100, T-47D, and BT-20. No ferritin H transcript was detected in the mortal line S-130 or in other primary HBEC cultures. Increased levels of mRNA transcript signals were also detected in total RNA from breast cancer tissue samples. Tissue with ductal hyperplasia had higher expression levels than normal adjacent mammary tissue. In situ hybridization showed high levels of ferritin H transcript in mammary tissue areas with ductal hyperplasia, carcinoma in situ, and infiltrating ductal carcinoma. This is the first report of the differential expression and upregulation of human ferritin H chain gene in immortal HBECs. It may be an important factor in the process of immortalization, possibly an early stage of malignant transformation of HBECs, providing cells with iron necessary for growth and clonal expansion. Also, ferritin iron, once released, may increase the level of reactive iron, leading to an increase in oxygen free-radical generation, oxidative DNA damage, and mutation.10aBase Sequence10aBreast10aBreast Neoplasms10aCarcinoma, Ductal, Breast10aCarcinoma, Lobular10aCells, Cultured10aCloning, Molecular10aDNA, Complementary10aEpithelial Cells10aFemale10aFerritins10aGene Expression Regulation10aGene Expression Regulation, Neoplastic10aGene Library10aHumans10aModels, Biological10aMolecular Sequence Data10aNeoplasm Invasiveness10aTranscription, Genetic10aTumor Cells, Cultured1 aHiggy, N, A1 aSalicioni, A, M1 aRusso, I, H1 aZhang, P, L1 aRusso, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/differential-expression-of-human-ferritin-h-chain-gene-in-immortal01690nas a2200145 4500008004100000245004700041210004600088260000900134300001100143490000600154520122600160100001501386700001801401856012501419 1997 eng d00aGene regulation associated with apoptosis.0 aGene regulation associated with apoptosis c1997 a179-930 v73 aApoptosis, one of the best-studied forms of programmed cell death processes, plays an important role during the development and life-cycle of most multicellular organisms. The mechanisms underlying the initiation and manifestation of apoptotic cell death are the focus of the most recent cell death research. Generally, it is believed that cells are eliminated via a highly ordered and controlled program. This program might consist of the successive activation of unique apoptosis-specific genes, which are solely involved in the regulation of the programmed cell death. However, more and more evidence is accumulating that novel genes are not activated or induced during apoptosis, but rather many well-known genes previously described for their roles in processes such as proliferation and differentiation and belonging, for example, to the protein families of immediate-early genes and transcription factors become activated. The death-specific feature is achieved thereby by the extent, combination, and specific timing of gene expression. The involvement of the three different transcription factors glucocorticoid receptor (GR), nur77, and activator protein 1 (AP-1) in such a scenario is the focus of this review.1 aJehn, B, M1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/gene-regulation-associated-with-apoptosis00583nas a2200145 4500008004100000245010000041210006900141260001300210300000900223490000700232100001600239700001600255700001200271856015400283 1997 eng d00aIdentification of the cape buffalo serum trypanocidal protein: xanthine: oxygen oxidoreductase.0 aIdentification of the cape buffalo serum trypanocidal protein xa c1997 Aug a534S0 v251 aBlack, S, J1 aMuranjan, M1 aWang, Q uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-the-cape-buffalo-serum-trypanocidal-protein-xanthine02033nas a2200181 4500008004100000245005100041210005000092260000900142300001100151490000700162520147100169100001301640700001601653700001701669700001801686700001801704856012901722 1997 eng d00aImmunoglobulin gene diversification in cattle.0 aImmunoglobulin gene diversification in cattle c1997 a165-830 v153 aResearch in several species has revealed that different types of mammals have evolved divergent molecular and cellular strategies for generating immunoglobulin diversity. Other chapters in this text have highlighted the specific characteristics unique to chicken, rabbit, mouse, human and sheep B lymphocyte development; namely indicating differences in the mechanisms of diversity and the site of primary B cell development. Studies of the bovine system have indicated that like the sheep system, the ileal Peyer's patch (IPP) is a likely chicken bursal equivalent, and is a site of primary B lymphocyte development. Substantial investigation in sheep has indicated that Ig diversity is created by untemplated somatic mutation and intense selective pressure (Reynaud et al., 1991). The frequency of alteration in the sheep Ig light chain gene locus also is characteristic of the bovine system, however, recent evidence from sequencing of bovine lambda light chain genes indicates that one mechanism that contributes to diversity is gene conversion, utilizing several pseudogenes located in the Ig locus (Parng et al., 1996). The mechanism by which this hyperalteration of Ig genes occurs in both sheep and cattle is poorly understood and is thus the focus of considerable investigation. The study of events in the IPP may also have informative ramifications for secondary diversification of the Ig repertoire by somatic hyperalteration in germinal centers.
1 aMeyer, A1 aParng, C, L1 aHansal, S, A1 aOsborne, B, A1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/immunoglobulin-gene-diversification-in-cattle04106nas a2200457 4500008004100000245012700041210006900168260001600237300001000253490000800263520262500271653001902896653004402915653004402959653003303003653004303036653004303079653003203122653001203154653001403166653001303180653001103193653002403204653001303228653002403241653001503265653001703280653001303297653001603310653001303326653001703339653001903356653000903375653001703384653002603401653002203427100002003449700001703469700001503486856014703501 1997 eng d00aParticipation of both adrenergic and opioidergic systems in the negative feedback of adrenal progesterone on LH secretion.0 aParticipation of both adrenergic and opioidergic systems in the c1997 Aug 13 a283-70 v3323 aIt has been shown that adrenal progesterone plays an important role in regulating the negative feedback of oestrogen on luteinizing hormone (LH) release in ovariectomized and oestrogen-treated rats. The purpose of the present study was to determine whether adrenal progesterone modulation of LH secretion is mediated by adrenergic and opioidergic systems in ovariectomized and oestrogen-treated rats. Oestradiol benzoate (20 micrograms/rat) was given s.c. to ovariectomized rats on day 0. Control animals were injected with the vehicle alone. The specific adrenoceptor antagonists prazosin (10 mg/kg), idazoxan (100 micrograms/kg), metoprolol (10 mg/kg) or ICI 118,551 (200 micrograms/kg) were injected at 12.00 and 20.00 h on day 2 and at 08.00 h on day 3 to oestrogen-primed rats treated or not with RU486. Control animals were injected with saline. RU486 (10 mg/kg) was administered s.c. at 08.00 h on day 3 to oestradiol-treated animals receiving adrenoceptor antagonists or saline. Naloxone (2 mg/kg) was administered i.p. 30 min before blood-sampling to oestrogen-primed rats treated or not with RU486. All groups were blood-sampled at 13.00 and 18.00 h on day 3, and LH concentration was measured by radioimmunoassay. The administration of oestradiol to ovariectomized rats decreased serum LH levels at 13.00 and 18.00 h on day 3. Prazosin or idazoxan partially prevented the effect of oestradiol at 13.00 h, while metoprolol, ICI 118,551 or naloxone totally blocked the inhibitory effect of oestradiol on LH secretion; both adrenoceptor and opioid receptor antagonists also prevented the effect of oestrogen on LH concentration at 18.00 h. RU486 increased serum LH concentration at 18.00 h in oestrogen-primed rats to values higher than in ovariectomized control rats, with no effect at noon. The administration of prazosin to ovariectomized and oestrogen-primed rats treated with RU486 prevented this increase while the other adrenoceptor antagonists or naloxone increased serum LH concentrations at 18.00 h. The present study shows that RU486 switches the feedback of oestradiol on LH secretion from negative to positive in ovariectomized and oestradiol-primed rats, activating a stimulatory alpha 1-adrenergic pathway during the afternoon, and gives strong evidence about the participation of adrenal progesterone modulating neurotransmitter systems involved in the secretion of LH. It also supports the participation of endogenous opioid peptides in the negative feedback of oestradiol, suggesting that the inhibitory tone of endogenous opioid peptide is active regardless the action of adrenal progesterone.10aAdrenal Glands10aAdrenergic alpha-1 Receptor Antagonists10aAdrenergic alpha-2 Receptor Antagonists10aAdrenergic alpha-Antagonists10aAdrenergic beta-1 Receptor Antagonists10aAdrenergic beta-2 Receptor Antagonists10aAdrenergic beta-Antagonists10aAnimals10aEstradiol10aFeedback10aFemale10aHormone Antagonists10aIdazoxan10aLuteinizing Hormone10aMetoprolol10aMifepristone10aNaloxone10aOvariectomy10aPrazosin10aProgesterone10aPropanolamines10aRats10aRats, Wistar10aReceptors, Adrenergic10aReceptors, Opioid1 aSalicioni, A, M1 aCarón, R, W1 aDeis, R, P uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/participation-of-both-adrenergic-and-opioidergic-systems-in-the02928nas a2200325 4500008004100000245014500041210006900186260001300255300001100268490000700279520188200286653001202168653002902180653001402209653001102223653002402234653001902258653001702277653001302294653002002307653001602327653001702343653001402360653000902374653001702383100001702400700002002417700001502437856015002452 1997 eng d00aRegulation of prolactin secretion by adrenal steroids in oestrogen-treated ovariectomized rats: participation of endogenous opioid peptides.0 aRegulation of prolactin secretion by adrenal steroids in oestrog c1997 Oct a1433-80 v363 aThe purpose of the present study was to determine whether glucocorticoid inhibition of prolactin (PRL) release in oestrogen-treated ovariectomized (OVX) rats is mediated by endogenous opioid peptides (EOPs). All the animals were OVX and given oestradiol benzoate (OB, 20 microg/rat, s.c.) 2 weeks later (day 0). On day 3 they received vehicle, mifepristone (MIF, 10 mg/kg, s.c.) or hydrocortisone (HYD, 2 mg/rat, s.c.), in combination with the opioid antagonist naloxone (NAL, 2 mg/kg, i.p.) or vehicle. Serum PRL concentration was then measured by RIA at 13.00 and 18.00 hr, to include assessment of diurnal variation of PRL secretion. At 13.00 hr either MIF or NAL alone increased PRL secretion with no additional effect when NAL was combined with MIF. HYD had no significant inhibitory effect, but NAL with HYD increased PRL secretion. At 18.00 hr serum PRL concentration was higher than at 13.00 hr, and not affected significantly by MIF or NAL alone, although PRL secretion was increased by treatment with both. HYD inhibited PRL secretion and this inhibition was prevented by NAL. In a second experiment to distinguish antiglucocorticoid and antiprogesterone effects of MIF, we administered progesterone (2 mg/rat, s.c.) or a specific progesterone antiserum. In contrast with MIF, the progesterone antibody had no effect on PRL secretion at 13.00 hr, nor on the stimulation by NAL, while progesterone (unlike HYD) increased PRL secretion and NAL attenuated this response; this was opposite to the effect of NAL with HYD. Similarly, at 18.00 hr the interaction of MIF and NAL was not explained by antagonism of progesterone. Together, these results indicate inhibition of PRL by glucocorticoids but not progesterone, mediated in part by EOPs. At 18.00 hr endogenous glucocorticoids do not regulate oestrogen-stimulated PRL release, although HYD is inhibitory through EOPs.10aAnimals10aAnti-Inflammatory Agents10aEstradiol10aFemale10aHormone Antagonists10aHydrocortisone10aMifepristone10aNaloxone10aOpioid Peptides10aOvariectomy10aProgesterone10aProlactin10aRats10aRats, Wistar1 aCarón, R, W1 aSalicioni, A, M1 aDeis, R, P uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/regulation-of-prolactin-secretion-by-adrenal-steroids-in-oestrogen02698nas a2200349 4500008004100000245010200041210006900143260001500212300001100227490000700238520158700245653001901832653003601851653001801887653001101905653002101916653002001937653001101957653003101968653001101999653002802010653001902038653001402057653003102071100001602102700002002118700001602138700001602154700001702170700001302187856014802200 1997 eng d00aTamoxifen down-regulates CD36 messenger RNA levels in normal and neoplastic human breast tissues.0 aTamoxifen downregulates CD36 messenger RNA levels in normal and c1997 Feb 1 a378-810 v573 aTamoxifen (TAM) exerts a long-term suppressive effect on human breast cancer cell proliferation. To determine whether the effects of TAM are mediated by specific gene activation or repression, normal and tumoral human breast tissues obtained before and during TAM treatment were analyzed by differential display technique. Total RNA for differential display analysis was obtained from breast tissues from two women with the diagnosis of estrogen receptor-positive stage II (T2N1M0) infiltrating ductal carcinoma, made by incisional biopsy, followed by modified radical mastectomy performed after a 30-day treatment with TAM (20 mg/day). One 202-bp cDNA band, AP5-1, was present in normal and tumoral biopsy samples, but was absent in breast tissue obtained during TAM treatment, and was confirmed by Northern hybridization, which showed a 2.7-kb band in both patients. The differentially expressed cDNA fragment showed 99% homology to Homo sapiens CD36 gene, a glycoprotein that acts as a receptor for the extracellular matrix proteins thrombospondin-1, collagen types I and IV, and oxidized low-density lipoprotein. These results indicate that the down-regulation of CD36 induced by TAM might represent alternative or additional mechanisms of action of this drug affecting the functions of thrombospondin-1, which is involved in hematogenous tumor spread, invasion and angiogenesis, and oxidized low-density lipoprotein, playing a role in inhibition of arteriosclerosis. The multiple functions affected by the down-regulation of CD36 by TAM warrant the need for additional studies.10aAntigens, CD3610aAntineoplastic Agents, Hormonal10aBase Sequence10aBreast10aBreast Neoplasms10aDown-Regulation10aFemale10aGene Expression Regulation10aHumans10aMolecular Sequence Data10aRNA, Messenger10aTamoxifen10aTranscriptional Activation1 aSilva, I, D1 aSalicioni, A, M1 aRusso, I, H1 aHiggy, N, A1 aGebrim, L, H1 aRusso, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/tamoxifen-down-regulates-cd36-messenger-rna-levels-in-normal-and00572nas a2200133 4500008004100000245012000041210006900161260001300230300001300243490000700256100001400263700001600277856014500293 1996 eng d00aAddition of a competitive primer can dramatically improve the specificity of PCR amplification of specific alleles.0 aAddition of a competitive primer can dramatically improve the sp c1996 Oct a586, 5900 v211 aZhu, K, Y1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/addition-of-a-competitive-primer-can-dramatically-improve-the01164nas a2200133 4500008004100000245007200041210006900113260001300182300001100195490000600206520065000212100001800862856015000880 1996 eng d00aApoptosis and the maintenance of homoeostasis in the immune system.0 aApoptosis and the maintenance of homoeostasis in the immune syst c1996 Apr a245-540 v83 aThe regulation of cell proliferation and the selection against autoreactive cells in the lymphoid system both occur through the induction of apoptosis. Many of the signals that induce apoptosis in lymphocytes are now well defined. Interactions between Fas and its ligand have emerged as a major mechanism for the deletion of activated peripheral T cells and autoreactive B cells. Although the signal-transduction pathway leading from engagement of Fas to apoptosis is not entirely clear, significant advances have been made recently. There has also been progress in the elucidation of the mechanisms that regulate apoptosis in the immune system.1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/apoptosis-and-the-maintenance-of-homoeostasis-in-the-immune-system02536nas a2200169 4500008004100000245011300041210006900154260001300223300001000236490000700246520190400253100001402157700001602171700001802187700001802205856014302223 1996 eng d00aApplication of random amplified polymorphic DNA analysis to differentiate strains of Salmonella enteritidis.0 aApplication of random amplified polymorphic DNA analysis to diff c1996 Apr a870-60 v343 aA random amplified polymorphic DNA (RAPD) fingerprinting method has been developed to differentiate Salmonella enteritidis isolates. A total of 65 arbitrary primers were screened with S. enteritidis isolates of different phage types. This allowed selection of a panel of primers capable of detecting DNA polymorphisms among S. enteritidis isolates. This panel was used to examine a panel of 29 isolates of S. enteritidis which had been previously characterized by other subtyping methods, including phage typing (PT) (n = 7), ribotyping (RT) (n = 13), and pulsed-field gel electrophoresis (PFGE). Applied collectively, these three methods resolved the collection into 20 different subtypes. However, by the RAPD fingerprinting method alone, 14 RAPD subtypes were revealed. Eight isolates of S. enteritidis phage type 8 that failed to be discriminated by other typing methods (PT, RT, and PFGE) were resolved into three different subtypes by RAPD analysis. In contrast, isolates that were derived from the same sources were not differentiated by any of the subtyping methods employed, including PT, RT, PFGE, and RAPD analysis. This RAPD approach to S. enteritidis subtyping provided more discriminatory power than did any of several other subtyping methods applied individually. Once the challenging step of primer identification was accomplished, determinations of the appropriate concentrations of arbitrary primer, DNA template, and MG2+ ion were also necessary for optimal discriminatory power. The bacterial DNA used in this RAPD protocol was obtained by boiling the bacterial sample. This simple procedure yielded DNA that produced fingerprint patterns as consistent as those obtained from phenol-chloroform-extracted DNA. Clearly, when appropriately constituted primer sets are identified and employed, RAPD analysis provides a simple, rapid, and powerful subtyping method for S. enteritidis.1 aLin, A, W1 aUsera, M, A1 aBarrett, T, J1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/application-of-random-amplified-polymorphic-dna-analysis-to00449nas a2200121 4500008004100000245005400041210005200095260001300147300001100160490000700171100001800178856013100196 1996 eng d00aCell death in vertebrates: lessons from the worm.0 aCell death in vertebrates lessons from the worm c1996 Dec a489-910 v121 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cell-death-in-vertebrates-lessons-from-the-worm02459nas a2200169 4500008004100000245012200041210006900163260001600232300001200248490000700260520180800267100002002075700001302095700001502108700001802123856014802141 1996 eng d00aComparison of T cell cytokines in resistant and susceptible mice infected with virulent Brucella abortus strain 2308.0 aComparison of T cell cytokines in resistant and susceptible mice c1996 Dec 31 a193-2030 v163 aC57BL/10 and BALB/c mice differ in their abilities to clear infections with the intracellular bacterium Brucella abortus strain 2308. We have previously reported that in vivo of IL-10 in the susceptible BALB/c mice results in significantly fewer bacteria in their spleens 1 week after infection with 5 x 10(3) colony forming units (CFU) of 2308. Here we extend those studies and report a similar effect when IL-4 is neutralized. In contrast, in the more resistant C57BL/10 mice infected with 5 x 10(3) CFU, neither neutralization of IL-4 significantly decreased the level of infection nor did it in either BALB/c or C57BL/10 mice infected with a 1000-fold higher dose of strain 2308. While splenocytes from the later mentioned groups of mice produced IL-10 in response to stimulation with brucella antigen, they also produced higher levels of interferon (IFN)-gamma than those from BALB/c mice with the low challenge dose of 5 x 10(3) CFU. Results of in vivo neutralization of IFN-gamma by monoclonal antibodies (MAb) reported here and elsewhere indicated that IFN-gamma is important for control; thus, we postulate that the higher levels of IFN-gamma override the detrimental effects of Th2 cytokines. In vitro studies also showed that macrophages from the more resistant C57BL/10 mice were less susceptible to the ability of IL-10 to decrease anti-brucella activities than were BALB/c macrophages. CD4+ T cells were principally responsible for the production of IL-10 in BALB/c but not C57BL/10 splenocyte populations. C57BL/10 splenocytes produced more IFN-gamma than those from BALB/c mice in response to stimulation with brucella antigens. These differences between BALB/c and C57BL/10 mice may contribute to the superior capacity of C57BL/10 mice to control infections with B. abortus strain 2308.1 aFernandes, D, M1 aJiang, X1 aJung, J, H1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/comparison-of-t-cell-cytokines-in-resistant-and-susceptible-mice02285nas a2200169 4500008004100000245007100041210006900112260001600181300001200197490000800209520168300217100001601900700001401916700001801930700001801948856014901966 1996 eng d00aGene conversion contributes to Ig light chain diversity in cattle.0 aGene conversion contributes to Ig light chain diversity in cattl c1996 Dec 15 a5478-860 v1573 aIn humans and mice, extensive gene rearrangement is the major mechanism of diversification of the primary Ig repertoire. This study shows that cattle depart from this pattern because rearrangement in the light chain locus is sharply limited. Furthermore, in cattle, gene conversion contributes to the diversification of the primary light chain repertoire. Sequencing of germ-line and expressed Vlambda genes revealed three important features. First, the germ line contained a number of Vlambda pseudogenes. In fact, 14 (70%) of the 20 germ-line genes identified and sequenced were pseudogenes, because they had one or more of the following defects: lack of recombination signal sequences at the 3' end, stop codons within the reading frame or truncations, and/or insertions or deletions that resulted in loss of reading frame. Second, Vlambda cDNA from ileal Peyer's patch B cells demonstrated that the light chain repertoire arises from only a small number of V(J) rearrangements. Even though two J genes were identified in the germ line, all of the expressed Vlambda genes examined contained the same J segment, indicating that only a single J gene participates in rearrangement at the lambda locus. Third, a significant number of departures from the germ-line sequences of rearranged Vlambda can be traced to donor sequences of one or more Vlambda pseudogenes. We conclude that a limited number of rearrangements and gene conversion play a role in contributing to the diversification of the primary lambda repertoire. Furthermore, while clear indications of a role for somatic mutation in lambda diversification was seen, V gene rearrangement was not a major factor.
1 aParng, C, L1 aHansal, S1 aGoldsby, R, A1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/gene-conversion-contributes-to-ig-light-chain-diversity-in-cattle01318nas a2200205 4500008004100000245005500041210005400096260001300150300001000163490000700173520068500180100001800865700001600883700002100899700001300920700001600933700001100949700001900960856013300979 1996 eng d00aGenes that regulate apoptosis in the mouse thymus.0 aGenes that regulate apoptosis in the mouse thymus c1996 Jan a18-220 v603 aElimination of self-reactive T lymphocytes occurs during T-cell development in the thymus by a process known as negative selection. The mechanism that drives negative selection is apoptosis. To identify genes that regulate apoptosis in the mouse thymus, a library of negatively selected T cells was constructed and, by subtractive screening, several differentially regulated genes were isolated. Two transcripts that are repressed during cell death were identified, in addition to two induced transcripts. Further experiments demonstrated that cell death in thymocytes can occur via several induction pathways and each pathway appears to be regulated by a unique cascade of genes.1 aOsborne, B, A1 aSmith, S, W1 aMcLaughlin, K, A1 aGrimm, L1 aKallinch, T1 aLiu, Z1 aSchwartz, L, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genes-that-regulate-apoptosis-in-the-mouse-thymus00600nas a2200181 4500008004100000245005700041210005600098260000900154300001200163490000800175100001800183700001600201700002100217700001300238700001400251700001800265856013500283 1996 eng d00aGenetic regulation of apoptosis in the mouse thymus.0 aGenetic regulation of apoptosis in the mouse thymus c1996 a199-2070 v4061 aOsborne, B, A1 aSmith, S, W1 aMcLaughlin, K, A1 aGrimm, L1 aMorgan, G1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/genetic-regulation-of-apoptosis-in-the-mouse-thymus02165nas a2200157 4500008004100000245013300041210006900174260001300243300001000256490000700266520155000273100001401823700001401837700001601851856014001867 1996 eng d00aA Point Mutation of Acetylcholinesterase Associated with Azinphosmethyl Resistance and Reduced Fitness in Colorado Potato Beetle0 aPoint Mutation of Acetylcholinesterase Associated with Azinphosm c1996 Jun a100-80 v553 aA serine to glycine point mutation of acetylcholinesterase (AChE, EC 1.1.1.7) was identified in an azinphosmethyl-resistant strain of Colorado potato beetle [Leptinotarsa decemlineata (Say)]. The position of the mutation corresponds to Val 238 of the Torpedo AChE and represents the first amino acid residue to form the alpha-helix, alpha-E'1. The predicted secondary structure of the mutation-containing region of AChE suggested that the transition from the turn to the alpha-helix occurs sooner in the sequence when serine is replaced by glycine. Thus, conformational changes in the AChE due to the alpha-helix deformation were expected to impinge upon both the catalytic and the peripheral binding sites, resulting in the modification of the bindings of organophosphorus insecticides and other ligands to these sites. The mutation appeared to be associated with the fitness of the beetle. The intrinsic rate of increase of the azinphosmethyl-resistant (AZ-R) strain was relatively low when the beetles were reared on the Russet Burbank potato cultivar, but was relatively high when they were reared on the NDA 1725-1 potato cultivar. Because these two potato cultivars contain different amounts of steroidal glycoalkaloids (e.g., alpha-solanine and alpha-chaconine), the different fitness of the AZ-R strain on different potato cultivars may be partially attributed to the increased sensitivity of the azinphosmethyl-resistant form of AChE to the inhibition by alpha-solanine and reduced sensitivity to alpha-chaconine as previously reported.1 aZhu, K, Y1 aLee, S, H1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/a-point-mutation-of-acetylcholinesterase-associated-with01457nas a2200181 4500008004100000245006300041210006200104260001500166300001200181490000700193520084400200100001601044700001901060700001801079700001901097700001801116856014101134 1996 eng d00aProteasomes play an essential role in thymocyte apoptosis.0 aProteasomes play an essential role in thymocyte apoptosis c1996 Aug 1 a3835-440 v153 aCell death in many different organisms requires the activation of proteolytic cascades involving cytosolic proteases. Here we describe a novel requirement in thymocyte cell death for the 20S proteasome, a highly conserved multicatalytic protease found in all eukaryotes. Specific inhibitors of proteasome function blocked cell death induced by ionizing radiation, glucocorticoids or phorbol ester. In addition to inhibiting apoptosis, these signals prevented the cleavage of poly(ADP-ribose) polymerase that accompanies many cell deaths. Since overall rates of protein degradation were not altered significantly during cell death in thymocytes, these results suggest that the proteasome may either degrade regulatory protein(s) that normally inhibit the apoptotic pathway or may proteolytically activate protein(s) than promote cell death.1 aGrimm, L, M1 aGoldberg, A, L1 aPoirier, G, G1 aSchwartz, L, M1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/proteasomes-play-an-essential-role-in-thymocyte-apoptosis01902nas a2200157 4500008004100000245004700041210004200088260001300130300001100143490000700154520140100161100002101562700001801583700001801601856012501619 1996 eng d00aThe role of oxygen in thymocyte apoptosis.0 arole of oxygen in thymocyte apoptosis c1996 May a1170-40 v263 aSignals generated by T cell receptor (TCR) cross-linking (or phorbol 12-myristate-13-acetate + Ca2+ ionophore), glucocorticoids or ionizing radiation all stimulate apoptotic cell death in thymocytes by signals that are initially distinct from each other. However, when these stimuli were administered to thymocyte cultures that were maintained under an atmosphere containing less than 20 ppm oxygen as opposed to one that contained 18.5% molecular oxygen, cell death was inhibited or abrogated, suggesting that the induction of death by all three different stimuli depend on the presence of molecular oxygen. Studies of the effects of the cysteine analog N-acetyl cysteine (NAC) with normal thymocytes demonstrated that this antioxidant inhibited the induction of death by each of the different stimuli in a manner the paralleled anaerobiosis. Furthermore, studies with thymocytes demonstrated that the induction of nur77, a gene shown to be involved in thymocyte apoptosis signaled through the TCR or its surrogates, is not inhibited by NAC or dependent upon molecular oxygen. The possible implications for negative selection of NAC-mediated inhibition of TCR-signaled thymocyte cell death was examined in thymic organ culture. Treatment of these cultures with NAC provided significant protection against staphylococcal enterotoxin B-mediated deletion of V beta 8-expressing thymocytes.
1 aMcLaughlin, K, A1 aOsborne, B, A1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-role-of-oxygen-in-thymocyte-apoptosis01335nas a2200145 4500008004100000245003000041210002900071260001300100300001000113490000700123520091400130100001801044700001901062856010801081 1995 eng d00aCell death suffers a TKO.0 aCell death suffers a TKO c1995 Jun a557-90 v173 aThe cytokine interferon-gamma (IFN-gamma), initiates both cell cycle arrest and cell death in certain cell lines. Through a novel strategy of cell transfection with episomal vectors expressing antisense cDNAs, Deiss et al. have demonstrated that it is possible to isolate genes that are required for the initiation of cell death by the cytokine IFN-gamma. This approach, referred to as TKO, for Technical Knock Out, has identified several genes whose activity appears to be essential for the induction of apoptosis by IFN-gamma in HeLa cells. Interestingly, these genes appear to mediate IFN-gamma-induced apoptosis in HeLa cells, but their inhibition by antisense does not ameliorate the antiproliferative effects of IFN-gamma in these cells. The clever strategy employed by these authors holds promise for others who wish to isolate genes required for other differentiative processes in cultured cell lines.1 aOsborne, B, A1 aSchwartz, L, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cell-death-suffers-a-tko01896nas a2200145 4500008004100000245012700041210006900168260001300237300001200250490000700262520130200269100001401571700001601585856014901601 1995 eng d00aCloning and sequencing of a cDNA encoding acetylcholinesterase in Colorado potato beetle, Leptinotarsa decemlineata (Say).0 aCloning and sequencing of a cDNA encoding acetylcholinesterase i c1995 Dec a1129-380 v253 aA cDNA encoding acetylcholinesterase (AChE, EC 1.1.1.7) was cloned from a cDNA library constructed from an insecticide-susceptible strain of Colorado potato beetle, Leptinotarsa decemlineata (Say). The complete amino acid sequence of AChE deduced from the cDNA consisted of 29 residues for the putative signal peptide and 600 residues for the mature protein with a predicted molecular weight of 67,994. Northern blot analysis of poly(A) RNA showed an approx 13.1-kb transcript. The mature protein sequence had 57 and 61% of amino acid residues identical to those of Drosophila melanogaster and Anopheles stephensi, respectively, and produced a remarkably similar hydropathy profile when compared to those of the two dipterous species. The three residues (Ser, Glu and His) that putatively form the catalytic triad and the six Cys that form intra-subunit disulfide bonds were completely conserved when compared to the other seven AChEs from a broad range of animal species reported to date. Other properties of the deduced protein of AChE, including molecular weight and amino acid composition, agreed well with those of a previously reported study on the purified AChE from the same insect species. All these features firmly established that the cloned cDNA encodes AChE in Colorado potato beetle.1 aZhu, K, Y1 aClark, J, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cloning-and-sequencing-of-a-cdna-encoding-acetylcholinesterase-in00543nas a2200157 4500008004100000245005600041210005400097260001600151300001000167490000800177100001600185700001400201700001800215700001800233856013400251 1995 eng d00aDiversification of bovine lambda-light chain genes.0 aDiversification of bovine lambdalight chain genes c1995 Sep 29 a155-70 v7641 aParng, C, L1 aHansal, S1 aGoldsby, R, A1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/diversification-of-bovine-lambda-light-chain-genes01169nas a2200133 4500008004100000245007400041210006900115260001300184300001000197490000600207520065300213100001800866856015100884 1995 eng d00aInduction of genes during apoptosis: examples from the immune system.0 aInduction of genes during apoptosis examples from the immune sys c1995 Feb a27-330 v63 aThe immune system provides many examples of cell populations that are susceptible to the induction of apoptosis. Self-reactive immature cells are deleted by triggering apoptosis. Additionally, mature peripheral lymphocytes are induced to undergo apoptosis, particularly when hyperactivated. In the past few years, several genes have been linked to cell death in lymphoid cells. However, only a handful of these genes has been shown to be required for cell death to occur in the immune system. This review focuses on signals known to mediate apoptosis in the immune system and those genes demonstrated to be required for the induction of cell death.1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/induction-of-genes-during-apoptosis-examples-from-the-immune-system02073nas a2200157 4500008004100000245010600041210006900147260001300216300001200229490000700241520146300248100002001711700001401731700001801745856015201763 1995 eng d00aLack of a role for natural killer cells in early control of Brucella abortus 2308 infections in mice.0 aLack of a role for natural killer cells in early control of Bruc c1995 Oct a4029-330 v633 aStudies were conducted to determine if natural killer (NK) cells are important for early control of the virulent strain Brucella abortus 2308 following infection of mice with high or low challenge doses. Splenocytes from C57BL/10 and BALB/c mice that had been infected with the lower dose of B. abortus displayed increased cytotoxicity against YAC-1 cells during the first week after infection, while infection of C57BL/10 mice with the higher challenge dose either did not alter the level of NK cytotoxic activity or decreased it, depending upon the time postinfection. In vivo depletion of NK cells by monoclonal antibody anti-NK1.1 or polyclonal anti-asialoGM1 antiserum did not result in an increase in the number of brucellae recovered from the spleens or livers of the brucella-resistant C57BL/10 mice or from the spleens of the susceptible BALB/c mice during the first week after infection. Treatment of control mice with the NK-reactive antibodies, however, decreased killing of the NK-sensitive target YAC-1, indicating that the NK cell depletion regimes were effective. Our results suggest that NK cells are not crucial for early control of B. abortus 2308 even though they may be activated following infection. Further experiments indicated that treatment of C57BL/10 mice with poly(A:U) did not decrease the number of brucellae recovered from their spleens although it did decrease the CFU in livers of mice infected with the high challenge dose.1 aFernandes, D, M1 aBenson, R1 aBaldwin, C, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/lack-of-a-role-for-natural-killer-cells-in-early-control-of-brucella00482nas a2200145 4500008004100000245004500041210004400086260000900130300001100139490000800150100001600158700002100174700001800195856012300213 1995 eng d00aMolecular events in thymocyte apoptosis.0 aMolecular events in thymocyte apoptosis c1995 a147-620 v2001 aSmith, S, W1 aMcLaughlin, K, A1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/molecular-events-in-thymocyte-apoptosis00544nas a2200145 4500008004100000245006700041210006600108260001300174300001000187490000700197100001700204700001600221700001600237856014500253 1995 eng d00aPreliminary study of synergism of acid rain and diflubenzuron.0 aPreliminary study of synergism of acid rain and diflubenzuron c1995 Jun a833-60 v541 aMartin, P, J1 aClark, J, M1 aEdman, J, D uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/preliminary-study-of-synergism-of-acid-rain-and-diflubenzuron01802nas a2200169 4500008004100000245008000041210006900121260000900190300000900199490000700208520120800215100001601423700001601439700001401455700002101469856014201490 1995 eng d00aResistance to avermectins: extent, mechanisms, and management implications.0 aResistance to avermectins extent mechanisms and management impli c1995 a1-300 v403 aThe avermectins represent a group of natural compounds with potent pesticidal activities. Because of their novel mode of action, they represent an important resource for pest control and resistance management. In the Colorado potato beetle, the house fly, and the two-spotted spider mite, resistance to abamectin is usually autosomal, recessive, and polygenic. Although these aspects are beneficial in resistance management, the fact that resistance could be readily selected for suggests that abamectin needs to be used in moderation. Furthermore, several major resistance mechanisms (e.g. excretion, oxidative metabolism, penetration) and minor factors (e.g. altered target site, conjugation, hydrolysis/sequestration) have been implicated in abamectin resistance. Thus, the question is not whether resistance to abamectin will occur but is simply when and how it will occur. To address this problem, we have gathered information on the genetics, biochemical mechanisms, effectiveness of synergists, and cross-resistances to other insecticides from three abamectin-resistant insects. Judicious implementation of this information may prove useful in the resistance management of this natural pesticide.1 aClark, J, M1 aScott, J, G1 aCampos, F1 aBloomquist, J, R uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/resistance-to-avermectins-extent-mechanisms-and-management01209nas a2200181 4500008004100000245009100041210006900132260000900201300001100210490000700221520055900228100001800787700001600805700001400821700002100835700001900856856015200875 1995 eng d00aTransient transfection assays to examine the requirement of putative cell death genes.0 aTransient transfection assays to examine the requirement of puta c1995 a99-1060 v463 aIn conclusion, this chapter provides a convenient and efficient method for the detection and analysis of transiently transfected cells. Such strategies allow a fast and simple analysis of the requirement for particular genes that have been identified as being induced during apoptosis. Our experience has been that, when screening for "cell death genes," it is easy to isolate genes induced during apoptosis but far more difficult to determine the requirement for any given gene. These protocols have rendered such determinations much simpler to perform.1 aOsborne, B, A1 aSmith, S, W1 aLiu, Z, G1 aMcLaughlin, K, A1 aSchwartz, L, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/transient-transfection-assays-to-examine-the-requirement-of-putative01565nas a2200181 4500008004100000245011900041210006900160260001600229300001000245490000800255520088100263100001401144700001601158700002101174700001901195700001801214856015101232 1994 eng d00aApoptotic signals delivered through the T-cell receptor of a T-cell hybrid require the immediate-early gene nur77.0 aApoptotic signals delivered through the Tcell receptor of a Tcel c1994 Jan 20 a281-40 v3673 aEngagement of the T-cell antigen receptor (TCR) on immature thymic T cells induces death by apoptosis. Although several lines of evidence indicate that apoptosis requires de novo gene expression, little is known about the molecular pathways that mediate this response. Here we show that nur77 (refs 4-7), a zinc-finger transcription factor, is expressed in response to TCR engagement in immature T cells and T-cell hybrids. Antisense inhibition of nur77 expression prevents apoptosis in TCR-stimulated cells. nur77 is also expressed in response to mitogens, but in this case transcription is regulated by 5' upstream elements that are distinct from those used for induction of apoptosis. In addition, polyadenylation is only observed on nur77 transcripts found in condemned cells. These data support a role for nur77 in cell death that may be distinct from that of activation.1 aLiu, Z, G1 aSmith, S, W1 aMcLaughlin, K, A1 aSchwartz, L, M1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/apoptotic-signals-delivered-through-the-t-cell-receptor-of-a-t-cell00513nas a2200133 4500008004100000245006600041210006200107260001300169300001000182490000700192100001900199700001800218856014300236 1994 eng d00aCed-3/ICE: evolutionarily conserved regulation of cell death.0 aCed3ICE evolutionarily conserved regulation of cell death c1994 Jun a387-90 v161 aSchwartz, L, M1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/ced-3-ice-evolutionarily-conserved-regulation-of-cell-death00851nas a2200145 4500008004100000245004500041210004400086260001300130300001000143490000600153520038600159100001800545700001900563856012300582 1994 eng d00aEssential genes that regulate apoptosis.0 aEssential genes that regulate apoptosis c1994 Nov a394-90 v43 aThe expression of several genes has been associated with the induction of apoptosis in a wide variety of vertebrate and invertebrate organisms. However, relatively few gene products have been demonstrated to be required for cell death. This review highlights genes that are required for apoptosis and proposes mechanisms by which the proteins encoded by these genes might function.1 aOsborne, B, A1 aSchwartz, L, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/essential-genes-that-regulate-apoptosis00645nas a2200181 4500008004100000245007100041210006900112260001300181300001100194490000800205100001800213700001600231700001400247700002100261700001300282700001900295856014900314 1994 eng d00aIdentification of genes induced during apoptosis in T lymphocytes.0 aIdentification of genes induced during apoptosis in T lymphocyte c1994 Dec a301-200 v1421 aOsborne, B, A1 aSmith, S, W1 aLiu, Z, G1 aMcLaughlin, K, A1 aGrimm, L1 aSchwartz, L, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/identification-of-genes-induced-during-apoptosis-in-t-lymphocytes02861nas a2200337 4500008004100000245016800041210006900209260001300278300001000291490000700301520178800308653001902096653001802115653001202133653001402145653001402159653001102173653002002184653001602204653001302220653001702233653001602250653001702266653001402283653000902297653001702306100001702323700002002340700001502360856014802375 1994 eng d00aMifepristone treatment demonstrates the participation of adrenal glucocorticoids in the regulation of oestrogen-induced prolactin secretion in ovariectomized rats.0 aMifepristone treatment demonstrates the participation of adrenal c1994 Mar a385-90 v483 aAccumulated evidence indicates that the adrenal cortex is able to regulate prolactin (PRL) secretion in rats. The aim of this study was to determine the participation of adrenal steroids on the regulation of PRL release in ovariectomized (OVX) and oestrogen-treated rats, by using mifepristone or a specific progesterone antiserum. Blood samples were obtained at 13:00 and 18:00 h 3 days after priming with oestradiol benzoate (OB). A significant increase in serum PRL at 13:00 and 18:00 h was induced by OB treatment. The administration of mifepristone to OVX and oestrogen-primed rats enhanced serum PRL increase at 13:00 h, without modifying the values at 18:00 h; while the administration of progesterone antiserum did not modify PRL levels, indicating that the effect of mifepristone on PRL secretion is due to its antiglucocorticoid action. Adrenalectomy induced a release of PRL at 13:00 h similar to that observed in the OVX and oestrogen-primed rats after mifepristone administration. Treatment with a low dose of progesterone (0.1 mg/rat) to OVX, adrenalectomized and oestrogen-primed rats did not modify the effect of adrenalectomy in serum PRL. Progesterone (2 mg/rat) given at 08:00 h to OVX and oestrogen-primed rats increased serum PRL 5 h later. Mifepristone treatment partially reverted the PRL increase induced by progesterone. These results suggest that after a previous sensitization of the pituitary by oestrogen, circulating glucocorticoids may exert a direct inhibitory effect on PRL release. This inhibition takes place at 13:00 h on day 3. On the other hand, the lack of effect of mifepristone or adrenalectomy on the PRL release at 18:00 h may also indicate that neither progesterone nor glucocorticoids modify PRL release induced by oestrogen at this time.10aAdrenal Cortex10aAdrenalectomy10aAnimals10aEstradiol10aEstrogens10aFemale10aGlucocorticoids10aImmune Sera10aKinetics10aMifepristone10aOvariectomy10aProgesterone10aProlactin10aRats10aRats, Wistar1 aCarón, R, W1 aSalicioni, A, M1 aDeis, R, P uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/mifepristone-treatment-demonstrates-the-participation-of-adrenal02212nas a2200157 4500008004100000245012000041210006900161260001500230300001100245490000800256520158000264100002301844700001801867700001801885856015101903 1994 eng d00aQuantitative analysis of the B cell repertoire by limiting dilution analysis and fluorescent in situ hybridization.0 aQuantitative analysis of the B cell repertoire by limiting dilut c1994 Apr 1 a309-270 v1543 aWe have made a quantitative and concurrent analysis of B cell frequencies and VH gene family expression to study the influence of tissue type and age on the development and establishment of the primary B cell repertoire. Using LPS-mediated limiting dilution analysis and a panel of antigens we show that the newly generated B cell specificities from bone marrow get distributed without a bias to peripheral tissues such as spleen and Peyer's patches throughout the lifetime of the animal. Comparison of the B cell frequencies in animals of four different age groups (2-4 days old, 3, 12, and 18 months old) reveals that while the neonatal repertoire is comparable to that of adults, there was a selective twofold increase in the generation and distribution of B cells reactive with autologous mouse red blood cells in older mice compared to young ones. By means of a novel technique that employs fluorescent in situ hybridization and flow cytometry, we have also compared the VH gene family usage in large numbers of single B cells from these mice. Analysis of the same cell population (surface Ig+, functional B lineage cells) for expression of 7183, J558, and S107 VH families shows a preferential twofold increase in the use of VH 7183 in neonates compared to adults, while all three families show no significant difference in levels of expression during adult life or between primary and secondary lymphoid tissues. Taken together, our data indicate that specific and selective changes occur in both VH gene usage and antibody frequencies during murine ontogeny.
1 aRavichandran, K, S1 aOsborne, B, A1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/quantitative-analysis-of-the-b-cell-repertoire-by-limiting-dilution02512nas a2200337 4500008004100000245011000041210006900151260001300220300001000233490000800243520147900251653001901730653001801749653001201767653001401779653001401793653001301807653001101820653001601831653002401847653001701871653001601888653001701904653002101921653000901942653001701951100002001968700001701988700001502005856015402020 1993 eng d00aAdrenal progesterone facilitates the negative feedback of oestrogen on LH release in ovariectomized rats.0 aAdrenal progesterone facilitates the negative feedback of oestro c1993 Nov a253-80 v1393 aThere is evidence that the adrenals play a role in the regulation of the synthesis and release of gonadotrophins in various vertebrates. The aim of this study was to determine the part played by adrenal steroids, with special reference to progesterone, on the concentration of LH in ovariectomized (OVX) and oestrogen-primed rats. OVX rats received a single s.c. injection of vehicle or oestradiol benzoate (OB, 20 micrograms/rat). This day was designated as day 0. Three or four days later (day 3-day 4), the rats were treated with mifepristone (10 mg/kg) or with two doses of progesterone antiserum and blood samples were obtained at 13.00 and 18.00 h. OB treatment of OVX rats reduced serum LH at 13.00 h and 18.00 h on day 3 but only at 13.00 h on day 4. The administration of mifepristone at 08.00 h to OVX and oestrogen-treated rats induced a significant increase in serum LH at 18.00 h on days 3 and 4, without modifying the values at 13.00 h. When mifepristone was given at 13.00 h a much larger increase in serum LH was obtained at 18.00 h. In OVX and oestrogen-treated rats, adrenalectomy on day 2 (08.00-09.00 h) induced an increase in serum LH at 18.00 h similar to that observed in the OVX and oestrogen-primed rats after mifepristone treatment. In order to determine the specificity of the effect of mifepristone, a group of OVX and oestrogen-treated rats was injected with progesterone antiserum at 08.00 and 13.00 h on day 3.(ABSTRACT TRUNCATED AT 250 WORDS)10aAdrenal Glands10aAdrenalectomy10aAnimals10aEstradiol10aEstrogens10aFeedback10aFemale10aImmune Sera10aLuteinizing Hormone10aMifepristone10aOvariectomy10aProgesterone10aRadioimmunoassay10aRats10aRats, Wistar1 aSalicioni, A, M1 aCarón, R, W1 aDeis, R, P uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/adrenal-progesterone-facilitates-the-negative-feedback-of-oestrogen-on01661nas a2200169 4500008004100000245005500041210005400096260001500150300001000165490000700175520110700182100001901289700001601308700001601324700001801340856013301358 1993 eng d00aDo all programmed cell deaths occur via apoptosis?0 aDo all programmed cell deaths occur via apoptosis c1993 Feb 1 a980-40 v903 aDuring development, large numbers of cells die by a nonpathological process referred to as programmed cell death. In many tissues, dying cells display similar changes in morphology and chromosomal DNA organization, which has been termed apoptosis. Apoptosis is such a widely documented phenomenon that many authors have assumed all programmed cell deaths occur by this process. Two well-characterized model systems for programmed cell death are (i) the death of T cells during negative selection in the mouse thymus and (ii) the loss of intersegmental muscles of the moth Manduca sexta at the end of metamorphosis. In this report we compare the patterns of cell death displayed by T cells and the intersegmental muscles and find that they differ in terms of cell-surface morphology, nuclear ultrastructure, DNA fragmentation, and polyubiquitin gene expression. Unlike the T cells, which are known to die via apoptosis, we find that the intersegmental muscles display few of the features that characterize apoptosis. These data suggest that more than one cell death mechanism is used during development.1 aSchwartz, L, M1 aSmith, S, W1 aJones, M, E1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/do-all-programmed-cell-deaths-occur-via-apoptosis01056nas a2200145 4500008004100000245005500041210005300096260001300149300001100162490000700173520056100180100001900741700001800760856013200778 1993 eng d00aProgrammed cell death, apoptosis and killer genes.0 aProgrammed cell death apoptosis and killer genes c1993 Dec a582-900 v143 aA cursory examination of the literature reveals that the study of programmed cell death and apoptosis is increasing exponentially. Most contributors to this field have come either from developmental biology or immunology and view programmed cell death from different perspectives, leading both to confusion and an inability to fully appreciate the literature from other disciplines. Here, Lawrence Schwartz and Barbara Osborne define the terms and ideas relevant to the study of cell death in a way that will be accessible to investigators from all fields.1 aSchwartz, L, M1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/programmed-cell-death-apoptosis-and-killer-genes01920nas a2200169 4500008004100000245009400041210006900135260001600204300001100220490000800231520128000239100002301519700001901542700001801561700001801579856015301597 1992 eng d00aImmunoglobulin VH usage analysis by fluorescent in situ hybridization and flow cytometry.0 aImmunoglobulin VH usage analysis by fluorescent in situ hybridiz c1992 Aug 30 a249-590 v1533 aWe have devised a flow cytometry-based fluorescent in situ hybridization assay that permits analysis of gene expression in a large number of single cells. In this technique, fixed and permeabilized cells are incubated with biotinylated single-stranded RNA probes and by means of a fluorescently labelled second-step reagent, the cells are analyzed by flow cytometry. This is a rapid and simple method that allows all of the steps in the procedure to be performed on cells in suspension. Using this approach, we demonstrate here that immunoglobulin heavy chain variable region (VH) gene expression can be analyzed among individual cells using particular VH family-specific probes. This technique has a high degree of accuracy (greater than 97%) in detecting the fraction of cells expressing a specific message in a population and is sensitive enough to detect immunoglobulin message in LPS activated B cells. The technique has been applied successfully to monitor gene expression in homogeneous and heterogeneous populations. It also allows concurrent analysis of cell surface proteins and gene expression through two-color flow cytometry. This method of monitoring gene expression in individual cells may have a number of applications in immunology and cell biology.
1 aRavichandran, K, S1 aSemproni, A, R1 aGoldsby, R, A1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/immunoglobulin-vh-usage-analysis-by-fluorescent-in-situ-hybridization01550nas a2200157 4500008004100000245005900041210005700100260001700157300001000174490000700184520101100191100001401202700001801216700002101234856013701255 1992 eng d00aSalmonella enteritidis-specific monoclonal antibodies.0 aSalmonella enteritidisspecific monoclonal antibodies c1992 Apr-Jun a455-80 v363 aThree monoclonal antibodies (MAbs) were derived that are specific for Salmonella enteritidis. Such antibodies are of interest because reagents that specifically identify S. enteritidis are potentially useful for the diagnosis and detection of this pathogen. Immunization of BALB/c mice with intact, unfixed, ultraviolet-killed S. enteritidis permitted the derivation of a collection of hybridomas among which were found three MAbs: 1053, 1110, and 1170. Each MAb reacted with six independent field isolates of S. enteritidis, including phage type 4. However, none of these S. enteritidis-specific MAbs reacted with any of the following members of a broad diversity of Salmonella species: S. typhimurium, S. pullorum, S. berta, S. agona, S. dublin, S. miami, S. heidelberg, S. montevideo, S. senftenberg, and S. schwarzengrund. The S. enteritidis-specific determinant recognized by these MAbs is heat-labile, and preliminary experiments indicate that at least two of the MAbs recognize the same determinant.1 aLin, A, W1 aGoldsby, R, A1 aSnoeyenbos, G, H uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/salmonella-enteritidis-specific-monoclonal-antibodies00661nas a2200169 4500008004100000245011400041210006900155260001300224300001100237490000700248100001600255700001700271700001800288700001700306700001500323856015300338 1991 eng d00aAirborne drift residues collected near apple orchard environments due to application of insecticide mixtures.0 aAirborne drift residues collected near apple orchard environment c1991 Jun a829-360 v461 aClark, J, M1 aMarion, J, R1 aTessier, D, M1 aBrooks, M, W1 aColi, W, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/airborne-drift-residues-collected-near-apple-orchard-environments-due01887nas a2200205 4500008004100000245011600041210006900157260001300226300001100239490000600250520115600256100001901412700001601431700001601447700001701463700001701480700001701497700001401514856015301528 1990 eng d00aCuticular lipid differences between the malaria vector and non-vector forms of the Anopheles maculatus complex.0 aCuticular lipid differences between the malaria vector and nonve c1990 Oct a405-130 v43 aTwo chromosomal forms (E and F) of the Anopheles maculatus Theobald complex were distinguished by gas-liquid chromatographic (GC) analysis of cuticular lipids in association with a multivariate principal component analysis. The GC chromatogram obtained from n-hexane extracts of individual specimens showed no consistent qualitative differences in normalized peak areas between forms. Of the seventeen consistent peaks, five were found to be quantitatively different between forms at a high (99.5-99.95%) level of statistical confidence. Relative ratios of these five quantitatively different GC peaks were used as criteria to distinguish single specimens as either form E or form F. Chemical structures of the five GC peaks were assigned by both electron impact and chemical ionization gas chromatography/mass spectrometry analysis. The first three peaks, which were always doublets, were partially resolved saturated and mono-unsaturated free fatty acids; the other two peaks were n-alkanes. Principal component analysis substantiated that the vector form E has very similar cuticular lipid profiles and is well separated from the non-vector form F.1 aKittayapong, P1 aClark, J, M1 aEdman, J, D1 aPotter, T, L1 aLavine, B, K1 aMarion, J, R1 aBrooks, M uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/cuticular-lipid-differences-between-the-malaria-vector-and-non-vector01980nas a2200157 4500008004100000245013400041210006900175260001500244300001200259490000800271520133800279100001901617700001701636700001801653856015101671 1989 eng d00aAnalysis of a novel VHS107 haplotype in CLA-2 and WSA mice. Evidence for gene conversion among IgVH genes in outbred populations.0 aAnalysis of a novel VHS107 haplotype in CLA2 and WSA mice Eviden c1989 Dec 1 a1811-230 v1703 aGene conversion has been suggested as the basis for many VH allelic differences, particularly in the murine VHS107 family. Whether conversion among IgVH genes is likely to have occurred in outbred populations has not been directly addressed. The CLA-2/Cn and WSA strains, which were recently and independently derived from a feral population exhibiting low responsiveness to PC, provide the opportunity to approach this question. In previous studies, the heavy chain cDNA sequence of a PC-specific hybridoma derived from CLA-2/Cn suggested gene conversion events within the VHS107 family. Accordingly, we have examined the germline VHS107 genes of CLA-2/Cn and WSA. The results indicate that: (a) The CLA-2 and WSA strains bear an identical but novel VHS107 family haplotype, which lacks a V3 element and contains a V1, a V13, and two V11 genes; (b) low PC responsiveness in these populations is unlikely due to an inability to express the V1 member of the VHS107 gene family; and (c) when compared with the other known VHS107 haplotypes, the proportion of differences consistent with gene conversion greatly exceeds that expected by random base substitution. Thus, gene conversion events appear to have occurred with considerable frequency in the evolution of the murine VHS107 family, especially among the V3, V13, and V11 members.1 aFerguson, S, E1 aCancro, M, P1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/analysis-of-a-novel-vhs107-haplotype-in-cla-2-and-wsa-mice-evidence02420nas a2200145 4500008004100000245012500041210006900166260001600235300001200251490000700263520182200270100001602092700001702108856014902125 1989 eng d00aRole of ion channels and intraterminal calcium homeostasis in the action of deltamethrin at presynaptic nerve terminals.0 aRole of ion channels and intraterminal calcium homeostasis in th c1989 Jul 15 a2233-450 v383 aUsing a continuous perfusion system, synaptosomes prepared from rat brain released [3H]norepinephrine in a Ca2+-dependent manner when pulse depolarized by briefly elevating external potassium concentrations. Tetrodotoxin (10(-7) M), a sodium channel blocker, inhibited 48% of this pulsed release, and D595 (10(-5) M), a phenethylamine-type calcium channel blocker, inhibited 21%. In combination, these two specific ion channel antagonists appear to function independently of each other in an additive fashion. Addition of deltamethrin to this preparation resulted in an enhanced release of [3H]norepinephrine which occurred in a biphasic fashion. At 10(-7) M, deltamethrin produced a 42% enhancement in the first or initial peak of [3H]norepinephrine release and a 100% enhancement in the second or tailing peak. Addition of deltamethrin to tetrodotoxin-pretreated synaptosomes resulted in a net 37% enhancement of the initial peak release and a net increase of 277% in the tailing peak. Addition of deltamethrin to D595-pretreated synaptosomes produced no significant effect on enhanced [3H]norepinephrine release from either peak. Since tetrodotoxin is a specific sodium channel blocker, deltamethrin may be enhancing [3H]norepinephrine release by increasing the uptake of Ca2 via other voltage-gated channels (e.g. calcium) or exchange mechanisms in addition to its action at voltage-gated sodium channels. To determine whether deltamethrin may also have an effect on intraterminal Ca2+ homeostasis, external Ca2+ was replaced with Ba2+ and synaptosomes were depolarized with pentylenetetrazole (PTZ). At 10(-5) M, deltamethrin produced a 66% increase in neurotransmitter release over that produced by PTZ alone. An estimated EC50 value of deltamethrin for PTZ-induced release was calculated to be 2.4 x 10(-10) M.1 aClark, J, M1 aBrooks, M, W uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/role-of-ion-channels-and-intraterminal-calcium-homeostasis-in-the02874nas a2200385 4500008004100000245013100041210006900172260001300241300001200254490000600266520164400272653001901916653001201935653003601947653002301983653002502006653003102031653002502062653001502087653001002102653000902112653002402121653000902145653002502154653001902179653002402198100002202222700001302244700001402257700001302271700001702284700002102301700001902322856014702341 1989 eng d00aTissue-specific expression, developmental regulation, and genetic mapping of the gene encoding CCAAT/enhancer binding protein.0 aTissuespecific expression developmental regulation and genetic m c1989 Aug a1146-560 v33 aThis paper presents the results of experiments that determine the chromosomal location of the mouse gene encoding CCAAT/enhancer binding protein (C/EBP) and measure its expression as a function of tissue type and temporal period of development in mice and rats. Three alleles of the C/EBP gene were identified according to restriction fragment length polymorphisms. The strain distribution pattern of the three alleles was determined in recombinant inbred mouse strains and compared to that of other mouse genes. These results mapped the gene to a position within 2.5 centimorgans (cM) of the structural gene encoding glucose phosphate isomerase on chromosome 7 of the mouse. The expression pattern of the C/EBP gene was studied by a combination of nucleic acid hybridization and antibody staining assays. High levels of C/EBP mRNA were observed in tissues known to metabolize lipid and cholesterol-related compounds at uncommonly high rates. These included liver, fat, intestine, lung, adrenal gland, and placenta. More detailed analysis of two of these tissues, liver and fat, showed that C/EBP expression was limited to fully differentiated cells. Moreover, analysis of the temporal pattern of expression of C/EBP mRNA in two tissues, liver and intestine, revealed a coordinated induction just prior to birth. These observations raise the possibility that the synthesis of C/EBP may be responsive to humoral factors and that modulation in C/EBP expression might mediate coordinated changes in gene expression that facilitate adaptive challenges met during development or during the fluctuating physiological states of adult life.
10aAdipose Tissue10aAnimals10aCCAAT-Enhancer-Binding Proteins10aChromosome Mapping10aDNA-Binding Proteins10aGene Expression Regulation10aImmunohistochemistry10aIntestines10aLiver10aMice10aMice, Inbred BALB C10aRats10aRats, Inbred Strains10aRNA, Messenger10aTissue Distribution1 aBirkenmeier, E, H1 aGwynn, B1 aHoward, S1 aJerry, J1 aGordon, J, I1 aLandschulz, W, H1 aMcKnight, S, L uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/tissue-specific-expression-developmental-regulation-and-genetic02773nas a2200145 4500008004100000245013000041210006900171260001600240300001100256490000700267520216100274100002202435700001802457856015202475 1988 eng d00aComparative idiotype network interactions: antigen mimicry by an anti-bovine idiotype monoclonal antibody in rats and cattle.0 aComparative idiotype network interactions antigen mimicry by an c1988 Summer a669-830 v123 aIdiotype/anti-idiotype networks have been extensively investigated in such conventional animal models as the mouse and the rabbit. However, systems of veterinary interest have remained largely unexplored. A monoclonal target idiotype, with which to begin such studies in cattle was provided by LHRB 19.17 an interspecific bovine x mouse hybridoma. This hybridoma was constructed by the fusion of supramammary lymph node cells from S. agalactiae-immunized lactating Holsteins with the Ig synthesis-permissive established cell line, SP 2/0. Two collections of monoclonal anti-idiotype antibodies were generated by fusion of spleen cells from LHRB 19.17-immunized Balb/c or A/J mice immunized with the monoclonal bovine idiotype, LHRB 19.17. Many of the anti-idiotypes inhibited binding of LHRB 19.17 to S. agalactiae, but only one, LHRAID 2.71, proved to be an internal image of a S. agalactiae epitope. Immunization of C/D outbred rats by priming with 100-300 micrograms of LHRAID 2.71 emulsified in CFA followed by a 300 micrograms boost at day 32 elicited anti- S. agalactiae antibody in 4/4 animals tested. Similarly, the injection of two lactating Holsteins with the anti-id resulted in the production of anti- S. agalactiae antibody in serum and milk. In both rats and cattle, the administration of the antigen-mimicking anti-idiotype induces the appearance of S. agalactiae-reactive horseradish peroxidase-streptavidin conjugate; LHRBs, interspecific bovine x mouse hybridomas secreting bovine Ig; LHRAID.X, monoclonal anti-bovine idiotype antibodies derived against LHRB 19.17; PBS, phosphate buffered saline; PBS/BSA, PBS containing 0.1% bovine serum albumin: antibody [AB3] that competes with LHRB 19.17 [AB1] for binding to LHRAID 2.71 [AB2]. It should also be noted that the immunization of C/D rats with S. agalactiae does not result in the appearance of idiotypes which compete with LHRB 19.17 for binding to LHRAID 2.71. We have concluded that immunization of two widely divergent species with the antigen mimicking LHRAID 2.71 induced a S. agalactiae-reactive idiotype which was not detectable in the immune response of rats to S. agalactiae.1 aArulanandam, A, R1 aGoldsby, R, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/comparative-idiotype-network-interactions-antigen-mimicry-by-an-anti01339nas a2200169 4500008004100000245006000041210005900101260001300160300001100173490000800184520077000192100001800962700001600980700001900996700001601015856013801031 1988 eng d00aEvolution of the IgA heavy chain gene in the genus Mus.0 aEvolution of the IgA heavy chain gene in the genus Mus c1988 Aug a925-310 v1193 aTo examine questions of immunoglobulin gene evolution, the IgA alpha heavy chain gene from Mus pahari, an evolutionarily distant relative to Mus musculus domesticus, was cloned and sequenced. The sequence, when compared to the IgA gene of BALB/c or human, demonstrated that the IgA gene is evolving in a mosaic fashion with the hinge region accumulating mutations most rapidly and the third domain at a considerably lower frequency. In spite of this pronounced accumulation of mutations, the hinge region appears to maintain the conformation of a random coil. A marked propensity to accumulate replacement over silent site changes in the coding regions was noted, as was a definite codon bias. The possibility that these two phenomena are interrelated is discussed.1 aOsborne, B, A1 aGolde, T, E1 aSchwartz, R, L1 aRudikoff, S uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/evolution-of-the-iga-heavy-chain-gene-in-the-genus-mus01596nas a2200157 4500008004100000245007300041210006900114260001500183300001200198490000800210520101600218100001901234700001601253700001801269856015101287 1988 eng d00aInteraction and sequence diversity among T15 VH genes in CBA/J mice.0 aInteraction and sequence diversity among T15 VH genes in CBAJ mi c1988 Oct 1 a1339-490 v1683 aNucleotide sequences of the four genes composing the T15 heavy chain variable region (VH) family of the CBA/J mouse have been determined. Comparison of these sequences with their published BALB/c and C57BL/10 homologues reveals that nucleotide differences found between given alleles of two strains, i.e., CBA/J and BALB/c, are observed in other family members of the same strain. We suggest that these patterns of sequence variation are most readily explained by gene interaction (conversion). Additionally, the sequence of a CBA/J hybridoma, 6G6, proposed to have been generated by gene conversion, is directly encoded by the CBA/J V11 gene indicating that the putative conversion has occurred meiotically in the germline. These results are consistent with the premise that gene correction is occurring frequently among members of this family and that such processes may contribute significantly to the evolution of Ig variable region genes even in the relatively short time frame of inbred strain derivation.1 aFerguson, S, E1 aRudikoff, S1 aOsborne, B, A uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/interaction-and-sequence-diversity-among-t15-vh-genes-in-cba-j-mice02127nas a2200205 4500008004100000245008400041210006900125260001300194300001000207490000700217520142600224100001801650700001801668700001901686700001301705700002501718700001501743700001701758856014601775 1987 eng d00aThe application of hybridoma technology to the study of bovine immunoglobulins.0 aapplication of hybridoma technology to the study of bovine immun c1987 Dec a25-350 v173 aStudies are described in which hybridoma technology is used to produce a variety of reagents for the characterization and manipulation of the bovine humoral immune system. Selected members of a set of murine monoclonal antibodies (MAb) specific for each of four major isotypes of bovine Ig constant regions, one specific for anti-bovine Ig constant regions as well as one specific for anti-bovine light chains are discussed. Interspecific fusion of bovine lymphocytes with the established mouse cell line, SP2/0 was used to produce a collection of stable hybridomas among which were found secretors of bovine IgG1, IgG2, IgM, IgA and bovine light chain. Interspecific fusion of SP2/0 with lymphocytes from a multiparous Holstein four days post immunization with Streptococcus agalactiae yielded MAb with specificity for the immunizing antigen. One of these hybridomas, LHRB 19.17, which displayed a particularly stable secretory phenotype, was used as an immunogen for the production of a library of murine monoclonal anti-idiotype antibodies. Competitive antigen binding analysis showed that 15 of the 24 anti-LHRB 19.17 idiotype antibodies isolated blocked the binding of the idiotype to its nominal antigen and so were candidates for evaluation as antigen mimics. Some of the ways in which monoclonal anti-idiotypes in particular, and monoclonal in general, might be of use in problems of animal disease are discussed.1 aGoldsby, R, A1 aSrikumaran, S1 aArulanandam, A1 aHague, B1 ade León, F, A Ponce1 aSevoian, M1 aGuidry, A, J uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/the-application-of-hybridoma-technology-to-the-study-of-bovine