%0 Journal Article %J Frontiers in Nutrition %D 2024 %T Mammary epithelium permeability during established lactation: associations with cytokine levels in human milk %A Kivlighan, Katie T. %A Schneider, Sallie S. %A Browne, Eva P. %A Pentecost, Brian T. %A Anderton, Douglas L. %A Arcaro, Kathleen F. %B Frontiers in Nutrition %V 11 %G eng %U http://dx.doi.org/10.3389/fnut.2024.1258905 %R 10.3389/fnut.2024.1258905 %0 Journal Article %J Lab on a Chip %D 2024 %T Rapid cell isolation in breastmilk in a non-clinical setting by a deterministic lateral displacement device and selective water and fat absorption %A Hawkins, Jamar %A Browne, Eva P. %A Arcaro, Kathleen F. %A Sun, Yubing %B Lab on a Chip %V 24 %P 604–614 %G eng %U http://dx.doi.org/10.1039/d3lc00899a %R 10.1039/d3lc00899a %0 Journal Article %J Nutrients %D 2023 %T Effect of Maternal Diet on Maternal Milk and Breastfed Infant Gut Microbiomes: A Scoping Review %A Rachel Taylor %A Deirdre Keane %A Paulina Borrego %A Kathleen Arcaro %B Nutrients %V 15 %P 1420 %G eng %U https://doi.org/10.3390/nu15061420 %R 10.3390/nu15061420 %0 Journal Article %J Biology of Reproduction %D 2022 %T Berberine alleviates LPS-induced apoptosis, oxidation, and skewed lineages during mouse preimplantation development %A Xiaosu Miao %A Wei Cui %B Biology of Reproduction %V 106 %P 699–709 %G eng %U https://doi.org/10.1093/biolre/ioac002 %R 10.1093/biolre/ioac002 %0 Journal Article %J Biol Reprod %D 2022 %T Berberine alleviates LPS-induced apoptosis, oxidation, and skewed lineages during mouse preimplantation development†. %A Miao, Xiaosu %A Cui, Wei %K Animals %K Apoptosis %K Berberine %K Cell Lineage %K Female %K Humans %K Lipopolysaccharides %K Mammals %K Mice %K Polycystic Ovary Syndrome %K Pregnancy %X

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.

%B Biol Reprod %V 106 %P 699-709 %8 2022 04 26 %G eng %N 4 %R 10.1093/biolre/ioac002 %0 Journal Article %J Mol Reprod Dev %D 2022 %T Borcs6 is required for endo-lysosomal degradation during early development. %A Bell, Charlotte J %A Gupta, Neha %A Tremblay, Kimberly D %A Mager, Jesse %K Animals %K Autophagy %K Endosomes %K Homeostasis %K Lysosomes %K Mammals %K Membrane Fusion %K Proteins %X

Early 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.

%B Mol Reprod Dev %V 89 %P 337-350 %8 2022 08 %G eng %N 8 %R 10.1002/mrd.23626 %0 Journal Article %J Dev Biol %D 2022 %T Deciphering the role of retinoic acid in hepatic patterning and induction in the mouse. %A Guertin, Taylor M %A Palaria, Amrita %A Mager, Jesse %A Sandell, Lisa L %A Trainor, Paul A %A Tremblay, Kimberly D %K Animals %K Endoderm %K Female %K Liver %K Mammals %K Mice %K Pregnancy %K Prospective Studies %K Tretinoin %K Vitamin A %X

Retinoic 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.

%B Dev Biol %V 491 %P 31-42 %8 2022 Nov %G eng %R 10.1016/j.ydbio.2022.08.003 %0 Journal Article %J Front Immunol %D 2022 %T Durable antibody and effector memory T cell responses in breastmilk from women with SARS-CoV-2. %A Narayanaswamy, Vignesh %A Pentecost, Brian T %A Telfer, Janice C %A Burnside, Amy S %A Schneider, Sallie S %A Alfandari, Dominique %A Baker, Ryan L %A Saiju, Aman %A Nodiff, Sam %A Arcaro, Kathleen F %K COVID-19 %K COVID-19 Vaccines %K Female %K Humans %K Immunoglobulin A %K Immunoglobulin G %K Immunoglobulin M %K Infant %K Lactation %K Memory T Cells %K Milk, Human %K Potassium %K Pregnancy %K Receptors, CCR7 %K SARS-CoV-2 %K Sodium %K Spike Glycoprotein, Coronavirus %X

Background: 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.

%B Front Immunol %V 13 %P 985226 %8 2022 %G eng %R 10.3389/fimmu.2022.985226 %0 Journal Article %J Molecular Reproduction and Development %D 2022 %T Early embryonic lethality of mice lacking POLD2 %A Xiaoqing Wu %A Yong Liu %A Wenying Wang %A Kate Crimmings %A Andrea Williams %A Jesse Mager %A Wei Cui %B Molecular Reproduction and Development %V 90 %P 98–108 %G eng %U https://doi.org/10.1002/mrd.23663 %R 10.1002/mrd.23663 %0 Journal Article %J Journal of Veterinary Internal Medicine %D 2022 %T Effect of early or late blood sampling on thyrotropin releasing hormone stimulation test results in horses %A Kristen Thane %A Cassandra Uricchio %A Nicholas Frank %B Journal of Veterinary Internal Medicine %I Wiley %G eng %U https://doi.org/10.1111/jvim.16362 %0 Journal Article %J Bioconjug Chem %D 2022 %T Evaluation of Cellular Targeting by Fab' vs Full-Length Antibodies in Antibody-Nanoparticle Conjugates (ANCs) Using CD4 T-cells. %A Singh, Khushboo %A Canakci, Mine %A Kanjilal, Pintu %A Williams, Natalie %A Shanthalingam, Sudarvili %A Osborne, Barbara A %A Thayumanavan, S %K Antibodies, Monoclonal %K CD4-Positive T-Lymphocytes %K Immunoconjugates %K Immunoglobulin Fab Fragments %K Nanoparticles %X

Targeted 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.

%B Bioconjug Chem %V 33 %P 486-495 %8 2022 03 16 %G eng %N 3 %R 10.1021/acs.bioconjchem.2c00024 %0 Journal Article %J Infect Immun %D 2022 %T Identification of Leptospiral Protein Antigens Recognized by WC1 γδ T Cell Subsets as Target for Development of Recombinant Vaccines. %A Teixeira, Aline F %A Gillespie, Alexandria %A Yirsaw, Alehegne %A Britton, Emily %A Telfer, Janice C %A Nascimento, Ana Lucia Tabet Oller %A Baldwin, Cynthia L %K Animals %K Antigens, Bacterial %K Bacterial Proteins %K Cattle %K Cattle Diseases %K Immunization %K Immunophenotyping %K Leptospira %K Leptospirosis %K Ligands %K Protein Binding %K Protein Interaction Domains and Motifs %K Receptors, Antigen, T-Cell, gamma-delta %K Recombinant Proteins %K T-Lymphocyte Subsets %K Vaccine Development %K Vaccines, Synthetic %X

Pathogenic 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.

%B Infect Immun %V 90 %P e0049221 %8 2022 01 25 %G eng %N 1 %R 10.1128/IAI.00492-21 %0 Journal Article %J J Mammary Gland Biol Neoplasia %D 2022 %T Induced mammary cancer in rat models: pathogenesis, genetics, and relevance to female breast cancer. %A Miller, James L %A Bartlett, Arianna P %A Harman, Rebecca M %A Majhi, Prabin Dhangada %A Jerry, D Joseph %A Van de Walle, Gerlinde R %K 9,10-Dimethyl-1,2-benzanthracene %K Animals %K Breast Neoplasms %K Carcinogens %K Estrogens %K Female %K Humans %K Mammary Neoplasms, Animal %K Mammary Neoplasms, Experimental %K Quantitative Trait Loci %K Rats %K Rats, Sprague-Dawley %X

Mammary 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.

%B J Mammary Gland Biol Neoplasia %V 27 %P 185-210 %8 2022 06 %G eng %N 2 %R 10.1007/s10911-022-09522-w %0 Journal Article %J Ecotoxicol Environ Saf %D 2022 %T Interindividual variation contributes to differential PCB 126 induced gene expression in primary breast epithelial cells and tissues. %A Morin, Stephanie M %A Majhi, Prabin Dhangada %A Crisi, Giovanna M %A Gregory, Kelly J %A Franca, Renata %A Schalet, Benjamin %A Mason, Holly %A Casaubon, Jesse Thomas %A Cao, Qing Jackie %A Haddad, Sandra %A Makari-Judson, Grace %A Jerry, D Joseph %A Schneider, Sallie S %K Cytochrome P-450 CYP1A1 %K Epithelial Cells %K Female %K Gene Expression %K Humans %K Polychlorinated Biphenyls %K Receptors, Aryl Hydrocarbon %X

PCB 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.

%B Ecotoxicol Environ Saf %V 241 %P 113722 %8 2022 Aug %G eng %R 10.1016/j.ecoenv.2022.113722 %0 Journal Article %J Dev Biol %D 2022 %T Mcrs1 is required for branchial arch and cranial cartilage development. %A Keer, Stephanie %A Cousin, Hélène %A Jourdeuil, Karyn %A Neilson, Karen M %A Tavares, Andre L P %A Alfandari, Dominique %A Moody, Sally A %K Branchial Region %K Branchio-Oto-Renal Syndrome %K Cartilage %K Gene Expression Regulation, Developmental %K Homeodomain Proteins %K Neural Crest %K Neural Plate %K RNA-Binding Proteins %X

Mcrs1 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.

%B Dev Biol %V 489 %P 62-75 %8 2022 09 %G eng %R 10.1016/j.ydbio.2022.06.002 %0 Journal Article %J Nat Commun %D 2022 %T Microbial enzymes induce colitis by reactivating triclosan in the mouse gastrointestinal tract. %A Zhang, Jianan %A Walker, Morgan E %A Sanidad, Katherine Z %A Zhang, Hongna %A Liang, Yanshan %A Zhao, Ermin %A Chacon-Vargas, Katherine %A Yeliseyev, Vladimir %A Parsonnet, Julie %A Haggerty, Thomas D %A Wang, Guangqiang %A Simpson, Joshua B %A Jariwala, Parth B %A Beaty, Violet V %A Yang, Jun %A Yang, Haixia %A Panigrahy, Anand %A Minter, Lisa M %A Kim, Daeyoung %A Gibbons, John G %A Liu, LinShu %A Li, Zhengze %A Xiao, Hang %A Borlandelli, Valentina %A Overkleeft, Hermen S %A Cloer, Erica W %A Major, Michael B %A Goldfarb, Dennis %A Cai, Zongwei %A Redinbo, Matthew R %A Zhang, Guodong %K Animals %K Anti-Infective Agents, Local %K Anticarcinogenic Agents %K Bacterial Proteins %K Binding Sites %K Biotransformation %K Carcinogenesis %K Carcinogens %K Colitis %K Colon %K Colorectal Neoplasms %K Gastrointestinal Microbiome %K Gene Expression %K Glucuronidase %K Glycoside Hydrolase Inhibitors %K Humans %K Mice %K Mice, Inbred C57BL %K Models, Molecular %K Protein Binding %K Protein Conformation, alpha-Helical %K Protein Conformation, beta-Strand %K Protein Interaction Domains and Motifs %K Recombinant Proteins %K Triclosan %X

Emerging 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.

%B Nat Commun %V 13 %P 136 %8 2022 01 10 %G eng %N 1 %R 10.1038/s41467-021-27762-y %0 Journal Article %J Reproduction %D 2022 %T Multidrug resistance transporter-1 dysfunction perturbs meiosis and Ca2+ homeostasis in oocytes. %A Nabi, Dalileh %A Bosi, Davide %A Gupta, Neha %A Thaker, Nidhi %A Fissore, Rafael A %A Brayboy, Lynae Maria %X

MDR-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.

%B Reproduction %8 2022 Oct 01 %G eng %R 10.1530/REP-22-0192 %0 Journal Article %J Obstet Gynecol %D 2022 %T Neutralizing Antibodies and Cytokines in Breast Milk After Coronavirus Disease 2019 (COVID-19) mRNA Vaccination. %A Narayanaswamy, Vignesh %A Pentecost, Brian T %A Schoen, Corina N %A Alfandari, Dominique %A Schneider, Sallie S %A Baker, Ryan %A Arcaro, Kathleen F %K Adult %K Antibodies, Neutralizing %K Antibodies, Viral %K Breast Feeding %K Cohort Studies %K COVID-19 Vaccines %K Cytokines %K Female %K Humans %K Immunoglobulin A %K Immunoglobulin G %K Infant %K Infant, Newborn %K Middle Aged %K Milk, Human %K SARS-CoV-2 %K Vaccination %X

OBJECTIVE: 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.

%B Obstet Gynecol %V 139 %P 181-191 %8 2022 02 01 %G eng %N 2 %R 10.1097/AOG.0000000000004661 %0 Journal Article %J Pestic Biochem Physiol %D 2022 %T New chemistries for the control of human head lice, Pediculus humanus capitis: A mini-review. %A Clark, John M %K Animals %K Humans %K Insecticides %K Lice Infestations %K Pediculus %X

Pediculus 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.

%B Pestic Biochem Physiol %V 181 %P 105013 %8 2022 Feb %G eng %R 10.1016/j.pestbp.2021.105013 %0 Journal Article %J BMC Womens Health %D 2022 %T New 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. %A Sturgeon, Susan R %A Sibeko, Lindiwe %A Balasubramanian, Raji %A Arcaro, Kathleen F %K Adult %K Biomarkers %K Breast Feeding %K Breast Neoplasms %K Counseling %K Diet %K Female %K Fruit %K Humans %K Infant %K Randomized Controlled Trials as Topic %K Vegetables %X

BACKGROUND: 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 .

%B BMC Womens Health %V 22 %P 389 %8 2022 09 24 %G eng %N 1 %R 10.1186/s12905-022-01967-9 %0 Journal Article %J Reproduction %D 2022 %T SPERM FACTORS AND EGG ACTIVATION: ICSI and the discovery of the sperm factor and PLCZ1. %A Gupta, Neha %A Akizawa, Hiroki %A Lee, Hoi Chang %A Fissore, Rafael A %K Animals %K Fertilization %K Humans %K Male %K Mammals %K Mice %K Oocytes %K Phosphoinositide Phospholipase C %K Sperm Injections, Intracytoplasmic %K Spermatozoa %X

The 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.

%B Reproduction %V 164 %P F9-F20 %8 2022 05 23 %G eng %N 1 %R 10.1530/REP-21-0487 %0 Journal Article %J J R Soc Interface %D 2022 %T Surface functionalization of poly(dimethylsiloxane) substrates facilitates culture of pre-implantation mouse embryos by blocking non-selective adsorption. %A Hawkins, Jamar %A Miao, Xiaosu %A Cui, Wei %A Sun, Yubing %K Adsorption %K Animals %K Dimethylpolysiloxanes %K Embryo, Mammalian %K Embryonic Development %K Mammals %K Mice %X

Poly(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.

%B J R Soc Interface %V 19 %P 20210929 %8 2022 04 %G eng %N 189 %R 10.1098/rsif.2021.0929 %0 Journal Article %J Front Immunol %D 2022 %T Targeting the Cbl-b-Notch1 axis as a novel immunotherapeutic strategy to boost CD8+ T-cell responses. %A Monticone, Giulia %A Huang, Zhi %A Csibi, Fred %A Leit, Silvana %A Ciccone, David %A Champhekar, Ameya S %A Austin, Jermaine E %A Ucar, Deniz A %A Hossain, Fokhrul %A Ibba, Salome V %A Boulares, A Hamid %A Carpino, Nicholas %A Xu, Keli %A Majumder, Samarpan %A Osborne, Barbara A %A Loh, Christine %A Miele, Lucio %K Adenosine %K CD8-Positive T-Lymphocytes %K Humans %K Immunotherapy %K Lymphoma %K Neoplasms %K Receptor, Notch1 %X

A 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.

%B Front Immunol %V 13 %P 987298 %8 2022 %G eng %R 10.3389/fimmu.2022.987298 %0 Journal Article %J J Infect Dis %D 2022 %T Use of a Contained Mycobacterium tuberculosis Mouse Infection Model to Predict Active Disease and Containment in Humans. %A Duffy, Fergal J %A Olson, Gregory S %A Gold, Elizabeth S %A Jahn, Ana %A Aderem, Alan %A Aitchison, John D %A Rothchild, Alissa C %A Diercks, Alan H %A Nemeth, Johannes %K Animals %K Biomarkers %K Humans %K Mice %K Mycobacterium tuberculosis %K Transcriptome %K Tuberculosis %X

Previous 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.

%B J Infect Dis %V 225 %P 1832-1840 %8 2022 05 16 %G eng %N 10 %R 10.1093/infdis/jiab130 %0 Journal Article %J Expert Reviews in Molecular Medicine %D 2021 %T Alveolar macrophages: novel therapeutic targets for respiratory diseases %A Pamelia N. Lim %A Maritza M. Cervantes %A Linh K. Pham %A Alissa C. Rothchild %B Expert Reviews in Molecular Medicine %I Cambridge University Press ({CUP}) %V 23 %G eng %U https://doi.org/10.1017/erm.2021.21 %0 Journal Article %J Expert Rev Mol Med %D 2021 %T Alveolar macrophages: novel therapeutic targets for respiratory diseases. %A Lim, Pamelia N %A Cervantes, Maritza M %A Pham, Linh K %A Rothchild, Alissa C %K COVID-19 %K Humans %K Lung %K Macrophages, Alveolar %K Pandemics %K SARS-CoV-2 %X

Alveolar 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.

%B Expert Rev Mol Med %V 23 %P e18 %8 2021 11 26 %G eng %R 10.1017/erm.2021.21 %0 Journal Article %J Mol Hum Reprod %D 2021 %T Biophysical optimization of preimplantation embryo culture: what mechanics can offer ART. %A Hawkins, Jamar %A Miao, Xiaosu %A Cui, Wei %A Sun, Yubing %K Animals %K Biomedical Engineering %K Biophysical Phenomena %K Blastocyst %K Embryo Culture Techniques %K Fallopian Tubes %K Female %K Humans %K Reproductive Techniques, Assisted %X

Owing 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.

%B Mol Hum Reprod %V 27 %8 2021 01 22 %G eng %N 1 %R 10.1093/molehr/gaaa087 %0 Journal Article %J International Journal of Molecular Sciences %D 2021 %T Caput Ligation Renders Immature Mouse Sperm Motile and Capable to Undergo cAMP-Dependent Phosphorylation %A Darya A. Tourzani %A Battistone, Maria A %A Salicioni, Ana M %A Breton, Sylvie %A Visconti, Pablo E %A Maria G. Gervasi %B International Journal of Molecular Sciences %I {MDPI} {AG} %V 22 %P 10241 %G eng %U https://doi.org/10.3390/ijms221910241 %0 Journal Article %J Australian Veterinary Journal %D 2021 %T The contraceptive efficacy of a self-assembling intra-uterine device in domestic mares %A CJ Joonè %A CM Gradil %A JA Picard %A JD Taylor %A D de Tonnerre %A J Cavalieri %B Australian Veterinary Journal %V 99 %P 130–136 %G eng %U https://doi.org/10.1111/avj.13055 %R 10.1111/avj.13055 %0 Journal Article %J Elife %D 2021 %T Deficient spermiogenesis in mice lacking . %A Wang, Feng %A Gervasi, Maria Gracia %A Bošković, Ana %A Sun, Fengyun %A Rinaldi, Vera D %A Yu, Jun %A Wallingford, Mary C %A Tourzani, Darya A %A Mager, Jesse %A Zhu, Lihua Julie %A Rando, Oliver J %A Visconti, Pablo E %A Strittmatter, Lara %A Bach, Ingolf %K Animals %K Genes, X-Linked %K Male %K Mice %K Mice, Knockout %K Sertoli Cells %K Spermatogenesis %K Ubiquitin-Protein Ligases %X

The 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.

%B Elife %V 10 %8 2021 02 23 %G eng %R 10.7554/eLife.63556 %0 Journal Article %J J Cell Sci %D 2021 %T Deletion of TRPV3 and CaV3.2 T-type channels in mice undermines fertility and Ca2+ homeostasis in oocytes and eggs. %A Mehregan, Aujan %A Ardestani, Goli %A Akizawa, Hiroki %A Carvacho, Ingrid %A Fissore, Rafael %K Animals %K Calcium %K Calcium Channels, T-Type %K Female %K Fertility %K Fertilization %K Gene Deletion %K Homeostasis %K Mice %K Mice, Knockout %K Oocytes %K TRPM Cation Channels %K TRPV Cation Channels %X

Ca2+ 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.

%B J Cell Sci %V 134 %8 2021 07 01 %G eng %N 13 %R 10.1242/jcs.257956 %0 Journal Article %J Front Cell Dev Biol %D 2021 %T Editorial: Targeting Developmental Pathways in Inflammation and Disease. %A Anastasiadou, Eleni %A Minter, Lisa M %A Felli, Maria Pia %B Front Cell Dev Biol %V 9 %P 791115 %8 2021 %G eng %R 10.3389/fcell.2021.791115 %0 Journal Article %J Front Cell Dev Biol %D 2021 %T Editorial: The Fertilization Success From the Oocyte's Perspective. %A Michaut, Marcela A %A Souza-Fabjan, Joanna M G %A Fissore, Rafael A %B Front Cell Dev Biol %V 9 %P 810420 %8 2021 %G eng %R 10.3389/fcell.2021.810420 %0 Journal Article %J Endocrinology %D 2021 %T Exposure to Propylparaben During Pregnancy and Lactation Induces Long-Term Alterations to the Mammary Gland in Mice. %A Mogus, Joshua P %A LaPlante, Charlotte D %A Bansal, Ruby %A Matouskova, Klara %A Schneider, Benjamin R %A Daniele, Elizabeth %A Silva, Shannon J %A Hagen, Mary J %A Dunphy, Karen A %A Jerry, D Joseph %A Schneider, Sallie S %A Vandenberg, Laura N %K Animals %K Cells, Cultured %K Endocrine Disruptors %K Female %K Lactation %K Male %K Mammary Glands, Animal %K Maternal Exposure %K Mice %K Mice, Inbred BALB C %K Parabens %K Pregnancy %K Prenatal Exposure Delayed Effects %X

The 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.

%B Endocrinology %V 162 %8 2021 06 01 %G eng %N 6 %R 10.1210/endocr/bqab041 %0 Journal Article %J Oncogene %D 2021 %T Genetic modifiers regulating DNA replication and double-strand break repair are associated with differences in mammary tumors in mouse models of Li-Fraumeni syndrome. %A Majhi, Prabin Dhangada %A Griner, Nicholas B %A Mayfield, Jacob A %A Compton, Shannon %A Kane, Jeffrey J %A Baptiste, Trevor A %A Dunphy, Karen A %A Roberts, Amy L %A Schneider, Sallie S %A Savage, Evan M %A Patel, Divyen %A Blackburn, Anneke C %A Maurus, Kim Joana %A Wiesmüller, Lisa %A Jerry, D Joseph %K Animals %K Breast Neoplasms %K Chromosome Mapping %K Disease Models, Animal %K Disease Susceptibility %K DNA Breaks, Double-Stranded %K DNA Repair %K DNA Replication %K Female %K Gene Expression Regulation %K Genes, Modifier %K Genetic Linkage %K Genetic Loci %K Li-Fraumeni Syndrome %K Mice %K Mice, Knockout %K Polymorphism, Single Nucleotide %K Recombinational DNA Repair %K Tumor Suppressor Protein p53 %X

Breast 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.

%B Oncogene %V 40 %P 5026-5037 %8 2021 08 %G eng %N 31 %R 10.1038/s41388-021-01892-5 %0 Journal Article %J Breastfeed Med %D 2021 %T Humoral and Cell-Mediated Immune Response in Colostrum from Women Diagnosed Positive for SARS-CoV-2. %A Narayanaswamy, Vignesh %A Pentecost, Brian %A Alfandari, Dominique %A Chin, Emily %A Minor, Kathleen %A Kastrinakis, Alyssa %A Lieberman, Tanya %A Arcaro, Kathleen F %A Leftwich, Heidi %K Breast Feeding %K Colostrum %K COVID-19 %K Female %K Humans %K Immunity, Cellular %K Immunity, Humoral %K Pregnancy %K Pregnancy Complications, Infectious %K Spike Glycoprotein, Coronavirus %X

To 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.

%B Breastfeed Med %V 16 %P 987-994 %8 2021 12 %G eng %N 12 %R 10.1089/bfm.2021.0082 %0 Journal Article %J Anim Reprod Sci %D 2021 %T An intrauterine device with potential to control fertility in feral equids. %A Gradil, Carlos %A Joone, Carolynne %A Haire, Teresa %A Fowler, Bradley %A Zinchuk, Jacquelyn %A Davies, Christopher J %A Ball, Barry %K Animals %K Contraception %K Estrous Cycle %K Female %K Horses %K Intrauterine Devices %K Pilot Projects %K Population Control %K Pregnancy %K Pregnancy Rate %X

Fertility 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.

%B Anim Reprod Sci %V 231 %P 106795 %8 2021 Aug %G eng %R 10.1016/j.anireprosci.2021.106795 %0 Journal Article %J Mater Horiz %D 2021 %T Nanotherapeutics using all-natural materials. Effective treatment of wound biofilm infections using crosslinked nanoemulsions. %A Li, Cheng-Hsuan %A Landis, Ryan F %A Makabenta, Jessa Marie %A Nabawy, Ahmed %A Tronchet, Tiphaine %A Archambault, Danielle %A Liu, Yuanchang %A Huang, Rui %A Golan, Morgane %A Cui, Wei %A Mager, Jesse %A Gupta, Akash %A Schmidt-Malan, Suzannah %A Patel, Robin %A Rotello, Vincent M %K Animals %K Anti-Bacterial Agents %K Anti-Infective Agents %K Bacterial Infections %K Biofilms %K Mice %K Wound Infection %X

Bacterial 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.

%B Mater Horiz %V 8 %P 1776-1782 %8 2021 06 01 %G eng %N 6 %R 10.1039/d0mh01826k %0 Journal Article %J Obstetrics &$\mathsemicolon$ Gynecology %D 2021 %T Neutralizing Antibodies and Cytokines in Breast Milk After Coronavirus Disease 2019 (COVID-19) mRNA Vaccination %A Vignesh Narayanaswamy %A Brian T. Pentecost %A Corina N. Schoen %A Dominique Alfandari %A Sallie S. Schneider %A Ryan Baker %A Kathleen F. Arcaro %B Obstetrics &$\mathsemicolon$ Gynecology %V 139 %P 181–191 %G eng %U https://doi.org/10.1097/aog.0000000000004661 %R 10.1097/aog.0000000000004661 %0 Journal Article %J Biomacromolecules %D 2021 %T Non-Covalent Carrier Hydrophobicity as a Universal Predictor of Intracellular Protein Activity. %A Hango, Christopher R %A Backlund, Coralie M %A Davis, Hazel C %A Posey, Nicholas D %A Minter, Lisa M %A Tew, Gregory N %K Biological Transport %K Cell-Penetrating Peptides %K Hydrophobic and Hydrophilic Interactions %K Polymers %K Protein Transport %X

Over 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.

%B Biomacromolecules %V 22 %P 2850-2863 %8 2021 07 12 %G eng %N 7 %R 10.1021/acs.biomac.1c00242 %0 Journal Article %J Front Cell Dev Biol %D 2021 %T Oocyte Spontaneous Activation: An Overlooked Cellular Event That Impairs Female Fertility in Mammals. %A Cui, Wei %X

In 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.

%B Front Cell Dev Biol %V 9 %P 648057 %8 2021 %G eng %R 10.3389/fcell.2021.648057 %0 Journal Article %J Environ Int %D 2021 %T Paternal preconception phthalate exposure alters sperm methylome and embryonic programming. %A Oluwayiose, Oladele A %A Marcho, Chelsea %A Wu, Haotian %A Houle, Emily %A Krawetz, Stephen A %A Suvorov, Alexander %A Mager, Jesse %A Richard Pilsner, J %K Animals %K Diethylhexyl Phthalate %K DNA Methylation %K Embryonic Development %K Epigenome %K Female %K Male %K Mice %K Mice, Inbred C57BL %K Phthalic Acids %K Pregnancy %K Spermatozoa %X

Preconception 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.

%B Environ Int %V 155 %P 106693 %8 2021 10 %G eng %R 10.1016/j.envint.2021.106693 %0 Journal Article %J Cancer Epidemiol Biomarkers Prev %D 2021 %T Prediagnostic White Blood Cell DNA Methylation and Risk of Breast Cancer in the Prostate Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) Cohort. %A Sturgeon, Susan R %A Sela, David A %A Browne, Eva P %A Einson, Jonah %A Rani, Asha %A Halabi, Mohamed %A Kania, Thomas %A Keezer, Andrew %A Balasubramanian, Raji %A Ziegler, Regina G %A Schairer, Catherine %A Kelsey, Karl T %A Arcaro, Kathleen F %K Aged %K Breast Neoplasms %K Case-Control Studies %K Cell Cycle Proteins %K CpG Islands %K DNA Methylation %K DNA-Binding Proteins %K Endonucleases %K Female %K Humans %K Intracellular Signaling Peptides and Proteins %K Leukocytes %K Membrane Proteins %K Membrane Transport Proteins %K Middle Aged %K Mitochondrial Proteins %K Prospective Studies %X

BACKGROUND: 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.

%B Cancer Epidemiol Biomarkers Prev %V 30 %P 1575-1581 %8 2021 08 %G eng %N 8 %R 10.1158/1055-9965.EPI-20-1717 %0 Journal Article %J Cancer Epidemiology, Biomarkers &$\mathsemicolon$ Prevention %D 2021 %T Prediagnostic White Blood Cell DNA Methylation and Risk of Breast Cancer in the Prostate Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) Cohort %A Susan R. Sturgeon %A David A. Sela %A Eva P. Browne %A Jonah Einson %A Asha Rani %A Mohamed Halabi %A Thomas Kania %A Andrew Keezer %A Raji Balasubramanian %A Regina G. Ziegler %A Catherine Schairer %A Karl T. Kelsey %A Kathleen F. Arcaro %B Cancer Epidemiology, Biomarkers &$\mathsemicolon$ Prevention %V 30 %P 1575–1581 %G eng %U https://doi.org/10.1158/1055-9965.epi-20-1717 %R 10.1158/1055-9965.epi-20-1717 %0 Journal Article %J Animal Reproduction Science %D 2021 %T Preliminary study of the contraceptive effect of a self-assembling intrauterine device (iUPODs) in mares maintained in a paddock with a fertile stallion %A Karl H. Hoopes %A Carlos M. Gradil %A Dirk K. Vanderwall %A Holly M. Mason %A Brendan A. Sarnecky %A Christopher J. Davies %B Animal Reproduction Science %V 235 %P 106881 %G eng %U https://doi.org/10.1016/j.anireprosci.2021.106881 %R 10.1016/j.anireprosci.2021.106881 %0 Journal Article %J Angew Chem Int Ed Engl %D 2021 %T Protein-Antibody Conjugates (PACs): A Plug-and-Play Strategy for Covalent Conjugation and Targeted Intracellular Delivery of Pristine Proteins. %A Liu, Bin %A Singh, Khushboo %A Gong, Shuai %A Canakci, Mine %A Osborne, Barbara A %A Thayumanavan, S %K Antibodies %K Cell Line, Tumor %K Fullerenes %K Humans %K Nanoparticles %K Particle Size %K Receptor, erbB-2 %X

We 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.

%B Angew Chem Int Ed Engl %V 60 %P 12813-12818 %8 2021 06 01 %G eng %N 23 %R 10.1002/anie.202103106 %0 Journal Article %J Nat Rev Drug Discov %D 2021 %T Targeting Notch in oncology: the path forward. %A Majumder, Samarpan %A Crabtree, Judy S %A Golde, Todd E %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %K Animals %K Antineoplastic Agents %K Drug Delivery Systems %K Humans %K Immunotherapy %K Neoplasms %K Neoplastic Stem Cells %K Receptors, Notch %K Signal Transduction %X

Notch 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.

%B Nat Rev Drug Discov %V 20 %P 125-144 %8 2021 02 %G eng %N 2 %R 10.1038/s41573-020-00091-3 %0 Journal Article %J Nat Rev Drug Discov %D 2021 %T Targeting Notch in oncology: the path forward. %A Majumder, Samarpan %A Crabtree, Judy S %A Golde, Todd E %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %K Animals %K Antineoplastic Agents %K Drug Delivery Systems %K Humans %K Immunotherapy %K Neoplasms %K Neoplastic Stem Cells %K Receptors, Notch %K Signal Transduction %X

Notch 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.

%B Nat Rev Drug Discov %V 20 %P 125-144 %8 2021 02 %G eng %N 2 %R 10.1038/s41573-020-00091-3 %0 Journal Article %J Developmental Biology %D 2021 %T Trim-Away mediated knock down uncovers a new function for Lbh during gastrulation of Xenopus laevis. %A Emma Weir %A Gretchen McLinden %A Alfandari, Dominique %A Cousin, Hélène %B Developmental Biology %I Elsevier {BV} %V 470 %P 74–83 %G eng %U https://doi.org/10.1016/j.ydbio.2020.10.014 %0 Journal Article %J Molecular Reproduction and Development %D 2021 %T TSSK3, a novel target for male contraception, is required for spermiogenesis %A Saman Nayyab %A M. G. Gervasi %A Darya A. Tourzani %A Diego A Caraballo %A Jha, Kula N %A Maria E. Teves %A Cui, Wei %A Gunda I. Georg %A Visconti, Pablo E %A Salicioni, Ana M %B Molecular Reproduction and Development %I Wiley %G eng %U https://doi.org/10.1002/mrd.23539 %0 Journal Article %J Front Cell Dev Biol %D 2021 %T When Viruses Cross Developmental Pathways. %A Trivedi, Pankaj %A Patel, Sandesh Kumar %A Bellavia, Diana %A Messina, Elena %A Palermo, Rocco %A Ceccarelli, Simona %A Marchese, Cinzia %A Anastasiadou, Eleni %A Minter, Lisa M %A Felli, Maria Pia %X

Aberrant 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.

%B Front Cell Dev Biol %V 9 %P 691644 %8 2021 %G eng %R 10.3389/fcell.2021.691644 %0 Journal Article %J Biol Reprod %D 2021 %T ZC3H4-a novel CCCH-type zinc finger protein-is essential for early embryogenesis in mice†. %A Su, Jianmin %A Miao, Xiaosu %A Archambault, Danielle %A Mager, Jesse %A Cui, Wei %K Animals %K Cell Proliferation %K DNA Breaks %K DNA-Binding Proteins %K Embryo Implantation %K Embryonic Development %K Female %K Gene Expression Regulation, Developmental %K Genotype %K Mice %K Mice, Knockout %K Mutation %X

Zinc 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.

%B Biol Reprod %V 104 %P 325-335 %8 2021 02 11 %G eng %N 2 %R 10.1093/biolre/ioaa215 %0 Journal Article %J Frontiers in Cell and Developmental Biology %D 2020 %T Acheron/Larp6 Is a Survival Protein That Protects Skeletal Muscle From Programmed Cell Death During Development %A Ankur Sheel %A Shao, Rong %A Christine Brown %A Joanne Johnson %A Alexandra Hamilton %A Danhui Sun %A Julia Oppenheimer %A Wendy Smith %A Visconti, Pablo E %A Michele Markstein %A Bigelow, Carol %A Lawrence M. Schwartz %B Frontiers in Cell and Developmental Biology %I Frontiers Media {SA} %V 8 %G eng %U https://doi.org/10.3389/fcell.2020.00622 %0 Journal Article %J Molecular Reproduction and Development %D 2020 %T Capacitation increases glucose consumption in murine sperm %A David M. Hidalgo %A Romarowski, Ana %A Gervasi, Maria Gracia %A Navarrete, Felipe %A Melanie Balbach %A Salicioni, Ana Maria %A Levin, Lonny R %A Buck, Jochen %A Visconti, Pablo E %B Molecular Reproduction and Development %I Wiley %G eng %U https://doi.org/10.1002/mrd.23421 %0 Journal Article %J Frontiers in Immunology %D 2020 %T CD28 Signaling Drives Notch Ligand Expression on CD4 T Cells %A Ankita Mitra %A Sudarvili Shanthalingam %A Heather L Sherman %A Khushboo Singh %A Canakci, Mine %A Joe A Torres %A Rebecca Lawlor %A Yong Ran %A Golde, Todd E %A Miele, Lucio %A Sankaran Thayumanavan %A Minter, Lisa M %A Osborne, Barbara A %B Frontiers in Immunology %I Frontiers Media {SA} %V 11 %G eng %U https://doi.org/10.3389/fimmu.2020.00735 %0 Journal Article %J Molecular Therapy %D 2020 %T Cell-Penetrating Anti-Protein Kinase C Theta Antibodies Act Intracellularly to Generate Stable, Highly Suppressive Regulatory T Cells %A E. Ilker Ozay %A Sudarvili Shanthalingam %A Heather L Sherman %A Joe A Torres %A Osborne, Barbara A %A Tew, Gregory N %A Minter, Lisa M %B Molecular Therapy %I Elsevier {BV} %V 28 %P 1987–2006 %G eng %U https://doi.org/10.1016/j.ymthe.2020.05.020 %0 Journal Article %J PLOS Pathogens %D 2020 %T Contained Mycobacterium tuberculosis infection induces concomitant and heterologous protection %A Johannes Nemeth %A Gregory S. Olson %A Alissa C. Rothchild %A Ana N. Jahn %A Dat Mai %A Fergal J. Duffy %A Jared L. Delahaye %A Sanjay Srivatsan %A Courtney R. Plumlee %A Kevin B. Urdahl %A Elizabeth S. Gold %A Alan Aderem %A Alan H. Diercks %E Christopher M. Sassetti %B PLOS Pathogens %I Public Library of Science ({PLoS}) %V 16 %P e1008655 %G eng %U https://doi.org/10.1371/journal.ppat.1008655 %0 Journal Article %J Biology of Reproduction %D 2020 %T CRISPR/CAS9-mediated amino acid substitution reveals phosphorylation residues of RSPH6A are not essential for male fertility in mice† %A Haruhiko Miyata %A Ferheen Abbasi %A Visconti, Pablo E %A Masahito Ikawa %B Biology of Reproduction %I Oxford University Press ({OUP}) %G eng %U https://doi.org/10.1093/biolre/ioaa161 %0 Journal Article %J Physiology %D 2020 %T Differences and Similarities: The Richness of Comparative Sperm Physiology %A Darszon, Alberto %A Takuya Nishigaki %A López-González, Ignacio %A Visconti, Pablo E %A Treviño, Claudia L %B Physiology %I American Physiological Society %V 35 %P 196–208 %G eng %U https://doi.org/10.1152/physiol.00033.2019 %0 Journal Article %J Frontiers in Immunology %D 2020 %T Differentiation of Pathogenic Th17 Cells Is Negatively Regulated by Let-7 MicroRNAs in a Mouse Model of Multiple Sclerosis %A Constance C Angelou %A Alexandria C Wells %A Jyothi Vijayaraghavan %A Carey E. Dougan %A Rebecca Lawlor %A Elizabeth Iverson %A Vanja Lazarevic %A Kimura, Motoko Y %A Shelly R. Peyton %A Minter, Lisa M %A Osborne, Barbara A %A Elena L Pobezinskaya %A Pobezinsky, Leonid A %B Frontiers in Immunology %I Frontiers Media {SA} %V 10 %G eng %U https://doi.org/10.3389/fimmu.2019.03125 %0 Journal Article %J Cell Calcium %D 2020 %T Divalent cation influx and calcium homeostasis in germinal vesicle mouse oocytes. %A Ardestani, Goli %A Mehregan, Aujan %A Fleig, Andrea %A Horgen, F David %A Carvacho, Ingrid %A Fissore, Rafael A %K Animals %K Calcium %K Calcium Channels %K Calcium Channels, T-Type %K Cations, Divalent %K Cell Membrane %K Fluorescence %K Homeostasis %K Manganese %K Metaphase %K Mice, Knockout %K Nickel %K Oocytes %K Ovum %K Strontium %K TRPM Cation Channels %X

Prior 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.

%B Cell Calcium %V 87 %P 102181 %8 2020 05 %G eng %R 10.1016/j.ceca.2020.102181 %0 Journal Article %J Environ Health Perspect %D 2020 %T Effects of Benzophenone-3 and Propylparaben on Estrogen Receptor-Dependent R-Loops and DNA Damage in Breast Epithelial Cells and Mice. %A Majhi, Prabin Dhangada %A Sharma, Aman %A Roberts, Amy L %A Daniele, Elizabeth %A Majewski, Aliza R %A Chuong, Lynn M %A Black, Amye L %A Vandenberg, Laura N %A Schneider, Sallie S %A Dunphy, Karen A %A Jerry, D Joseph %K Animals %K Benzophenones %K Cell Line, Tumor %K Environmental Pollutants %K Epithelial Cells %K Humans %K Mice %K Parabens %K R-Loop Structures %K Receptors, Estrogen %K Toxicity Tests %X

BACKGROUND: 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.

%B Environ Health Perspect %V 128 %P 17002 %8 2020 01 %G eng %N 1 %R 10.1289/EHP5221 %0 Journal Article %J Environmental Health Perspectives %D 2020 %T Effects of Benzophenone-3 and Propylparaben on Estrogen Receptor–Dependent R-Loops and DNA Damage in Breast Epithelial Cells and Mice %A Prabin Dhangada Majhi %A Aman Sharma %A Roberts, Amy L %A Elizabeth Daniele %A Aliza R Majewski %A Lynn M Chuong %A Amye L. Black %A Laura N. Vandenberg %A Schneider, Sallie S %A Dunphy, Karen A %A Jerry, D Joseph %B Environmental Health Perspectives %I Environmental Health Perspectives %V 128 %P 017002 %G eng %U https://doi.org/10.1289/ehp5221 %0 Journal Article %J Journal of Agricultural and Food Chemistry %D 2020 %T Effects of Linoleic Acid-Rich Diet on Plasma Profiles of Eicosanoids and Development of Colitis in Il-10–/– Mice %A Minhao Xie %A Jun Yang %A Jianan Zhang %A Heather L Sherman %A Zhenyu Zhang %A Minter, Lisa M %A Bruce D. Hammock %A Yeonhwa Park %A Guodong Zhang %B Journal of Agricultural and Food Chemistry %I American Chemical Society ({ACS}) %V 68 %P 7641–7647 %G eng %U https://doi.org/10.1021/acs.jafc.0c03024 %0 Journal Article %J British Journal of Nutrition %D 2020 %T Effects 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 Model %A M. B. Cicekdal %A B. T. Kazan %A B. G. Tuna %A U. Ozorhan %A I. D. Ekici %A F. Zhu %A O. Suakar %A A. Kuskucu %A O. F. Bayrak %A K. Arcaro %A M. P. Cleary %A S. Dogan %B British Journal of Nutrition %I Cambridge University Press ({CUP}) %V 123 %P 1079–1079 %G eng %U https://doi.org/10.1017/s0007114520000471 %0 Journal Article %J Molecular Biology of the Cell %D 2020 %T Endoplasmic reticulum transmembrane protein TMTC3 contributes to O-mannosylation of E-cadherin, cellular adherence, and embryonic gastrulation %A Jill B. Graham %A Johan C. Sunryd %A Ketan Mathavan %A Emma Weir %A Ida Signe Bohse Larsen %A Adnan Halim %A Henrik Clausen %A Cousin, Hélène %A Alfandari, Dominque %A Daniel N. Hebert %E Thomas Martin %B Molecular Biology of the Cell %I American Society for Cell Biology ({ASCB}) %V 31 %P 167–183 %G eng %U https://doi.org/10.1091/mbc.e19-07-0408 %0 Journal Article %J Epigenetics %D 2020 %T Enrichment of CpG island shore region hypermethylation in epigenetic breast field cancerization %A Meghan E. Muse %A Alexander J. Titus %A Lucas A. Salas %A Owen M. Wilkins %A Chelsey Mullen %A Kelly J. Gregory %A Schneider, Sallie S %A Crisi, Giovanna M %A M Jawale, Rahul %A Otis, Christopher N %A Brock C. Christensen %A Arcaro, Kathleen F %B Epigenetics %I Informa {UK} Limited %V 15 %P 1093–1106 %G eng %U https://doi.org/10.1080/15592294.2020.1747748 %0 Journal Article %J Reprod Toxicol %D 2020 %T Exposure to low doses of oxybenzone during perinatal development alters mammary gland morphology in male and female mice. %A Matouskova, Klara %A Jerry, D Joseph %A Vandenberg, Laura N %K Anal Canal %K Animals %K Benzophenones %K Cell Proliferation %K Epithelial Cells %K Estrogen Receptor alpha %K Female %K Genitalia, Female %K Genitalia, Male %K Lactation %K Male %K Mammary Glands, Animal %K Maternal-Fetal Exchange %K Mice, Inbred BALB C %K Pregnancy %K Prenatal Exposure Delayed Effects %K Sunscreening Agents %X

Oxybenzone (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.

%B Reprod Toxicol %V 92 %P 66-77 %8 2020 03 %G eng %R 10.1016/j.reprotox.2019.08.002 %0 Journal Article %J BIO-PROTOCOL %D 2020 %T Flow Cytometry Analysis and Fluorescence-activated Cell Sorting of Myeloid Cells from Lung and Bronchoalveolar Lavage Samples from Mycobacterium tuberculosis-infected Mice %A Alissa Rothchild %A Dat Mai %A Alan Aderem %A Alan Diercks %B BIO-PROTOCOL %I Bio-Protocol, {LLC} %V 10 %G eng %U https://doi.org/10.21769/bioprotoc.3630 %0 Journal Article %J J Mammary Gland Biol Neoplasia %D 2020 %T Inter-Individual Variation in Response to Estrogen in Human Breast Explants. %A Dunphy, Karen A %A Black, Amye L %A Roberts, Amy L %A Sharma, Aman %A Li, Zida %A Suresh, Sneha %A Browne, Eva P %A Arcaro, Kathleen F %A Ser-Dolansky, Jennifer %A Bigelow, Carol %A Troester, Melissa A %A Schneider, Sallie S %A Makari-Judson, Grace %A Crisi, Giovanna M %A Jerry, D Joseph %K Adolescent %K Adult %K Aged %K Apoptosis %K Biomarkers, Tumor %K Breast Neoplasms %K Cell Proliferation %K Estrogens %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K Middle Aged %K Prognosis %K Receptors, Estrogen %K Tumor Cells, Cultured %K Young Adult %X

Exposure 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.

%B J Mammary Gland Biol Neoplasia %V 25 %P 51-68 %8 2020 03 %G eng %N 1 %R 10.1007/s10911-020-09446-3 %0 Journal Article %J Journal of Mammary Gland Biology and Neoplasia %D 2020 %T Inter-Individual Variation in Response to Estrogen in Human Breast Explants %A Dunphy, Karen A %A Amye L. Black %A Roberts, Amy L %A Aman Sharma %A Zida Li %A Sneha Suresh %A Browne, Eva P %A Arcaro, Kathleen F %A Jennifer Ser-Dolansky %A Bigelow, Carol %A Troester, Melissa A %A Schneider, Sallie S %A Makari-Judson, Grace %A Crisi, Giovanna M %A Jerry, D Joseph %B Journal of Mammary Gland Biology and Neoplasia %I Springer Science and Business Media {LLC} %V 25 %P 51–68 %G eng %U https://doi.org/10.1007/s10911-020-09446-3 %0 Journal Article %J Molecular Reproduction and Development %D 2020 %T Loss of POLR1D results in embryonic lethality prior to blastocyst formation in mice %A Xiaosu Miao %A Tieqi Sun %A Morgane Golan %A Mager, Jesse %A Cui, Wei %B Molecular Reproduction and Development %I Wiley %G eng %U https://doi.org/10.1002/mrd.23427 %0 Journal Article %J Molecular Reproduction and Development %D 2020 %T Loss of POLR1D results in embryonic lethality prior to blastocyst formation in mice %A Xiaosu Miao %A Tieqi Sun %A Morgane Golan %A Jesse Mager %A Wei Cui %B Molecular Reproduction and Development %V 87 %P 1152–1158 %G eng %U https://doi.org/10.1002/mrd.23427 %R 10.1002/mrd.23427 %0 Journal Article %J Biology of Reproduction %D 2020 %T Loss of RBBP4 results in defective inner cell mass, severe apoptosis, hyperacetylated histones and preimplantation lethality in mice %A Xiaosu Miao %A Tieqi Sun %A Holly Barletta %A Mager, Jesse %A Cui, Wei %B Biology of Reproduction %I Oxford University Press ({OUP}) %G eng %U https://doi.org/10.1093/biolre/ioaa046 %0 Journal Article %J Biol Reprod %D 2020 %T Loss of RBBP4 results in defective inner cell mass, severe apoptosis, hyperacetylated histones and preimplantation lethality in mice†. %A Miao, Xiaosu %A Sun, Tieqi %A Barletta, Holly %A Mager, Jesse %A Cui, Wei %K Acetylation %K Animals %K Apoptosis %K Blastocyst %K Embryo Implantation %K Embryo Loss %K Embryonic Development %K Endoderm %K Female %K Gene Expression %K Histones %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Knockout %K Retinoblastoma-Binding Protein 4 %X

Retinoblastoma-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.

%B Biol Reprod %V 103 %P 13-23 %8 2020 06 23 %G eng %N 1 %R 10.1093/biolre/ioaa046 %0 Journal Article %J Developmental Biology %D 2020 %T Mcrs1 interacts with Six1 to influence early craniofacial and otic development %A Neilson, Karen M %A Stephanie Keer %A Nicole Bousquet %A Olivia Macrorie %A Himani D. Majumdar %A Kenyon, Kristy L %A Alfandari, Dominique %A Moody, Sally A %B Developmental Biology %I Elsevier {BV} %V 467 %P 39–50 %G eng %U https://doi.org/10.1016/j.ydbio.2020.08.013 %0 Journal Article %J Reproduction %D 2020 %T MCRS1 is essential for epiblast development during early mouse embryogenesis. %A Cui, Wei %A Cheong, Agnes %A Wang, Yongsheng %A Tsuchida, Yuran %A Liu, Yong %A Tremblay, Kimberly D %A Mager, Jesse %K Animals %K Blastocyst Inner Cell Mass %K Cell Differentiation %K Cell Lineage %K Embryo, Mammalian %K Embryonic Development %K Embryonic Stem Cells %K Female %K Gene Expression Regulation, Developmental %K Germ Layers %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Knockout %K Mutation %K RNA-Binding Proteins %X

Microspherule 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.

%B Reproduction %V 159 %P 1-13 %8 2020 01 %G eng %N 1 %R 10.1530/REP-19-0334 %0 Journal Article %J Biology of Reproduction %D 2020 %T Metabolic changes in mouse sperm during capacitation %A Melanie Balbach %A Gervasi, Maria Gracia %A David Martin Hidalgo %A Visconti, Pablo E %A Levin, Lonny R %A Buck, Jochen %B Biology of Reproduction %I Oxford University Press ({OUP}) %G eng %U https://doi.org/10.1093/biolre/ioaa114 %0 Journal Article %J Development %D 2020 %T MMP14 is required for delamination of chick neural crest cells independently of its catalytic activity %A Cyril Andrieu %A Audrey Montigny %A Anne Bibonne %A Evangeline Despin-Guitard %A Alfandari, Dominique %A Théveneau, Eric %B Development %I The Company of Biologists %V 147 %P dev183954 %G eng %U https://doi.org/10.1242/dev.183954 %0 Journal Article %J J Cancer Educ %D 2020 %T The Mouse Mammary Gland: a Tool to Inform Adolescents About Environmental Causes of Breast Cancer. %A Vandenberg, Laura N %A Kolla, SriDurgaDevi %A LaPlante, Charlotte D %A Jerry, D Joseph %K Adolescent %K Animals %K Breast Neoplasms %K Environmental Exposure %K Female %K Humans %K Laboratories %K Laboratory Personnel %K Mammary Glands, Animal %K Mice %X

Adolescence 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.

%B J Cancer Educ %V 35 %P 1094-1100 %8 2020 12 %G eng %N 6 %R 10.1007/s13187-019-01563-w %0 Journal Article %J Cell %D 2020 %T Multi-Omics Resolves a Sharp Disease-State Shift between Mild and Moderate COVID-19. %A Su, Yapeng %A Chen, Daniel %A Yuan, Dan %A Lausted, Christopher %A Choi, Jongchan %A Dai, Chengzhen L %A Voillet, Valentin %A Duvvuri, Venkata R %A Scherler, Kelsey %A Troisch, Pamela %A Baloni, Priyanka %A Qin, Guangrong %A Smith, Brett %A Kornilov, Sergey A %A Rostomily, Clifford %A Xu, Alex %A Li, Jing %A Dong, Shen %A Rothchild, Alissa %A Zhou, Jing %A Murray, Kim %A Edmark, Rick %A Hong, Sunga %A Heath, John E %A Earls, John %A Zhang, Rongyu %A Xie, Jingyi %A Li, Sarah %A Roper, Ryan %A Jones, Lesley %A Zhou, Yong %A Rowen, Lee %A Liu, Rachel %A Mackay, Sean %A O'Mahony, D Shane %A Dale, Christopher R %A Wallick, Julie A %A Algren, Heather A %A Zager, Michael A %A Wei, Wei %A Price, Nathan D %A Huang, Sui %A Subramanian, Naeha %A Wang, Kai %A Magis, Andrew T %A Hadlock, Jenn J %A Hood, Leroy %A Aderem, Alan %A Bluestone, Jeffrey A %A Lanier, Lewis L %A Greenberg, Philip D %A Gottardo, Raphael %A Davis, Mark M %A Goldman, Jason D %A Heath, James R %K Adolescent %K Adult %K Aged %K Aged, 80 and over %K COVID-19 %K Female %K Genomics %K Humans %K Male %K Middle Aged %K RNA-Seq %K SARS-CoV-2 %K Severity of Illness Index %K Single-Cell Analysis %X

We 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.

%B Cell %V 183 %P 1479-1495.e20 %8 2020 12 10 %G eng %N 6 %R 10.1016/j.cell.2020.10.037 %0 Journal Article %J Development %D 2020 %T Nuclear encoded mitochondrial ribosomal proteins are required to initiate gastrulation %A Agnes Cheong %A Danielle Archambault %A Rinat Degani %A Elizabeth Iverson %A Tremblay, Kimberly %A Mager, Jesse %B Development %I The Company of Biologists %G eng %U https://doi.org/10.1242/dev.188714 %0 Journal Article %J Analytical and Bioanalytical Chemistry %D 2020 %T Polymeric nanoassemblies for enrichment and detection of peptides and proteins in human breast milk %A Bo Zhao %A Jingjing Gao %A Mahalia A. C. Serrano %A Arcaro, Kathleen F %A Thayumanavan, S %A Vachet, Richard W %B Analytical and Bioanalytical Chemistry %I Springer Science and Business Media {LLC} %V 412 %P 1027–1035 %G eng %U https://doi.org/10.1007/s00216-019-02342-8 %0 Journal Article %J Human Molecular Genetics %D 2020 %T Prediagnostic breast milk DNA methylation alterations in women who develop breast cancer %A Lucas A. Salas %A Sara N Lundgren %A Browne, Eva P %A Punska, Elizabeth C %A Anderton, Douglas L %A Margaret R Karagas %A Arcaro, Kathleen F %A Brock C. Christensen %B Human Molecular Genetics %I Oxford University Press ({OUP}) %V 29 %P 662–673 %G eng %U https://doi.org/10.1093/hmg/ddz301 %0 Journal Article %J Mol Ther %D 2020 %T Protein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulate FOXP3 Stability in Regulatory T Cells. %A Ozay, E Ilker %A Shanthalingam, Sudarvili %A Torres, Joe A %A Osborne, Barbara A %A Tew, Gregory N %A Minter, Lisa M %K Cell-Penetrating Peptides %K Forkhead Transcription Factors %K Gene Expression Regulation %K Heterogeneous-Nuclear Ribonucleoprotein L %K Promoter Regions, Genetic %K Protein Binding %K Protein D-Aspartate-L-Isoaspartate Methyltransferase %K Protein Kinase C-theta %K Protein Stability %K Signal Transduction %K T-Lymphocytes, Regulatory %X

T 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.

%B Mol Ther %V 28 %P 2220-2236 %8 2020 10 07 %G eng %N 10 %R 10.1016/j.ymthe.2020.06.012 %0 Journal Article %J Mol Ther %D 2020 %T Protein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulate FOXP3 Stability in Regulatory T Cells. %A Ozay, E Ilker %A Shanthalingam, Sudarvili %A Torres, Joe A %A Osborne, Barbara A %A Tew, Gregory N %A Minter, Lisa M %K Cell-Penetrating Peptides %K Forkhead Transcription Factors %K Gene Expression Regulation %K Heterogeneous-Nuclear Ribonucleoprotein L %K Promoter Regions, Genetic %K Protein Binding %K Protein D-Aspartate-L-Isoaspartate Methyltransferase %K Protein Kinase C-theta %K Protein Stability %K Signal Transduction %K T-Lymphocytes, Regulatory %X

T 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.

%B Mol Ther %V 28 %P 2220-2236 %8 2020 10 07 %G eng %N 10 %R 10.1016/j.ymthe.2020.06.012 %0 Journal Article %J Molecular Therapy %D 2020 %T Protein Kinase C Theta Modulates PCMT1 through hnRNPL to Regulate FOXP3 Stability in Regulatory T Cells %A E. Ilker Ozay %A Sudarvili Shanthalingam %A Joe A Torres %A Osborne, Barbara A %A Tew, Gregory N %A Minter, Lisa M %B Molecular Therapy %I Elsevier {BV} %V 28 %P 2220–2236 %G eng %U https://doi.org/10.1016/j.ymthe.2020.06.012 %0 Journal Article %J Developmental Biology %D 2020 %T Protein phosphatase 1 regulatory subunit 35 is required for ciliogenesis, notochord morphogenesis, and cell-cycle progression during murine development %A Danielle Archambault %A Agnes Cheong %A Elizabeth Iverson %A Tremblay, Kimberly D %A Mager, Jesse %B Developmental Biology %I Elsevier {BV} %V 465 %P 1–10 %G eng %U https://doi.org/10.1016/j.ydbio.2020.06.011 %0 Journal Article %J Fertil Steril %D 2020 %T SOHLHs are essential for fertility regardless of gender or population. %A Cui, Wei %K Basic Helix-Loop-Helix Transcription Factors %K Fertility %K Gender Identity %K Humans %K Male %K Reproduction %B Fertil Steril %V 114 %P 283-284 %8 2020 08 %G eng %N 2 %R 10.1016/j.fertnstert.2020.06.010 %0 Journal Article %J Biology of Reproduction %D 2020 %T Testis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception† %A Salicioni, Ana M %A Maria G. Gervasi %A Sosnik, Julian %A Darya A. Tourzani %A Saman Nayyab %A Diego A Caraballo %A Visconti, Pablo E %B Biology of Reproduction %I Oxford University Press ({OUP}) %G eng %U https://doi.org/10.1093/biolre/ioaa064 %0 Journal Article %J Immunol Cell Biol %D 2020 %T The use of patient-derived breast tissue explants to study macrophage polarization and the effects of environmental chemical exposure. %A Gregory, Kelly J %A Morin, Stephanie M %A Kubosiak, Alex %A Ser-Dolansky, Jennifer %A Schalet, Benjamin J %A Jerry, D Joseph %A Schneider, Sallie S %K Benzophenones %K Breast %K Cell Polarity %K Endocrine Disruptors %K Environmental Exposure %K Female %K Humans %K Macrophage Activation %K Macrophages %K Tissue Culture Techniques %X

Ex 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.

%B Immunol Cell Biol %V 98 %P 883-896 %8 2020 11 %G eng %N 10 %R 10.1111/imcb.12381 %0 Journal Article %J Alzheimer{\textquotesingle}s Research {&} Therapy %D 2020 %T y-Secretase modulators exhibit selectivity for modulation of APP cleavage but inverse y-secretase modulators do not %A Christian B Lessard %A Edgardo Rodriguez %A Thomas B. Ladd %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %A Golde, Todd E %A Yong Ran %B Alzheimer{\textquotesingle}s Research {&} Therapy %I Springer Science and Business Media {LLC} %V 12 %G eng %U https://doi.org/10.1186/s13195-020-00622-5 %0 Journal Article %J Biology of Reproduction %D 2020 %T ZC3H4, a novel CCCH-type zinc finger protein, is essential for early embryogenesis in mice %A Jianmin Su %A Xiaosu Miao %A Danielle Archambault %A Mager, Jesse %A Cui, Wei %B Biology of Reproduction %I Oxford University Press ({OUP}) %G eng %U https://doi.org/10.1093/biolre/ioaa215 %0 Journal Article %J Frontiers in Immunology %D 2019 %T Adenosine A2A Receptor Stimulation Inhibits TCR-Induced Notch1 Activation in CD8+T-Cells %A Claudia Sorrentino %A Fokhrul Hossain %A Paulo C. Rodriguez %A Rosa A. Sierra %A Antonio Pannuti %A Stephen Hatfield %A Osborne, Barbara A %A Minter, Lisa M %A Miele, Lucio %A Silvana Morello %B Frontiers in Immunology %I Frontiers Media {SA} %V 10 %G eng %U https://doi.org/10.3389/fimmu.2019.00162 %0 Journal Article %J Molecular Therapy %D 2019 %T Alpha-1 Antitrypsin-Expressing Mesenchymal Stromal Cells Confer a Long-Term Survival Benefit in a Mouse Model of Lethal GvHD %A Sabine Geiger %A Emrah I. Ozay %A Ulf Geumann %A Marina K. Hereth %A Terese Magnusson %A Sudarvili Shanthalingam %A Daniela Hirsch %A Stefanie Kälin %A Christine Günther %A Osborne, Barbara A %A Tew, Gregory N %A Felix G. Hermann %A Minter, Lisa M %B Molecular Therapy %I Elsevier {BV} %V 27 %P 1436–1451 %G eng %U https://doi.org/10.1016/j.ymthe.2019.05.007 %0 Journal Article %J Science Immunology %D 2019 %T Alveolar macrophages generate a noncanonical NRF2-driven transcriptional response to Mycobacterium tuberculosis in vivo %A Alissa C. Rothchild %A Gregory S. Olson %A Johannes Nemeth %A Lynn M. Amon %A Dat Mai %A Elizabeth S. Gold %A Alan H. Diercks %A Alan Aderem %B Science Immunology %I American Association for the Advancement of Science ({AAAS}) %V 4 %P eaaw6693 %G eng %U https://doi.org/10.1126/sciimmunol.aaw6693 %0 Journal Article %J Frontiers in Immunology %D 2019 %T Corrigendum: Adenosine A2A Receptor Stimulation Inhibits TCR-Induced Notch1 Activation in CD8+T-Cells %A Claudia Sorrentino %A Fokhrul Hossain %A Paulo C. Rodriguez %A Rosa A. Sierra %A Antonio Pannuti %A Stephen Hatfield %A Osborne, Barbara A %A Minter, Lisa M %A Miele, Lucio %A Silvana Morello %B Frontiers in Immunology %I Frontiers Media {SA} %V 10 %G eng %U https://doi.org/10.3389/fimmu.2019.00935 %0 Journal Article %J Stem Cell Res %D 2019 %T Cymerus™ iPSC-MSCs significantly prolong survival in a pre-clinical, humanized mouse model of Graft-vs-host disease. %A Ozay, E Ilker %A Vijayaraghavan, Jyothi %A Gonzalez-Perez, Gabriela %A Shanthalingam, Sudarvili %A Sherman, Heather L %A Garrigan, Daniel T %A Chandiran, Karthik %A Torres, Joe A %A Osborne, Barbara A %A Tew, Gregory N %A Slukvin, Igor I %A Macdonald, Ross A %A Kelly, Kilian %A Minter, Lisa M %K Animals %K Disease Models, Animal %K Female %K Graft vs Host Disease %K Hematopoietic Stem Cell Transplantation %K Humans %K Induced Pluripotent Stem Cells %K Mesenchymal Stem Cell Transplantation %K Mesenchymal Stem Cells %K Mice %K Mice, Inbred NOD %X

The 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.

%B Stem Cell Res %V 35 %P 101401 %8 2019 03 %G eng %R 10.1016/j.scr.2019.101401 %0 Journal Article %J PLoS One %D 2019 %T The elongation factor Elof1 is required for mammalian gastrulation %A Adam P. Tellier %A Danielle Archambault %A Tremblay, Kimberly D %A Mager, Jesse %E Qiang Wu %B PLoS One %I Public Library of Science ({PLoS}) %V 14 %P e0219410 %G eng %U https://doi.org/10.1371/journal.pone.0219410 %0 Journal Article %J Breast Cancer Research %D 2019 %T Environmental exposures during windows of susceptibility for breast cancer: a framework for prevention research %A Mary Beth Terry %A and Karin B. Michels %A Julia Green Brody %A Celia Byrne %A Shiuan Chen %A Jerry, D Joseph %A Kristen M. C. Malecki %A Mary Beth Martin %A Rachel L. Miller %A Susan L. Neuhausen %A Kami Silk %A Amy Trentham-Dietz %B Breast Cancer Research %I Springer Science and Business Media {LLC} %V 21 %G eng %U https://doi.org/10.1186/s13058-019-1168-2 %0 Journal Article %J Breast Cancer Res %D 2019 %T Environmental exposures during windows of susceptibility for breast cancer: a framework for prevention research. %A Terry, Mary Beth %A Michels, Karin B %A Brody, Julia Green %A Byrne, Celia %A Chen, Shiuan %A Jerry, D Joseph %A Malecki, Kristen M C %A Martin, Mary Beth %A Miller, Rachel L %A Neuhausen, Susan L %A Silk, Kami %A Trentham-Dietz, Amy %K Animals %K Breast Neoplasms %K Disease Susceptibility %K Environmental Exposure %K Female %K Humans %K Maternal Exposure %K Menopause %K Pregnancy %K Puberty %K Research %K Risk Factors %K Time Factors %X

BACKGROUND: 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.

%B Breast Cancer Res %V 21 %P 96 %8 2019 08 20 %G eng %N 1 %R 10.1186/s13058-019-1168-2 %0 Journal Article %J Epigenetics %D 2019 %T Essential roles of HDAC1 and 2 in lineage development and genome-wide DNA methylation during mouse preimplantation development %A Panpan Zhao %A Huanan Wang %A Han Wang %A Yanna Dang %A Lei Luo %A Shuang Li %A Yan Shi %A Lefeng Wang %A Shaohua Wang %A Mager, Jesse %A Zhang, Kun %B Epigenetics %I Informa {UK} Limited %V 15 %P 369–385 %G eng %U https://doi.org/10.1080/15592294.2019.1669375 %0 Journal Article %J Reproductive Toxicology %D 2019 %T Exposure to low doses of oxybenzone during perinatal development alters mammary gland morphology in male and female mice %A Klara Matouskova %A Jerry, D Joseph %A Laura N. Vandenberg %B Reproductive Toxicology %I Elsevier {BV} %G eng %U https://doi.org/10.1016/j.reprotox.2019.08.002 %0 Journal Article %J Breast Cancer Res %D 2019 %T Gene expression signature of atypical breast hyperplasia and regulation by SFRP1. %A Gregory, Kelly J %A Roberts, Amy L %A Conlon, Erin M %A Mayfield, Jacob A %A Hagen, Mary J %A Crisi, Giovanna M %A Bentley, Brooke A %A Kane, Jeffrey J %A Makari-Judson, Grace %A Mason, Holly S %A Yu, Jun %A Zhu, Lihua Julie %A Simin, Karl %A Johnson, Jacob P S %A Khan, Ashraf %A Schneider, Ben R %A Schneider, Sallie S %A Jerry, D Joseph %K Adult %K Animals %K Biomarkers %K Biomarkers, Tumor %K Breast Neoplasms %K Disease Models, Animal %K Disease Progression %K Female %K Gene Expression Profiling %K Gene Expression Regulation %K Gene Regulatory Networks %K Humans %K Hyperplasia %K Intercellular Signaling Peptides and Proteins %K Mammary Glands, Human %K Membrane Proteins %K Mice %K Mice, Knockout %K Middle Aged %K Signal Transduction %K Transcriptome %X

BACKGROUND: 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.

%B Breast Cancer Res %V 21 %P 76 %8 2019 06 27 %G eng %N 1 %R 10.1186/s13058-019-1157-5 %0 Journal Article %J Breast Cancer Research %D 2019 %T Gene expression signature of atypical breast hyperplasia and regulation by SFRP1 %A Kelly J. Gregory %A Roberts, Amy L %A Erin M. Conlon %A Mayfield, Jacob A %A Hagen, Mary J %A Crisi, Giovanna M %A Bentley, Brooke A %A Jeffrey J. Kane %A Makari-Judson, Grace %A Holly S. Mason %A Jun Yu %A Lihua Julie Zhu %A Karl Simin %A Jacob P. S. Johnson %A Ashraf Khan %A Ben R. Schneider %A Schneider, Sallie S %A Jerry, D Joseph %B Breast Cancer Research %I Springer Science and Business Media {LLC} %V 21 %G eng %U https://doi.org/10.1186/s13058-019-1157-5 %0 Journal Article %J Journal of Biological Chemistry %D 2019 %T Individual and combined presenilin 1 and 2 knockouts reveal that both have highly overlapping functions in HEK293T cells %A Christian B Lessard %A Edgardo Rodriguez %A Thomas B. Ladd %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %A Golde, Todd E %A Yong Ran %B Journal of Biological Chemistry %I American Society for Biochemistry {&} Molecular Biology ({ASBMB}) %V 294 %P 11276–11285 %G eng %U https://doi.org/10.1074/jbc.ra119.008041 %0 Journal Article %J J Biol Chem %D 2019 %T Individual and combined presenilin 1 and 2 knockouts reveal that both have highly overlapping functions in HEK293T cells. %A Lessard, Christian B %A Rodriguez, Edgardo %A Ladd, Thomas B %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %A Golde, Todd E %A Ran, Yong %K Alzheimer Disease %K Amyloid Precursor Protein Secretases %K Animals %K CRISPR-Cas Systems %K Gene Knockdown Techniques %K HEK293 Cells %K Humans %K Hydrolysis %K Mice %K Presenilin-1 %K Presenilin-2 %K Substrate Specificity %X

Presenilins 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.

%B J Biol Chem %V 294 %P 11276-11285 %8 2019 07 19 %G eng %N 29 %R 10.1074/jbc.RA119.008041 %0 Journal Article %J Journal of Animal Science and Biotechnology %D 2019 %T Manipulation of bicarbonate concentration in sperm capacitation media improves in vitro fertilization output in porcine species %A Cristina Soriano-Úbeda %A Jon Romero-Aguirregomezcorta %A Carmen Matás %A Visconti, Pablo E %A Francisco A. García-Vázquez %B Journal of Animal Science and Biotechnology %I Springer Science and Business Media {LLC} %V 10 %G eng %U https://doi.org/10.1186/s40104-019-0324-y %0 Journal Article %J Journal of Cancer Education %D 2019 %T The Mouse Mammary Gland: a Tool to Inform Adolescents About Environmental Causes of Breast Cancer %A Laura N. Vandenberg %A SriDurgaDevi Kolla %A Charlotte D LaPlante %A Jerry, D Joseph %B Journal of Cancer Education %I Springer Science and Business Media {LLC} %G eng %U https://doi.org/10.1007/s13187-019-01563-w %0 Journal Article %J Human Molecular Genetics %D 2019 %T A null allele of Dnaaf2 displays embryonic lethality and mimics human ciliary dyskinesia %A Agnes Cheong %A Rinat Degani %A Tremblay, Kimberly D %A Mager, Jesse %B Human Molecular Genetics %I Oxford University Press ({OUP}) %G eng %U https://doi.org/10.1093/hmg/ddz106 %0 Journal Article %J Molecular Systems Biology %D 2019 %T Path-seq identifies an essential mycolate remodeling program for mycobacterial host adaptation %A Eliza JR Peterson %A Rebeca Bailo %A Alissa C. Rothchild %A Mario L Arrieta-Ortiz %A Amardeep Kaur %A Min Pan %A Dat Mai %A Abrar A Abidi %A Charlotte Cooper %A Alan Aderem %A Apoorva Bhatt %A Nitin S Baliga %B Molecular Systems Biology %I {EMBO} %V 15 %G eng %U https://doi.org/10.15252/msb.20188584 %0 Journal Article %J Industrial & Engineering Chemistry Research %D 2019 %T Postfunctionalization of Noncationic RNA–Polymer Complexes for RNA Delivery %A Ziwen Jiang %A Cui, Wei %A Mager, Jesse %A Thayumanavan, S %B Industrial & Engineering Chemistry Research %I American Chemical Society ({ACS}) %V 58 %P 6982–6991 %G eng %U https://doi.org/10.1021/acs.iecr.9b00666 %0 Journal Article %J Molecular Pharmaceutics %D 2019 %T Protein Transduction Domain Mimics Facilitate Rapid Antigen Delivery into Monocytes %A Coralie M. Backlund %A Ladan Parhamifar %A Minter, Lisa %A Tew, Gregory N %A Thomas L. Andresen %B Molecular Pharmaceutics %I American Chemical Society ({ACS}) %V 16 %P 2462–2469 %G eng %U https://doi.org/10.1021/acs.molpharmaceut.9b00070 %0 Journal Article %J Journal of Equine Veterinary Science %D 2019 %T Self-Assembling Intrauterine Device (Upod) Modulation of the Reproductive Cycle in Mares %A Gradil, Carlos M %A Cassandra K. Uricchio %A Allison Schwarz %B Journal of Equine Veterinary Science %I Elsevier {BV} %V 83 %P 102690 %G eng %U https://doi.org/10.1016/j.jevs.2019.02.009 %0 Journal Article %J ACS Applied Materials & Interfaces %D 2019 %T Symbiotic Self-Assembly Strategy toward Lipid-Encased Cross-Linked Polymer Nanoparticles for Efficient Gene Silencing %A Kingshuk Dutta %A Davide Bochicchio %A Alexander E. Ribbe %A Alfandari, Dominique %A Mager, Jesse %A Giovanni M. Pavan %A Thayumanavan, S %B ACS Applied Materials & Interfaces %I American Chemical Society ({ACS}) %V 11 %P 24971–24983 %G eng %U https://doi.org/10.1021/acsami.9b04731 %0 Journal Article %J Cancer Research %D 2019 %T Targeted Metabolomics Identifies the Cytochrome P450 Monooxygenase Eicosanoid Pathway as a Novel Therapeutic Target of Colon Tumorigenesis %A Weicang Wang %A Jun Yang %A Matthew L. Edin %A Yuxin Wang %A Ying Luo %A Debin Wan %A Haixia Yang %A Chun-Qing Song %A Wen Xue %A Katherine Z. Sanidad %A Mingyue Song %A Heather A. Bisbee %A Jennifer A. Bradbury %A Guanjun Nan %A Jianan Zhang %A Pei-an Betty Shih %A Kin Sing Stephen Lee %A Minter, Lisa M %A Daeyoung Kim %A Hang Xiao %A Jun-Yan Liu %A Bruce D. Hammock %A Darryl C. Zeldin %A Guodong Zhang %B Cancer Research %I American Association for Cancer Research ({AACR}) %V 79 %P 1822–1830 %G eng %U https://doi.org/10.1158/0008-5472.can-18-3221 %0 Journal Article %J Reproduction, Fertility and Development %D 2019 %T Tissue plasminogen activator (tPA) of paternal origin is necessary for the success of in vitro but not of in vivo fertilization in the mouse %A Francisco A. García-Vázquez %A C. Soriano-Úbeda %A R. Laguna-Barraza %A M José Izquierdo-Rico %A Navarrete, Felipe A %A Visconti, Pablo E %A A. Gutiérrez-Adán %A P. Coy %B Reproduction, Fertility and Development %I {CSIRO} Publishing %V 31 %P 433 %G eng %U https://doi.org/10.1071/rd18175 %0 Journal Article %J Frontiers in Cell and Developmental Biology %D 2019 %T Transient Sperm Starvation Improves the Outcome of Assisted Reproductive Technologies %A Navarrete, Felipe A %A Luis Aguila %A David Martin-Hidalgo %A Darya A. Tourzani %A Luque, Guillermina M %A Goli Ardestani %A Francisco A. García-Vázquez %A Levin, Lonny R %A Buck, Jochen %A Darszon, Alberto %A Buffone, Mariano G %A Mager, Jesse %A Fissore, Rafael A %A Salicioni, Ana Maria %A Gervasi, Maria Gracia %A Visconti, Pablo E %B Frontiers in Cell and Developmental Biology %I Frontiers Media {SA} %V 7 %G eng %U https://doi.org/10.3389/fcell.2019.00262 %0 Journal Article %J Toxicological Sciences %D 2019 %T Triclocarban Exposure Exaggerates Spontaneous Colonic Inflammation in Il-10-/- Mice %A Minhao Xie %A Hongna Zhang %A Weicang Wang %A Heather L Sherman %A Minter, Lisa M %A Zongwei Cai %A Guodong Zhang %B Toxicological Sciences %I Oxford University Press ({OUP}) %V 174 %P 92–99 %G eng %U https://doi.org/10.1093/toxsci/kfz248 %0 Journal Article %J Biomacromolecules %D 2018 %T Bait-and-Switch Supramolecular Strategy To Generate Noncationic RNA–Polymer Complexes for RNA Delivery %A Ziwen Jiang %A Cui, Wei %A Prasad, Priyaa %A Mollie A. Touve %A Nathan C. Gianneschi %A Mager, Jesse %A Thayumanavan, S %B Biomacromolecules %I American Chemical Society ({ACS}) %8 dec %G eng %U https://doi.org/10.1021/acs.biomac.8b01321 %0 Journal Article %J Frontiers in Cell and Developmental Biology %D 2018 %T Changes in Protein O-GlcNAcylation During Mouse Epididymal Sperm Maturation %A Darya A. Tourzani %A Paudel, Bidur %A Miranda, Patricia V %A Visconti, Pablo E %A Maria G. Gervasi %B Frontiers in Cell and Developmental Biology %I Frontiers Media {SA} %V 6 %8 jun %G eng %U https://doi.org/10.3389/fcell.2018.00060 %0 Journal Article %J Science Translational Medicine %D 2018 %T A common antimicrobial additive increases colonic inflammation and colitis-associated colon tumorigenesis in mice %A Haixia Yang %A Weicang Wang %A Kymberleigh A. Romano %A Min Gu %A Katherine Z. Sanidad %A Daeyoung Kim %A Jun Yang %A Birgitta Schmidt %A Dipak Panigrahy %A Ruisong Pei %A Derek A. Martin %A E. Ilker Ozay %A Yuxin Wang %A Mingyue Song %A Bradley W. Bolling %A Hang Xiao %A Minter, Lisa M %A Guang-Yu Yang %A Zhenhua Liu %A Federico E. Rey %A Guodong Zhang %B Science Translational Medicine %I American Association for the Advancement of Science ({AAAS}) %V 10 %P eaan4116 %G eng %U https://doi.org/10.1126/scitranslmed.aan4116 %0 Journal Article %J Pest Manag Sci %D 2018 %T Comparative CYP-omic analysis between the DDT-susceptible and -resistant Drosophila melanogaster strains 91-C and 91-R. %A Seong, Keon Mook %A Coates, Brad S %A Berenbaum, May R %A Clark, John M %A Pittendrigh, Barry R %K Animals %K Cytochrome P-450 Enzyme System %K DDT %K Drosophila melanogaster %K Insect Proteins %K Insecticide Resistance %K Insecticides %K Proteomics %X

BACKGROUND: 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.

%B Pest Manag Sci %V 74 %P 2530-2543 %8 2018 Nov %G eng %N 11 %R 10.1002/ps.4936 %0 Journal Article %D 2018 %T Comparative two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of human milk to identify dysregulated proteins in breast cancer %A Aslebagh, Roshanak %A Devika Channaveerappa %A Arcaro, Kathleen F %A Darie, Costel C %I Wiley %V 39 %P 1723–1734 %8 jun %G eng %U https://doi.org/10.1002/elps.201800025 %0 Journal Article %J Cold Spring Harbor Protocols %D 2018 %T Cranial Neural Crest Explants %A Cousin, Hélène %A Alfandari, Dominique %B Cold Spring Harbor Protocols %I Cold Spring Harbor Laboratory %V 2018 %P pdb.prot097394 %G eng %U https://doi.org/10.1101/pdb.prot097394 %0 Journal Article %J Cold Spring Harbor Protocols %D 2018 %T Cranial Neural Crest Transplants %A Cousin, Hélène %B Cold Spring Harbor Protocols %I Cold Spring Harbor Laboratory %V 2018 %P pdb.prot097402 %8 jan %G eng %U https://doi.org/10.1101/pdb.prot097402 %0 Journal Article %J genesis %D 2018 %T Cut loose and run: The complex role of ADAM proteases during neural crest cell development %A Alfandari, Dominique %A Lisa A. Taneyhill %B genesis %I Wiley %V 56 %P e23095 %G eng %U https://doi.org/10.1002/dvg.23095 %0 Journal Article %D 2018 %T Development of a genetically encoded sensor for endogenous CaMKII activity %A Goli Ardestani %A Megan West %A Thomas J Maresca %A Fissore, Rafael A %A Margaret M Stratton %I Cold Spring Harbor Laboratory %8 jun %G eng %U https://doi.org/10.1101/359398 %0 Journal Article %J Journal of the Academy of Nutrition and Dietetics %D 2018 %T Dietary Intervention to Increase Fruit and Vegetable Consumption in Breastfeeding Women: A Pilot Randomized Trial Measuring Inflammatory Markers in Breast Milk %A Angela R. Essa %A Browne, Eva P %A Punska, Elizabeth C %A Katelyn Perkins %A Emily Boudreau %A Hilary Wiggins %A Anderton, Douglas L %A Lindiwe Sibeko %A Sturgeon, Susan R %A Arcaro, Kathleen F %B Journal of the Academy of Nutrition and Dietetics %I Elsevier {BV} %8 sep %G eng %U https://doi.org/10.1016/j.jand.2018.06.015 %0 Journal Article %J Journal of Cellular Biochemistry %D 2018 %T DNA methylation in breast cancers: Differences based on estrogen receptor status and recurrence %A Williams, Kristin E %A M Jawale, Rahul %A Schneider, Sallie S %A Otis, Christopher N %A Pentecost, Brian T %A Arcaro, Kathleen F %B Journal of Cellular Biochemistry %I Wiley %8 sep %G eng %U https://doi.org/10.1002/jcb.27431 %0 Journal Article %J Toxicological Sciences %D 2018 %T Effects of Consumer Antimicrobials Benzalkonium Chloride, Benzethonium Chloride, and Chloroxylenol on Colonic Inflammation and Colitis-Associated Colon Tumorigenesis in Mice %A Katherine Z. Sanidad %A Haixia Yang %A Weicang Wang %A E. Ilker Ozay %A Jun Yang %A Min Gu %A Emmet Karner %A Jianan Zhang %A Daeyoung Kim %A Minter, Lisa M %A Hang Xiao %A Guodong Zhang %B Toxicological Sciences %I Oxford University Press ({OUP}) %V 163 %P 490–499 %G eng %U https://doi.org/10.1093/toxsci/kfy045 %0 Journal Article %J Food Chem Toxicol %D 2018 %T Exposure to permethrin promotes high fat diet-induced weight gain and insulin resistance in male C57BL/6J mice. %A Xiao, Xiao %A Sun, Quancai %A Kim, Yoo %A Yang, Szu-Hao %A Qi, Weipeng %A Kim, Daeyoung %A Yoon, Kyong Sup %A Clark, John M %A Park, Yeonhwa %K Animals %K Diet, High-Fat %K Dose-Response Relationship, Drug %K Glucose %K Homeostasis %K Insecticides %K Insulin Resistance %K Lipid Metabolism %K Male %K Mice %K Mice, Inbred C57BL %K Permethrin %X

Permethrin 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.

%B Food Chem Toxicol %V 111 %P 405-416 %8 2018 Jan %G eng %R 10.1016/j.fct.2017.11.047 %0 Journal Article %J Mamm Genome %D 2018 %T Genetic variation in sensitivity to estrogens and breast cancer risk. %A Jerry, D Joseph %A Shull, James D %A Hadsell, Darryl L %A Rijnkels, Monique %A Dunphy, Karen A %A Schneider, Sallie S %A Vandenberg, Laura N %A Majhi, Prabin Dhangada %A Byrne, Celia %A Trentham-Dietz, Amy %K Animals %K Breast Neoplasms %K Estradiol %K Estrogens %K Female %K Genetic Predisposition to Disease %K Genetic Variation %K Humans %K Mammary Glands, Human %K Mammary Neoplasms, Animal %K Mice %K Quantitative Trait Loci %K Risk Factors %X

Breast 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.

%B Mamm Genome %V 29 %P 24-37 %8 2018 02 %G eng %N 1-2 %R 10.1007/s00335-018-9741-z %0 Journal Article %J Mammalian Genome %D 2018 %T Genetic variation in sensitivity to estrogens and breast cancer risk %A Jerry, D Joseph %A James D. Shull %A Darryl L. Hadsell %A Monique Rijnkels %A Dunphy, Karen A %A Schneider, Sallie S %A Laura N. Vandenberg %A Prabin Dhangada Majhi %A Celia Byrne %A Amy Trentham-Dietz %B Mammalian Genome %I Springer Nature %V 29 %P 24–37 %8 feb %G eng %U https://doi.org/10.1007/s00335-018-9741-z %0 Journal Article %J Future Oncology %D 2018 %T Let's fight cancer: let-7 is a tool to enhance antitumor immune responses %A Pobezinsky, Leonid A %A Alexandria C Wells %B Future Oncology %I Future Medicine Ltd %V 14 %P 1141–1145 %8 may %G eng %U https://doi.org/10.2217/fon-2018-0037 %0 Journal Article %J Cell Reprogram %D 2018 %T MC1568 Enhances Histone Acetylation During Oocyte Meiosis and Improves Development of Somatic Cell Nuclear Transfer Embryos in Pig. %A Wang, Huili %A Cui, Wei %A Meng, Chunhua %A Zhang, Jun %A Li, Yinxia %A Qian, Yong %A Xing, Guangdong %A Zhao, Dongmin %A Cao, Shaoxian %K Acetylation %K Animals %K Blastocyst %K Cellular Reprogramming %K Cloning, Organism %K Embryonic Development %K Female %K Histone Code %K Histone Deacetylase Inhibitors %K Histones %K Hydroxamic Acids %K Meiosis %K Nuclear Transfer Techniques %K Oocytes %K Pregnancy %K Pyrroles %K Sus scrofa %X

An 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.

%B Cell Reprogram %V 20 %P 55-65 %8 2018 02 %G eng %N 1 %R 10.1089/cell.2017.0023 %0 Journal Article %J Reproduction %D 2018 %T Med20 is essential for early embryogenesis and regulates Nanog expression %A Cui, Wei %A Marcho, Chelsea %A Yongsheng Wang %A Rinat Degani %A Morgane Golan %A Tremblay, Kimberly D %A Jaime Rivera-Pérez %A Mager, Jesse %B Reproduction %I Bioscientifica %8 dec %G eng %U https://doi.org/10.1530/rep-18-0508 %0 Journal Article %J Frontiers in Immunology %D 2018 %T Notch Signaling in Myeloid Cells as a Regulator of Tumor Immune Responses %A Fokhrul Hossain %A Samarpan Majumder %A Deniz A. Ucar %A Paulo C. Rodriguez %A Golde, Todd E %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %B Frontiers in Immunology %I Frontiers Media {SA} %V 9 %G eng %U https://doi.org/10.3389/fimmu.2018.01288 %0 Journal Article %J Frontiers in Oncology %D 2018 %T Notch Signaling Regulates Mitochondrial Metabolism and NF-kB Activity in Triple-Negative Breast Cancer Cells via IKKa-Dependent Non-canonical Pathways %A Fokhrul Hossain %A Claudia Sorrentino %A Deniz A. Ucar %A Yin Peng %A Margarite Matossian %A Dorota Wyczechowska %A Judy Crabtree %A Jovanny Zabaleta %A Silvana Morello %A Luis Del Valle %A Matthew Burow %A Bridgette Collins-Burow %A Antonio Pannuti %A Minter, Lisa M %A Golde, Todd E %A Osborne, Barbara A %A Miele, Lucio %B Frontiers in Oncology %I Frontiers Media {SA} %V 8 %G eng %U https://doi.org/10.3389/fonc.2018.00575 %0 Journal Article %J Mol Immunol %D 2018 %T Notch1 primes CD4 T cells for T helper type I differentiation through its early effects on miR-29. %A Chandiran, Karthik %A Lawlor, Rebecca %A Pannuti, Antonio %A Perez, Gabriela Gonzalez %A Srinivasan, Janani %A Golde, Todd E %A Miele, Lucio %A Osborne, Barbara A %A Minter, Lisa M %K Animals %K CD4-Positive T-Lymphocytes %K Cell Differentiation %K Interferon-gamma %K Mice %K Mice, Inbred C57BL %K MicroRNAs %K NF-kappa B %K NIH 3T3 Cells %K Receptor, Notch1 %K Signal Transduction %K Th1 Cells %K Transcription Factors %K Transcription, Genetic %X

The 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.

%B Mol Immunol %V 99 %P 191-198 %8 2018 07 %G eng %R 10.1016/j.molimm.2018.05.002 %0 Journal Article %J Molecular Immunology %D 2018 %T Notch1 primes CD4 T cells for T helper type I differentiation through its early effects on miR-29 %A Chandiran, Karthik %A Rebecca Lawlor %A Antonio Pannuti %A Gabriela Gonzalez Perez %A Srinivasan, Janani %A Golde, Todd E %A Miele, Lucio %A Osborne, Barbara A %A Minter, Lisa M %B Molecular Immunology %I Elsevier {BV} %V 99 %P 191–198 %G eng %U https://doi.org/10.1016/j.molimm.2018.05.002 %0 Journal Article %J Journal of Cellular Physiology %D 2018 %T Only a subpopulation of mouse sperm displays a rapid increase in intracellular calcium during capacitation %A Luque, Guillermina M %A Tomas Dalotto-Moreno %A David Martin-Hidalgo %A Carla Ritagliati %A Lis C. Puga Molina %A Romarowski, Ana %A Paula A. Balestrini %A Liza J. Schiavi-Ehrenhaus %A Nicolas Gilio %A Krapf, Dario %A Visconti, Pablo E %A Buffone, Mariano G %B Journal of Cellular Physiology %I Wiley %8 jun %G eng %U https://doi.org/10.1002/jcp.26883 %0 Journal Article %J Journal of the Endocrine Society %D 2018 %T Oxybenzone Alters Mammary Gland Morphology in Mice Exposed During Pregnancy and Lactation %A Charlotte D LaPlante %A Ruby Bansal %A Dunphy, Karen A %A Jerry, D Joseph %A Laura N. Vandenberg %B Journal of the Endocrine Society %I The Endocrine Society %V 2 %P 903–921 %8 may %G eng %U https://doi.org/10.1210/js.2018-00024 %0 Journal Article %J Hepatology %D 2018 %T Patterning of the hepato-pancreatobiliary boundary by BMP reveals heterogeneity within the murine liver bud %A Amrita Palaria %A Angelo, Jesse R %A Taylor M. Guertin %A Mager, Jesse %A Tremblay, Kimberly D %B Hepatology %I Wiley %V 68 %P 274–288 %8 may %G eng %U https://doi.org/10.1002/hep.29769 %0 Journal Article %J Breast Cancer Research and Treatment %D 2018 %T Pro-inflammatory cytokines and growth factors in human milk: an exploratory analysis of racial differences to inform breast cancer etiology %A Murphy, Jeanne %A Pfeiffer, Ruth M %A Brittny C. Davis Lynn %A Caballero, Ana I %A Browne, Eva P %A Punska, Elizabeth C %A Yang, Hannah P %A Roni T. Falk %A Anderton, Douglas L %A Gierach, Gretchen L %A Arcaro, Kathleen F %A Sherman, Mark E %B Breast Cancer Research and Treatment %I Springer Nature America, Inc %8 aug %G eng %U https://doi.org/10.1007/s10549-018-4907-7 %0 Journal Article %J ELECTROPHORESIS %D 2018 %T Proteomics analysis of human breast milk to assess breast cancer risk %A Aslebagh, Roshanak %A Devika Channaveerappa %A Arcaro, Kathleen F %A Darie, Costel C %B ELECTROPHORESIS %I Wiley %V 39 %P 653–665 %8 feb %G eng %U https://doi.org/10.1002/elps.201700123 %0 Journal Article %J Insect Mol Biol %D 2018 %T RNA interference validation of detoxification genes involved in ivermectin tolerance in Drosophila melanogaster. %A Kim, J H %A Moreau, J A %A Ali, Y %A Razo, P %A Hong, K B %A Yoon, K S %A Clark, J M %K Animals %K Drosophila melanogaster %K Drug Tolerance %K Female %K Inactivation, Metabolic %K Insecticide Resistance %K Insecticides %K Ivermectin %K RNA Interference %X

Previously, 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.

%B Insect Mol Biol %V 27 %P 651-660 %8 2018 10 %G eng %N 5 %R 10.1111/imb.12512 %0 Journal Article %J Bioconjugate Chemistry %D 2018 %T The Role of Cargo Binding Strength in Polymer-Mediated Intracellular Protein Delivery %A Nicholas D. Posey %A Christopher R. Hango %A Minter, Lisa M %A Tew, Gregory N %B Bioconjugate Chemistry %I American Chemical Society ({ACS}) %V 29 %P 2679–2690 %G eng %U https://doi.org/10.1021/acs.bioconjchem.8b00363 %0 Journal Article %J Frontiers in Immunology %D 2018 %T Rotenone Treatment Reveals a Role for Electron Transport Complex I in the Subcellular Localization of Key Transcriptional Regulators During T Helper Cell Differentiation %A Emrah Ilker Ozay %A Heather L Sherman %A Victoria Mello %A Grace Trombley %A Adam Lerman %A Tew, Gregory N %A Yadava, Nagendra %A Minter, Lisa M %B Frontiers in Immunology %I Frontiers Media {SA} %V 9 %G eng %U https://doi.org/10.3389/fimmu.2018.01284 %0 Journal Article %J Development %D 2018 %T Single-cell murine genetic fate mapping reveals bipotential hepatoblasts and novel multi-organ endoderm progenitors %A Gabriel K. El Sebae %A Joseph M. Malatos %A Mary-Kate E. Cone %A Rhee, Siyeon %A Angelo, Jesse R %A Mager, Jesse %A Tremblay, Kimberly D %B Development %I The Company of Biologists %V 145 %P dev168658 %8 sep %G eng %U https://doi.org/10.1242/dev.168658 %0 Journal Article %J Biology of Reproduction %D 2018 %T Sperm capacitation is associated with phosphorylation of the testis-specific radial spoke protein Rsph6a† %A Paudel, Bidur %A Gervasi, Maria Gracia %A James Porambo %A Diego Caraballo %A Darya Tourzani %A Mager, Jesse %A Platt, Mark D %A Salicioni, Ana Maria %A Visconti, Pablo E %B Biology of Reproduction %I Oxford University Press ({OUP}) %8 sep %G eng %U https://doi.org/10.1093/biolre/ioy202 %0 Journal Article %J Biology of Reproduction %D 2018 %T Sperm capacitation is associated with phosphorylation of the testis-specific radial spoke protein Rsph6a %A Paudel, Bidur %A Gervasi, Maria Gracia %A James Porambo %A Diego A Caraballo %A Darya A. Tourzani %A Mager, Jesse %A Platt, Mark D %A Salicioni, Ana Maria %A Visconti, Pablo E %B Biology of Reproduction %I Oxford University Press ({OUP}) %8 sep %G eng %U https://doi.org/10.1093/biolre/ioy202 %0 Journal Article %J J Am Assoc Lab Anim Sci %D 2018 %T Sterilization of Silastic Capsules Containing 17β-Estradiol for Effective Hormone Delivery in Mus musculus. %A Majewski, Aliza R %A Chuong, Lynn M %A Neill, Hannah M %A Roberts, Amy L %A Jerry, D Joseph %A Dunphy, Karen A %X

Silastic 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.

%B J Am Assoc Lab Anim Sci %8 2018 Oct 12 %G eng %R 10.30802/AALAS-JAALAS-18-000030 %0 Journal Article %J Journal of the American Association for Laboratory Animal Science %D 2018 %T Sterilization of Silastic Capsules Containing 17β-Estradiol for Effective Hormone Delivery in Mus musculus %A Aliza R Majewski %A Lynn M Chuong %A Hannah M Neill %A Roberts, Amy L %A Jerry, D Joseph %A Dunphy, Karen A %B Journal of the American Association for Laboratory Animal Science %I American Association for Laboratory Animal Science %G eng %U https://doi.org/10.30802/aalas-jaalas-18-000030 %0 Journal Article %J Journal of Cell Science %D 2018 %T Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis %A Romarowski, Ana %A Ángel G. Velasco Félix %A Paulina Torres Rodri'guez %A Mari'a G. Gervasi %A Xu, Xinran %A Luque, Guillermina M %A Gastón Contreras-Jiménez %A Sánchez-Cárdenas, Claudia %A Héctor V. Rami'rez-Gómez %A Krapf, Diego %A Visconti, Pablo E %A Krapf, Dario %A Adán Guerrero %A Darszon, Alberto %A Buffone, Mariano G %B Journal of Cell Science %I The Company of Biologists %V 131 %P jcs218958 %G eng %U https://doi.org/10.1242/jcs.218958 %0 Journal Article %J PLoS One %D 2018 %T Tauroursodeoxycholic acid (TUDCA) alleviates endoplasmic reticulum stress of nuclear donor cells under serum starvation. %A Zhang, Ying %A Qu, Pengxiang %A Ma, Xiaonan %A Qiao, Fang %A Ma, Yefei %A Qing, Suzhu %A Zhang, Yong %A Wang, Yongsheng %A Cui, Wei %K Apoptosis %K Cell Cycle %K Endoplasmic Reticulum Stress %K Food Deprivation %K Humans %K Stress, Physiological %K Taurochenodeoxycholic Acid %X

Serum 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.

%B PLoS One %V 13 %P e0196785 %8 2018 %G eng %N 5 %R 10.1371/journal.pone.0196785 %0 Book Section %B Chromatin Regulation of Early Embryonic Lineage Specification %D 2018 %T Transcriptional Regulation and Genes Involved in First Lineage Specification During Preimplantation Development %A Cui, Wei %A Mager, Jesse %B Chromatin Regulation of Early Embryonic Lineage Specification %I Springer International Publishing %P 31–46 %8 nov %G eng %U https://doi.org/10.1007/978-3-319-63187-5_4 %0 Journal Article %J Cancer Cell International %D 2018 %T TROP2 methylation and expression in tamoxifen-resistant breast cancer %A Zimmers, Stephanie M %A Browne, Eva P %A Williams, Kristin E %A M Jawale, Rahul %A Otis, Christopher N %A Schneider, Sallie S %A Arcaro, Kathleen F %B Cancer Cell International %I Springer Nature %V 18 %8 jul %G eng %U https://doi.org/10.1186/s12935-018-0589-9 %0 Journal Article %J Development %D 2018 %T XenopusADAM19 regulates Wnt signaling and neural crest specification by stabilizing ADAM13 %A Jiejing Li %A Mark Perfetto %A Neuner, Russell %A Harinath Bahudhanapati %A Laura Christian %A Ketan Mathavan %A Lance C. Bridges %A Alfandari, Dominique %A Shuo Wei %B Development %I The Company of Biologists %V 145 %P dev158154 %G eng %U https://doi.org/10.1242/dev.158154 %0 Journal Article %J J Sci Food Agric %D 2017 %T 4,4'-Dichlorodiphenyltrichloroethane (DDT) and 4,4'-dichlorodiphenyldichloroethylene (DDE) inhibit myogenesis in C2C12 myoblasts. %A Kim, Jonggun %A Park, Min Young %A Kim, Yoo %A Yoon, Kyong Sup %A Clark, John Marshall %A Park, Yeonhwa %A Whang, Kwang-Youn %K Animals %K Cell Line %K DDT %K Dichlorodiphenyl Dichloroethylene %K Gene Expression %K Insecticides %K Mice %K Muscle Development %K Myoblasts %K MyoD Protein %K Myogenic Regulatory Factor 5 %X

BACKGROUND: 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.

%B J Sci Food Agric %V 97 %P 5176-5185 %8 2017 Dec %G eng %N 15 %R 10.1002/jsfa.8399 %0 Journal Article %J Journal of Biological Chemistry %D 2017 %T Active site–adjacent phosphorylation at Tyr-397 by c-Abl kinase inactivates caspase-9 %A Banyuhay P. Serrano %A Hannah S. Szydlo %A Alfandari, Dominique %A Hardy, Jeanne A %B Journal of Biological Chemistry %I American Society for Biochemistry {&} Molecular Biology ({ASBMB}) %V 292 %P 21352–21365 %G eng %U https://doi.org/10.1074/jbc.m117.811976 %0 Journal Article %J Journal of Veterinary Internal Medicine %D 2017 %T Blood Glucose and Insulin Concentrations after Octreotide Administration in Horses With Insulin Dysregulation %A N. Frank %A P. Hermida %A A. Sanchez-Londoño %A R. Singh %A C.M. Gradil %A C.K. Uricchio %B Journal of Veterinary Internal Medicine %I Wiley %V 31 %P 1188–1192 %8 may %G eng %U https://doi.org/10.1111/jvim.14718 %0 Journal Article %J Mechanisms of Development %D 2017 %T Cadherins function during the collective cell migration of Xenopus Cranial Neural Crest cells: revisiting the role of E-cadherin %A Cousin, Hélène %B Mechanisms of Development %I Elsevier {BV} %V 148 %P 79–88 %8 dec %G eng %U https://doi.org/10.1016/j.mod.2017.04.006 %0 Journal Article %J J Am Chem Soc %D 2017 %T Cancer Cell Discrimination Using Host-Guest "Doubled" Arrays. %A Le, Ngoc D B %A Yesilbag Tonga, Gulen %A Mout, Rubul %A Kim, Sung-Tae %A Wille, Marcos E %A Rana, Subinoy %A Dunphy, Karen A %A Jerry, D Joseph %A Yazdani, Mahdieh %A Ramanathan, Rajesh %A Rotello, Caren M %A Rotello, Vincent M %K Binding Sites %K Biosensing Techniques %K Bridged-Ring Compounds %K Cell Line, Tumor %K Gold %K Humans %K Imidazoles %K Luminescent Proteins %K Metal Nanoparticles %K Molecular Structure %K Nanotechnology %K Neoplasms %X

We 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.

%B J Am Chem Soc %V 139 %P 8008-8012 %8 2017 06 14 %G eng %N 23 %R 10.1021/jacs.7b03657 %0 Journal Article %J Journal of the American Chemical Society %D 2017 %T Cancer Cell Discrimination Using Host–Guest “Doubled” Arrays %A Ngoc D. B. Le %A Gulen Yesilbag Tonga %A Rubul Mout %A Sung-Tae Kim %A Marcos E. Wille %A Subinoy Rana %A Dunphy, Karen A %A Jerry, D Joseph %A Mahdieh Yazdani %A Rajesh Ramanathan %A Caren M. Rotello %A Rotello, Vincent M %B Journal of the American Chemical Society %I American Chemical Society ({ACS}) %V 139 %P 8008–8012 %8 jun/2017 %G eng %U https://doi.org/10.1021/jacs.7b03657 %0 Journal Article %J Genome Biol Evol %D 2017 %T Changes in Neuronal Signaling and Cell Stress Response Pathways are Associated with a Multigenic Response of Drosophila melanogaster to DDT Selection. %A Seong, Keon Mook %A Coates, Brad S %A Sun, Weilin %A Clark, John M %A Pittendrigh, Barry R %K Animals %K DDT %K Drosophila melanogaster %K Drosophila Proteins %K Female %K Gene Expression Profiling %K Gene Expression Regulation %K Genome, Insect %K Insecticide Resistance %K Insecticides %K Male %K Signal Transduction %K Stress, Physiological %X

The 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.

%B Genome Biol Evol %V 9 %P 3356-3372 %8 2017 12 01 %G eng %N 12 %R 10.1093/gbe/evx252 %0 Journal Article %J Insect Mol Biol %D 2017 %T Comparison of the proliferation and excretion of Bartonella quintana between body and head lice following oral challenge. %A Kim, J H %A Previte, D J %A Yoon, K S %A Murenzi, E %A Koehler, J E %A Pittendrigh, B R %A Lee, S H %A Clark, J M %K Animals %K Bartonella quintana %K Gastrointestinal Tract %K Green Fluorescent Proteins %K Humans %K Insect Vectors %K Pediculus %K Reactive Oxygen Species %K Trench Fever %X

Human 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.

%B Insect Mol Biol %V 26 %P 266-276 %8 2017 06 %G eng %N 3 %R 10.1111/imb.12292 %0 Journal Article %J Reproduction %D 2017 %T Defective sperm head decondensation undermines the success of ICSI in the bovine %A Luis Aguila %A Ricardo Felmer %A Maria Elena Arias %A Navarrete, Felipe %A David Martin-Hidalgo %A Lee, Hoi Chang %A Visconti, Pablo %A Fissore, Rafael %B Reproduction %I {BioScientifica} %V 154 %P 207–218 %8 jul %G eng %U https://doi.org/10.1530/rep-17-0270 %0 Journal Article %J Food Chem Toxicol %D 2017 %T Deltamethrin increases the fat accumulation in 3T3-L1 adipocytes and Caenorhabditis elegans. %A Shen, Peiyi %A Hsieh, Tsung-Hsiu %A Yue, Yiren %A Sun, Quancai %A Clark, John M %A Park, Yeonhwa %K 3T3-L1 Cells %K Acetyl-CoA Carboxylase %K Adipocytes %K Adipogenesis %K AMP-Activated Protein Kinases %K Animals %K Caenorhabditis elegans %K CCAAT-Enhancer-Binding Proteins %K Insecticides %K Locomotion %K Mice %K Nitriles %K Phosphorylation %K Pyrethrins %K Triglycerides %X

Research 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.

%B Food Chem Toxicol %V 101 %P 149-156 %8 2017 Mar %G eng %R 10.1016/j.fct.2017.01.015 %0 Journal Article %J {eLife} %D 2017 %T Dual control of pcdh8l/PCNS expression and function in Xenopus laevis neural crest cells by adam13/33 via the transcription factors tfap2alpha and arid3a. %A Vikram Khedgikar %A Abbruzzese, Genevieve %A Ketan Mathavan %A Hannah Szydlo %A Helene Cousin %A Alfandari, Dominique %B {eLife} %I {eLife} Sciences Organisation, Ltd. %V 6 %8 aug %G eng %U https://doi.org/10.7554/elife.26898 %0 Journal Article %J {PLOS} {ONE} %D 2017 %T The ectodomain of cadherin-11 binds to erbB2 and stimulates Akt phosphorylation to promote cranial neural crest cell migration %A Ketan Mathavan %A Vikram Khedgikar %A Vanessa Bartolo %A Alfandari, Dominique %E Aamir Ahmad %B {PLOS} {ONE} %I Public Library of Science ({PLoS}) %V 12 %P e0188963 %G eng %U https://doi.org/10.1371/journal.pone.0188963 %0 Journal Article %J J Agric Food Chem %D 2017 %T Effectiveness of Commercial and Homemade Washing Agents in Removing Pesticide Residues on and in Apples. %A Yang, Tianxi %A Doherty, Jeffery %A Zhao, Bin %A Kinchla, Amanda J %A Clark, John M %A He, Lili %K Detergents %K Food Contamination %K Food Handling %K Fruit %K Malus %K Pesticide Residues %K Pesticides %K Phosmet %K Thiabendazole %X

Removal 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.

%B J Agric Food Chem %V 65 %P 9744-9752 %8 2017 Nov 08 %G eng %N 44 %R 10.1021/acs.jafc.7b03118 %0 Journal Article %J PLoS ONE %D 2017 %T Effects of embryo-derived exosomes on the development of bovine cloned embryos %A Pengxiang Qu %A Suzhu Qing %A Ruiqi Liu %A Hongyu Qin %A Weiwei Wang %A Fang Qiao %A Hui Ge %A Jun Liu %A Yong Zhang %A Cui, Wei %A Yongsheng Wang %E Wei Shen %B PLoS ONE %I Public Library of Science ({PLoS}) %V 12 %P e0174535 %8 mar %G eng %U https://doi.org/10.1371/journal.pone.0174535 %0 Journal Article %J Neurotoxicology %D 2017 %T Evaluation 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. %A Murenzi, Edwin %A Toltin, Abigail C %A Symington, Steven B %A Morgan, Molly M %A Clark, John M %K Animals %K Brain Tissue Transplantation %K Drug Evaluation, Preclinical %K Female %K Ion Channels %K Neurilemma %K Oocytes %K Rats, Sprague-Dawley %K Toxicology %K Transplantation, Heterologous %K Voltage-Gated Sodium Channels %K Xenopus laevis %X

Microtransplantation 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.

%B Neurotoxicology %V 60 %P 260-273 %8 2017 May %G eng %R 10.1016/j.neuro.2016.04.004 %0 Journal Article %J J Agric Food Chem %D 2017 %T Imidacloprid Promotes High Fat Diet-Induced Adiposity in Female C57BL/6J Mice and Enhances Adipogenesis in 3T3-L1 Adipocytes via the AMPKα-Mediated Pathway. %A Sun, Quancai %A Qi, Weipeng %A Xiao, Xiao %A Yang, Szu-Hao %A Kim, Daeyoung %A Yoon, Kyong Sup %A Clark, John M %A Park, Yeonhwa %K Adipocytes %K Adipogenesis %K Adiposity %K AMP-Activated Protein Kinases %K Animals %K Diet, High-Fat %K Female %K Humans %K Imidazoles %K Insecticides %K Male %K Mice %K Mice, Inbred C57BL %K Neonicotinoids %K Nitro Compounds %K Obesity %K Proto-Oncogene Proteins c-akt %K Signal Transduction %X

Imidacloprid, 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.

%B J Agric Food Chem %V 65 %P 6572-6581 %8 2017 Aug 09 %G eng %N 31 %R 10.1021/acs.jafc.7b02584 %0 Journal Article %J Journal of Cellular Physiology %D 2017 %T Individual-specific variation in the respiratory activities of HMECs and their bioenergetic response to IGF1 and TNFa %A Schneider, Sallie S %A Elizabeth M. Henchey %A Nazneen Sultana %A Stephanie M. Morin %A Jerry, D Joseph %A Makari-Judson, Grace %A Crisi, Giovanna M %A Arenas, Richard B %A Melissa Johnson %A Holly S. Mason %A Yadava, Nagendra %B Journal of Cellular Physiology %I Wiley-Blackwell %V 232 %P 2750–2765 %8 may/2017 %G eng %U https://doi.org/10.1002/jcp.25932 %0 Journal Article %J J Cell Physiol %D 2017 %T Individual-specific variation in the respiratory activities of HMECs and their bioenergetic response to IGF1 and TNFα. %A Schneider, Sallie S %A Henchey, Elizabeth M %A Sultana, Nazneen %A Morin, Stephanie M %A Jerry, D Joseph %A Makari-Judson, Grace %A Crisi, Giovanna M %A Arenas, Richard B %A Johnson, Melissa %A Mason, Holly S %A Yadava, Nagendra %K Adult %K Aged %K Breast Neoplasms %K Cell Respiration %K Energy Metabolism %K Epithelial Cells %K Female %K Humans %K Insulin-Like Growth Factor I %K Mammary Glands, Human %K Metabolomics %K Middle Aged %K Oxidation-Reduction %K Phenotype %K Pyruvic Acid %K Time Factors %K Tumor Cells, Cultured %K Tumor Necrosis Factor-alpha %K Young Adult %X

Metabolic 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.

%B J Cell Physiol %V 232 %P 2750-2765 %8 2017 Oct %G eng %N 10 %R 10.1002/jcp.25932 %0 Journal Article %J J Agric Food Chem %D 2017 %T Investigation of Pesticide Penetration and Persistence on Harvested and Live Basil Leaves Using Surface-Enhanced Raman Scattering Mapping. %A Yang, Tianxi %A Zhao, Bin %A Kinchla, Amanda J %A Clark, John M %A He, Lili %K Gold %K Metal Nanoparticles %K Ocimum basilicum %K Pesticide Residues %K Plant Leaves %K Spectrum Analysis, Raman %K Thiabendazole %X

Understanding 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.

%B J Agric Food Chem %V 65 %P 3541-3550 %8 2017 May 03 %G eng %N 17 %R 10.1021/acs.jafc.7b00548 %0 Journal Article %J eLife %D 2017 %T Modulation of let-7 miRNAs controls the differentiation of effector CD8 T cells %A Alexandria C Wells %A Keith A Daniels %A Constance C Angelou %A Eric Fagerberg %A Amy S Burnside %A Michele Markstein %A Alfandari, Dominique %A Raymond M Welsh %A Elena L Pobezinskaya %A Pobezinsky, Leonid A %B eLife %I {eLife} Sciences Organisation, Ltd. %V 6 %8 jul %G eng %U https://doi.org/10.7554/elife.26398 %0 Journal Article %J Andrology %D 2017 %T Molecular changes and signaling events occurring in spermatozoa during epididymal maturation %A M. G. Gervasi %A Visconti, P E %B Andrology %I Wiley-Blackwell %V 5 %P 204–218 %8 mar %G eng %U https://doi.org/10.1111/andr.12320 %0 Journal Article %J Animal Biotechnology %D 2017 %T Molecular Characterization and Comparison of Phospholipase C zeta (PLCZ1) Gene Between Swamp (Bubalus carabanensis) and Riverine (Bubalus bubalis) Buffaloes: Its Implications and Future Perspectives %A Eufrocina P. Atabay %A Roseline D. Tadeo %A Edwin C. Atabay %A Emma V. Venturina %A Fissore, Rafael A %A Claro N. Mingala %B Animal Biotechnology %I Informa {UK} Limited %P 1–9 %8 aug %G eng %U https://doi.org/10.1080/10495398.2017.1350689 %0 Journal Article %J Developmental Biology %D 2017 %T Pa2G4 is a novel Six1 co-factor that is required for neural crest and otic development %A Neilson, Karen M %A Genevieve Abbruzzesse %A Kristy Kenyon %A Vanessa Bartolo %A Patrick Krohn %A Alfandari, Dominique %A Moody, Sally A %B Developmental Biology %I Elsevier {BV} %V 421 %P 171–182 %8 jan %G eng %U https://doi.org/10.1016/j.ydbio.2016.11.021 %0 Journal Article %J Virulence %D 2017 %T Passive therapy with humanized anti-staphylococcal enterotoxin B antibodies attenuates systemic inflammatory response and protects from lethal pneumonia caused by staphylococcal enterotoxin B-producing Staphylococcus aureus %A Melissa J. Karau %A Tilahun, Mulualem E %A Ashton Krogman %A Osborne, Barbara A %A Goldsby, Richard A %A Chella S. David %A Jayawant N. Mandrekar %A Robin Patel %A Rajagopalan, Govindarajan %B Virulence %I Informa {UK} Limited %V 8 %P 1148–1159 %8 jan %G eng %U https://doi.org/10.1080/21505594.2016.1267894 %0 Journal Article %J Food Chem Toxicol %D 2017 %T Permethrin alters glucose metabolism in conjunction with high fat diet by potentiating insulin resistance and decreases voluntary activities in female C57BL/6J mice. %A Xiao, Xiao %A Kim, Yoo %A Kim, Daeyoung %A Yoon, Kyong Sup %A Clark, John M %A Park, Yeonhwa %K Animals %K Body Weight %K Dietary Fats %K Energy Intake %K Female %K Glucose %K Homeostasis %K Insecticides %K Insulin Resistance %K Mice %K Mice, Inbred C57BL %K Motor Activity %K Permethrin %X

Permethrin, 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.

%B Food Chem Toxicol %V 108 %P 161-170 %8 2017 Oct %G eng %N Pt A %R 10.1016/j.fct.2017.07.053 %0 Journal Article %J Food Chem Toxicol %D 2017 %T Permethrin decreased insulin-stimulated AKT phosphorylation dependent on extracellular signal-regulated kinase-1 (ERK), but not AMP-activated protein kinase α (AMPKα), in C2C12 myotubes. %A Sun, Quancai %A Peng, Ye %A Qi, Weipeng %A Kim, Yoo %A Clark, John M %A Kim, Daeyoung %A Park, Yeonhwa %K AMP-Activated Protein Kinases %K Animals %K Glucose Transporter Type 4 %K Insecticides %K Insulin %K Mice %K Mitogen-Activated Protein Kinase 3 %K Muscle Fibers, Skeletal %K Permethrin %K Phosphorylation %K Proto-Oncogene Proteins c-akt %K Signal Transduction %X

Previously 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.

%B Food Chem Toxicol %V 109 %P 95-101 %8 2017 Nov %G eng %N Pt 1 %R 10.1016/j.fct.2017.08.046 %0 Journal Article %J Food Chem Toxicol %D 2017 %T Permethrin potentiates adipogenesis via intracellular calcium and endoplasmic reticulum stress-mediated mechanisms in 3T3-L1 adipocytes. %A Xiao, Xiao %A Qi, Weipeng %A Clark, John M %A Park, Yeonhwa %K 3T3-L1 Cells %K Adipocytes %K Adipogenesis %K Animals %K Calcium %K Cell Differentiation %K Endoplasmic Reticulum %K Endoplasmic Reticulum Stress %K Insecticides %K Mice %K Permethrin %X

Permethrin, 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.

%B Food Chem Toxicol %V 109 %P 123-129 %8 2017 Nov %G eng %N Pt 1 %R 10.1016/j.fct.2017.08.049 %0 Journal Article %J Equine Veterinary Journal %D 2017 %T Pharmacokinetics of intravenous lithium chloride and assessment of agreement between two methods of lithium concentration measurement in the horse %A L. M. Martin %A A. D. Bukoski %A D. D. Whelchel %A T. J. Evans %A C. E. Wiedmeyer %A Black, S J %A P. J. Johnson %B Equine Veterinary Journal %I Wiley-Blackwell %8 dec %G eng %U https://doi.org/10.1111/evj.12778 %0 Journal Article %J Food Chem Toxicol %D 2017 %T Potential contribution of insecticide exposure and development of obesity and type 2 diabetes. %A Xiao, Xiao %A Clark, John M %A Park, Yeonhwa %K Animals %K Diabetes Mellitus, Type 2 %K Environmental Exposure %K Humans %K Insecticides %K Obesity %X

The 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.

%B Food Chem Toxicol %V 105 %P 456-474 %8 2017 Jul %G eng %R 10.1016/j.fct.2017.05.003 %0 Journal Article %J Journal of Cancer Epidemiology %D 2017 %T Relationships between Global DNA Methylation in Circulating White Blood Cells and Breast Cancer Risk Factors %A Nayha Chopra-Tandon %A Haotian Wu %A Arcaro, Kathleen F %A Sturgeon, Susan R %B Journal of Cancer Epidemiology %I Hindawi Limited %V 2017 %P 1–25 %G eng %U https://doi.org/10.1155/2017/2705860 %0 Journal Article %J {mBio} %D 2017 %T Role of Granulocyte-Macrophage Colony-Stimulating Factor Production by T Cells during Mycobacterium tuberculosis Infection %A Alissa C. Rothchild %A Britni Stowell %A Girija Goyal %A Cláudio Nunes-Alves %A Qianting Yang %A Kadamba Papavinasasundaram %A Christopher M. Sassetti %A Glenn Dranoff %A Xinchun Chen %A Jinhee Lee %A Samuel M. Behar %E Liise-anne Pirofski %B {mBio} %I American Society for Microbiology %V 8 %G eng %U https://doi.org/10.1128/mbio.01514-17 %0 Journal Article %J Chemistry - A European Journal %D 2017 %T ROMP- and RAFT-Based Guanidinium-Containing Polymers as Scaffolds for Protein Mimic Synthesis %A Joel M. Sarapas %A Coralie M. Backlund %A Brittany M. deRonde %A Minter, Lisa M %A Tew, Gregory N %B Chemistry - A European Journal %I Wiley %V 23 %P 6858–6863 %G eng %U https://doi.org/10.1002/chem.201700423 %0 Journal Article %J Journal of Control Release %D 2017 %T Sequence segregation improves non-covalent protein delivery %A Sgolastra, Federica %A Coralie M. Backlund %A E. I. Ozay %A Brittany M. deRonde %A Minter, Lisa M %A Tew, Gregory N %B Journal of Control Release %I Elsevier {BV} %V 254 %P 131–136 %8 may/2017 %G eng %U https://doi.org/10.1016/j.jconrel.2017.03.387 %0 Journal Article %J Scientific Reports %D 2017 %T Sperm-borne miR-449b influences cleavage, epigenetic reprogramming and apoptosis of SCNT embryos in bovine %A Mengyun Wang %A Yang Gao %A Pengxiang Qu %A Suzhu Qing %A Fang Qiao %A Yong Zhang %A Mager, Jesse %A Yongsheng Wang %B Scientific Reports %I Springer Nature %V 7 %8 oct %G eng %U https://doi.org/10.1038/s41598-017-13899-8 %0 Journal Article %J Environmental Toxicology and Chemistry %D 2017 %T Tools to minimize interlaboratory variability in vitellogenin gene expression monitoring programs %A Aaron Jastrow %A Denise A. Gordon %A Kasie M. Auger %A Punska, Elizabeth C %A Arcaro, Kathleen F %A Kristen Keteles %A Dana Winkelman %A David Lattier %A Adam Biales %A James M. Lazorchak %B Environmental Toxicology and Chemistry %I Wiley-Blackwell %V 36 %P 3102–3107 %8 aug %G eng %U https://doi.org/10.1002/etc.3885 %0 Journal Article %J J Am Heart Assoc %D 2017 %T Use of p53-Silenced Endothelial Progenitor Cells to Treat Ischemia in Diabetic Peripheral Vascular Disease. %A Kundu, Nabanita %A Domingues, Cleyton C %A Chou, Cyril %A Ahmadi, Neeki %A Houston, Sara %A Jerry, D Joseph %A Sen, Sabyasachi %K Animals %K Cyclin-Dependent Kinase Inhibitor p21 %K Diabetes Mellitus, Experimental %K Diabetes Mellitus, Type 1 %K Diabetes Mellitus, Type 2 %K Diabetic Angiopathies %K Disease Models, Animal %K Endothelial Progenitor Cells %K Gene Silencing %K Hindlimb %K Ischemia %K Mice %K Mice, Inbred C57BL %K Mice, Inbred NOD %K Mice, Knockout %K Muscle, Skeletal %K Neovascularization, Physiologic %K Nitric Oxide Synthase Type III %K Peripheral Vascular Diseases %K Platelet Endothelial Cell Adhesion Molecule-1 %K Regional Blood Flow %K Tumor Suppressor Protein p53 %K Vascular Endothelial Growth Factor A %X

BACKGROUND: 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.

%B J Am Heart Assoc %V 6 %8 2017 Apr 01 %G eng %N 4 %R 10.1161/JAHA.116.005146 %0 Journal Article %J Journal of the American Heart Association %D 2017 %T Use of p53-Silenced Endothelial Progenitor Cells to Treat Ischemia in Diabetic Peripheral Vascular Disease %A Nabanita Kundu %A Cleyton C. Domingues %A Cyril Chou %A Neeki Ahmadi %A Sara Houston %A Jerry, D Joseph %A Sen, Sabyasachi %B Journal of the American Heart Association %I Ovid Technologies (Wolters Kluwer Health) %V 6 %P e005146 %8 apr/2017 %G eng %U https://doi.org/10.1161/jaha.116.005146 %0 Journal Article %J Breast Cancer Research %D 2017 %T White blood cell DNA methylation and risk of breast cancer in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) %A Sturgeon, Susan R %A J. Richard Pilsner %A Arcaro, Kathleen F %A Kaoru Ikuma %A Haotian Wu %A Soon-Mi Kim %A Nayha Chopra-Tandon %A Adam R. Karpf %A Ziegler, Regina G %A Schairer, Catherine %A Balasubramanian, Raji %A Reckhow, David A %B Breast Cancer Research %I Springer Nature %V 19 %8 aug %G eng %U https://doi.org/10.1186/s13058-017-0886-6 %0 Journal Article %J Cellular Reprogramming %D 2017 %T YY1 Is Required for Posttranscriptional Stability of SOX2 and OCT4Proteins %A Wallingford, Mary C %A Hiller, Jacob %A Zhang, Kun %A Mager, Jesse %B Cellular Reprogramming %I Mary Ann Liebert Inc %V 19 %P 263–269 %8 aug/2017 %G eng %U https://doi.org/10.1089/cell.2017.0002 %0 Journal Article %J {EMBO} Molecular Medicine %D 2017 %T γ-Secretase inhibitors in cancer clinical trials are pharmacologically and functionally distinct. %A Yong Ran %A Fokhrul Hossain %A Antonio Pannuti %A Christian B Lessard %A Gabriela Z Ladd %A Joo In Jung %A Minter, Lisa M %A Osborne, Barbara A %A Miele, Lucio %A Golde, Todd E %B {EMBO} Molecular Medicine %I {EMBO} %V 9 %P 950–966 %8 may %G eng %U https://doi.org/10.15252/emmm.201607265 %0 Journal Article %J Developmental Biology %D 2016 %T ADAM13 cleavage of cadherin-11 promotes CNC migration independently of the homophilic binding site %A Abbruzzese, Genevieve %A Becker, Sarah F %A Kashef, Jubin %A Alfandari, Dominique %B Developmental Biology %I Elsevier {BV} %V 415 %P 383–390 %8 jul %G eng %U https://doi.org/10.1016/j.ydbio.2015.07.018 %0 Journal Article %J Environ Sci Technol %D 2016 %T Alteration of the Nonsystemic Behavior of the Pesticide Ferbam on Tea Leaves by Engineered Gold Nanoparticles. %A Hou, Ruyan %A Zhang, Zhiyun %A Pang, Shintaro %A Yang, Tianxi %A Clark, John M %A He, Lili %K Gold %K Metal Nanoparticles %K Pesticides %K Spectrum Analysis, Raman %K Tea %X

A 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.

%B Environ Sci Technol %V 50 %P 6216-23 %8 2016 06 21 %G eng %N 12 %R 10.1021/acs.est.6b01336 %0 Journal Article %J Nature Communications %D 2016 %T Cadherin-11 localizes to focal adhesions and promotes cell–substrate adhesion %A Rahul P. Langhe %A Tetyana Gudzenko %A Michael Bachmann %A Becker, Sarah F %A Carina Gonnermann %A Claudia Winter %A Abbruzzese, Genevieve %A Alfandari, Dominique %A Marie-Claire Kratzer %A Clemens M. Franz %A Kashef, Jubin %B Nature Communications %I Springer Nature %V 7 %P 10909 %8 mar %G eng %U https://doi.org/10.1038/ncomms10909 %0 Journal Article %J The American Journal of Pathology %D 2016 %T Cellular Prion Protein Mediates Pancreatic Cancer Cell Survival and Invasion through Association with and Enhanced Signaling of Notch1 %A Yiwei Wang %A Shuiliang Yu %A Dan Huang %A Min Cui %A Huankai Hu %A Lihua Zhang %A Weihuan Wang %A Neetha Parameswaran %A Mark Jackson %A Osborne, Barbara %A Barbara Bedogni %A Chaoyang Li %A Man-Sun Sy %A Wei Xin %A Lan Zhou %B The American Journal of Pathology %I Elsevier {BV} %V 186 %P 2945–2956 %8 nov %G eng %U https://doi.org/10.1016/j.ajpath.2016.07.010 %0 Journal Article %J Mol Reprod Dev %D 2016 %T Chang's meaning of capacitation: A molecular perspective. %A Gervasi, Maria Gracia %A Visconti, Pablo E %X

Dr. 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.

%B Mol Reprod Dev %8 2016 Jun 3 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/27256723?dopt=Abstract

%0 Journal Article %J Cancer Epidemiol Biomarkers Prev %D 2016 %T Depression, Antidepressant Use, and Postmenopausal Breast Cancer Risk. %A Brown, Susan B %A Hankinson, Susan E %A Arcaro, Kathleen F %A Qian, Jing %A Reeves, Katherine W %X

BACKGROUND: 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.

%B Cancer Epidemiol Biomarkers Prev %V 25 %P 158-64 %8 2016 Jan %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/26578537?dopt=Abstract

%0 Journal Article %J Cancer Cell Int %D 2016 %T ELF5 and DOK7 regulation in anti-estrogen treated cells and tumors. %A Fitzgerald, Lily M %A Browne, Eva P %A Christie, Kevin D %A Punska, Elizabeth C %A Simmons, Leo O %A Williams, Kristin E %A Pentecost, Brian T %A M Jawale, Rahul %A Otis, Christopher N %A Arcaro, Kathleen F %X

BACKGROUND: 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.

%B Cancer Cell Int %V 16 %P 8 %8 2016 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/26884724?dopt=Abstract

%0 Journal Article %J Journal of Cellular Physiology %D 2016 %T Essential Role of CFTR in PKA-Dependent Phosphorylation, Alkalinization, and Hyperpolarization During Human Sperm Capacitation %A Lis C. Puga Molina %A Nicolás A. Pinto %A Paulina Torres Rodriguez %A Romarowski, Ana %A Alberto Vicens Sanchez %A Visconti, Pablo E %A Darszon, Alberto %A Treviño, Claudia L %A Buffone, Mariano G %B Journal of Cellular Physiology %I Wiley-Blackwell %V 232 %P 1404–1414 %8 oct %G eng %U https://doi.org/10.1002/jcp.25634 %0 Journal Article %J J Food Sci %D 2016 %T Evaluation of the Penetration of Multiple Classes of Pesticides in Fresh Produce Using Surface-Enhanced Raman Scattering Mapping. %A Yang, Tianxi %A Zhao, Bin %A Hou, Ruyan %A Zhang, Zhiyun %A Kinchla, Amanda J %A Clark, John M %A He, Lili %X

Understanding 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.

%B J Food Sci %V 81 %P T2891-T2901 %8 2016 Nov %G eng %N 11 %R 10.1111/1750-3841.13520 %0 Journal Article %J Food Chem Toxicol %D 2016 %T Fipronil promotes adipogenesis via AMPKα-mediated pathway in 3T3-L1 adipocytes. %A Sun, Quancai %A Qi, Weipeng %A Yang, Jeremy J %A Yoon, Kyong Sup %A Clark, John M %A Park, Yeonhwa %K 3T3-L1 Cells %K Acetyl-CoA Carboxylase %K Adipocytes %K Adipogenesis %K Aminoimidazole Carboxamide %K AMP-Activated Protein Kinases %K Animals %K CCAAT-Enhancer-Binding Protein-alpha %K Cell Differentiation %K Cells, Cultured %K Gene Expression Regulation %K Immunoblotting %K Insecticides %K Mice %K PPAR gamma %K Pyrazoles %K Real-Time Polymerase Chain Reaction %K Reverse Transcriptase Polymerase Chain Reaction %K Ribonucleotides %K RNA, Messenger %K Signal Transduction %K Triglycerides %X

Emerging 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.

%B Food Chem Toxicol %V 92 %P 217-23 %8 2016 Jun %G eng %R 10.1016/j.fct.2016.04.011 %0 Journal Article %J Proceedings of the National Academy of Sciences %D 2016 %T HIF-KDM3A-MMP12 regulatory circuit ensures trophoblast plasticity and placental adaptations to hypoxia %A Damayanti Chakraborty %A Cui, Wei %A Gracy X. Rosario %A Regan L. Scott %A Pramod Dhakal %A Stephen J. Renaud %A Makoto Tachibana %A M. A. Karim Rumi %A Clifford W. Mason %A Adam J. Krieg %A Michael J. Soares %B Proceedings of the National Academy of Sciences %I Proceedings of the National Academy of Sciences %V 113 %P E7212–E7221 %8 nov %G eng %U https://doi.org/10.1073/pnas.1612626113 %0 Journal Article %J J Agric Food Chem %D 2016 %T Imidacloprid Promotes High Fat Diet-Induced Adiposity and Insulin Resistance in Male C57BL/6J Mice. %A Sun, Quancai %A Xiao, Xiao %A Kim, Yoo %A Kim, Daeyoung %A Yoon, Kyoon Sup %A Clark, John M %A Park, Yeonhwa %K Adipose Tissue, White %K Adiposity %K AMP-Activated Protein Kinases %K Animals %K Blood Glucose %K Diet, High-Fat %K Humans %K Imidazoles %K Insecticides %K Insulin %K Insulin Resistance %K Liver %K Male %K Mice %K Mice, Inbred C57BL %K Neonicotinoids %K Nitro Compounds %K Obesity %K Weight Gain %X

Imidacloprid, 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.

%B J Agric Food Chem %V 64 %P 9293-9306 %8 2016 Dec 14 %G eng %N 49 %R 10.1021/acs.jafc.6b04322 %0 Journal Article %J Chem %D 2016 %T Immunomodulatory effects of coated gold nanoparticles in LPS-stimulated and murine model systems. %A Moyano, Daniel F %A Liu, Yuanchang %A Ayaz, Furkan %A Hou, Singyuk %A Puangploy, Premsak %A Duncan, Bradley %A Osborne, Barbara A %A Rotello, Vincent M %X

The 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.

%B Chem %V 1 %P 320-327 %8 2016 %G eng %N 2 %R 10.1016/j.chempr.2016.07.007 %0 Journal Article %J Polymer Chemistry %D 2016 %T Increased hydrophobic block length of PTDMs promotes protein internalization %A Coralie M. Backlund %A Sgolastra, Federica %A Ronja Otter %A Minter, Lisa M %A Toshihide Takeuchi %A Shiroh Futaki %A Tew, Gregory N %B Polymer Chemistry %I Royal Society of Chemistry ({RSC}) %V 7 %P 7514–7521 %G eng %U https://doi.org/10.1039/c6py01615d %0 Journal Article %J Molecular Therapy %D 2016 %T Intracellular Delivery of Anti-pPKC0 (Thr538) via Protein Transduction Domain Mimics for Immunomodulation %A E. I. Ozay %A Gonzalez-Perez, Gabriela %A Joe A Torres %A Jyothi Vijayaraghavan %A Rebecca Lawlor %A Heather L Sherman %A Daniel T Garrigan %A Amy S Burnside %A Osborne, Barbara A %A Tew, Gregory N %A Minter, Lisa M %B Molecular Therapy %I Elsevier {BV} %V 24 %P 2118–2130 %8 dec %G eng %U https://doi.org/10.1038/mt.2016.177 %0 Journal Article %J Mol Ther %D 2016 %T Intracellular Delivery of Anti-pPKCθ (Thr538) via Protein Transduction Domain Mimics for Immunomodulation. %A Ozay, E Ilker %A Gonzalez-Perez, Gabriela %A Torres, Joe A %A Vijayaraghavan, Jyothi %A Lawlor, Rebecca %A Sherman, Heather L %A Garrigan, Daniel T %A Burnside, Amy S %A Osborne, Barbara A %A Tew, Gregory N %A Minter, Lisa M %K Animals %K Antibodies, Monoclonal, Humanized %K Cell Differentiation %K Cell Proliferation %K Cell-Penetrating Peptides %K Graft vs Host Disease %K Humans %K Immunomodulation %K Isoenzymes %K Leukocytes, Mononuclear %K Lymphocyte Activation %K Mice %K Phosphorylation %K Protein Kinase C %K Protein Kinase C-theta %K Signal Transduction %K Th1 Cells %X

Targeting 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.

%B Mol Ther %V 24 %P 2118-2130 %8 2016 Dec %G eng %N 12 %R 10.1038/mt.2016.177 %0 Journal Article %J BioMed Research International %D 2016 %T Male Infertility: Genetics, Mechanism, and Therapies %A Charles Coutton %A Fissore, Rafael A %A Gianpiero D. Palermo %A Katrien Stouffs %A Aminata Touré %B BioMed Research International %I Hindawi Limited %V 2016 %P 1–1 %G eng %U https://doi.org/10.1155/2016/7372362 %0 Journal Article %J Pediatr Dermatol %D 2016 %T Management of Head Louse Infestations in the United States-A Literature Review. %A Koch, Ellen %A Clark, John Marshall %A Cohen, Bernard %A Meinking, Terri L %A Ryan, William G %A Stevenson, Audrey %A Yetman, Robert %A Yoon, Kyong Sup %K Administration, Topical %K Animals %K Databases, Factual %K Drug Combinations %K Female %K Hexachlorocyclohexane %K Humans %K Incidence %K Insecticides %K Ivermectin %K Lice Infestations %K Macrolides %K Male %K Pediculus %K Risk Assessment %K Treatment Outcome %K United States %K United States Food and Drug Administration %X

UNLABELLED: 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.

%B Pediatr Dermatol %V 33 %P 466-72 %8 2016 Sep %G eng %N 5 %R 10.1111/pde.12982 %0 Journal Article %J Biomacromolecules %D 2016 %T Mapping Optimal Charge Density and Length of ROMP-Based PTDMs for siRNA Internalization %A Leah M. Caffrey %A Brittany M. deRonde %A Minter, Lisa M %A Tew, Gregory N %B Biomacromolecules %I American Chemical Society ({ACS}) %V 17 %P 3205–3212 %G eng %U https://doi.org/10.1021/acs.biomac.6b00900 %0 Journal Article %J Proceedings of the National Academy of Sciences %D 2016 %T MiR-155–regulated molecular network orchestrates cell fate in the innate and adaptive immune response to Mycobacterium tuberculosis %A Alissa C. Rothchild %A James R. Sissons %A Shahin Shafiani %A Christopher Plaisier %A Deborah Min %A Dat Mai %A Mark Gilchrist %A Jacques Peschon %A Ryan P. Larson %A Andreas Bergthaler %A Nitin S Baliga %A Kevin B. Urdahl %A Alan Aderem %B Proceedings of the National Academy of Sciences %I Proceedings of the National Academy of Sciences %V 113 %P E6172–E6181 %G eng %U https://doi.org/10.1073/pnas.1608255113 %0 Journal Article %J Biomacromolecules %D 2016 %T Optimal Hydrophobicity in Ring-Opening Metathesis Polymerization-Based Protein Mimics Required for siRNA Internalization %A Brittany M. deRonde %A Nicholas D. Posey %A Ronja Otter %A Leah M. Caffrey %A Minter, Lisa M %A Tew, Gregory N %B Biomacromolecules %I American Chemical Society ({ACS}) %V 17 %P 1969–1977 %G eng %U https://doi.org/10.1021/acs.biomac.6b00138 %0 Journal Article %J Biomacromolecules %D 2016 %T Optimal Hydrophobicity in Ring-Opening Metathesis Polymerization-Based Protein Mimics Required for siRNA Internalization %A Brittany M. deRonde %A Nicholas D. Posey %A Ronja Otter %A Leah M. Caffrey %A Minter, Lisa M %A Tew, Gregory N %B Biomacromolecules %I American Chemical Society ({ACS}) %V 17 %P 1969–1977 %8 jun %G eng %U https://doi.org/10.1021/acs.biomac.6b00138 %0 Journal Article %J Breast Cancer Res Treat %D 2016 %T Potential of breastmilk analysis to inform early events in breast carcinogenesis: rationale and considerations. %A Murphy, Jeanne %A Sherman, Mark E %A Browne, Eva P %A Caballero, Ana I %A Punska, Elizabeth C %A Pfeiffer, Ruth M %A Yang, Hannah P %A Lee, Maxwell %A Yang, Howard %A Gierach, Gretchen L %A Arcaro, Kathleen F %X

This 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.

%B Breast Cancer Res Treat %V 157 %P 13-22 %8 2016 May %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/27107568?dopt=Abstract

%0 Journal Article %J Parasite Immunology %D 2016 %T Prospects for vaccination against pathogenic African trypanosomes %A Black, S J %A J. M. Mansfield %B Parasite Immunology %I Wiley-Blackwell %V 38 %P 735–743 %8 dec %G eng %U https://doi.org/10.1111/pim.12387 %0 Journal Article %J Experimental Cell Research %D 2016 %T Rab3A, a possible marker of cortical granules, participates in cortical granule exocytosis in mouse eggs %A Oscar Daniel Bello %A Andrea Isabel Cappa %A Matilde de Paola %A Maria Natalia Zanetti %A Mitsunori Fukuda %A Fissore, Rafael A %A Luis S. Mayorga %A Marcela A. Michaut %B Experimental Cell Research %I Elsevier {BV} %V 347 %P 42–51 %8 sep %G eng %U https://doi.org/10.1016/j.yexcr.2016.07.005 %0 Journal Article %J Anal Chem %D 2016 %T Real-Time and in Situ Monitoring of Pesticide Penetration in Edible Leaves by Surface-Enhanced Raman Scattering Mapping. %A Yang, Tianxi %A Zhang, Zhiyun %A Zhao, Bin %A Hou, Ruyan %A Kinchla, Amanda %A Clark, John M %A He, Lili %K Gold %K Metal Nanoparticles %K Pesticides %K Plant Leaves %K Spectrum Analysis, Raman %K Spinacia oleracea %X

Understanding 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.

%B Anal Chem %V 88 %P 5243-50 %8 2016 05 17 %G eng %N 10 %R 10.1021/acs.analchem.6b00320 %0 Journal Article %J Proceedings of the National Academy of Sciences %D 2016 %T Rethinking progesterone regulation of female reproductive cyclicity %A Kaiyu Kubota %A Cui, Wei %A Pramod Dhakal %A Michael W. Wolfe %A M. A. Karim Rumi %A Jay L. Vivian %A Katherine F. Roby %A Michael J. Soares %B Proceedings of the National Academy of Sciences %I Proceedings of the National Academy of Sciences %V 113 %P 4212–4217 %8 mar %G eng %U https://doi.org/10.1073/pnas.1601825113 %0 Journal Article %J Adv Anat Embryol Cell Biol %D 2016 %T Sperm Capacitation and Acrosome Reaction in Mammalian Sperm. %A Stival, Cintia %A Puga Molina, Lis Del C %A Paudel, Bidur %A Buffone, Mariano G %A Visconti, Pablo E %A Krapf, Dario %X

Physiological 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.

%B Adv Anat Embryol Cell Biol %V 220 %P 93-106 %8 2016 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/27194351?dopt=Abstract

%0 Journal Article %J Sci Rep %D 2016 %T Splice form variant and amino acid changes in MDR49 confers DDT resistance in transgenic Drosophila. %A Seong, Keon Mook %A Sun, Weilin %A Clark, John M %A Pittendrigh, Barry R %K Alternative Splicing %K Amino Acid Sequence %K Amino Acids %K Animals %K Animals, Genetically Modified %K ATP Binding Cassette Transporter, Subfamily B %K DDT %K Drosophila melanogaster %K Drosophila Proteins %K Insecticide Resistance %K Mutation %K Open Reading Frames %K Protein Isoforms %K Sequence Analysis, RNA %X

The 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.

%B Sci Rep %V 6 %P 23355 %8 2016 Mar 22 %G eng %R 10.1038/srep23355 %0 Journal Article %J Scientific Reports %D 2016 %T Towards Functional Annotation of the Preimplantation Transcriptome: An RNAi Screen in Mammalian Embryos %A Cui, Wei %A Dai, Xiangpeng %A Marcho, Chelsea %A Han, Zhengbin %A Zhang, Kun %A Tremblay, Kimberly D %A Mager, Jesse %B Scientific Reports %I Springer Nature %V 6 %8 nov %G eng %U https://doi.org/10.1038/srep37396 %0 Journal Article %J Scientific Reports %D 2016 %T Transient exposure to calcium ionophore enables in vitro fertilization in sterile mouse models %A Navarrete, Felipe A %A Alvau, Antonio %A Lee, Hoi Chang %A Levin, Lonny R %A Buck, Jochen %A Patricia Martin-De Leon %A Celia M. Santi %A Krapf, Dario %A Mager, Jesse %A Fissore, Rafael A %A Salicioni, Ana M %A Darszon, Alberto %A Visconti, Pablo E %B Scientific Reports %I Springer Nature %V 6 %8 sep %G eng %U https://doi.org/10.1038/srep33589 %0 Journal Article %J Scientific Reports %D 2016 %T TRPM7-like channels are functionally expressed in oocytes and modulate post-fertilization embryo development in mouse %A Ingrid Carvacho %A Goli Ardestani %A Lee, Hoi Chang %A Kaitlyn McGarvey %A Fissore, Rafael A %A Karin Lykke-Hartmann %B Scientific Reports %I Springer Nature %V 6 %8 sep %G eng %U https://doi.org/10.1038/srep34236 %0 Journal Article %J Cell Calcium %D 2016 %T TRPV3 channels mediate Ca2+ influx induced by 2-APB in mouse eggs %A Lee, Hoi Chang %A Yoon, Sook-Young %A Karin Lykke-Hartmann %A Fissore, Rafael A %A Ingrid Carvacho %B Cell Calcium %I Elsevier {BV} %V 59 %P 21–31 %8 jan %G eng %U https://doi.org/10.1016/j.ceca.2015.12.001 %0 Journal Article %J {PLOS} Pathogens %D 2016 %T Trypanosoma brucei Co-opts NK Cells to Kill Splenic B2 B Cells %A Frenkel, Deborah %A Fengqiu Zhang %A Guirnalda, Patrick %A Carole Haynes %A Bockstal, Viki %A Radwanska, Magdalena %A Magez, Stefan %A Black, Samuel J %E Jude Ezeh Uzonna %B {PLOS} Pathogens %I Public Library of Science ({PLoS}) %V 12 %P e1005733 %8 jul %G eng %U https://doi.org/10.1371/journal.ppat.1005733 %0 Journal Article %J Development %D 2016 %T The tyrosine kinase FER is responsible for the capacitation-associated increase in tyrosine phosphorylation in murine sperm. %A Alvau, Antonio %A Battistone, Maria Agustina %A Gervasi, Maria Gracia %A Navarrete, Felipe A %A Xu, Xinran %A Sánchez-Cárdenas, Claudia %A De la Vega-Beltran, Jose Luis %A Da Ros, Vanina G %A Greer, Peter %A Darszon, Alberto %A Krapf, Diego %A Salicioni, Ana Maria %A Cuasnicu, Patricia %A Visconti, Pablo E %X

Sperm 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.

%B Development %8 2016 May 25 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/27226326?dopt=Abstract

%0 Journal Article %J Dev Biol %D 2015 %T ADAM13 cleavage of cadherin-11 promotes CNC migration independently of the homophilic binding site. %A Abbruzzese, Genevieve %A Becker, Sarah F %A Kashef, Jubin %A Alfandari, Dominique %X

The 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.

%B Dev Biol %8 2015 Jul 21 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/26206614?dopt=Abstract

%0 Journal Article %J Mol Biol Evol %D 2015 %T Alternative Splice in Alternative Lice. %A Tovar-Corona, Jaime M %A Castillo-Morales, Atahualpa %A Chen, Lu %A Olds, Brett P %A Clark, John M %A Reynolds, Stuart E %A Pittendrigh, Barry R %A Feil, Edward J %A Urrutia, Araxi O %K Alternative Splicing %K Animals %K Gene Ontology %K Genes, Insect %K Humans %K Pediculus %K Phthiraptera %X

Genomic 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.

%B Mol Biol Evol %V 32 %P 2749-59 %8 2015 Oct %G eng %N 10 %R 10.1093/molbev/msv151 %0 Journal Article %J Epigenetics %D 2015 %T Analysis of DNA methylation and gene expression in radiation-resistant head and neck tumors. %A Chen, Xiaofei %A Liu, Liang %A Mims, Jade %A Punska, Elizabeth C %A Williams, Kristin E %A Zhao, Weiling %A Arcaro, Kathleen F %A Tsang, Allen W %A Zhou, Xiaobo %A Furdui, Cristina M %K Azacitidine %K Cell Line, Tumor %K CpG Islands %K Cyclin D2 %K DNA Methylation %K Gene Expression Regulation, Neoplastic %K Head and Neck Neoplasms %K Humans %K Promoter Regions, Genetic %K Radiation Tolerance %X

Resistance 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.

%B Epigenetics %V 10 %P 545-61 %8 2015 %G eng %N 6 %1

http://www.ncbi.nlm.nih.gov/pubmed/25961636?dopt=Abstract

%0 Journal Article %J Cancer Causes Control %D 2015 %T Association of TGF-β2 levels in breast milk with severity of breast biopsy diagnosis. %A Yang, Hannah P %A Schneider, Sallie Smith %A Chisholm, Christina M %A Browne, Eva P %A Mahmood, Sidra %A Gierach, Gretchen L %A Lenington, Sarah %A Anderton, Douglas L %A Sherman, Mark E %A Arcaro, Kathleen F %X

PURPOSE: 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.

%B Cancer Causes Control %V 26 %P 345-54 %8 2015 Mar %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/25604865?dopt=Abstract

%0 Journal Article %J J Cell Physiol %D 2015 %T Biphasic Role of Calcium in Mouse Sperm Capacitation Signaling Pathways. %A Navarrete, Felipe A %A Francisco A. García-Vázquez %A Alvau, Antonio %A Escoffier, Jessica %A Krapf, Dario %A Sánchez-Cárdenas, Claudia %A Salicioni, Ana M %A Darszon, Alberto %A Visconti, Pablo E %X

Mammalian 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.

%B J Cell Physiol %8 2015 Jan 17 %G ENG %1 http://www.ncbi.nlm.nih.gov/pubmed/25597298?dopt=Abstract %0 Journal Article %J Developmental Dynamics %D 2015 %T cis-regulatory control of Mesp1 expression by YY1 and SP1 during mouse embryogenesis %A Beketaev, Ilimbek %A Zhang, Yi %A Weng, Kuo-Chan %A Rhee, Siyeon %A Yu, Wei %A Liu, Yu %A Mager, Jesse %A Wang, Jun %B Developmental Dynamics %I Wiley-Blackwell %V 245 %P 379–387 %8 nov %G eng %U https://doi.org/10.1002/dvdy.24349 %0 Journal Article %J Dev Dyn %D 2015 %T cis-regulatory control of Mesp1 expression by YY1 and SP1 during mouse embryogenesis. %A Beketaev, Ilimbek %A Zhang, Yi %A Weng, Kuo-Chan %A Rhee, Siyeon %A Yu, Wei %A Liu, Yu %A Mager, Jesse %A Wang, Jun %X

BACKGROUND: 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.

%B Dev Dyn %8 2015 Sep 18 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/26384464?dopt=Abstract

%0 Journal Article %J Blood %D 2015 %T CXCR4 expression on pathogenic T cells facilitates their bone marrow infiltration in a mouse model of aplastic anemia %A C. Arieta Kuksin %A G. Gonzalez-Perez %A L. M. Minter %B Blood %I American Society of Hematology %V 125 %P 2087–2094 %8 feb %G eng %U https://doi.org/10.1182/blood-2014-08-594796 %0 Journal Article %J Biomacromolecules %D 2015 %T Development of Guanidinium-Rich Protein Mimics for Efficient siRNA Delivery into Human T Cells %A Brittany M. deRonde %A Joe A Torres %A Minter, Lisa M %A Tew, Gregory N %B Biomacromolecules %I American Chemical Society ({ACS}) %V 16 %P 3172–3179 %8 oct %G eng %U https://doi.org/10.1021/acs.biomac.5b00795 %0 Journal Article %J Applied and Environmental Microbiology %D 2015 %T Direct Image-Based Enumeration of Clostridium phytofermentans Cells on Insoluble Plant Biomass Growth Substrates %A Jesús G. Alvelo-Maurosa %A Scott J. Lee %A Samuel P. Hazen %A Susan B. Leschine %E R. M. Kelly %B Applied and Environmental Microbiology %I American Society for Microbiology %V 82 %P 972–978 %8 dec %G eng %U https://doi.org/10.1128/aem.03119-15 %0 Journal Article %J Dev Biol %D 2015 %T Editorial: Cell adhesion in development. %A Bajanca, Fernanda %A Alfandari, Dominique %A Thorsteinsdóttir, Sólveig %A Théveneau, Eric %B Dev Biol %V 401 %P 1 %8 2015 May 1 %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/25882001?dopt=Abstract

%0 Journal Article %J Reproduction %D 2015 %T Epigenetic dynamics during preimplantation development %A Marcho, Chelsea %A Cui, Wei %A Mager, Jesse %B Reproduction %I {BioScientifica} %V 150 %P R109–R120 %8 jun %G eng %U https://doi.org/10.1530/rep-15-0180 %0 Journal Article %J Biochem Biophys Res Commun %D 2015 %T Expression patterns of long noncoding RNAs from Dlk1-Dio3 imprinted region and the potential mechanisms of Gtl2 activation during blastocyst development. %A Han, Zhengbin %A Yu, Changwei %A Tian, Yijun %A Zeng, Tiebo %A Cui, Wei %A Mager, Jesse %A Wu, Qiong %X

The 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.

%B Biochem Biophys Res Commun %V 463 %P 167-73 %8 2015 Jul 31 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/26005002?dopt=Abstract

%0 Journal Article %J PLOS ONE %D 2015 %T Genome and Transcriptome of Clostridium phytofermentans, Catalyst for the Direct Conversion of Plant Feedstocks to Fuels %A Elsa Petit %A Maddalena V. Coppi %A James C. Hayes %A Andrew C. Tolonen %A Thomas Warnick %A William G. Latouf %A Danielle Amisano %A Amy Biddle %A Supratim Mukherjee %A Natalia Ivanova %A Athanassios Lykidis %A Miriam Land %A Loren Hauser %A Nikos Kyrpides %A Bernard Henrissat %A Joanne Lau %A Danny J. Schnell %A George M. Church %A Susan B. Leschine %A Blanchard, Jeffrey L %E Willem J.H. van Berkel %B PLOS ONE %I Public Library of Science ({PLoS}) %V 10 %P e0118285 %8 jun %G eng %U https://doi.org/10.1371/journal.pone.0118285 %0 Journal Article %J PLoS One %D 2015 %T A glycine insertion in the estrogen-related receptor (ERR) is associated with enhanced expression of three cytochrome P450 genes in transgenic Drosophila melanogaster. %A Sun, Weilin %A Valero, M Carmen %A Seong, Keon Mook %A Steele, Laura D %A Huang, I-Ting %A Lee, Chien-Hui %A Clark, John M %A Qiu, Xinghui %A Pittendrigh, Barry R %K Amino Acid Sequence %K Animals %K Animals, Genetically Modified %K Conserved Sequence %K Cytochrome P-450 Enzyme System %K Drosophila melanogaster %K Drosophila Proteins %K Enzyme Induction %K Female %K Gene Expression %K Glycine %K Male %K Molecular Sequence Data %K Mutagenesis, Insertional %K Phylogeny %K Receptors, Estrogen %X

Insecticide-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.

%B PLoS One %V 10 %P e0118779 %8 2015 %G eng %N 3 %R 10.1371/journal.pone.0118779 %0 Journal Article %J PLoS Pathogens %D 2015 %T Human and Murine Clonal CD8+ T Cell Expansions Arise during Tuberculosis Because of TCR Selection %A Cláudio Nunes-Alves %A Matthew G. Booty %A Stephen M. Carpenter %A Alissa C. Rothchild %A Constance J. Martin %A Danielle Desjardins %A Katherine Steblenko %A Henrik N. Kløverpris %A Rajhmun Madansein %A Duran Ramsuran %A Alasdair Leslie %A Margarida Correia-Neves %A Samuel M. Behar %E David M. Lewinsohn %B PLoS Pathogens %I Public Library of Science ({PLoS}) %V 11 %P e1004849 %G eng %U https://doi.org/10.1371/journal.ppat.1004849 %0 Journal Article %J The Journal of Immunology %D 2015 %T Impact of Notch1 Deletion in Macrophages on Proinflammatory Cytokine Production and the Outcome of Experimental Autoimmune Encephalomyelitis %A W. Wongchana %A R. G. Lawlor %A Osborne, B A %A T. Palaga %B The Journal of Immunology %I The American Association of Immunologists %V 195 %P 5337–5346 %8 oct %G eng %U https://doi.org/10.4049/jimmunol.1401770 %0 Journal Article %J Nat Immunol %D 2015 %T Let-7 microRNAs target the lineage-specific transcription factor PLZF to regulate terminal NKT cell differentiation and effector function. %A Pobezinsky, Leonid A %A Etzensperger, Ruth %A Jeurling, Susanna %A Alag, Amala %A Kadakia, Tejas %A McCaughtry, Tom M %A Kimura, Motoko Y %A Sharrow, Susan O %A Guinter, Terry I %A Feigenbaum, Lionel %A Singer, Alfred %X

Lethal-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.

%B Nat Immunol %V 16 %P 517-24 %8 2015 May %G eng %N 5 %1

http://www.ncbi.nlm.nih.gov/pubmed/25848867?dopt=Abstract

%0 Journal Article %J Frontiers in Oncology %D 2015 %T The Link between Autoimmunity and Lymphoma: Does NOTCH Signaling Play a Contributing Role? %A Kuksin, Christina Arieta %A Minter, Lisa M %B Frontiers in Oncology %I Frontiers Media {SA} %V 5 %8 feb %G eng %U https://doi.org/10.3389/fonc.2015.00051 %0 Journal Article %J Development %D 2015 %T Mga is essential for the survival of pluripotent cells during peri-implantation development. %A Washkowitz, Andrew J %A Schall, Caroline %A Zhang, Kun %A Wurst, Wolfgang %A Floss, Thomas %A Mager, Jesse %A Papaioannou, Virginia E %K Alleles %K Animals %K Apoptosis %K Blastocyst Inner Cell Mass %K Cell Differentiation %K Cell Proliferation %K Cell Survival %K Cells, Cultured %K Crosses, Genetic %K Embryo Implantation %K Embryonic Stem Cells %K Female %K Gene Knockdown Techniques %K Gene Targeting %K Genotype %K Germ Layers %K Male %K Mice %K Mutagenesis %K Mutation %K Ornithine Decarboxylase %K Pluripotent Stem Cells %K Polyamines %K Transcription Factors %X

The 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.

%B Development %V 142 %P 31-40 %8 2015 Jan 1 %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/25516968?dopt=Abstract

%0 Journal Article %J Neuroscience %D 2015 %T Microarray analysis of neonatal rat anteroventral periventricular transcriptomes identifies the proapoptotic Cugbp2 gene as sex-specific and regulated by estradiol. %A Del Pino Sans, J %A Krishnan, S %A Aggison, L K %A Adams, H L %A Shrikant, M M %A López-Giráldez, F %A Petersen, S L %K Analysis of Variance %K Animals %K Animals, Newborn %K CELF Proteins %K Estradiol %K Female %K Gene Expression Regulation, Developmental %K Hypothalamus, Anterior %K Male %K Nerve Tissue Proteins %K Oligonucleotide Array Sequence Analysis %K Pregnancy %K Rats %K Rats, Sprague-Dawley %K Receptors, GABA %K Receptors, Glutamate %K RNA, Messenger %K Sex Differentiation %K Transcriptome %X

Sexually 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.

%B Neuroscience %V 303 %P 312-22 %8 2015 Sep 10 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/26166732?dopt=Abstract

%0 Journal Article %J Mol Reprod Dev %D 2015 %T Nop2 is required for mammalian preimplantation development. %A Cui, Wei %A Pizzollo, Jason %A Han, Zhengbin %A Marcho, Chelsea %A Zhang, Kun %A Mager, Jesse %X

Nucleolar 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.

%B Mol Reprod Dev %8 2015 Dec 3 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/26632338?dopt=Abstract

%0 Journal Article %J Insect Biochem Mol Biol %D 2015 %T Odorant receptor-based discovery of natural repellents of human lice. %A Pelletier, Julien %A Xu, Pingxi %A Yoon, Kyong S %A Clark, John M %A Leal, Walter S %K Animals %K Electrophysiological Phenomena %K Female %K Insect Repellents %K Insect Vectors %K Oocytes %K Pediculus %K Receptors, Odorant %K Smell %K Xenopus laevis %X

The 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.

%B Insect Biochem Mol Biol %V 66 %P 103-9 %8 2015 Nov %G eng %R 10.1016/j.ibmb.2015.10.009 %0 Journal Article %J Cellular Reprogramming %D 2015 %T Optimized Protocols for In Vitro Maturation of Rat Oocytes Dramatically Improve Their Developmental Competence to a Level Similar to That of Ovulated Oocytes %A Guang-Zhong Jiao %A Cui, Wei %A Rui Yang %A Juan Lin %A Shuai Gong %A Hua-Yu Lian %A Ming-Ju Sun %A Jing-He Tan %B Cellular Reprogramming %I Mary Ann Liebert Inc %8 dec %G eng %U https://doi.org/10.1089/cell.2015.0055 %0 Journal Article %J Dev Biol %D 2015 %T PKA-dependent phosphorylation of LIMK1 and Cofilin is essential for mouse sperm acrosomal exocytosis. %A Romarowski, Ana %A Battistone, Maria A %A La Spina, Florenza A %A Puga Molina, Lis Del C %A Luque, Guillermina M %A Vitale, Alejandra M %A Cuasnicu, Patricia S %A Visconti, Pablo E %A Krapf, Dario %A Buffone, Mariano G %K Acrosome Reaction %K Actins %K Animals %K Cofilin 1 %K Crosses, Genetic %K Cyclic AMP-Dependent Protein Kinases %K Exocytosis %K Gene Expression Regulation %K Lim Kinases %K Male %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Mice, Transgenic %K Microscopy, Fluorescence %K Phosphorylation %K Signal Transduction %K Sperm Capacitation %K Spermatozoa %X

Mammalian 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.

%B Dev Biol %V 405 %P 237-49 %8 2015 Sep 15 %G eng %N 2 %1

http://www.ncbi.nlm.nih.gov/pubmed/26169470?dopt=Abstract

%0 Journal Article %J Biomacromolecules %D 2015 %T Polyamide Nanogels from Generally Recognized as Safe Components and Their Toxicity in Mouse Preimplantation Embryos. %A Prasad, Priyaa %A Molla, Mijanur Rahaman %A Cui, Wei %A Canakci, Mine %A Osborne, Barbara %A Mager, Jesse %A Thayumanavan, S %X

Safe 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.

%B Biomacromolecules %V 16 %P 3491-8 %8 2015 Nov 9 %G eng %N 11 %1

http://www.ncbi.nlm.nih.gov/pubmed/26367020?dopt=Abstract

%0 Journal Article %J Cell Rep %D 2015 %T Prostate tumorigenesis induced by PTEN deletion involves estrogen receptor β repression. %A Mak, Paul %A Li, Jiarong %A Samanta, Sanjoy %A Chang, Cheng %A Jerry, D Joseph %A Davis, Roger J %A Leav, Irwin %A Mercurio, Arthur M %X

The 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.

%B Cell Rep %V 10 %P 1982-91 %8 2015 Mar 31 %G eng %N 12 %1

http://www.ncbi.nlm.nih.gov/pubmed/25818291?dopt=Abstract

%0 Journal Article %J Cell Rep %D 2015 %T Prostate tumorigenesis induced by PTEN deletion involves estrogen receptor β repression. %A Mak, Paul %A Li, Jiarong %A Samanta, Sanjoy %A Chang, Cheng %A Jerry, D Joseph %A Davis, Roger J %A Leav, Irwin %A Mercurio, Arthur M %K Animals %K Cell Line, Tumor %K Cell Transformation, Neoplastic %K Estrogen Receptor beta %K Gene Expression Regulation, Neoplastic %K Humans %K Male %K Mice %K Prostatic Neoplasms %K PTEN Phosphohydrolase %K Signal Transduction %X

The 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.

%B Cell Rep %V 10 %P 1982-91 %8 2015 Mar 31 %G eng %N 12 %R 10.1016/j.celrep.2015.02.063 %0 Journal Article %J PLoS One %D 2015 %T Selective 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. %A Steele, Laura D %A Coates, Brad %A Valero, M Carmen %A Sun, Weilin %A Seong, Keon Mook %A Muir, William M %A Clark, John M %A Pittendrigh, Barry R %K Animals %K DDT %K Drosophila melanogaster %K Genome %K Insecticide Resistance %X

Adaptation 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.

%B PLoS One %V 10 %P e0123066 %8 2015 %G eng %N 3 %R 10.1371/journal.pone.0123066 %0 Journal Article %J Epigenetics Chromatin %D 2015 %T Tissue-specific regulation of Igf2r/Airn imprinting during gastrulation. %A Marcho, Chelsea %A Bevilacqua, Ariana %A Tremblay, Kimberly D %A Mager, Jesse %X

BACKGROUND: 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.

%B Epigenetics Chromatin %V 8 %P 10 %8 2015 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/25918552?dopt=Abstract

%0 Journal Article %J Am J Epidemiol %D 2015 %T Urinary melatonin concentration and the risk of breast cancer in Nurses' Health Study II. %A Brown, Susan B %A Hankinson, Susan E %A Eliassen, A Heather %A Reeves, Katherine W %A Qian, Jing %A Arcaro, Kathleen F %A Wegrzyn, Lani R %A Willett, Walter C %A Schernhammer, Eva S %X Experimental 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. %B Am J Epidemiol %V 181 %P 155-62 %8 2015 Feb 1 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/25587174?dopt=Abstract %0 Journal Article %J Comparative Biochemistry and Physiology Part C: Toxicology {&} Pharmacology %D 2015 %T Using Xenopus to discover new genes involved in branchiootorenal spectrum disorders %A Moody, Sally A %A Neilson, Karen M %A Kenyon, Kristy L %A Alfandari, Dominique %A Pignoni, Francesca %B Comparative Biochemistry and Physiology Part C: Toxicology {&} Pharmacology %I Elsevier {BV} %V 178 %P 16–24 %8 dec %G eng %U https://doi.org/10.1016/j.cbpc.2015.06.007 %0 Journal Article %J Comp Biochem Physiol C Toxicol Pharmacol %D 2015 %T Using Xenopus to discover new genes involved in Branchiootorenal spectrum disorders. %A Moody, Sally A %A Neilson, Karen M %A Kenyon, Kristy L %A Alfandari, Dominique %A Pignoni, Francesca %X

Congenital 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.

%B Comp Biochem Physiol C Toxicol Pharmacol %8 2015 Jun 24 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/26117063?dopt=Abstract

%0 Journal Article %J Pestic Biochem Physiol %D 2015 %T Utilization of the human louse genome to study insecticide resistance and innate immune response. %A Clark, J Marshall %A Yoon, Kyong Sup %A Kim, Ju Hyeon %A Lee, Si Hyeock %A Pittendrigh, Barry R %K Animals %K Biological Assay %K Genome, Insect %K Humans %K Immunity, Innate %K Insecticide Resistance %K Insecticides %K Pediculus %K Pyrethrins %X

Since 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.

%B Pestic Biochem Physiol %V 120 %P 125-32 %8 2015 May %G eng %R 10.1016/j.pestbp.2014.11.008 %0 Journal Article %J J Cell Sci %D 2015 %T The Wnt receptor Frizzled-4 modulates ADAM13 metalloprotease activity. %A Abbruzzese, Genevieve %A Gorny, Anne-Kathrin %A Kaufmann, Lilian T %A Cousin, Hélène %A Kleino, Iivari %A Steinbeisser, Herbert %A Alfandari, Dominique %X

Cranial 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.

%B J Cell Sci %V 128 %P 1139-49 %8 2015 Mar 15 %G eng %N 6 %1

http://www.ncbi.nlm.nih.gov/pubmed/25616895?dopt=Abstract

%0 Journal Article %J BMC Microbiol %D 2014 %T Activation of epidermal growth factor receptor is required for Chlamydia trachomatis development. %A Patel, Achchhe L %A Chen, Xiaofei %A Wood, Scott T %A Stuart, Elizabeth S %A Arcaro, Kathleen F %A Molina, Doris P %A Petrovic, Snezana %A Furdui, Cristina M %A Tsang, Allen W %X Background 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. %B BMC Microbiol %V 14 %P 277 %8 2014 Dec 4 %G ENG %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/25471819?dopt=Abstract %0 Journal Article %J Chemosphere %D 2014 %T Determination of free Bisphenol A (BPA) concentrations in breast milk of U.S. women using a sensitive LC/MS/MS method. %A Zimmers, Stephanie M %A Browne, Eva P %A O'Keefe, Patrick W %A Anderton, Douglas L %A Kramer, Lawrence %A Reckhow, David A %A Arcaro, Kathleen F %X

Bisphenol 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.

%B Chemosphere %V 104 %P 237-43 %8 2014 Jun %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/24507723?dopt=Abstract

%0 Journal Article %J Methods Mol Biol %D 2014 %T Dissecting DR3 signaling. %A Pobezinskaya, Yelena L %A Liu, Zhenggang %A Choksi, Swati %X

Receptor 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.

%B Methods Mol Biol %V 1155 %P 15-22 %8 2014 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/24788169?dopt=Abstract

%0 Journal Article %J Anticancer Res %D 2014 %T DNA methylation in paired breast epithelial and white blood cells from women undergoing reduction mammoplasty. %A Sturgeon, Susan R %A Arcaro, Kathleen F %A Johnson, Melissa A %A Balasubramanian, Raji %A Zorn, Martha %A Jerry, D Joseph %A Schneider, Sallie Smith %K Adolescent %K Adult %K Breast Neoplasms %K DNA Methylation %K DNA, Neoplasm %K Female %K Genes, Tumor Suppressor %K Humans %K Leukocytes %K Long Interspersed Nucleotide Elements %K Mammaplasty %K Middle Aged %K Neoplasms, Glandular and Epithelial %K Polymerase Chain Reaction %K Prognosis %K Young Adult %X

BACKGROUND: 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.

%B Anticancer Res %V 34 %P 2985-90 %8 2014 Jun %G eng %N 6 %1

http://www.ncbi.nlm.nih.gov/pubmed/24922663?dopt=Abstract

%0 Journal Article %J Nat Commun %D 2014 %T EphrinB2 affects apical constriction in Xenopus embryos and is regulated by ADAM10 and flotillin-1. %A Ji, Yon Ju %A Hwang, Yoo-Seok %A Mood, Kathleen %A Cho, Hee-Jun %A Lee, Hyun-Shik %A Winterbottom, Emily %A Cousin, Hélène %A Daar, Ira O %X

The 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.

%B Nat Commun %V 5 %P 3516 %8 2014 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/24662724?dopt=Abstract

%0 Journal Article %J Mol Hum Reprod %D 2014 %T Evidence for the involvement of proline-rich tyrosine kinase 2 in tyrosine phosphorylation downstream of protein kinase A activation during human sperm capacitation. %A Battistone, M A %A Alvau, A %A Salicioni, A M %A Visconti, P E %A Da Ros, V G %A Cuasnicú, P S %X

Sperm 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.

%B Mol Hum Reprod %8 2014 Sep 1 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/25180269?dopt=Abstract

%0 Journal Article %J PLoS One %D 2014 %T Genome-wide sequencing and an open reading frame analysis of dichlorodiphenyltrichloroethane (DDT) susceptible (91-C) and resistant (91-R) Drosophila melanogaster laboratory populations. %A Steele, Laura D %A Muir, William M %A Seong, Keon Mook %A Valero, M Carmen %A Rangesa, Madhumitha %A Sun, Weilin %A Clark, John M %A Coates, Brad %A Pittendrigh, Barry R %K Alleles %K Amino Acid Substitution %K Animals %K Chromosome Mapping %K Chromosomes, Insect %K DDT %K Drosophila melanogaster %K Drosophila Proteins %K Drug Resistance %K Female %K Genome, Insect %K Genome-Wide Association Study %K High-Throughput Nucleotide Sequencing %K Male %K Mutation %K Open Reading Frames %K Polymorphism, Single Nucleotide %X

The 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.

%B PLoS One %V 9 %P e98584 %8 2014 %G eng %N 6 %R 10.1371/journal.pone.0098584 %0 Journal Article %J Mol Biol Cell %D 2014 %T GSK3 and Polo-like kinase regulate ADAM13 function during cranial neural crest cell migration. %A Abbruzzese, Genevieve %A Cousin, Hélène %A Salicioni, Ana Maria %A Alfandari, Dominique %X

ADAMs 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.

%B Mol Biol Cell %8 2014 Oct 8 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/25298404?dopt=Abstract

%0 Journal Article %J PLoS Negl Trop Dis %D 2014 %T Heterogeneous feeding patterns of the dengue vector, Aedes aegypti, on individual human hosts in rural Thailand. %A Harrington, Laura C %A Fleisher, Andrew %A Ruiz-Moreno, Diego %A Vermeylen, Francoise %A Wa, Chrystal V %A Poulson, Rebecca L %A Edman, John D %A Clark, John M %A Jones, James W %A Kitthawee, Sangvorn %A Scott, Thomas W %K Adolescent %K Adult %K Aedes %K Animals %K Child %K Dengue %K Feeding Behavior %K Female %K Humans %K Insect Bites and Stings %K Insect Vectors %K Male %K Thailand %X

BACKGROUND: 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.

%B PLoS Negl Trop Dis %V 8 %P e3048 %8 2014 Aug %G eng %N 8 %R 10.1371/journal.pntd.0003048 %0 Journal Article %J Epigenetics %D 2014 %T High-density array analysis of DNA methylation in tamoxifen-resistant breast cancer cell lines. %A Williams, Kristin E %A Anderton, Douglas L %A Lee, Maxwell P %A Pentecost, Brian T %A Arcaro, Kathleen F %X

Roughly 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.

%B Epigenetics %V 9 %8 2014 Feb %G ENG %N 2 %1

http://www.ncbi.nlm.nih.gov/pubmed/24225485?dopt=Abstract

%0 Journal Article %J Infect Agent Cancer %D 2014 %T Human Mammary Tumor Virus (HMTV) sequences in human milk. %A Nartey, Teiko %A Moran, Heberth %A Marin, Tania %A Arcaro, Kathleen F %A Anderton, Douglas L %A Etkind, Polly %A Holland, James F %A Melana, Stella M %A Pogo, Beatriz G-T %X

BACKGROUND: 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.

%B Infect Agent Cancer %V 9 %P 20 %8 2014 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/25120582?dopt=Abstract

%0 Journal Article %J Gene Expr Patterns %D 2014 %T Identification of a novel isoform of the leukemia-associated MLLT1 (ENL/LTG19) protein. %A Wallingford, Mary C %A Filkins, Rachel %A Adams, Danielle %A Walentuk, Melanie %A Salicioni, Ana Maria %A Visconti, Pablo E %A Mager, Jesse %X

Genome 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.

%B Gene Expr Patterns %V 17 %P 11-15 %8 2014 Dec 3 %G ENG %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/25481096?dopt=Abstract

%0 Journal Article %J Biol Reprod %D 2014 %T Impaired Sperm Maturation in Rnase9 Knockout Mice. %A Westmuckett, Andrew D %A Nguyen, Edward B %A Herlea-Pana, Oana M %A Alvau, Antonio %A Salicioni, Ana M %A Moore, Kevin Lee %X

Ribonuclease, 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.

%B Biol Reprod %8 2014 Apr 9 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/24719258?dopt=Abstract

%0 Journal Article %J Biomacromolecules %D 2014 %T Importance of sequence specific hydrophobicity in synthetic protein transduction domain mimics. %A Sgolastra, Federica %A Minter, Lisa M %A Osborne, Barbara A %A Tew, Gregory N %X

A 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.

%B Biomacromolecules %V 15 %P 812-20 %8 2014 Mar 10 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/24506414?dopt=Abstract

%0 Journal Article %J Nature Reviews Microbiology %D 2014 %T In search of a new paradigm for protective immunity to TB %A Cláudio Nunes-Alves %A Matthew G. Booty %A Stephen M. Carpenter %A Pushpa Jayaraman %A Alissa C. Rothchild %A Samuel M. Behar %B Nature Reviews Microbiology %I Springer Science and Business Media {LLC} %V 12 %P 289–299 %G eng %U https://doi.org/10.1038/nrmicro3230 %0 Journal Article %J PLoS Pathogens %D 2014 %T iNKT Cell Production of GM-CSF Controls Mycobacterium tuberculosis %A Alissa C. Rothchild %A Pushpa Jayaraman %A Cláudio Nunes-Alves %A Samuel M. Behar %E David M. Lewinsohn %B PLoS Pathogens %I Public Library of Science ({PLoS}) %V 10 %P e1003805 %G eng %U https://doi.org/10.1371/journal.ppat.1003805 %0 Journal Article %J J Med Entomol %D 2014 %T Knockdown resistance allele frequencies in North American head louse (Anoplura: Pediculidae) populations. %A Yoon, Kyong Sup %A Previte, Domenic J %A Hodgdon, Hilliary E %A Poole, Bryan C %A Kwon, Deok Ho %A El-Ghar, Gamal E Abo %A Lee, Si Hyeock %A Clark, J Marshall %K Animals %K Canada %K Gene Frequency %K Genotyping Techniques %K Insecticide Resistance %K Insecticides %K Mutation %K Pediculus %K Permethrin %K Sodium Channels %K United States %X

The 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.

%B J Med Entomol %V 51 %P 450-7 %8 2014 Mar %G eng %N 2 %R 10.1603/me13139 %0 Journal Article %J PlosOne %D 2014 %T Measuring cytotoxicity by bioluminescence imaging outperforms the standard chromium-51 release assay. %A Kambayashi, Taku %A Behrens, Edward M. %A Bachmann, Michael H. %A Salicioni, Ana Maria %A Baldwin, Cynthia L %A Karimi, Mobin A. %A Lee, Eric %K cytotoxicity %B PlosOne %V 10.1371/journal.pone.0089357 %8 02/2014 %G eng %U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0089357 %0 Journal Article %J Front Immunol %D 2014 %T Non-Canonical Notch Signaling Drives Activation and Differentiation of Peripheral CD4(+) T Cells. %A Dongre, Anushka %A Surampudi, Lalitha %A Lawlor, Rebecca G %A Fauq, Abdul H %A Miele, Lucio %A Golde, Todd E %A Minter, Lisa M %A Osborne, Barbara A %X

Cleavage 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.

%B Front Immunol %V 5 %P 54 %8 2014 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/24611064?dopt=Abstract

%0 Journal Article %J Mol Biol Evol %D 2014 %T A nonsynonymous mutation in the transcriptional regulator lbh is associated with cichlid craniofacial adaptation and neural crest cell development. %A Powder, Kara E %A Cousin, Hélène %A McLinden, Gretchen P %A Craig Albertson, R %X

Since 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.

%B Mol Biol Evol %V 31 %P 3113-24 %8 2014 Dec %G eng %N 12 %1

http://www.ncbi.nlm.nih.gov/pubmed/25234704?dopt=Abstract

%0 Journal Article %J Front Immunol %D 2014 %T NOTCH1 Can Initiate NF-κB Activation via Cytosolic Interactions with Components of the T Cell Signalosome. %A Shin, Hyun Mu %A Tilahun, Mulualem E %A Cho, Ok Hyun %A Chandiran, Karthik %A Kuksin, Christina Arieta %A Keerthivasan, Shilpa %A Fauq, Abdul H %A Golde, Todd E %A Miele, Lucio %A Thome, Margot %A Osborne, Barbara A %A Minter, Lisa M %X

T 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.

%B Front Immunol %V 5 %P 249 %8 2014 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/24904593?dopt=Abstract

%0 Journal Article %J Breast Cancer Res %D 2014 %T Parity-related molecular signatures and breast cancer subtypes by estrogen receptor status. %A Rotunno, Melissa %A Sun, Xuezheng %A Figueroa, Jonine %A Sherman, Mark E %A Garcia-Closas, Montserrat %A Meltzer, Paul %A Williams, Tyisha %A Schneider, Sallie Smith %A Jerry, D Joseph %A Yang, Xiaohong R %A Troester, Melissa A %K Adolescent %K Adult %K Aged %K Breast Neoplasms %K Case-Control Studies %K Cluster Analysis %K Female %K Gene Expression Profiling %K Humans %K Middle Aged %K Odds Ratio %K Parity %K Pregnancy %K Receptors, Estrogen %K Risk Factors %K Transcriptome %K Young Adult %X

INTRODUCTION: 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.

%B Breast Cancer Res %V 16 %P R74 %8 2014 %G eng %N 4 %1

http://www.ncbi.nlm.nih.gov/pubmed/25005139?dopt=Abstract

%0 Journal Article %J Breast Cancer Res %D 2014 %T Parity-related molecular signatures and breast cancer subtypes by estrogen receptor status. %A Rotunno, Melissa %A Sun, Xuezheng %A Figueroa, Jonine %A Sherman, Mark E %A Garcia-Closas, Montserrat %A Meltzer, Paul %A Williams, Tyisha %A Schneider, Sallie Smith %A Jerry, D Joseph %A Yang, Xiaohong R %A Troester, Melissa A %K Adolescent %K Adult %K Aged %K Breast Neoplasms %K Case-Control Studies %K Cluster Analysis %K Female %K Gene Expression Profiling %K Humans %K Middle Aged %K Odds Ratio %K Parity %K Pregnancy %K Receptors, Estrogen %K Risk Factors %K Transcriptome %K Young Adult %X

INTRODUCTION: 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.

%B Breast Cancer Res %V 16 %P R74 %8 2014 Jul 08 %G eng %N 4 %R 10.1186/bcr3689 %0 Journal Article %J J Hum Lact %D 2014 %T Promoter Methylation in Epithelial-Enriched and Epithelial-Depleted Cell Populations Isolated from Breast Milk. %A Browne, Eva P %A Dinc, Signem E %A Punska, Elizabeth C %A Agus, Sami %A Vitrinel, Ayca %A Erdag, Gulay Ciler %A Anderton, Douglas L %A Arcaro, Kathleen F %A Yilmaz, Bayram %X

BACKGROUND: 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.

%B J Hum Lact %8 2014 Aug 27 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/25164041?dopt=Abstract

%0 Journal Article %J Journal of Assisted Reproduction and Genetics %D 2014 %T Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters %A Lee, Hoi Chang %A Margaret Arny %A Grow, Daniel %A Daniel Dumesic %A Fissore, Rafael A %A Teru Jellerette-Nolan %B Journal of Assisted Reproduction and Genetics %I Springer Nature %V 31 %P 749–756 %8 apr %G eng %U https://doi.org/10.1007/s10815-014-0229-9 %0 Journal Article %J Nature %D 2014 %T RLIM is dispensable for X-chromosome inactivation in the mouse embryonic epiblast. %A Shin, JongDae %A Wallingford, Mary C %A Gallant, Judith %A Marcho, Chelsea %A Jiao, Baowei %A Byron, Meg %A Bossenz, Michael %A Lawrence, Jeanne B %A Jones, Stephen N %A Mager, Jesse %A Bach, Ingolf %K Animals %K Down-Regulation %K Embryo Implantation %K Embryo, Mammalian %K Female %K Germ Layers %K Histones %K In Situ Hybridization, Fluorescence %K Lysine %K Methylation %K Mice %K Mice, Knockout %K RNA, Long Noncoding %K Ubiquitin-Protein Ligases %K X Chromosome Inactivation %X

In 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.

%B Nature %V 511 %P 86-9 %8 2014 Jul 3 %G eng %N 7507 %1

http://www.ncbi.nlm.nih.gov/pubmed/24870238?dopt=Abstract

%0 Journal Article %J Journal of Cellular Physiology %D 2014 %T Role of Caspase-3 Cleaved IP3R1 on Ca2+ Homeostasis and Developmental Competence of Mouse Oocytes and Eggs %A Zhang, Nan %A Fissore, Rafael A %B Journal of Cellular Physiology %I Wiley-Blackwell %V 229 %P 1842–1854 %8 jul %G eng %U https://doi.org/10.1002/jcp.24638 %0 Journal Article %J {PLoS} {ONE} %D 2014 %T The Role of Lipolysis Stimulated Lipoprotein Receptor in Breast Cancer and Directing Breast Cancer Cell Behavior %A Denise K. Reaves %A Fagan-Solis, Katerina D %A Karen Dunphy %A Shannon D. Oliver %A David W. Scott %A Jodie M. Fleming %E Alan S. Fanning %B {PLoS} {ONE} %I Public Library of Science ({PLoS}) %V 9 %P e91747 %8 mar %G eng %U https://doi.org/10.1371/journal.pone.0091747 %0 Journal Article %J PLoS ONE %D 2014 %T Role of Na+/Ca2+ Exchanger (NCX) in Modulating Postovulatory Aging of Mouse and Rat Oocytes %A Chuan-Xin Zhang %A Cui, Wei %A Min Zhang %A Jie Zhang %A Tian-Yang Wang %A Jiang Zhu %A Guang-Zhong Jiao %A Jing-He Tan %E Qing-Yuan Sun %B PLoS ONE %I Public Library of Science ({PLoS}) %V 9 %P e93446 %8 apr %G eng %U https://doi.org/10.1371/journal.pone.0093446 %0 Journal Article %J J Cell Physiol %D 2014 %T SKP2 overexpression is associated with increased serine 10 phosphorylation of p27 (pSer10p27) in triple-negative breast cancer. %A Fagan-Solis, Katerina D %A Pentecost, Brian T %A Gozgit, Joseph M %A Bentley, Brooke A %A Marconi, Sharon M %A Otis, Christopher N %A Anderton, Douglas L %A Schneider, Sallie Smith %A Arcaro, Kathleen F %K Adult %K Age Factors %K Aged %K Aged, 80 and over %K Cell Cycle %K Cell Proliferation %K Cyclin D1 %K Cyclin-Dependent Kinase 2 %K Cyclin-Dependent Kinase Inhibitor p27 %K Estrogen Receptor alpha %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K MCF-7 Cells %K Middle Aged %K Neoplasm Grading %K Neoplasm Invasiveness %K Phosphorylation %K RNA Interference %K S-Phase Kinase-Associated Proteins %K Serine %K Signal Transduction %K Transfection %K Triple Negative Breast Neoplasms %K Tumor Markers, Biological %K Up-Regulation %X

S-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.

%B J Cell Physiol %V 229 %P 1160-9 %8 2014 Sep %G eng %N 9 %1

http://www.ncbi.nlm.nih.gov/pubmed/24443386?dopt=Abstract

%0 Journal Article %J J Immunol %D 2014 %T Specific Recognition of Mycobacterial Protein and Peptide Antigens by γδ T Cell Subsets following Infection with Virulent Mycobacterium bovis. %A McGill, Jodi L %A Sacco, Randy E %A Baldwin, Cynthia L %A Telfer, Janice C %A Palmer, Mitchell V %A Waters, Ray W %X

Promoting 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.

%B J Immunol %8 2014 Feb 14 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/24532582?dopt=Abstract

%0 Journal Article %J Sci Signal %D 2014 %T T cell receptor stimulation impairs IL-7 receptor signaling by inducing expression of the microRNA miR-17 to target Janus kinase 1. %A Katz, Gil %A Pobezinsky, Leonid A %A Jeurling, Susanna %A Shinzawa, Miho %A Van Laethem, Francois %A Singer, Alfred %X

T 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.

%B Sci Signal %V 7 %P ra83 %8 2014 %G eng %N 340 %1

http://www.ncbi.nlm.nih.gov/pubmed/25161318?dopt=Abstract

%0 Journal Article %J Journal of Biological Chemistry %D 2014 %T TMTC1 and TMTC2 Are Novel Endoplasmic Reticulum Tetratricopeptide Repeat-containing Adapter Proteins Involved in Calcium Homeostasis %A Johan C. Sunryd %A Cheon, Banyoon %A Jill B. Graham %A Kristina M. Giorda %A Fissore, Rafael A %A Daniel N. Hebert %B Journal of Biological Chemistry %I American Society for Biochemistry {&} Molecular Biology ({ASBMB}) %V 289 %P 16085–16099 %8 apr %G eng %U https://doi.org/10.1074/jbc.m114.554071 %0 Journal Article %J Adv Exp Med Biol %D 2014 %T Using breast milk to assess breast cancer risk: the role of mass spectrometry-based proteomics. %A Schneider, Sallie S %A Aslebagh, Roshanak %A Wetie, Armand G Ngounou %A Sturgeon, Susan R %A Darie, Costel C %A Arcaro, Kathleen F %X

Although 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.

%B Adv Exp Med Biol %V 806 %P 399-408 %8 2014 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/24952194?dopt=Abstract

%0 Journal Article %J World J Clin Oncol %D 2014 %T Weight gain following breast cancer diagnosis: Implication and proposed mechanisms. %A Makari-Judson, Grace %A Braun, Barry %A Jerry, D Joseph %A Mertens, Wilson C %X

Weight 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.

%B World J Clin Oncol %V 5 %P 272-82 %8 2014 Aug 10 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/25114844?dopt=Abstract

%0 Journal Article %J World J Clin Oncol %D 2014 %T Weight gain following breast cancer diagnosis: Implication and proposed mechanisms. %A Makari-Judson, Grace %A Braun, Barry %A Jerry, D Joseph %A Mertens, Wilson C %X

Weight 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.

%B World J Clin Oncol %V 5 %P 272-82 %8 2014 Aug 10 %G eng %N 3 %R 10.5306/wjco.v5.i3.272 %0 Journal Article %J Cell Calcium %D 2013 %T Ca2 homeostas+is and regulation of ER Ca2+ in mammalian oocytes/eggs %A Wakai, Takuya %A Fissore, Rafael A %B Cell Calcium %I Elsevier {BV} %V 53 %P 63–67 %8 jan %G eng %U https://doi.org/10.1016/j.ceca.2012.11.010 %0 Book Section %B Energy and Environment Series %D 2013 %T CHAPTER 7. Technologies to Study Plant Biomass Fermentation Using the Model Bacterium Clostridium Phytofermentans %A Andrew C. Tolonen %A Elsa Petit %A Blanchard, Jeffrey L %A Tom Warnick %A Susan B. Leschine %B Energy and Environment Series %I Royal Society of Chemistry %P 114–139 %G eng %U https://doi.org/10.1039/9781849734738-00114 %0 Journal Article %J J Biol Chem %D 2013 %T Compartmentalization of Distinct cAMP Signaling Pathways in Mammalian Sperm. %A Wertheimer, Eva %A Krapf, Dario %A Vega-Beltran, José Luis %A Sánchez-Cárdenas, Claudia %A Navarrete, Felipe %A Haddad, Douglas %A Escoffier, Jessica %A Salicioni, Ana M %A Levin, Lonny R %A Buck, Jochen %A Mager, Jesse %A Darszon, Alberto %A Visconti, Pablo E %X

Fertilization 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.

%B J Biol Chem %8 2013 Oct 15 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/24129574?dopt=Abstract

%0 Journal Article %J Biology of Reproduction %D 2013 %T Control of Spontaneous Activation of Rat Oocytes by Regulating Plasma Membrane Na+/Ca2+ Exchanger Activities %A W. Cui %A J. Zhang %A C.-X. Zhang %A G.-Z. Jiao %A M. Zhang %A T.-Y. Wang %A M.-J. Luo %A J.-H. Tan %B Biology of Reproduction %I Oxford University Press ({OUP}) %V 88 %P 160–160 %8 may/2013 %G eng %U https://doi.org/10.1095/biolreprod.113.108266 %0 Journal Article %J Dev Biol %D 2013 %T CTR9/PAF1c regulates molecular lineage identity, histone H3K36 trimethylation and genomic imprinting during preimplantation development. %A Zhang, Kun %A Haversat, Jocelyn M %A Mager, Jesse %X

Genome-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.

%B Dev Biol %V 383 %P 15-27 %8 2013 Nov 1 %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/24036311?dopt=Abstract

%0 Journal Article %J Mol Hum Reprod %D 2013 %T Functional human sperm capacitation requires both bicarbonate-dependent PKA activation and down-regulation of Ser/Thr phosphatases by Src family kinases. %A Battistone, M A %A Da Ros, V G %A Salicioni, A M %A Navarrete, F A %A Krapf, D %A Visconti, P E %A Cuasnicú, P S %X

In 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.

%B Mol Hum Reprod %8 2013 May 19 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/23630234?dopt=Abstract

%0 Journal Article %J Cancer Cell %D 2013 %T A Genome-wide siRNA Screen Identifies Proteasome Addiction as a Vulnerability of Basal-like Triple-Negative Breast Cancer Cells. %A Petrocca, Fabio %A Altschuler, Gabriel %A Tan, Shen Mynn %A Mendillo, Marc L %A Yan, Haoheng %A Jerry, D Joseph %A Kung, Andrew L %A Hide, Winston %A Ince, Tan A %A Lieberman, Judy %X

Basal-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.

%B Cancer Cell %V 24 %P 182-96 %8 2013 Aug 12 %G eng %N 2 %1

http://www.ncbi.nlm.nih.gov/pubmed/23948298?dopt=Abstract

%0 Journal Article %J J Biol Chem %D 2013 %T Heat shock protein 90 functions to stabilize and activate the testis-specific serine/threonine kinases, a family of kinases essential for male fertility. %A Jha, Kula N %A Coleman, Alyssa R %A Wong, Lily %A Salicioni, Ana M %A Howcroft, Elizabeth %A Johnson, Gibbes R %K Animals %K Cercopithecus aethiops %K COS Cells %K Enzyme Stability %K Fertility %K HSP90 Heat-Shock Proteins %K Humans %K Male %K Mice %K Mice, Mutant Strains %K Proteasome Endopeptidase Complex %K Protein Kinase Inhibitors %K Protein-Serine-Threonine Kinases %K Proteolysis %K Spermatids %K Ubiquitination %X

Spermiogenesis 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.

%B J Biol Chem %V 288 %P 16308-20 %8 2013 Jun 7 %G eng %N 23 %1

http://www.ncbi.nlm.nih.gov/pubmed/23599433?dopt=Abstract

%0 Journal Article %J Theriogenology %D 2013 %T Identification of phospholipase C zeta in normospermic and teratospermic domestic cat sperm %A Ana Izabel S. Balbin Villaverde %A Eduardo G. Fioratti %A Fissore, Rafael A %A He, Changli %A Lee, Hoi Chang %A Fabiana F. Souza %A Fernanda C. Landim-Alvarenga %A Maria Denise Lopes %B Theriogenology %I Elsevier {BV} %V 80 %P 722–729 %8 oct %G eng %U https://doi.org/10.1016/j.theriogenology.2013.06.005 %0 Journal Article %J Nat Immunol %D 2013 %T IL-7 signaling must be intermittent, not continuous, during CD8⁺ T cell homeostasis to promote cell survival instead of cell death. %A Kimura, Motoko Y %A Pobezinsky, Leonid A %A Guinter, Terry I %A Thomas, Julien %A Adams, Anthony %A Park, Jung-Hyun %A Tai, Xuguang %A Singer, Alfred %K Animals %K CD8-Positive T-Lymphocytes %K Cell Death %K Cell Proliferation %K Cell Survival %K Gene Expression Regulation %K Homeostasis %K Interferon-gamma %K Interleukin-7 %K Lymphocyte Activation %K Mice %K Mice, Transgenic %K Receptors, Antigen, T-Cell %K Signal Transduction %K Time Factors %X

The 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.

%B Nat Immunol %V 14 %P 143-51 %8 2013 Feb %G eng %N 2 %1

http://www.ncbi.nlm.nih.gov/pubmed/23242416?dopt=Abstract

%0 Journal Article %J {PLoS} {ONE} %D 2013 %T Impact of Laminitis on the Canonical Wnt Signaling Pathway in Basal Epithelial Cells of the Equine Digital Laminae %A Wang, Le %A Erica A. Pawlak %A Johnson, Philip J %A Belknap, James K %A Susan Eades %A Sharon Stack %A Helene Cousin %A Black, Samuel J %E Elizabeth G. Laird %B {PLoS} {ONE} %I Public Library of Science ({PLoS}) %V 8 %P e56025 %8 feb %G eng %U https://doi.org/10.1371/journal.pone.0056025 %0 Journal Article %J J Mammary Gland Biol Neoplasia %D 2013 %T Impaired mitochondrial metabolism and mammary carcinogenesis. %A Yadava, Nagendra %A Schneider, Sallie S %A Jerry, D Joseph %A Kim, Chul %K Animals %K Breast Neoplasms %K Carcinogens, Environmental %K Disease Susceptibility %K Environmental Exposure %K Female %K Humans %K Mammary Glands, Animal %K Mammary Glands, Human %K Mitochondria %K Oxidative Phosphorylation %X

Mitochondrial 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.

%B J Mammary Gland Biol Neoplasia %V 18 %P 75-87 %8 2013 Mar %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/23269521?dopt=Abstract

%0 Journal Article %J {PLoS} {ONE} %D 2013 %T An In Vitro Model of the Horse Gut Microbiome Enables Identification of Lactate-Utilizing Bacteria that Differentially Respond to Starch Induction %A Amy S. Biddle %A Black, Samuel J %A Blanchard, Jeffrey L %E Stefan Bereswill %B {PLoS} {ONE} %I Public Library of Science ({PLoS}) %V 8 %P e77599 %8 oct %G eng %U https://doi.org/10.1371/journal.pone.0077599 %0 Journal Article %J PLOS ONE %D 2013 %T Involvement of a Bacterial Microcompartment in the Metabolism of Fucose and Rhamnose by Clostridium phytofermentans %A Elsa Petit %A W. Greg LaTouf %A Maddalena V. Coppi %A Thomas A. Warnick %A Devin Currie %A Igor Romashko %A Supriya Deshpande %A Kelly Haas %A Jesús G. Alvelo-Maurosa %A Colin Wardman %A Danny J. Schnell %A Susan B. Leschine %A Blanchard, Jeffrey L %E Valerie de Crécy-Lagard %B PLOS ONE %I Public Library of Science ({PLoS}) %V 8 %P e54337 %8 jan %G eng %U https://doi.org/10.1371/journal.pone.0054337 %0 Journal Article %J Cell %D 2013 %T Lck availability during thymic selection determines the recognition specificity of the T cell repertoire. %A Van Laethem, Francois %A Tikhonova, Anastasia N %A Pobezinsky, Leonid A %A Tai, Xuguang %A Kimura, Motoko Y %A Le Saout, Cécile %A Guinter, Terry I %A Adams, Anthony %A Sharrow, Susan O %A Bernhardt, Günter %A Feigenbaum, Lionel %A Singer, Alfred %K Animals %K Lymphocyte Specific Protein Tyrosine Kinase p56(lck) %K Major Histocompatibility Complex %K Mice %K Receptors, Antigen, T-Cell, alpha-beta %K Receptors, Virus %K Signal Transduction %K T-Lymphocytes %K Thymocytes %K Thymus Gland %X

Thymic 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.

%B Cell %V 154 %P 1326-41 %8 2013 Sep 12 %G eng %N 6 %1

http://www.ncbi.nlm.nih.gov/pubmed/24034254?dopt=Abstract

%0 Journal Article %J Equine Veterinary Journal %D 2013 %T Manual corneal thickness measurements of healthy equine eyes using a portable spectral-domain optical coherence tomography device %A C. G. Pirie %A A. F. Alario %A C. M Barysauskas %A C. Gradil %A C.K. Uricchio %B Equine Veterinary Journal %I Wiley %V 46 %P 631–634 %8 dec %G eng %U https://doi.org/10.1111/evj.12198 %0 Journal Article %J Biology of Reproduction %D 2013 %T Mechanisms by which a Lack of Germinal Vesicle (GV) Material Causes Oocyte Meiotic Defects: A Study Using Oocytes Manipulated to Replace GV with Primary Spermatocyte Nuclei %A Jie Zhang %A Cui, Wei %A Qing Li %A Tian-Yang Wang %A Hong-Shu Sui %A Jun-Zuo Wang %A Ming-Jiu Luo %A Jing-He Tan %B Biology of Reproduction %I Oxford University Press ({OUP}) %V 89 %8 oct/2013 %G eng %U https://doi.org/10.1095/biolreprod.113.111500 %0 Journal Article %J Cell Death Dis %D 2013 %T MiR203 mediates subversion of stem cell properties during mammary epithelial differentiation via repression of ΔNP63α and promotes mesenchymal-to-epithelial transition. %A DeCastro, A J %A Dunphy, K A %A Hutchinson, J %A Balboni, A L %A Cherukuri, P %A Jerry, D J %A DiRenzo, J %K Animals %K Cell Differentiation %K Cell Line %K Epithelial-Mesenchymal Transition %K G1 Phase Cell Cycle Checkpoints %K Homeodomain Proteins %K Humans %K Mammary Glands, Animal %K Mammary Glands, Human %K MCF-7 Cells %K Mice %K MicroRNAs %K Neoplastic Stem Cells %K Protein Isoforms %K RNA Interference %K RNA, Small Interfering %K Stem Cells %K Transcription Factors %K Tumor Suppressor Proteins %X

During 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.

%B Cell Death Dis %V 4 %P e514 %8 2013 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/23449450?dopt=Abstract

%0 Journal Article %J Animal Science Journal %D 2013 %T Molecular characteristics of horse phospholipase C zeta (PLCzeta) %A Kana Sato %A Wakai, Takuya %A Yasunari Seita %A Akiko Takizawa %A Fissore, Rafael A %A Ito, Junya %A Kashiwazaki, Naomi %B Animal Science Journal %I Wiley-Blackwell %V 84 %P 359–368 %8 feb %G eng %U https://doi.org/10.1111/asj.12044 %0 Journal Article %J Dev Dyn %D 2013 %T Morphogenetic analysis of peri-implantation development. %A Wallingford, Mary C %A Angelo, Jesse R %A Mager, Jesse %X

BACKGROUND: 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.

%B Dev Dyn %V 242 %P 1110-20 %8 2013 Sep %G eng %N 9 %1

http://www.ncbi.nlm.nih.gov/pubmed/23728800?dopt=Abstract

%0 Journal Article %J J Neuroendocrinol %D 2013 %T Nonclassical progesterone signalling molecules in the nervous system. %A Petersen, S L %A Intlekofer, K A %A Moura-Conlon, P J %A Brewer, D N %A Del Pino Sans, J %A Lopez, J A %K Central Nervous System %K Humans %K Progesterone %K Receptors, Progesterone %K Signal Transduction %X

Progesterone (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.

%B J Neuroendocrinol %V 25 %P 991-1001 %8 2013 Nov %G eng %N 11 %1

http://www.ncbi.nlm.nih.gov/pubmed/23763432?dopt=Abstract

%0 Journal Article %J Rare Dis %D 2013 %T NOTCH signaling in immune-mediated bone marrow failure of aplastic anemia. %A Minter, Lisa M %X

Severe 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.

%B Rare Dis %V 1 %P e26764 %8 2013 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/25003012?dopt=Abstract

%0 Journal Article %J Mol Ther %D 2013 %T Novel protein transduction domain mimics as nonviral delivery vectors for siRNA targeting NOTCH1 in primary human T cells. %A Tezgel, A Özgül %A Gonzalez-Perez, Gabriela %A Telfer, Janice C %A Osborne, Barbara A %A Minter, Lisa M %A Tew, Gregory N %K CD4-Positive T-Lymphocytes %K Cell Differentiation %K Gene Knockdown Techniques %K Humans %K Jurkat Cells %K Receptor, Notch1 %K RNA Interference %K RNA, Small Interfering %K Transduction, Genetic %X

RNA 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.

%B Mol Ther %V 21 %P 201-9 %8 2013 Jan %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/23070119?dopt=Abstract

%0 Journal Article %J Cancer Cell Int %D 2013 %T Oncogenic transformation of mammary epithelial cells by transforming growth factor beta independent of mammary stem cell regulation. %A Dunphy, Karen A %A Seo, Jae-Hong %A Kim, Daniel J %A Roberts, Amy L %A Tao, Luwei %A Direnzo, James %A Balboni, Amanda L %A Crisi, Giovanna M %A Hagen, Mary J %A Chandrasekaran, Thiruppavai %A Gauger, Kelly J %A Schneider, Sallie Smith %A Jerry, D Joseph %X

BACKGROUND: 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.

%B Cancer Cell Int %V 13 %P 74 %8 2013 Jul 25 %G ENG %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/23883065?dopt=Abstract

%0 Journal Article %J J Natl Cancer Inst %D 2013 %T Postpartum Remodeling, Lactation, and Breast Cancer Risk: Summary of a National Cancer Institute-Sponsored Workshop. %A Faupel-Badger, Jessica M %A Arcaro, Kathleen F %A Balkam, Jane J %A Eliassen, A Heather %A Hassiotou, Foteini %A Lebrilla, Carlito B %A Michels, Karin B %A Palmer, Julie R %A Schedin, Pepper %A Stuebe, Alison M %A Watson, Christine J %A Sherman, Mark E %X

The 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.

%B J Natl Cancer Inst %8 2013 Nov 13 %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/23264680?dopt=Abstract

%0 Journal Article %J Breast Cancer Res %D 2013 %T Pregnancy offers new insights into mechanisms of breast cancer risk and resistance. %A Jerry, D Joseph %A Makari-Judson, Grace %A Crisi, Giovanna M %A Dunphy, Karen A %K Breast Neoplasms %K Cell Lineage %K Cyclin-Dependent Kinase Inhibitor p27 %K Female %K Gene Expression Profiling %K Humans %K Mammary Glands, Human %K Parity %K Pregnancy %K Stem Cells %X

Pregnancy 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.

%B Breast Cancer Res %V 15 %P 312 %8 2013 %G eng %N 5 %1

http://www.ncbi.nlm.nih.gov/pubmed/24060354?dopt=Abstract

%0 Journal Article %J J Cell Biochem %D 2013 %T The 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. %A Fagan-Solis, Katerina D %A Schneider, Sallie Smith %A Pentecost, Brian T %A Bentley, Brook A %A Otis, Christopher N %A Gierthy, John F %A Arcaro, Kathleen F %X

Breast 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.

%B J Cell Biochem %8 2013 Jun %G ENG %1

http://www.ncbi.nlm.nih.gov/pubmed/23255405?dopt=Abstract

%0 Journal Article %J Proceedings of the National Academy of Sciences %D 2013 %T Recognition of microbial and mammalian phospholipid antigens by NKT cells with diverse TCRs %A R. V. V. Tatituri %A G. F. M. Watts %A V. Bhowruth %A N. Barton %A A. Rothchild %A F.-F. Hsu %A C. F. Almeida %A L. R. Cox %A L. Eggeling %A S. Cardell %A J. Rossjohn %A D. I. Godfrey %A S. M. Behar %A G. S. Besra %A M. B. Brenner %A M. Brigl %B Proceedings of the National Academy of Sciences %I Proceedings of the National Academy of Sciences %V 110 %P 1827–1832 %G eng %U https://doi.org/10.1073/pnas.1220601110 %0 Journal Article %J Journal of Cell Science %D 2013 %T Regulation of endoplasmic reticulum Ca 2+ oscillations in mammalian eggs %A Wakai, Takuya %A Zhang, Nan %A Peter Vangheluwe %A Fissore, Rafael A %B Journal of Cell Science %I The Company of Biologists %V 126 %P 5714–5724 %8 oct %G eng %U https://doi.org/10.1242/jcs.136549 %0 Journal Article %J Equine Veterinary Journal %D 2013 %T Regulation of hypoxia-inducible factor-1-alpha and HIF-1 alpha-related genes in equine digital lamellae and in cultured keratinocytes %A E. A. Pawlak %A R. J. Geor %A M. R. Watts %A Black, S J %A P. J. Johnson %A Belknap, J K %B Equine Veterinary Journal %I Wiley-Blackwell %V 46 %P 203–209 %8 jul %G eng %U https://doi.org/10.1111/evj.12092 %0 Journal Article %J J Exp Med %D 2013 %T Therapeutic targeting of NOTCH signaling ameliorates immune-mediated bone marrow failure of aplastic anemia. %A Roderick, Justine E %A Gonzalez-Perez, Gabriela %A Kuksin, Christina Arieta %A Dongre, Anushka %A Roberts, Emily R %A Srinivasan, Janani %A Andrzejewski, Chester %A Fauq, Abdul H %A Golde, Todd E %A Miele, Lucio %A Minter, Lisa M %K Amyloid Precursor Protein Secretases %K Anemia, Aplastic %K Animals %K Bone Marrow %K Disease Models, Animal %K Enzyme Inhibitors %K Female %K Hematopoietic Stem Cell Transplantation %K Humans %K Male %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Mice, Knockout %K Mice, Transgenic %K Receptor, Notch1 %K Signal Transduction %X

Severe 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.

%B J Exp Med %V 210 %P 1311-29 %8 2013 Jul 1 %G eng %N 7 %1

http://www.ncbi.nlm.nih.gov/pubmed/23733784?dopt=Abstract

%0 Journal Article %J Cell %D 2013 %T Tryptophan Biosynthesis Protects Mycobacteria from CD4 T-Cell-Mediated Killing %A Yanjia J. Zhang %A Manchi C. Reddy %A Thomas R. Ioerger %A Alissa C. Rothchild %A Veronique Dartois %A Brian M. Schuster %A Andrej Trauner %A Deeann Wallis %A Stacy Galaviz %A Curtis Huttenhower %A James C. Sacchettini %A Samuel M. Behar %A Eric J. Rubin %B Cell %I Elsevier {BV} %V 155 %P 1296–1308 %G eng %U https://doi.org/10.1016/j.cell.2013.10.045 %0 Journal Article %J PLoS One %D 2013 %T Visceral endoderm expression of Yin-Yang1 (YY1) is required for VEGFA maintenance and yolk sac development. %A Rhee, Siyeon %A Guerrero-Zayas, Mara-Isel %A Wallingford, Mary C %A Ortiz-Pineda, Pablo %A Mager, Jesse %A Tremblay, Kimberly D %K Animals %K Cell Death %K Cell Proliferation %K Endoderm %K Female %K Gene Expression %K Gene Knockout Techniques %K Male %K Mesoderm %K Signal Transduction %K Vascular Endothelial Growth Factor A %K Yolk Sac %K YY1 Transcription Factor %X

Mouse 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.

%B PLoS One %V 8 %P e58828 %8 2013 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/23554936?dopt=Abstract

%0 Journal Article %J Developmental biology %D 2012 %T ADAM13 function is required in the 3 dimensional context of the embryo during cranial neural crest cell migration in Xenopus laevis. %A Cousin, Hélène %A Abbruzzese, Genevieve %A McCusker, Catherine %A Alfandari, Dominique %X

The 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.

%B Developmental biology %8 2012 Jun 7 %G ENG %1 http://www.ncbi.nlm.nih.gov/pubmed/22683825?dopt=Abstract %0 Journal Article %J Topics in current chemistry %D 2012 %T Advances in the mode of action of pyrethroids. %A Clark, J Marshall %A Symington, Steven B %X The 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. %B Topics in current chemistry %V 314 %P 49-72 %8 2012 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/22025067?dopt=Abstract %0 Journal Article %J J Infect Dis %D 2012 %T B Lymphocytes provide an infection niche for intracellular bacterium Brucella abortus. %A Goenka, Radhika %A Guirnalda, Patrick D %A Black, Samuel J %A Baldwin, Cynthia L %K Animals %K Antibodies, Bacterial %K B-Lymphocytes %K Brucella abortus %K Brucellosis %K Complement System Proteins %K DNA Replication %K Endosomes %K Immunoglobulin M %K Lysosomes %K Macrophages %K Mice %K Mice, Inbred BALB C %K Monocytes %K Phagocytosis %K Stem Cells %K Survival %K Transforming Growth Factor beta1 %X

BACKGROUND: 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.

%B J Infect Dis %V 206 %P 91-8 %8 2012 Jul 1 %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/22561364?dopt=Abstract

%0 Journal Article %J Blood %D 2012 %T Basis of CTLA-4 function in regulatory and conventional CD4(+) T cells. %A Tai, Xuguang %A Van Laethem, Francois %A Pobezinsky, Leonid %A Guinter, Terry %A Sharrow, Susan O %A Adams, Anthony %A Granger, Larry %A Kruhlak, Michael %A Lindsten, Tullia %A Thompson, Craig B %A Feigenbaum, Lionel %A Singer, Alfred %K Animals %K Cell Membrane %K Clonal Anergy %K CTLA-4 Antigen %K Golgi Apparatus %K Lymphocyte Activation %K Mice %K Mice, Knockout %K Receptors, Antigen, T-Cell %K Signal Transduction %K T-Lymphocytes, Regulatory %X

CTLA-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.

%B Blood %V 119 %P 5155-63 %8 2012 May 31 %G eng %N 22 %1

http://www.ncbi.nlm.nih.gov/pubmed/22403258?dopt=Abstract

%0 Journal Article %J Biotechnology for Biofuels %D 2012 %T Biological conversion assay using Clostridium phytofermentans to estimate plant feedstock quality %A Scott J. Lee %A Thomas A. Warnick %A Sivakumar Pattathil %A Jesús G. Alvelo-Maurosa %A Michelle J Serapiglia %A Heather McCormick %A Virginia Brown %A Naomi F Young %A Danny J. Schnell %A Lawrence B Smart %A Michael G Hahn %A Jeffrey F Pedersen %A Susan B. Leschine %A Samuel P. Hazen %B Biotechnology for Biofuels %I Springer Nature %V 5 %P 5 %G eng %U https://doi.org/10.1186/1754-6834-5-5 %0 Journal Article %J Current topics in microbiology and immunology %D 2012 %T Canonical and Non-Canonical Notch Signaling in CD4(+) T Cells. %A Minter, Lisa M %A Osborne, Barbara A %X For 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. %B Current topics in microbiology and immunology %8 2012 Jun 15 %G ENG %1 http://www.ncbi.nlm.nih.gov/pubmed/22695917?dopt=Abstract %0 Journal Article %J Nat Immunol %D 2012 %T Clonal deletion and the fate of autoreactive thymocytes that survive negative selection. %A Pobezinsky, Leonid A %A Angelov, Georgi S %A Tai, Xuguang %A Jeurling, Susanna %A Van Laethem, Francois %A Feigenbaum, Lionel %A Park, Jung-Hyun %A Singer, Alfred %K Animals %K Antigens, CD28 %K Antigens, CD4 %K Antigens, CD8 %K Cell Differentiation %K Clonal Deletion %K Flow Cytometry %K Immune Tolerance %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Mice, Knockout %K Mice, Transgenic %K Receptors, Antigen, T-Cell %K Signal Transduction %K Thymocytes %K Thymus Gland %X

Clonal 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.

%B Nat Immunol %V 13 %P 569-78 %8 2012 Jun %G eng %N 6 %1

http://www.ncbi.nlm.nih.gov/pubmed/22544394?dopt=Abstract

%0 Journal Article %J Developmental biology %D 2012 %T cSrc is necessary for epididymal development and is incorporated into sperm during epididymal transit. %A Krapf, Dario %A Chun Ruan, Ye %A Wertheimer, Eva V %A Battistone, Maria A %A Pawlak, John B %A Sanjay, Archana %A Pilder, Stephen H %A Cuasnicu, Patricia %A Breton, Sylvie %A Visconti, Pablo E %X Changes 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. %B Developmental biology %V 369 %P 43-53 %8 2012 Sep 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/22750823?dopt=Abstract %0 Journal Article %J Developmental biology %D 2012 %T Depletion of Suds3 reveals an essential role in early lineage specification. %A Zhang, Kun %A Dai, Xiangpeng %A Wallingford, Mary C %A Mager, Jesse %X Preimplantation 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. %B Developmental biology %8 2012 Nov 1 %G ENG %1 http://www.ncbi.nlm.nih.gov/pubmed/23123966?dopt=Abstract %0 Journal Article %J Epigenetics : official journal of the DNA Methylation Society %D 2012 %T Detection of promoter methylation of tumor suppressor genes in serum DNA of breast cancer cases and benign breast disease controls. %A Sturgeon, Susan R %A Balasubramanian, Raji %A Schairer, Catherine %A Muss, Hyman B %A Ziegler, Regina G %A Arcaro, Kathleen F %X Tumors 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. %B Epigenetics : official journal of the DNA Methylation Society %V 7 %P 1258-67 %8 2012 Nov 1 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/22986510?dopt=Abstract %0 Journal Article %J BMC cancer %D 2012 %T Differential expression of cancer associated proteins in breast milk based on age at first full term pregnancy. %A Qin, Wenyi %A Zhang, Ke %A Kliethermes, Beth %A Ruhlen, Rachel L %A Browne, Eva P %A Arcaro, Kathleen F %A Sauter, Edward R %X BACKGROUND: 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. %B BMC cancer %V 12 %P 100 %8 2012 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/22436421?dopt=Abstract %0 Journal Article %J Journal of human lactation : official journal of International Lactation Consultant Association %D 2012 %T Differential expression of cancer-related proteins in paired breast milk samples from women with breast cancer. %A Arcaro, Kathleen F %A Browne, Eva P %A Qin, Wenyi %A Zhang, Ke %A Anderton, Douglas L %A Sauter, Edward R %X Background: 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. %B Journal of human lactation : official journal of International Lactation Consultant Association %V 28 %P 543-6 %8 2012 Nov %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/22914689?dopt=Abstract %0 Journal Article %J American journal of veterinary research %D 2012 %T Distribution and processing of a disintegrin and metalloproteinase with thrombospondin motifs-4, aggrecan, versican, and hyaluronan in equine digital laminae. %A Pawlak, Erica %A Wang, Le %A Johnson, Philip J %A Nuovo, Gerard %A Taye, Almaz %A Belknap, James K %A Alfandari, Dominique %A Black, Samuel J %X

OBJECTIVE: 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.

%B American journal of veterinary research %V 73 %P 1035-46 %8 2012 Jul %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/22738056?dopt=Abstract %0 Journal Article %J American journal of veterinary research %D 2012 %T Effects 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. %A Wang, Le %A Pawlak, Erica %A Johnson, Philip J %A Belknap, James K %A Alfandari, Dominique %A Black, Samuel J %X

OBJECTIVE: 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.

%B American journal of veterinary research %V 73 %P 1047-56 %8 2012 Jul %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/22738057?dopt=Abstract %0 Journal Article %J Journal of cellular physiology %D 2012 %T Electrophysiological evidence for the presence of cystic fibrosis transmembrane conductance regulator (CFTR) in mouse sperm. %A Fierro, Dulce Figueiras %A Acevedo, Juan José %A Martínez, Pablo %A Escoffier, Jessica %A Sepúlveda, Francisco V %A Balderas, Enrique %A Orta, Gerardo %A Visconti, Pablo %A Darszon, Alberto %X Mammalian 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. %B Journal of cellular physiology %8 2012 Jul 25 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/22833409?dopt=Abstract %0 Journal Article %J Journal of cell science %D 2012 %T Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation. %A Escoffier, Jessica %A Krapf, Dario %A Navarrete, Felipe %A Darszon, Alberto %A Visconti, Pablo E %X Mammalian 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. %B Journal of cell science %8 2012 Feb 2 %G ENG %1 http://www.ncbi.nlm.nih.gov/pubmed/22302997?dopt=Abstract %0 Journal Article %J BMC Genet %D 2012 %T Gene number determination and genetic polymorphism of the gamma delta T cell co-receptor WC1 genes. %A Chen, Chuang %A Herzig, Carolyn T A %A Alexander, Leeson J %A Keele, John W %A McDaneld, Tara G %A Telfer, Janice C %A Baldwin, Cynthia L %K Animals %K Base Sequence %K Cattle %K Databases, Genetic %K DNA, Complementary %K Gene Dosage %K Genome %K Membrane Glycoproteins %K Molecular Sequence Data %K Multigene Family %K Phylogeny %K Polymorphism, Genetic %K Protein Structure, Tertiary %K Real-Time Polymerase Chain Reaction %K Receptors, Antigen, T-Cell, gamma-delta %X

BACKGROUND: 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.

%B BMC Genet %V 13 %P 86 %8 2012 %G eng %1

http://www.ncbi.nlm.nih.gov/pubmed/23072335?dopt=Abstract

%0 Journal Article %J Zygote %D 2012 %T Identification of 4 genes required for mammalian blastocyst formation. %A Maserati, Marc %A Dai, Xiangpeng %A Walentuk, Melanie %A Mager, Jesse %B Zygote %V In Press %G eng %0 Journal Article %J Human reproduction (Oxford, England) %D 2012 %T A maternally inherited autosomal point mutation in human phospholipase C zeta (PLCζ) leads to male infertility. %A Kashir, Junaid %A Konstantinidis, Michalis %A Jones, Celine %A Lemmon, Bernadette %A Lee, Hoi Chang %A Hamer, Rebecca %A Heindryckx, Bjorn %A Deane, Charlotte M %A De Sutter, Petra %A Fissore, Rafael A %A Parrington, John %A Wells, Dagan %A Coward, Kevin %X BACKGROUND: 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. %B Human reproduction (Oxford, England) %V 27 %P 222-31 %8 2012 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/22095789?dopt=Abstract %0 Journal Article %J Breast Cancer Res Treat %D 2012 %T Normal breast tissue of obese women is enriched for macrophage markers and macrophage-associated gene expression. %A Sun, Xuezheng %A Casbas-Hernandez, Patricia %A Bigelow, Carol %A Makowski, Liza %A Joseph Jerry, D %A Schneider, Sallie Smith %A Troester, Melissa A %K Adipose Tissue %K Adolescent %K Adult %K Biological Markers %K Body Mass Index %K Breast %K Cluster Analysis %K Female %K Gene Expression Profiling %K Gene Expression Regulation %K Humans %K Inflammation %K Macrophages %K Mammaplasty %K Middle Aged %K Obesity %K Young Adult %X

Activation 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.

%B Breast Cancer Res Treat %V 131 %P 1003-12 %8 2012 Feb %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/22002519?dopt=Abstract

%0 Journal Article %J Science signaling %D 2012 %T Notch and the survival of regulatory T cells: location is everything! %A Minter, Lisa M %A Osborne, Barbara A %X Signaling 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. %B Science signaling %V 5 %P pe31 %8 2012 %G eng %N 234 %1 http://www.ncbi.nlm.nih.gov/pubmed/22827995?dopt=Abstract %0 Journal Article %J J Med Entomol %D 2012 %T Ovicidal response of NYDA formulations on the human head louse (Anoplura: Pediculidae) using a hair tuft bioassay. %A Strycharz, Joseph P %A Lao, Alice R %A Alves, Anna-Maria %A Clark, J Marshall %K Animals %K Biological Assay %K Dimethylpolysiloxanes %K Female %K Hair %K Humans %K Larva %K Ovum %K Pediculus %K Toxicity Tests %X

Using 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.

%B J Med Entomol %V 49 %P 336-42 %8 2012 Mar %G eng %N 2 %R 10.1603/me11178 %0 Journal Article %J Cancer cell international %D 2012 %T Paralemmin-1 is over-expressed in estrogen-receptor positive breast cancers. %A Turk, Casey M %A Fagan-Solis, Katerina D %A Williams, Kristin E %A Gozgit, Joseph M %A Smith-Schneider, Sallie %A Marconi, Sharon A %A Otis, Christopher N %A Crisi, Giovanna M %A Anderton, Douglas L %A Kilimann, Manfred W %A Arcaro, Kathleen F %X UNLABELLED: 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. %B Cancer cell international %V 12 %P 17 %8 2012 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/22574838?dopt=Abstract %0 Journal Article %J Biology of reproduction %D 2012 %T Participation of the Cl-/HCO3- Exchangers SLC26A3 and SLC26A6, the Cl- Channel CFTR, and the Regulatory Factor SLC9A3R1 in Mouse Sperm Capacitation. %A Chávez, Julio C %A Hernández-González, Enrique O %A Wertheimer, Eva %A Visconti, Pablo E %A Darszon, Alberto %A Treviño, Claudia L %X Sperm 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. %B Biology of reproduction %V 86 %P 1-14 %8 2012 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/21976599?dopt=Abstract %0 Journal Article %J Journal of cellular physiology %D 2012 %T Regulation of inositol 1,4,5-trisphosphate receptor function during mouse oocyte maturation. %A Wakai, Takuya %A Vanderheyden, Veerle %A Yoon, Sook-Young %A Cheon, Banyoon %A Zhang, Nan %A Parys, Jan B %A Fissore, Rafael A %K Animals %K Calcium Signaling %K Cyclin-Dependent Kinase Inhibitor Proteins %K Female %K Gene Expression Regulation %K Inositol 1,4,5-Trisphosphate Receptors %K Mice %K Oocytes %K Phosphorylation %K Protein Transport %X At 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. %B Journal of cellular physiology %V 227 %P 705-17 %8 2012 Feb %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/21465476?dopt=Abstract %0 Journal Article %J Cell Cycle %D 2012 %T The role of TRADD in death receptor signaling. %A Pobezinskaya, Yelena L %A Liu, Zhenggang %K Apoptosis %K Humans %K Mitogen-Activated Protein Kinases %K NF-kappa B %K Receptors, Death Domain %K Receptors, TNF-Related Apoptosis-Inducing Ligand %K Receptors, Tumor Necrosis Factor, Type I %K Signal Transduction %K TNF Receptor-Associated Death Domain Protein %K TNF-Related Apoptosis-Inducing Ligand %K Tumor Necrosis Factor Ligand Superfamily Member 15 %X

TRADD (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.

%B Cell Cycle %V 11 %P 871-6 %8 2012 Mar 1 %G eng %N 5 %1

http://www.ncbi.nlm.nih.gov/pubmed/22333735?dopt=Abstract

%0 Journal Article %J PLoS ONE %D 2012 %T Roles of MAPK and Spindle Assembly Checkpoint in Spontaneous Activation and MIII Arrest of Rat Oocytes %A Cui, Wei %A Jie Zhang %A Hua-Yu Lian %A Hui-Li Wang %A De-Qiang Miao %A Chuan-Xin Zhang %A Ming-Jiu Luo %A Jing-He Tan %E Jay M. Baltz %B PLoS ONE %I Public Library of Science ({PLoS}) %V 7 %P e32044 %8 feb %G eng %U https://doi.org/10.1371/journal.pone.0032044 %0 Journal Article %J Biology of reproduction %D 2012 %T Sperm Bioenergetics in a Nutshell. %A Visconti, P %X In 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. %B Biology of reproduction %8 2012 Aug 22 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/22914312?dopt=Abstract %0 Journal Article %J Expert Rev Proteomics %D 2012 %T Sperm phosphoproteomics: historical perspectives and current methodologies. %A Porambo, James R. %A Salicioni, Ana M %A Visconti, Pablo E %A Platt, Mark D %K Fertilization %K Gene Expression Regulation %K Humans %K Infertility, Male %K Male %K Mass Spectrometry %K Phosphoproteins %K Phosphorylation %K Proteomics %K Spermatozoa %X

Mammalian 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.

%B Expert Rev Proteomics %V 9 %P 533-48 %8 2012 Oct %G eng %N 5 %1

http://www.ncbi.nlm.nih.gov/pubmed/23194270?dopt=Abstract

%0 Magazine Article %D 2012 %T Testis-Specific Kinases in male Fertility and as Targets for Contraception %A Salicioni, Ana M %A Romano, Fabian B %A Visconti, Pablo E %B American Pharmaceutical Review %V 15 %8 09/2012 %G eng %N 5 %9 Review %0 Journal Article %J Molecular neurodegeneration %D 2012 %T Transient pharmacologic lowering of Aß production prior to deposition results in sustained reduction of amyloid plaque pathology. %A Das, Pritam %A Verbeeck, Christophe %A Minter, Lisa %A Chakrabarty, Paramita %A Felsenstein, Kevin %A Kukar, Thomas %A Maharvi, Ghulam %A Fauq, Abdul %A Osborne, Barbara A %A Golde, Todd E %X ABSTRACT: 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. %B Molecular neurodegeneration %V 7 %P 39 %8 2012 Aug 14 %G ENG %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/22892055?dopt=Abstract %0 Journal Article %J Developmental biology %D 2012 %T Yin-Yang1 is required for epithelial-to-mesenchymal transition and regulation of Nodal signaling during mammalian gastrulation. %A Trask, Mary C %A Tremblay, Kimberly D %A Mager, Jesse %X

The 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.

%B Developmental biology %V 368 %P 273-82 %8 2012 Aug 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/22669107?dopt=Abstract %0 Journal Article %J Immunity %D 2012 %T αβ T cell receptors that do not undergo major histocompatibility complex-specific thymic selection possess antibody-like recognition specificities. %A Tikhonova, Anastasia N %A Van Laethem, Francois %A Hanada, Ken-ichi %A Lu, Jinghua %A Pobezinsky, Leonid A %A Hong, Changwan %A Guinter, Terry I %A Jeurling, Susanna K %A Bernhardt, Günter %A Park, Jung-Hyun %A Yang, James C %A Sun, Peter D %A Singer, Alfred %K Amino Acid Sequence %K Animals %K Antibody Specificity %K Epitopes, T-Lymphocyte %K Gene Deletion %K Ligands %K Major Histocompatibility Complex %K Mice %K Molecular Sequence Data %K Protein Binding %K Receptors, Antigen, T-Cell, alpha-beta %K Receptors, Virus %K T-Lymphocytes %K Thymus Gland %X

Major 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.

%B Immunity %V 36 %P 79-91 %8 2012 Jan 27 %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/22209676?dopt=Abstract

%0 Journal Article %J Neuroscience %D 2011 %T 17β-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. %A Intlekofer, K A %A Petersen, S L %X Recent 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. %B Neuroscience %V 176 %P 86-92 %8 2011 Mar 10 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/21185909?dopt=Abstract %0 Journal Article %J Médecine sciences : M/S %D 2011 %T ADAM and cell migration: the unexpected role of the cytoplasmic domain %A Cousin, Hélène %A Alfandari, Dominique %B Médecine sciences : M/S %V 27 %P 1069-71 %8 2011 Dec %G fre %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/22192744?dopt=Abstract %0 Journal Article %J J Immunol %D 2011 %T The adaptor protein TRADD is essential for TNF-like ligand 1A/death receptor 3 signaling. %A Pobezinskaya, Yelena L %A Choksi, Swati %A Morgan, Michael J %A Cao, Xiumei %A Liu, Zheng-Gang %K Animals %K Blotting, Western %K Cell Separation %K Electrophoretic Mobility Shift Assay %K Flow Cytometry %K Immunoprecipitation %K Lymphocyte Activation %K Mice %K Mice, Knockout %K Receptors, Tumor Necrosis Factor, Member 25 %K Signal Transduction %K T-Lymphocytes %K TNF Receptor-Associated Death Domain Protein %K Tumor Necrosis Factor Ligand Superfamily Member 15 %X

TNFR-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.

%B J Immunol %V 186 %P 5212-6 %8 2011 May 1 %G eng %N 9 %1

http://www.ncbi.nlm.nih.gov/pubmed/21421854?dopt=Abstract

%0 Journal Article %J Pflügers Archiv : European journal of physiology %D 2011 %T Alterations in calcium oscillatory activity in vitrified mouse eggs impact on egg quality and subsequent embryonic development. %A Kim, Bo Yeun %A Yoon, Sook-Young %A Cha, Soo Kyoung %A Kwak, Ki Hoon %A Fissore, Rafael A %A Parys, Jan B %A Yoon, Tae Ki %A Lee, Dong Ryul %K Animals %K Calcium Signaling %K Cryoprotective Agents %K Dimethyl Sulfoxide %K Embryonic Development %K Ethylene Glycol %K Female %K Fertilization in Vitro %K Inositol 1,4,5-Trisphosphate Receptors %K Male %K Mice %K Ovum %K Sperm Injections, Intracytoplasmic %K Vitrification %X Cryopreservation 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. %B Pflügers Archiv : European journal of physiology %V 461 %P 515-26 %8 2011 May %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/21431324?dopt=Abstract %0 Journal Article %J The Journal of infectious diseases %D 2011 %T B cell-deficient mice display markedly enhanced resistance to the intracellular bacterium Brucella abortus. %A Goenka, Radhika %A Parent, Michelle A %A Elzer, Philip H %A Baldwin, Cynthia L %X Brucella species are facultative intracellular bacteria that cause lifelong infections in humans and livestock. %B The Journal of infectious diseases %V 203 %P 1136-46 %8 2011 Apr 15 %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/21451002?dopt=Abstract %0 Journal Article %J Insect molecular biology %D 2011 %T Brief exposures of human body lice to sublethal amounts of ivermectin over-transcribes detoxification genes involved in tolerance. %A Yoon, K S %A Strycharz, J P %A Baek, J H %A Sun, W %A Kim, J H %A Kang, J S %A Pittendrigh, B R %A Lee, S H %A Clark, J M %X Transcriptional profiling results, using our non-invasive induction assay {short exposure intervals (2-5 h) to sublethal amounts of insecticides [Transcriptional profiling results, using our non-invasive induction assay {short exposure intervals (2-5 h) to sublethal amounts of insecticides [< lethal concentration 3% (LC(3)) at 24 h] administered by stress-reducing means (contact vs. immersion screen) and with induction assessed in a time frame when tolerance is still present [~lethal concentration 90% (LC(90)) in 2-4 h]}, showed that ivermectin-induced detoxification genes from body lice are identified by quantitative real-time PCR analyses. Of the cytochrome P450 monooxygenase and ATP binding cassette transporter genes induced by ivermectin, CYP6CJ1, CYP9AG1, CYP9AG2 and PhABCC4 were respectively most significantly over-expressed, had high basal expression levels and were most closely related to genes from other organisms that metabolized insecticides, including ivermectin. Injection of double-stranded RNAs (dsRNAs) against either CYP9AG2 or PhABCC4 into non-induced female lice reduced their respective transcript level and resulted in increased sensitivity to ivermectin, indicating that these two genes are involved in the xenobiotic metabolism of ivermectin and in the production of tolerance.

%B Insect Mol Biol %V 20 %P 687-99 %8 2011 Dec %G eng %N 6 %R 10.1111/j.1365-2583.2011.01097.x %0 Journal Article %J Cold Spring Harbor perspectives in biology %D 2011 %T Ca2+ signaling during mammalian fertilization: requirements, players, and adaptations. %A Wakai, Takuya %A Vanderheyden, Veerle %A Fissore, Rafael A %K Animals %K Calcium Signaling %K Fertilization %K Humans %K Male %K Mammals %K Ovum %K Spermatozoa %X Changes in the intracellular concentration of calcium ([Ca(2+)](i)) represent a vital signaling mechanism enabling communication among cells and between cells and the environment. The initiation of embryo development depends on a [Ca(2+)](i) increase(s) in the egg, which is generally induced during fertilization. The [Ca(2+)](i) increase signals egg activation, which is the first stage in embryo development, and that consist of biochemical and structural changes that transform eggs into zygotes. The spatiotemporal patterns of [Ca(2+)](i) at fertilization show variability, most likely reflecting adaptations to fertilizing conditions and to the duration of embryonic cell cycles. In mammals, the focus of this review, the fertilization [Ca(2+)](i) signal displays unique properties in that it is initiated after gamete fusion by release of a sperm-derived factor and by periodic and extended [Ca(2+)](i) responses. Here, we will discuss the events of egg activation regulated by increases in [Ca(2+)](i), the possible downstream targets that effect these egg activation events, and the property and identity of molecules both in sperm and eggs that underpin the initiation and persistence of the [Ca(2+)](i) responses in these species. %B Cold Spring Harbor perspectives in biology %V 3 %8 2011 Apr %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/21441584?dopt=Abstract %0 Journal Article %J Molecular Reproduction and Development %D 2011 %T Caffeine alleviates the deterioration of Ca2+ release mechanisms and fragmentation of in vitro-aged mouse eggs %A Zhang, Nan %A Wakai, Takuya %A Fissore, Rafael A %B Molecular Reproduction and Development %V 78 %P 684-701 %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/737?dopt=Abstract %0 Journal Article %J PloS one %D 2011 %T Chimeric anti-staphylococcal enterotoxin B antibodies and lovastatin act synergistically to provide in vivo protection against lethal doses of SEB. %A Tilahun, Mulualem E %A Kwan, Alan %A Natarajan, Kannan %A Quinn, Megan %A Tilahun, Ashenafi Y %A Xie, Chen %A Margulies, David H %A Osborne, Barbara A %A Goldsby, Richard A %A Rajagopalan, Govindarajan %X Staphylococcal enterotoxin B (SEB) is one of a family of toxins secreted by Staphylococcus aureus that act as superantigens, activating a large fraction of the T-cell population and inducing production of high levels of inflammatory cytokines that can cause toxic shock syndrome (TSS) and death. Extracellular engagement of the TCR of T-cells and class II MHC of antigen presenting cells by SEB triggers the activation of many intracellular signaling processes. We engineered chimeric antibodies to block the extracellular engagement of cellular receptors by SEB and used a statin to inhibit intracellular signaling. Chimeric human-mouse antibodies directed against different neutralizing epitopes of SEB synergistically inhibited its activation of human T-cells in vitro. In the in vivo model of lethal toxic shock syndrome (TSS) in HLA-DR3 transgenic mice, two of these antibodies conferred significant partial protection when administered individually, but offered complete protection in a synergistic manner when given together. Similarly, in vivo, lovastatin alone conferred only partial protection from TSS similar to single anti-SEB antibodies. However, used in combination with one chimeric neutralizing anti-SEB antibody, lovastatin provided complete protection against lethal TSS in HLA-DR3 transgenic mice. These experiments demonstrate that in vivo protection against lethal doses of SEB can be achieved by a statin of proven clinical safety and chimeric human-mouse antibodies, agents now widely used and known to be of low immunogenicity in human hosts. %B PloS one %V 6 %P e27203 %8 2011 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/22102880?dopt=Abstract %0 Journal Article %J Insect biochemistry and molecular biology %D 2011 %T Comparison of the humoral and cellular immune responses between body and head lice following bacterial challenge. %A Kim, Ju Hyeon %A Min, Jee Sun %A Kang, Jae Soon %A Kwon, Deok Ho %A Yoon, Kyong Sup %A Strycharz, Joseph %A Koh, Young Ho %A Pittendrigh, Barry Robert %A Clark, J Marshall %A Lee, Si Hyeock %K Animals %K Escherichia coli %K Female %K Fluorescein-5-isothiocyanate %K Gene Expression Profiling %K Genes, Insect %K Genome %K Immunity, Cellular %K Immunity, Humoral %K Microscopy, Fluorescence %K Pediculus %K Phagocytosis %K Signal Transduction %K Species Specificity %K Staphylococcus aureus %K Transcription, Genetic %X The differences in the immune response between body lice, Pediculus humanus humanus, and head lice, Pediculus humanus capitis, were investigated initially by measuring the proliferation rates of two model bacteria, a Gram-positive Staphylococcus aureus and a Gram-negative Escherichia coli, following challenge by injection. Body lice showed a significantly reduced immune response compared to head lice particularly to E. coli at the early stage of the immune challenge. Annotation of the body louse genome identified substantially fewer immune-related genes compared with other insects. Nevertheless, all required genetic components of the major immune pathways, except for the immune deficiency (Imd) pathway, are still retained in the body louse genome. Transcriptional profiling of representative genes involved in the humoral immune response, following bacterial challenge, revealed that both body and head lice, regardless of their developmental stages, exhibited an increased immune response to S. aureus but little to E. coli. Head lice, however, exhibited a significantly higher phagocytotic activity against E. coli than body lice, whereas the phagocytosis against S. aureus differed only slightly between body and head lice. These findings suggest that the greater immune response in head lice against E. coli is largely due to enhanced phagocytosis and not due to differences in the humoral immune response. The reduced phagocytotic activity in body lice could be responsible, in part, for their increased vector competence. %B Insect biochemistry and molecular biology %V 41 %P 332-9 %8 2011 May %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/21296152?dopt=Abstract %0 Journal Article %J Journal of cellular physiology %D 2011 %T Complexity of polycomb group function: diverse mechanisms of target specificity. %A Trask, Mary C %A Mager, Jesse %K Animals %K Binding Sites %K DNA %K Embryo, Mammalian %K Epigenesis, Genetic %K Gene Expression Regulation, Developmental %K Gene Silencing %K Humans %K Protein Binding %K Repressor Proteins %K RNA, Untranslated %K Signal Transduction %X Epigenetic regulation of gene expression has become relevant to nearly all areas of biomedical research. The emergence of technologies that allow for examination of the epigenome combined with identification of key protein complexes that mediate the myriad chromatin modifications that occur have greatly enhanced the versatility and efficacy of tools with which to study normal development and disease states. The evolutionarily conserved polycomb group genes (PcG) have been identified as a predominant mechanism by which gene silencing occurs during development, differentiation, and disease. While molecular events that target PcG complexes have been well defined in some non-vertebrate models, the details of locus specificity and functional diversity of mammalian PcG proteins have not yet unresolved. Here we discuss recent findings that offer novel mechanistic events and add complexity to our understanding of PcG function in vertebrates. %B Journal of cellular physiology %V 226 %P 1719-21 %8 2011 Jul %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/20799281?dopt=Abstract %0 Journal Article %J Biomacromolecules %D 2011 %T De novo designed protein transduction domain mimics from simple synthetic polymers. %A Tezgel, A Özgül %A Telfer, Janice C %A Tew, Gregory N %X Protein transduction domains (PTDs) that readily transverse cellular membranes are of great interest and are attractive tools for the intracellular delivery of bioactive molecules. Learning to program synthetic polymers and oligomers with the appropriate chemical information to capture adequately the biological activity of proteins is critical to our improved understanding of how these natural molecules work. In addition, the versatility of these synthetic mimics provides the opportunity to discover analogs with superior properties compared with their native sequences. Here we report the first detailed structure-activity relationship of a new PTD family of polymers based on a completely abiotic backbone. The synthetic approach easily allows doubling the density of guanidine functional groups, which increases the transduction efficiency of the sequences. Cellular uptake studies on three different cell lines (HEK 293T, CHO, and Jurkat T cells) confirm that these synthetic analogs are highly efficient novel protein transduction domain mimics (PTDMs), which are more effective than TAT(49-57) and nonaarginine (R9) and also highlight the usefulness of polymer chemistry at the chemistry-biology interface. %B Biomacromolecules %V 12 %P 3078-83 %8 2011 Aug 8 %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/21714570?dopt=Abstract %0 Journal Article %J Neuroscience %D 2011 %T Distribution of mRNAs encoding classical progestin receptor, progesterone membrane components 1 and 2, serpine mRNA binding protein 1, and progestin and ADIPOQ receptor family members 7 and 8 in rat forebrain. %A Intlekofer, K A %A Petersen, S L %X Several lines of evidence suggest the existence of multiple progestin receptors that may account for rapid and delayed effects of progesterone in the CNS. The delayed effects have been long attributed to activation of the classical progestin receptor (Pgr). Recent studies have discovered novel progestin signaling molecules that may be responsible for rapid effects. These include progesterone receptor membrane component 1 (Pgrmc1), Pgrmc2, progestin and adipoQ receptor 7 (Paqr7) and Paqr8. The functions of these molecules have been investigated extensively in non-neural, but not in neural tissues, partly because it is unclear which are expressed in the brain and where they are expressed. To address these issues, we compared the distributions of mRNAs encoding Pgr, Pgrmc1, Pgrmc2, Paqr7 and Paqr8 using in situ hybridization with radiolabeled oligodeoxynucleotidyl probes in forebrain tissues of estradiol-treated female rats. We also examined the distribution of serpine mRNA binding protein 1 (Serbp1), a putative binding partner of Pgrmc1. Analyses of adjacent brain sections showed that the highest expression of mRNAs encoding Pgr, Pgrmc1, Pgrmc2 and Serbp1 was detected in several hypothalamic nuclei important for female reproduction. In contrast, expression patterns of Paqr7 and Paqr8 were low and homogeneous in the hypothalamus, and more abundant in thalamic nuclei. The neuroanatomical distributions of these putative progestin signaling molecules suggest that Pgrmc1 and Pgrmc2 may play roles in neuroendocrine functions while Paqr7 and Paqr8 are more likely to regulate sensory and cognitive functions. %B Neuroscience %V 172 %P 55-65 %8 2011 Jan 13 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/20977928?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2011 %T Expressed gene sequence of the IFNγ-response chemokine CXCL9 of cattle, horses, and swine. %A Hudgens, Edward %A Tompkins, Dannielle %A Boyd, Patricia %A Lunney, Joan K %A Horohov, David %A Baldwin, Cynthia L %X This report describes the cloning and characterization of expressed gene sequences of bovine, equine, and swine CXCL9 from RNA obtained from peripheral blood mononuclear cells (PBMC) and other tissues. The bovine coding region was 378 nucleotides in length, while the equine and swine coding regions were 381 nucleotides. Mapping showed that all three sequences were coded for in four exons in the genome, as are the human and mouse genes. The bovine, equine, and swine coding regions shared 83%, 86%, and 84% homology with human CXCL9, respectively, and all three were 74% homologous with mouse CXCL9. Cladogram comparison of the nucleotide sequences of CXCL9 showed that the bovine, equine and swine sequences were more closely related to one another than to either the human or the mouse sequences. However, the human sequence was more closely related to them than it was to the mouse sequence. These relationships were preserved when the deduced amino acid sequences were evaluated and all sequences showed conservation of the characteristic four cysteines. This work sets the stage for further work with these molecules; an integral goal of the U.S. Veterinary Immune Reagent Network is to develop reagents for investigating diseases in livestock species, poultry, and fish. %B Veterinary immunology and immunopathology %V 141 %P 317-21 %8 2011 Jun 15 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/21497916?dopt=Abstract %0 Journal Article %J Molec Hum Reprod %D 2011 %T Expression and localization of five members of the testis-specific serine kinase (Tssk) family in mouse and human sperm and testis %A Li, Y %A Sosnik, J %A Brassard, L %A Reese, M %A Spiridonov, NA %A Bates, TC %A Johnson, GR %A Anguita, J %A Visconti, P E %A Salicioni, A M %X

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.

%B Molec Hum Reprod %V 17 %P 42-56 %8 2011 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/749?dopt=Abstract %0 Journal Article %J Biology of reproduction %D 2011 %T Guanine-nucleotide exchange factors (RAPGEF3/RAPGEF4) induce sperm membrane depolarization and acrosomal exocytosis in capacitated stallion sperm. %A McPartlin, L A %A Visconti, P E %A Bedford-Guaus, S J %K Acrosome %K Animals %K Cyclic AMP %K Cyclic AMP-Dependent Protein Kinase Catalytic Subunits %K Dichlororibofuranosylbenzimidazole %K Exocytosis %K Guanine Nucleotide Exchange Factors %K Horses %K Male %K Membrane Potentials %K Mice %K Phosphorylation %K Protein-Tyrosine Kinases %K Signal Transduction %K Sperm Capacitation %K Thionucleotides %X Capacitation 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. %B Biology of reproduction %V 85 %P 179-88 %8 2011 Jul %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/21471298?dopt=Abstract %0 Journal Article %J Molecular reproduction and development %D 2011 %T Identification and functional analysis of an ovarian form of the egg activation factor phospholipase C zeta (PLCζ) in pufferfish. %A Coward, Kevin %A Ponting, Chris P %A Zhang, Nan %A Young, Claire %A Huang, Chang-Jen %A Chou, Chih-Ming %A Kashir, Junaid %A Fissore, Rafael A %A Parrington, John %K Animals %K Base Sequence %K Calcium %K Chickens %K Female %K Fish Proteins %K Gene Expression Regulation, Enzymologic %K Humans %K Male %K Mice %K Molecular Sequence Data %K Organ Specificity %K Ovary %K Ovum %K Phosphoinositide Phospholipase C %K Species Specificity %K Takifugu %X Recent 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. %B Molecular reproduction and development %V 78 %P 48-56 %8 2011 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/21268182?dopt=Abstract %0 Journal Article %J Epigenetics : official journal of the DNA Methylation Society %D 2011 %T Increased promoter methylation in exfoliated breast epithelial cells in women with a previous breast biopsy. %A Browne, Eva P %A Punska, Elizabeth C %A Lenington, Sarah %A Otis, Christopher N %A Anderton, Douglas L %A Arcaro, Kathleen F %X Accurately 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. %B Epigenetics : official journal of the DNA Methylation Society %V 6 %P 1425-35 %8 2011 Dec 1 %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/22139572?dopt=Abstract %0 Journal Article %J Journal of cellular physiology %D 2011 %T Inducing the liver: understanding the signals that promote murine liver budding. %A Tremblay, Kimberly D %X

The 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.

%B Journal of cellular physiology %V 226 %P 1727-31 %8 2011 Jul %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/20857423?dopt=Abstract %0 Journal Article %J Asian journal of andrology %D 2011 %T Ion channels, phosphorylation and mammalian sperm capacitation. %A Visconti, Pablo E %A Krapf, Dario %A De la Vega-Beltran, Jose Luis %A Acevedo, Juan José %A Darszon, Alberto %X Sexually 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. %B Asian journal of andrology %V 13 %P 395-405 %8 2011 May %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/21540868?dopt=Abstract %0 Journal Article %J Journal of medical entomology %D 2011 %T Ivermectin acts as a posteclosion nymphicide by reducing blood feeding of human head lice (Anoplura: Pediculidae) that hatched from treated eggs. %A Strycharz, Joseph P %A Berge, Noah M %A Alves, Anna-Maria %A Clark, J Marshall %X The 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. %B Journal of medical entomology %V 48 %P 1174-82 %8 2011 Nov %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/22238876?dopt=Abstract %0 Journal Article %J Human reproduction (Oxford, England) %D 2011 %T Loss of activity mutations in phospholipase C zeta (PLCζ) abolishes calcium oscillatory ability of human recombinant protein in mouse oocytes. %A Kashir, Junaid %A Jones, Celine %A Lee, Hoi Chang %A Rietdorf, Katja %A Nikiforaki, Dimitra %A Durrans, Claire %A Ruas, Margarida %A Tee, Sze Tian %A Heindryckx, Bjorn %A Galione, Antony %A De Sutter, Petra %A Fissore, Rafael A %A Parrington, John %A Coward, Kevin %X BACKGROUND: 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. %B Human reproduction (Oxford, England) %V 26 %P 3372-87 %8 2011 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/22010140?dopt=Abstract %0 Journal Article %J J Mammary Gland Biol Neoplasia %D 2011 %T Map making in the 21st century: charting breast cancer susceptibility pathways in rodent models. %A Blackburn, Anneke C %A Jerry, D Joseph %K Animals %K Breast Neoplasms %K Female %K Genetic Predisposition to Disease %K Humans %K Mammary Neoplasms, Animal %K Mice %K Tumor Suppressor Protein p53 %X

Genetic 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.

%B J Mammary Gland Biol Neoplasia %V 16 %P 57-64 %8 2011 Apr %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/21380934?dopt=Abstract

%0 Journal Article %J The Journal of biological chemistry %D 2011 %T Mechanism of a genetically encoded dark-to-bright reporter for caspase activity. %A Nicholls, Samantha B %A Chu, Jun %A Abbruzzese, Genevieve %A Tremblay, Kimberly D %A Hardy, Jeanne A %K Animals %K Apoptosis %K Caspases %K Catalysis %K Genes, Reporter %K Green Fluorescent Proteins %K Mice %K Microscopy, Fluorescence %K NIH 3T3 Cells %K Proteolysis %X

Fluorescent 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.

%B The Journal of biological chemistry %V 286 %P 24977-86 %8 2011 Jul 15 %G eng %N 28 %1 http://www.ncbi.nlm.nih.gov/pubmed/21558267?dopt=Abstract %0 Journal Article %J J Biol Chem %D 2011 %T Mitochondrial dysfunction impairs tumor suppressor p53 expression/function. %A Compton, Shannon %A Kim, Chul %A Griner, Nicholas B %A Potluri, Prasanth %A Scheffler, Immo E %A Sen, Sabyasachi %A Jerry, D Joseph %A Schneider, Sallie %A Yadava, Nagendra %K Animals %K Base Sequence %K Cell Line %K Electron Transport Complex I %K Gamma Rays %K Gene Expression Regulation %K Mice %K Mice, Knockout %K Mitochondria %K Molecular Sequence Data %K Mutation %K Neoplasms %K Oxidative Phosphorylation %K Oxygen Consumption %K Tumor Suppressor Protein p53 %X

Recently, 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.

%B J Biol Chem %V 286 %P 20297-312 %8 2011 Jun 10 %G eng %N 23 %1

http://www.ncbi.nlm.nih.gov/pubmed/21502317?dopt=Abstract

%0 Journal Article %J J Immunol %D 2011 %T Notch signaling regulates mouse and human Th17 differentiation. %A Keerthivasan, S %A Suleiman, R %A Lawlor, R %A Roderick, J %A Bates, T %A Minter, L %A Anguita, J %A Juncadella, I %A Nickoloff, BJ %A Le Poole, IC %A Miele, L %A Osborne, B A %B J Immunol %V 187 %P 692-701 %8 07/2011 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/739?dopt=Abstract %0 Journal Article %J Molecular reproduction and development %D 2011 %T PLCζ and its role as a trigger of development in vertebrates. %A Ito, Junya %A Parrington, John %A Fissore, Rafael A %X A 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. %B Molecular reproduction and development %V 78 %P 846-53 %8 2011 Oct-Nov %G eng %N 10-11 %1 http://www.ncbi.nlm.nih.gov/pubmed/21823187?dopt=Abstract %0 Journal Article %J J Agric Food Chem %D 2011 %T Pyrethroid mode(s) of action in the context of Food Quality Protection Act (FQPA) regulation. %A Gammon, Derek W %A Leggett, Michael F %A Clark, John M %K Animals %K Cockroaches %K Food Contamination %K Insecticides %K Ion Channels %K Nervous System %K Poisoning %K Pyrethrins %K Rats %K United States %K United States Environmental Protection Agency %X

A 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.

%B J Agric Food Chem %V 59 %P 2773-85 %8 2011 Apr 13 %G eng %N 7 %R 10.1021/jf103901k %0 Journal Article %J Cancer cell %D 2011 %T Radiation acts on the microenvironment to affect breast carcinogenesis by distinct mechanisms that decrease cancer latency and affect tumor type. %A Nguyen, David H %A Oketch-Rabah, Hellen A %A Illa-Bochaca, Irineu %A Geyer, Felipe C %A Reis-Filho, Jorge S %A Mao, Jian-Hua %A Ravani, Shraddha A %A Zavadil, Jiri %A Borowsky, Alexander D %A Jerry, D Joseph %A Dunphy, Karen A %A Seo, Jae Hong %A Haslam, Sandra %A Medina, Daniel %A Barcellos-Hoff, Mary Helen %K Animals %K Breast Neoplasms %K Cell Transformation, Neoplastic %K Dose-Response Relationship, Radiation %K Epithelial Cells %K Female %K Gene Expression Profiling %K Gene Expression Regulation, Neoplastic %K Gene Regulatory Networks %K Mammary Glands, Animal %K Mice %K Mice, Inbred BALB C %K Mice, Knockout %K Neoplasms, Radiation-Induced %K Radiation Chimera %K Reaction Time %K Receptors, Estrogen %K Time Factors %K Transforming Growth Factor beta1 %K Tumor Burden %K Tumor Microenvironment %K Tumor Suppressor Protein p53 %K Whole-Body Irradiation %X

Tissue 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.

%B Cancer cell %V 19 %P 640-51 %8 2011 May 17 %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/21575864?dopt=Abstract %0 Journal Article %J Stem cells (Dayton, Ohio) %D 2011 %T Repression of mammary stem/progenitor cells by p53 is mediated by Notch and separable from apoptotic activity. %A Tao, Luwei %A Roberts, Amy L %A Dunphy, Karen A %A Bigelow, Carol %A Yan, Haoheng %A Jerry, D Joseph %X

Breast 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.

%B Stem cells (Dayton, Ohio) %V 29 %P 119-27 %8 2011 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/21280161?dopt=Abstract %0 Journal Article %J J Mammary Gland Biol Neoplasia %D 2011 %T The role of activin in mammary gland development and oncogenesis. %A Dunphy, Karen A %A Schneyer, Alan L %A Hagen, Mary J %A Jerry, D Joseph %K Activins %K Animals %K Breast Neoplasms %K Female %K Humans %K Mammary Glands, Animal %K Mammary Glands, Human %K Mammary Neoplasms, Experimental %K Signal Transduction %K Transforming Growth Factor beta %X

TGFβ 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.

%B J Mammary Gland Biol Neoplasia %V 16 %P 117-26 %8 2011 Jun %G eng %N 2 %1

http://www.ncbi.nlm.nih.gov/pubmed/21475961?dopt=Abstract

%0 Journal Article %J Journal of mammary gland biology and neoplasia %D 2011 %T The role of activin in mammary gland development and oncogenesis. %A Dunphy, Karen A %A Schneyer, Alan L %A Hagen, Mary J %A Jerry, D Joseph %X

TGFβ 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.

%B Journal of mammary gland biology and neoplasia %V 16 %P 117-26 %8 2011 Jun %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/21475961?dopt=Abstract %0 Journal Article %J FASEB J %D 2011 %T The role of TRADD in TRAIL-induced apoptosis and signaling. %A Cao, Xiumei %A Pobezinskaya, Yelena L %A Morgan, Michael J %A Liu, Zheng-Gang %K Animals %K Apoptosis %K Fas-Associated Death Domain Protein %K GTPase-Activating Proteins %K Humans %K Mice %K Receptors, TNF-Related Apoptosis-Inducing Ligand %K Signal Transduction %K TNF Receptor-Associated Death Domain Protein %K TNF-Related Apoptosis-Inducing Ligand %X

Tumor 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.

%B FASEB J %V 25 %P 1353-8 %8 2011 Apr %G eng %N 4 %1

http://www.ncbi.nlm.nih.gov/pubmed/21187341?dopt=Abstract

%0 Journal Article %J Molecular immunology %D 2011 %T Scavenger receptor WC1 contributes to the γδ T cell response to Leptospira. %A Wang, Fei %A Herzig, Carolyn T A %A Chen, Chuang %A Hsu, Haoting %A Baldwin, Cynthia L %A Telfer, Janice C %X

WC1 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.

%B Molecular immunology %V 48 %P 801-9 %8 2011 Mar %G eng %N 6-7 %1 http://www.ncbi.nlm.nih.gov/pubmed/21227509?dopt=Abstract %0 Journal Article %J Commun Integr Biol %D 2011 %T Simplify, simplify: Lifestyle and compact genome of the body louse provide a unique functional genomics opportunity. %A Pittendrigh, Barry R %A Berenbaum, May R %A Seufferheld, Manfredo J %A Margam, Venu M %A Strycharz, Joseph P %A Yoon, Kyong S %A Sun, Weilin %A Reenan, Robert %A Lee, Si Hyeock %A Clark, John M %X

The 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.

%B Commun Integr Biol %V 4 %P 188-91 %8 2011 Mar %G eng %N 2 %R 10.4161/cib.4.2.14279 %0 Journal Article %J Communicative & integrative biology %D 2011 %T Simplify, simplify: Lifestyle and compact genome of the body louse provide a unique functional genomics opportunity. %A Pittendrigh, Barry R %A Berenbaum, May R %A Seufferheld, Manfredo J %A Margam, Venu M %A Strycharz, Joseph P %A Yoon, Kyong S %A Sun, Weilin %A Reenan, Robert %A Lee, Si Hyeock %A Clark, John M %X The 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. %B Communicative & integrative biology %V 4 %P 188-91 %8 2011 Mar %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/21655436?dopt=Abstract %0 Journal Article %J PLoS pathogens %D 2011 %T T. brucei infection reduces B lymphopoiesis in bone marrow and truncates compensatory splenic lymphopoiesis through transitional B-cell apoptosis. %A Bockstal, Viki %A Guirnalda, Patrick %A Caljon, Guy %A Goenka, Radhika %A Telfer, Janice C %A Frenkel, Deborah %A Radwanska, Magdalena %A Magez, Stefan %A Black, Samuel J %X

African 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.

%B PLoS pathogens %V 7 %P e1002089 %8 2011 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/21738467?dopt=Abstract %0 Journal Article %J Developmental cell %D 2011 %T Translocation of the cytoplasmic domain of ADAM13 to the nucleus is essential for Calpain8-a expression and cranial neural crest cell migration. %A Cousin, Hélène %A Abbruzzese, Genevieve %A Kerdavid, Erin %A Gaultier, Alban %A Alfandari, Dominique %K ADAM Proteins %K Amyloid Precursor Protein Secretases %K Animals %K Calpain %K Cell Line %K Cell Movement %K Cell Nucleus %K Conserved Sequence %K Embryo, Nonmammalian %K Evolution, Molecular %K Gene Expression Regulation, Developmental %K Humans %K Membrane Proteins %K Neural Crest %K Protein Structure, Tertiary %K Protein Transport %K Skull %K Structure-Activity Relationship %K Xenopus laevis %K Xenopus Proteins %X

ADAMs 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.

%B Developmental cell %V 20 %P 256-63 %8 2011 Feb 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/21316592?dopt=Abstract %0 Journal Article %J Developmental biology %D 2011 %T Transmembrane adenylyl cyclase regulates amphibian sperm motility through protein kinase A activation. %A O'Brien, Emma D %A Krapf, Dario %A Cabada, Marcelo O %A Visconti, Pablo E %A Arranz, Silvia E %K Adenylate Cyclase %K Animals %K Bufo arenarum %K Cell Membrane %K Cyclic AMP %K Cyclic AMP-Dependent Protein Kinases %K Enzyme Activation %K Hypotonic Solutions %K Male %K Phosphorylation %K Sperm Motility %K Spermatozoa %X Sperm 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. %B Developmental biology %V 350 %P 80-8 %8 2011 Feb 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/21126515?dopt=Abstract %0 Journal Article %J PloS one %D 2011 %T Wdr74 is required for blastocyst formation in the mouse. %A Maserati, Marc %A Walentuk, Melanie %A Dai, Xiangpeng %A Holston, Olivia %A Adams, Danielle %A Mager, Jesse %K Animals %K Apoptosis %K Blastocyst %K Cell Differentiation %K Female %K Gene Expression Regulation, Developmental %K Injections %K Male %K Mice %K Proteins %K RNA, Double-Stranded %K Tumor Suppressor Protein p53 %X Preimplantation 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. %B PloS one %V 6 %P e22516 %8 2011 %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/21799883?dopt=Abstract %0 Journal Article %J Biology of reproduction %D 2011 %T Yin-yang1 is required in the mammalian oocyte for follicle expansion. %A Griffith, Gillian J %A Trask, Mary C %A Hiller, Jacob %A Walentuk, Melanie %A Pawlak, John B %A Tremblay, Kimberly D %A Mager, Jesse %K Animals %K Base Sequence %K Bone Morphogenetic Protein 15 %K Cell Communication %K Female %K Gene Expression Regulation, Developmental %K Granulosa Cells %K Growth Differentiation Factor 9 %K Mice %K Mice, Inbred C57BL %K Mice, Knockout %K Models, Biological %K Oocytes %K Oogenesis %K RNA, Messenger %K YY1 Transcription Factor %X

The 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.

%B Biology of reproduction %V 84 %P 654-63 %8 2011 Apr %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/21123818?dopt=Abstract %0 Journal Article %J Journal of cellular physiology %D 2010 %T Analysis of CAPZA3 localization reveals temporally discrete events during the acrosome reaction. %A Sosnik, Julian %A Buffone, Mariano G %A Visconti, Pablo E %X In 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. %B Journal of cellular physiology %V 224 %P 575-80 %8 2010 Sep %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/20458735?dopt=Abstract %0 Journal Article %J BMC genomics %D 2010 %T Annotation and classification of the bovine T cell receptor delta genes. %A Herzig, Carolyn T A %A Lefranc, Marie-Paule %A Baldwin, Cynthia L %X gammadelta 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. %B BMC genomics %V 11 %P 100 %8 2010 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/20144200?dopt=Abstract %0 Journal Article %J J Am Chem Soc %D 2010 %T Array-based sensing of normal, cancerous, and metastatic cells using conjugated fluorescent polymers. %A Bajaj, Avinash %A Miranda, Oscar R %A Phillips, Ronnie %A Kim, Ik-Bum %A Jerry, D Joseph %A Bunz, Uwe H F %A Rotello, Vincent M %K Animals %K Biosensing Techniques %K Cell Line %K Discriminant Analysis %K Fluorescence %K HeLa Cells %K Humans %K Mice %K Mice, Inbred BALB C %K Molecular Structure %K Neoplasms %K Polymers %K Sensitivity and Specificity %X

A 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.

%B J Am Chem Soc %V 132 %P 1018-22 %8 2010 Jan 27 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/20039629?dopt=Abstract

%0 Journal Article %J Journal of the American Chemical Society %D 2010 %T Array-based sensing of normal, cancerous, and metastatic cells using conjugated fluorescent polymers. %A Bajaj, Avinash %A Miranda, Oscar R %A Phillips, Ronnie %A Kim, Ik-Bum %A Jerry, D Joseph %A Bunz, Uwe H F %A Rotello, Vincent M %X A 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. %B Journal of the American Chemical Society %V 132 %P 1018-22 %8 2010 Jan 27 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/20039629?dopt=Abstract %0 Journal Article %J Pest management science %D 2010 %T Determination of knockdown resistance allele frequencies in global human head louse populations using the serial invasive signal amplification reaction. %A Hodgdon, Hilliary E %A Yoon, Kyong Sup %A Previte, Domenic J %A Kim, Hyo Jeong %A Aboelghar, Gamal E %A Lee, Si Hyeock %A Clark, J Marshall %X Pediculosis 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. %B Pest management science %V 66 %P 1031-40 %8 2010 Sep %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/20564731?dopt=Abstract %0 Journal Article %J Molecular and cellular biology %D 2010 %T Domain-specific response of imprinted genes to reduced DNMT1. %A Weaver, Jamie R %A Sarkisian, Garnik %A Krapp, Christopher %A Mager, Jesse %A Mann, Mellissa R W %A Bartolomei, Marisa S %K Alleles %K Animals %K Base Sequence %K Basic Helix-Loop-Helix Transcription Factors %K Crosses, Genetic %K DNA (Cytosine-5-)-Methyltransferase %K DNA Methylation %K DNA Primers %K Female %K Gene Expression Regulation, Developmental %K Genomic Imprinting %K Insulin-Like Growth Factor II %K KCNQ1 Potassium Channel %K Kruppel-Like Transcription Factors %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Mutant Strains %K Multigene Family %K Mutant Proteins %K Placenta %K Pregnancy %K Protein Structure, Tertiary %X Imprinted 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. %B Molecular and cellular biology %V 30 %P 3916-28 %8 2010 Aug %G eng %N 16 %1 http://www.ncbi.nlm.nih.gov/pubmed/20547750?dopt=Abstract %0 Journal Article %J Ecotoxicology and environmental safety %D 2010 %T Effects of exposure water volume, depuration time, and feeding status on vitellogenin mRNA induction in male medaka (Oryzias latipes) exposed to 17 β-estradiol. %A Moffatt, Lauren T %A May, Chelsea L %A Studer, Kirsten E %A Reckhow, David A %A Arcaro, Kathleen F %X Bioassays 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. %B Ecotoxicology and environmental safety %V 73 %P 1835-41 %8 2010 Nov %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/20825989?dopt=Abstract %0 Journal Article %J Cell Mol Life Sci %D 2010 %T Estrogens, regulation of p53 and breast cancer risk: a balancing act. %A Jerry, D Joseph %A Dunphy, Karen A %A Hagen, Mary J %K Breast Neoplasms %K Chromatin Assembly and Disassembly %K Epigenesis, Genetic %K Estrogens %K Female %K Humans %K Receptors, Estrogen %K Risk Factors %K Tumor Suppressor Protein p53 %X

The 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.

%B Cell Mol Life Sci %V 67 %P 1017-23 %8 2010 Apr %G eng %N 7 %1

http://www.ncbi.nlm.nih.gov/pubmed/20238478?dopt=Abstract

%0 Journal Article %J Cellular and molecular life sciences : CMLS %D 2010 %T Estrogens, regulation of p53 and breast cancer risk: a balancing act. %A Jerry, D Joseph %A Dunphy, Karen A %A Hagen, Mary J %X

The 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.

%B Cellular and molecular life sciences : CMLS %V 67 %P 1017-23 %8 2010 Apr %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/20238478?dopt=Abstract %0 Journal Article %J BMC evolutionary biology %D 2010 %T Evolution of the CD163 family and its relationship to the bovine gamma delta T cell co-receptor WC1. %A Herzig, Carolyn T A %A Waters, Ray W %A Baldwin, Cynthia L %A Telfer, Janice C %X

The 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.

%B BMC evolutionary biology %V 10 %P 181 %8 2010 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/20550670?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2010 %T Expressed gene sequences of the equine cytokines interleukin-17 and interleukin-23. %A Tompkins, Dannielle %A Hudgens, Edward %A Horohov, David %A Baldwin, Cynthia L %X This 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. %B Veterinary immunology and immunopathology %V 133 %P 309-13 %8 2010 Feb 15 %G eng %N 2-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/19775756?dopt=Abstract %0 Journal Article %J Progress in molecular biology and translational science %D 2010 %T Formation of the murine endoderm: lessons from the mouse, frog, fish, and chick. %A Tremblay, Kimberly D %K Animals %K Anura %K Chick Embryo %K Endoderm %K Fishes %K Mice %K Morphogenesis %X The 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. %B Progress in molecular biology and translational science %V 96 %P 1-34 %8 2010 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/21075338?dopt=Abstract %0 Journal Article %J Proceedings of the National Academy of Sciences of the United States of America %D 2010 %T Genome sequences of the human body louse and its primary endosymbiont provide insights into the permanent parasitic lifestyle. %A Kirkness, Ewen F %A Haas, Brian J %A Sun, Weilin %A Braig, Henk R %A Perotti, M Alejandra %A Clark, John M %A Lee, Si Hyeock %A Robertson, Hugh M %A Kennedy, Ryan C %A Elhaik, Eran %A Gerlach, Daniel %A Kriventseva, Evgenia V %A Elsik, Christine G %A Graur, Dan %A Hill, Catherine A %A Veenstra, Jan A %A Walenz, Brian %A Tubío, José Manuel C %A Ribeiro, José M C %A Rozas, Julio %A Johnston, J Spencer %A Reese, Justin T %A Popadic, Aleksandar %A Tojo, Marta %A Raoult, Didier %A Reed, David L %A Tomoyasu, Yoshinori %A Kraus, Emily %A Krause, Emily %A Mittapalli, Omprakash %A Margam, Venu M %A Li, Hong-Mei %A Meyer, Jason M %A Johnson, Reed M %A Romero-Severson, Jeanne %A Vanzee, Janice Pagel %A Alvarez-Ponce, David %A Vieira, Filipe G %A Aguadé, Montserrat %A Guirao-Rico, Sara %A Anzola, Juan M %A Yoon, Kyong S %A Strycharz, Joseph P %A Unger, Maria F %A Christley, Scott %A Lobo, Neil F %A Seufferheld, Manfredo J %A Wang, Naikuan %A Dasch, Gregory A %A Struchiner, Claudio J %A Madey, Greg %A Hannick, Linda I %A Bidwell, Shelby %A Joardar, Vinita %A Caler, Elisabet %A Shao, Renfu %A Barker, Stephen C %A Cameron, Stephen %A Bruggner, Robert V %A Regier, Allison %A Johnson, Justin %A Viswanathan, Lakshmi %A Utterback, Terry R %A Sutton, Granger G %A Lawson, Daniel %A Waterhouse, Robert M %A Venter, J Craig %A Strausberg, Robert L %A Berenbaum, May R %A Collins, Frank H %A Zdobnov, Evgeny M %A Pittendrigh, Barry R %K Animals %K Enterobacteriaceae %K Genes, Bacterial %K Genes, Insect %K Genome, Bacterial %K Genome, Insect %K Genomics %K Humans %K Lice Infestations %K Molecular Sequence Data %K Pediculus %K Sequence Analysis, DNA %K Symbiosis %X As 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. %B Proceedings of the National Academy of Sciences of the United States of America %V 107 %P 12168-73 %8 2010 Jul 6 %G eng %N 27 %1 http://www.ncbi.nlm.nih.gov/pubmed/20566863?dopt=Abstract %0 Journal Article %J Molecular reproduction and development %D 2010 %T Identification and functional analysis of an ovarian form of the egg activation factor phospholipase C Zeta (PLCζ) in pufferfish. %A Coward, Kevin %A Ponting, Chris P %A Zhang, Nan %A Young, Claire %A Huang, Chang-Jen %A Chou, Chih-Ming %A Kashir, Junaid %A Fissore, Rafael A %A Parrington, John %X Recent 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. %B Molecular reproduction and development %8 2010 Dec 10 %G ENG %1 http://www.ncbi.nlm.nih.gov/pubmed/21240961?dopt=Abstract %0 Journal Article %J Hum Mutat %D 2010 %T Identification of a DMBT1 polymorphism associated with increased breast cancer risk and decreased promoter activity. %A Tchatchou, Sandrine %A Riedel, Angela %A Lyer, Stefan %A Schmutzhard, Julia %A Strobel-Freidekind, Olga %A Gronert-Sum, Sabine %A Mietag, Carola %A D'Amato, Mauro %A Schlehe, Bettina %A Hemminki, Kari %A Sutter, Christian %A Ditsch, Nina %A Blackburn, Anneke %A Hill, Linda Zhai %A Jerry, D Joseph %A Bugert, Peter %A Weber, Bernhard H F %A Niederacher, Dieter %A Arnold, Norbert %A Varon-Mateeva, Raymonda %A Wappenschmidt, Barbara %A Schmutzler, Rita K %A Engel, Christoph %A Meindl, Alfons %A Bartram, Claus R %A Mollenhauer, Jan %A Burwinkel, Barbara %K Adult %K Aged %K Breast Neoplasms %K Breast Neoplasms, Male %K Case-Control Studies %K Female %K Genetic Predisposition to Disease %K Humans %K Male %K Middle Aged %K Polymorphism, Single Nucleotide %K Promoter Regions, Genetic %K Receptors, Cell Surface %K Risk Factors %X

According 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.

%B Hum Mutat %V 31 %P 60-6 %8 2010 Jan %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/19830809?dopt=Abstract

%0 Journal Article %J Veterinary immunology and immunopathology %D 2010 %T In vivo priming and ex vivo activation of equine neutrophils in black walnut extract-induced equine laminitis is not attenuated by systemic lidocaine administration. %A Loftus, John P %A Williams, Jarred M %A Belknap, James K %A Black, Samuel J %X Laminitis 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. %B Veterinary immunology and immunopathology %V 138 %P 60-9 %8 2010 Nov 15 %G eng %N 1-2 %1 http://www.ncbi.nlm.nih.gov/pubmed/20667603?dopt=Abstract %0 Journal Article %J Parasitology %D 2010 %T Induction and regulation of Trypanosoma brucei VSG-specific antibody responses. %A Black, S J %A Guirnalda, P %A Frenkel, D %A Haynes, C %A Bockstal, V %X The 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. %B Parasitology %V 137 %P 2041-9 %8 2010 Dec %G eng %N 14 %1 http://www.ncbi.nlm.nih.gov/pubmed/20025827?dopt=Abstract %0 Journal Article %J The Journal of biological chemistry %D 2010 %T Inhibition of Ser/Thr phosphatases induces capacitation-associated signaling in the presence of Src kinase inhibitors. %A Krapf, Dario %A Arcelay, Enid %A Wertheimer, Eva V %A Sanjay, Archana %A Pilder, Stephen H %A Salicioni, Ana M %A Visconti, Pablo E %K Aniline Compounds %K Animals %K Cyclic AMP %K Cyclic AMP-Dependent Protein Kinases %K Enzyme Inhibitors %K Female %K Indoles %K Male %K Mice %K Mice, Inbred Strains %K Mice, Mutant Strains %K Nitriles %K Okadaic Acid %K Oxazoles %K Phosphorylation %K Protein-Serine-Threonine Kinases %K Quinolines %K Signal Transduction %K Sperm Capacitation %K Sperm-Ovum Interactions %K Spermatozoa %K src-Family Kinases %K Sulfonamides %K Tyrosine %X

Signaling 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.

%B The Journal of biological chemistry %V 285 %P 7977-85 %8 2010 Mar 12 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/20068039?dopt=Abstract %0 Journal Article %J Journal of cellular physiology %D 2010 %T Inositol 1,4,5-trisphosphate receptor 1 degradation in mouse eggs and impact on [Ca2+]i oscillations. %A Lee, Bora %A Yoon, Sook-Young %A Malcuit, Chris %A Parys, Jan B %A Fissore, Rafael A %K Adenosine %K Animals %K Calcium Signaling %K Down-Regulation %K Female %K Fertilization %K Humans %K Injections %K Inositol 1,4,5-Trisphosphate %K Inositol 1,4,5-Trisphosphate Receptors %K Mice %K Ovum %K Protein Processing, Post-Translational %K Time Factors %X The 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. %B Journal of cellular physiology %V 222 %P 238-47 %8 2010 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/19798695?dopt=Abstract %0 Journal Article %J Journal of visualized experiments : JoVE %D 2010 %T Mammary epithelial transplant procedure. %A Dunphy, Karen A %A Tao, Luwei %A Jerry, D Joseph %K Animals %K Epithelium %K Female %K Mammary Glands, Animal %K Mice %K Tissue Transplantation %X

This 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.

%B Journal of visualized experiments : JoVE %8 2010 %G eng %N 40 %1 http://www.ncbi.nlm.nih.gov/pubmed/20548284?dopt=Abstract %0 Journal Article %J Cell adhesion & migration %D 2010 %T Mechanism of Xenopus cranial neural crest cell migration. %A Alfandari, Dominque %A Cousin, Hélène %A Marsden, Mungo %K Animals %K Cell Adhesion %K Cell Movement %K Central Nervous System %K Gene Expression Regulation, Developmental %K Head %K Intracellular Signaling Peptides and Proteins %K Membrane Proteins %K Models, Biological %K Neural Crest %K Signal Transduction %K Wnt Proteins %K Xenopus laevis %X This 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. %B Cell adhesion & migration %V 4 %P 553-60 %8 2010 Oct-Dec %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/20505318?dopt=Abstract %0 Journal Article %J Science signaling %D 2010 %T Mechanisms of sperm-egg interactions: between sugars and broken bonds. %A Visconti, Pablo E %A Florman, Harvey M %X A 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. %B Science signaling %V 3 %P pe35 %8 2010 %G eng %N 142 %1 http://www.ncbi.nlm.nih.gov/pubmed/20923932?dopt=Abstract %0 Journal Article %J Am J Pathol %D 2010 %T Pathways contributing to development of spontaneous mammary tumors in BALB/c-Trp53+/- mice. %A Yan, Haoheng %A Blackburn, Anneke C %A McLary, S Christine %A Tao, Luwei %A Roberts, Amy L %A Xavier, Elizabeth A %A Dickinson, Ellen S %A Seo, Jae Hong %A Arenas, Richard B %A Otis, Christopher N %A Cao, Qing J %A Lawlor, Rebecca G %A Osborne, Barbara A %A Kittrell, Frances S %A Medina, Daniel %A Jerry, D Joseph %K Animals %K Female %K Gene Expression Regulation, Neoplastic %K Keratins %K Loss of Heterozygosity %K Mammary Neoplasms, Animal %K Mammary Neoplasms, Experimental %K Mice %K Mice, Inbred BALB C %K Neoplasm Transplantation %K Precancerous Conditions %K Receptor, erbB-2 %K Receptors, Estrogen %K Receptors, Notch %K Receptors, Progesterone %K Signal Transduction %K Tumor Suppressor Protein p53 %X

Mutation 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.

%B Am J Pathol %V 176 %P 1421-32 %8 2010 Mar %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/20110418?dopt=Abstract

%0 Journal Article %J The American journal of pathology %D 2010 %T Pathways contributing to development of spontaneous mammary tumors in BALB/c-Trp53+/- mice. %A Yan, Haoheng %A Blackburn, Anneke C %A McLary, S Christine %A Tao, Luwei %A Roberts, Amy L %A Xavier, Elizabeth A %A Dickinson, Ellen S %A Seo, Jae Hong %A Arenas, Richard B %A Otis, Christopher N %A Cao, Qing J %A Lawlor, Rebecca G %A Osborne, Barbara A %A Kittrell, Frances S %A Medina, Daniel %A Jerry, D Joseph %X Mutation 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. %B The American journal of pathology %V 176 %P 1421-32 %8 2010 Mar %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/20110418?dopt=Abstract %0 Journal Article %J Animal science journal = Nihon chikusan Gakkaihō %D 2010 %T Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes. %A Ito, Junya %A Yoshida, Tomoko %A Kasai, Yasushi %A Wakai, Takuya %A Parys, Jan B %A Fissore, Rafael A %A Kashiwazaki, Naomi %K Animals %K Calcium Signaling %K CDC2 Protein Kinase %K Cells, Cultured %K Epitopes %K Female %K Inositol 1,4,5-Trisphosphate Receptors %K Meiosis %K Mice %K Mitogen-Activated Protein Kinases %K Oocytes %K Phosphoproteins %K Phosphorylation %K Sus scrofa %X During 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. %B Animal science journal = Nihon chikusan Gakkaihō %V 81 %P 34-41 %8 2010 Feb %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/20163670?dopt=Abstract %0 Journal Article %J Infection and immunity %D 2010 %T Potent neutralization of staphylococcal enterotoxin B by synergistic action of chimeric antibodies. %A Tilahun, Mulualem E %A Rajagopalan, Govindarajan %A Shah-Mahoney, Nalini %A Lawlor, Rebecca G %A Tilahun, Ashenafi Y %A Xie, Chen %A Natarajan, Kannan %A Margulies, David H %A Ratner, David I %A Osborne, Barbara A %A Goldsby, Richard A %X Staphylococcal 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. %B Infection and immunity %V 78 %P 2801-11 %8 2010 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/20308304?dopt=Abstract %0 Journal Article %J Epigenetics : official journal of the DNA Methylation Society %D 2010 %T Quantitative analysis of promoter methylation in exfoliated epithelial cells isolated from breast milk of healthy women. %A Wong, Chung M %A Anderton, Douglas L %A Smith-Schneider, Sallie %A Wing, Megan A %A Greven, Melissa C %A Arcaro, Kathleen F %K Adult %K Age Factors %K Breast %K Breast Neoplasms %K Cadherins %K CpG Islands %K Cytoskeletal Proteins %K DNA Methylation %K Epithelial Cells %K Female %K Genes, Tumor Suppressor %K Glutathione S-Transferase pi %K Humans %K Intercellular Signaling Peptides and Proteins %K Lactation %K Membrane Proteins %K Middle Aged %K Milk, Human %K Promoter Regions, Genetic %K Retinol-Binding Proteins, Cellular %K Risk Factors %K Tumor Suppressor Proteins %K Young Adult %X Promoter 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. %B Epigenetics : official journal of the DNA Methylation Society %V 5 %P 645-55 %8 2010 Oct 1 %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/20716965?dopt=Abstract %0 Journal Article %J Epigenetics : official journal of the DNA Methylation Society %D 2010 %T Quantitative analysis of promoter methylation in exfoliated epithelial cells isolated from breast milk of healthy women. %A Wong, Chung M %A Anderton, Douglas L %A Smith-Schneider, Sallie %A Wing, Megan A %A Greven, Melissa C %A Arcaro, Kathleen F %X Promoter 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. %B Epigenetics : official journal of the DNA Methylation Society %V 5 %P 645-55 %8 2010 Oct 1 %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/20716965?dopt=Abstract %0 Journal Article %J PLoS One %D 2010 %T Role of JNK in a Trp53-dependent mouse model of breast cancer. %A Cellurale, Cristina %A Weston, Claire R %A Reilly, Judith %A Garlick, David S %A Jerry, D Joseph %A Sluss, Hayla K %A Davis, Roger J %K Animals %K Breast Neoplasms %K Disease Models, Animal %K Female %K Mammary Glands, Animal %K Mice %K Mice, Inbred BALB C %K Mitogen-Activated Protein Kinase 8 %K Mitogen-Activated Protein Kinase 9 %K Survival Analysis %K Tumor Suppressor Protein p53 %X

The 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.

%B PLoS One %V 5 %P e12469 %8 2010 %G eng %N 8 %1

http://www.ncbi.nlm.nih.gov/pubmed/20814571?dopt=Abstract

%0 Journal Article %J Small (Weinheim an der Bergstrasse, Germany) %D 2010 %T The role of surface functionality on acute cytotoxicity, ROS generation and DNA damage by cationic gold nanoparticles. %A Chompoosor, Apiwat %A Saha, Krishnendu %A Ghosh, Partha S %A Macarthy, Dylan J %A Miranda, Oscar R %A Zhu, Zheng-Jiang %A Arcaro, Kathleen F %A Rotello, Vincent M %B Small (Weinheim an der Bergstrasse, Germany) %V 6 %P 2246-9 %8 2010 Oct 18 %G eng %N 20 %1 http://www.ncbi.nlm.nih.gov/pubmed/20818619?dopt=Abstract %0 Journal Article %J Biology of reproduction %D 2010 %T Species-specific differences in the activity and nuclear localization of murine and bovine phospholipase C zeta 1. %A Cooney, Melissa A %A Malcuit, Christopher %A Cheon, Banyoon %A Holland, Michael K %A Fissore, Rafael A %A D'Cruz, Nancy T %K Animals %K Calcium Signaling %K Cattle %K Down-Regulation %K Inositol 1,4,5-Trisphosphate Receptors %K Male %K Mice %K Microinjections %K Oocytes %K Phosphoinositide Phospholipase C %K Recombinant Proteins %K RNA, Complementary %K Species Specificity %K Spermatozoa %X Injection 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. %B Biology of reproduction %V 83 %P 92-101 %8 2010 Jul %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/20357268?dopt=Abstract %0 Journal Article %J Small (Weinheim an der Bergstrasse, Germany) %D 2010 %T Surface properties dictate uptake, distribution, excretion, and toxicity of nanoparticles in fish. %A Zhu, Zheng-Jiang %A Carboni, Rachel %A Quercio, Michael J %A Yan, Bo %A Miranda, Oscar R %A Anderton, Douglas L %A Arcaro, Kathleen F %A Rotello, Vincent M %A Vachet, Richard W %B Small (Weinheim an der Bergstrasse, Germany) %V 6 %P 2261-5 %8 2010 Oct 18 %G eng %N 20 %1 http://www.ncbi.nlm.nih.gov/pubmed/20842664?dopt=Abstract %0 Journal Article %J Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology %D 2010 %T Vitellogenesis in Bufo arenarum: identification, characterization and immunolocalization of high molecular mass lipovitellin during oogenesis. %A O'Brien, Emma D %A Salicioni, Ana M %A Cabada, Marcelo O %A Arranz, Silvia E %K Amino Acid Sequence %K Animals %K Bufo arenarum %K Egg Proteins %K Female %K Immunohistochemistry %K Molecular Sequence Data %K Molecular Weight %K Oocytes %K Protein Transport %K Sequence Analysis, DNA %K Time Factors %K Vitellogenesis %X Vitellogenin (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). %B Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology %V 155 %P 256-65 %8 2010 Mar %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/19932187?dopt=Abstract %0 Journal Article %J Clin Cancer Res %D 2009 %T Activation of host wound responses in breast cancer microenvironment. %A Troester, Melissa A %A Lee, Myung Hee %A Carter, Matthew %A Fan, Cheng %A Cowan, David W %A Perez, Erick Roman %A Pirone, Jason R %A Perou, Charles M %A Jerry, D Joseph %A Schneider, Sallie Smith %K Breast %K Breast Neoplasms %K Cyclooxygenase 2 %K Cysteine-Rich Protein 61 %K Disease Progression %K Female %K Gene Expression Profiling %K Gene Expression Regulation, Neoplastic %K Genome, Human %K Humans %K Immunohistochemistry %K Mammaplasty %K Neovascularization, Pathologic %K Reproducibility of Results %K Wound Healing %X

PURPOSE: 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.

%B Clin Cancer Res %V 15 %P 7020-8 %8 2009 Nov 15 %G eng %N 22 %1

http://www.ncbi.nlm.nih.gov/pubmed/19887484?dopt=Abstract

%0 Journal Article %J Seminars in cell & developmental biology %D 2009 %T ADAM function in embryogenesis. %A Alfandari, Dominique %A McCusker, Catherine %A Cousin, Hélène %X

Cleavage 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.

%B Seminars in cell & developmental biology %V 20 %P 153-63 %8 2009 Apr %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/18935966?dopt=Abstract %0 Journal Article %J Molecular immunology %D 2009 %T Antigenic basis of diversity in the gammadelta T cell co-receptor WC1 family. %A Chen, Chuang %A Herzig, Carolyn T A %A Telfer, Janice C %A Baldwin, Cynthia L %X

WC1 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.

%B Molecular immunology %V 46 %P 2565-75 %8 2009 Aug %G eng %N 13 %1 http://www.ncbi.nlm.nih.gov/pubmed/19539374?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2009 %T Cloning and expression of ADAM-related metalloproteases in equine laminitis. %A Coyne, Michael J %A Cousin, Hélène %A Loftus, John P %A Johnson, Philip J %A Belknap, James K %A Gradil, Carlos M %A Black, Samuel J %A Alfandari, Dominique %X

Equine 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.

%B Veterinary immunology and immunopathology %V 129 %P 231-41 %8 2009 Jun 15 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/19131116?dopt=Abstract %0 Journal Article %J Proceedings of the National Academy of Sciences of the United States of America %D 2009 %T Detection and differentiation of normal, cancerous, and metastatic cells using nanoparticle-polymer sensor arrays. %A Bajaj, Avinash %A Miranda, Oscar R %A Kim, Ik-Bum %A Phillips, Ronnie L %A Jerry, D Joseph %A Bunz, Uwe H F %A Rotello, Vincent M %X Rapid 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. %B Proceedings of the National Academy of Sciences of the United States of America %V 106 %P 10912-6 %8 2009 Jul 7 %G eng %N 27 %1 http://www.ncbi.nlm.nih.gov/pubmed/19549846?dopt=Abstract %0 Journal Article %J Proc Natl Acad Sci U S A %D 2009 %T Detection and differentiation of normal, cancerous, and metastatic cells using nanoparticle-polymer sensor arrays. %A Bajaj, Avinash %A Miranda, Oscar R %A Kim, Ik-Bum %A Phillips, Ronnie L %A Jerry, D Joseph %A Bunz, Uwe H F %A Rotello, Vincent M %K Animals %K Biosensing Techniques %K Cell Line, Tumor %K Humans %K Mice %K Nanoparticles %K Neoplasm Metastasis %K Neoplasms %K Polymers %X

Rapid 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.

%B Proc Natl Acad Sci U S A %V 106 %P 10912-6 %8 2009 Jul 7 %G eng %N 27 %1

http://www.ncbi.nlm.nih.gov/pubmed/19549846?dopt=Abstract

%0 Journal Article %J Journal of Asia-Pacific entomology %D 2009 %T Determination, mechanism and monitoring of knockdown resistance in permethrin-resistant human head lice, Pediculus humanus capitis. %A Clark, J Marshall %X Permethrin 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. %B Journal of Asia-Pacific entomology %V 12 %P 1-7 %8 2009 Mar 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/20161186?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2009 %T Expressed gene sequence of bovine IL23A and IL23R. %A Chen, Chuang %A Herzig, Carolyn T A %A Baldwin, Cynthia L %X The 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. %B Veterinary immunology and immunopathology %V 128 %P 425-30 %8 2009 Apr 15 %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/19117612?dopt=Abstract %0 Journal Article %J Molecular biology of the cell %D 2009 %T Extracellular cleavage of cadherin-11 by ADAM metalloproteases is essential for Xenopus cranial neural crest cell migration. %A McCusker, Catherine %A Cousin, Hélène %A Neuner, Russell %A Alfandari, Dominique %X

Cell 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.

%B Molecular biology of the cell %V 20 %P 78-89 %8 2009 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/18946084?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2009 %T Extracellular matrix, leukocyte migration and laminitis. %A Black, S J %X The structure and dynamic nature of extracellular matrix is discussed in the context of healthy and diseased tissues particularly the equine digital laminae. %B Veterinary immunology and immunopathology %V 129 %P 161-3 %8 2009 Jun 15 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/19110317?dopt=Abstract %0 Journal Article %J BMC genomics %D 2009 %T Genomic organization and classification of the bovine WC1 genes and expression by peripheral blood gamma delta T cells. %A Herzig, Carolyn T A %A Baldwin, Cynthia L %X WC1 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. %B BMC genomics %V 10 %P 191 %8 2009 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/19393067?dopt=Abstract %0 Journal Article %J Gene expression patterns : GEP %D 2009 %T Identification of novel oocyte and granulosa cell markers. %A Malcuit, Christopher %A Trask, Mary C %A Santiago, Laurelis %A Beaudoin, Emily %A Tremblay, Kimberly D %A Mager, Jesse %X

Here 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.

%B Gene expression patterns : GEP %V 9 %P 404-10 %8 2009 Sep %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/19539053?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2009 %T Leukocyte-derived and endogenous matrix metalloproteinases in the lamellae of horses with naturally acquired and experimentally induced laminitis. %A Loftus, John P %A Johnson, Philip J %A Belknap, James K %A Pettigrew, Amanda %A Black, Samuel J %X Inflammation 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. %B Veterinary immunology and immunopathology %V 129 %P 221-30 %8 2009 Jun 15 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/19101039?dopt=Abstract %0 Journal Article %J Communicative & integrative biology %D 2009 %T Life after proteolysis: Exploring the signaling capabilities of classical cadherin cleavage fragments. %A McCusker, Catherine D %A Alfandari, Dominique %X Classical 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. %B Communicative & integrative biology %V 2 %P 155-7 %8 2009 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/19513270?dopt=Abstract %0 Journal Article %J Biology of reproduction %D 2009 %T Localization of low-density detergent-resistant membrane proteins in intact and acrosome-reacted mouse sperm. %A Miranda, Patricia V %A Allaire, Alicia %A Sosnik, Julian %A Visconti, Pablo E %X Mammalian 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. %B Biology of reproduction %V 80 %P 897-904 %8 2009 May %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/19144954?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 2009 %T Notch regulates cytolytic effector function in CD8+ T cells. %A Cho, Ok Hyun %A Shin, Hyun Mu %A Miele, Lucio %A Golde, Todd E %A Fauq, Abdul %A Minter, Lisa M %A Osborne, Barbara A %X The 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. %B Journal of immunology (Baltimore, Md. : 1950) %V 182 %P 3380-9 %8 2009 Mar 15 %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/19265115?dopt=Abstract %0 Journal Article %J Blood %D 2009 %T Notch signaling mediates G1/S cell-cycle progression in T cells via cyclin D3 and its dependent kinases. %A Joshi, Ila %A Minter, Lisa M %A Telfer, Janice %A Demarest, Renée M %A Capobianco, Anthony J %A Aster, Jon C %A Sicinski, Piotr %A Fauq, Abdul %A Golde, Todd E %A Osborne, Barbara A %X Notch 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. %B Blood %V 113 %P 1689-98 %8 2009 Feb 19 %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/19001083?dopt=Abstract %0 Journal Article %J Cell calcium %D 2009 %T Regulation of inositol 1,4,5-trisphosphate receptor type 1 function during oocyte maturation by MPM-2 phosphorylation. %A Vanderheyden, Veerle %A Wakai, Takuya %A Bultynck, Geert %A De Smedt, Humbert %A Parys, Jan B %A Fissore, Rafael A %K Amino Acid Sequence %K Animals %K Antibodies, Monoclonal %K Calcium %K Cell Cycle Proteins %K Cell Line %K Cells, Cultured %K Computer Simulation %K Consensus Sequence %K Epitopes %K Female %K Inositol 1,4,5-Trisphosphate Receptors %K Mice %K Oocytes %K Phosphorylation %K Protein-Serine-Threonine Kinases %K Proto-Oncogene Proteins %X Egg 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). %B Cell calcium %V 46 %P 56-64 %8 2009 Jul %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/19482353?dopt=Abstract %0 Journal Article %J The Journal of experimental medicine %D 2009 %T Runx proteins regulate Foxp3 expression. %A Bruno, Ludovica %A Mazzarella, Luca %A Hoogenkamp, Maarten %A Hertweck, Arnulf %A Cobb, Bradley S %A Sauer, Stephan %A Hadjur, Suzana %A Leleu, Marion %A Naoe, Yoshinori %A Telfer, Janice C %A Bonifer, Constanze %A Taniuchi, Ichiro %A Fisher, Amanda G %A Merkenschlager, Matthias %K Animals %K Core Binding Factor Alpha 3 Subunit %K Core Binding Factor alpha Subunits %K Core Binding Factor beta Subunit %K Feedback, Physiological %K Forkhead Transcription Factors %K Genes, Dominant %K Mice %K Protein Structure, Tertiary %K T-Lymphocytes, Regulatory %X Runx 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. %B The Journal of experimental medicine %V 206 %P 2329-37 %8 2009 Oct 26 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/19841090?dopt=Abstract %0 Journal Article %J Journal of cell science %D 2009 %T Tssk6 is required for Izumo relocalization and gamete fusion in the mouse. %A Sosnik, Julian %A Miranda, Patricia V %A Spiridonov, Nikolay A %A Yoon, Sook-Young %A Fissore, Rafael A %A Johnson, Gibbes R %A Visconti, Pablo E %X

One 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.

%B Journal of cell science %V 122 %P 2741-9 %8 2009 Aug 1 %G eng %N Pt 15 %1 http://www.ncbi.nlm.nih.gov/pubmed/19596796?dopt=Abstract %0 Journal Article %J European journal of immunology %D 2009 %T Tyrosine phosphorylation of scavenger receptor cysteine-rich WC1 is required for the WC1-mediated potentiation of TCR-induced T-cell proliferation. %A Wang, Fei %A Herzig, Carolyn %A Ozer, Dar %A Baldwin, Cynthia L %A Telfer, Janice C %X

Workshop 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.

%B European journal of immunology %V 39 %P 254-66 %8 2009 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/19130552?dopt=Abstract %0 Journal Article %J Proceedings of the National Academy of Sciences of the United States of America %D 2009 %T Understanding the molecular basis of sperm capacitation through kinase design. %A Visconti, Pablo E %B Proceedings of the National Academy of Sciences of the United States of America %V 106 %P 667-8 %8 2009 Jan 20 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/19144927?dopt=Abstract %0 Journal Article %J Journal of proteome research %D 2009 %T Use of differential isotopic labeling and mass spectrometry to analyze capacitation-associated changes in the phosphorylation status of mouse sperm proteins. %A Platt, Mark D %A Salicioni, Ana M %A Hunt, Donald F %A Visconti, Pablo E %K Amino Acid Sequence %K Animals %K Chromatography, Affinity %K Fourier Analysis %K Isotope Labeling %K Male %K Mass Spectrometry %K Mice %K Molecular Sequence Data %K Phosphopeptides %K Phosphorylation %K Proteome %K Sperm Capacitation %K Spermatozoa %X

Mammalian 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.

%B Journal of proteome research %V 8 %P 1431-40 %8 2009 Mar %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/19186949?dopt=Abstract %0 Journal Article %J Biomacromolecules %D 2009 %T Virus-inspired approach to nonviral gene delivery vehicles. %A Roy, Raghunath %A Jerry, D Joseph %A Thayumanavan, S %X The 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. %B Biomacromolecules %V 10 %P 2189-93 %8 2009 Aug 10 %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/19722558?dopt=Abstract %0 Journal Article %J Biomacromolecules %D 2009 %T Virus-inspired approach to nonviral gene delivery vehicles. %A Roy, Raghunath %A Jerry, D Joseph %A Thayumanavan, S %K Breast Neoplasms %K Cells, Cultured %K Female %K Gene Products, tat %K Gene Transfer Techniques %K Genetic Vectors %K Humans %K Kidney %K Peptide Fragments %K Polymers %K Transfection %X

The 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.

%B Biomacromolecules %V 10 %P 2189-93 %8 2009 Aug 10 %G eng %N 8 %1

http://www.ncbi.nlm.nih.gov/pubmed/19722558?dopt=Abstract

%0 Journal Article %J Mechanisms of development %D 2009 %T Xenopus ADAM19 is involved in neural, neural crest and muscle development. %A Neuner, Russell %A Cousin, Hélène %A McCusker, Catherine %A Coyne, Michael %A Alfandari, Dominique %X

ADAM19 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.

%B Mechanisms of development %V 126 %P 240-55 %8 2009 Mar-Apr %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/19027850?dopt=Abstract %0 Journal Article %J Journal of medical entomology %D 2008 %T Biochemical and molecular analysis of deltamethrin resistance in the common bed bug (Hemiptera: Cimicidae). %A Yoon, Kyong Sup %A Kwon, Deok Ho %A Strycharz, Joseph P %A Hollingsworth, Craig S %A Lee, Si Hyeock %A Clark, J Marshall %X This 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. %B Journal of medical entomology %V 45 %P 1092-101 %8 2008 Nov %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/19058634?dopt=Abstract %0 Journal Article %J The Journal of biological chemistry %D 2008 %T Chloride Is essential for capacitation and for the capacitation-associated increase in tyrosine phosphorylation. %A Wertheimer, Eva V %A Salicioni, Ana M %A Liu, Weimin %A Treviño, Claudia L %A Chavez, Julio %A Hernández-González, Enrique O %A Darszon, Alberto %A Visconti, Pablo E %K Acrosome Reaction %K Animals %K Bumetanide %K Cell Survival %K Chlorides %K Cyclic AMP %K Cyclic AMP-Dependent Protein Kinases %K Female %K Furosemide %K Male %K Mice %K Phosphorylation %K Signal Transduction %K Sodium Potassium Chloride Symporter Inhibitors %K Sodium-Potassium-Chloride Symporters %K Sperm Capacitation %K Sperm Motility %K Spermatozoa %K Tyrosine %K Zona Pellucida %X

After 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.

%B The Journal of biological chemistry %V 283 %P 35539-50 %8 2008 Dec 19 %G eng %N 51 %1 http://www.ncbi.nlm.nih.gov/pubmed/18957426?dopt=Abstract %0 Journal Article %J Expert Opinion on Biological Therapy %D 2008 %T Elite control of HIV infection: implications for vaccine design %A BM Baker %A BL Block %A AC Rothchild %A BD Walker %B Expert Opinion on Biological Therapy %I Informa Healthcare %V 9 %P 55–69 %G eng %U https://doi.org/10.1517/14712590802571928 %0 Journal Article %J Breast cancer research : BCR %D 2008 %T Estrogen and progesterone induce persistent increases in p53-dependent apoptosis and suppress mammary tumors in BALB/c-Trp53+/- mice. %A Dunphy, Karen A %A Blackburn, Anneke C %A Yan, Haoheng %A O'Connell, Lauren R %A Jerry, D Joseph %X

Treatment 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.

%B Breast cancer research : BCR %V 10 %P R43 %8 2008 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/18471300?dopt=Abstract %0 Journal Article %J Nat Immunol %D 2008 %T The function of TRADD in signaling through tumor necrosis factor receptor 1 and TRIF-dependent Toll-like receptors. %A Pobezinskaya, Yelena L %A Kim, You-Sun %A Choksi, Swati %A Morgan, Michael J %A Li, Tao %A Liu, Chengyu %A Liu, Zhenggang %K Animals %K Fibroblasts %K Macrophages %K Mice %K Mitogen-Activated Protein Kinases %K Signal Transduction %K TNF Receptor-Associated Death Domain Protein %K TNF Receptor-Associated Factor 1 %K Toll-Like Receptors %K Tumor Necrosis Factor Receptor-Associated Peptides and Proteins %K Ubiquitin %X

The 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.

%B Nat Immunol %V 9 %P 1047-54 %8 2008 Sep %G eng %N 9 %1

http://www.ncbi.nlm.nih.gov/pubmed/18641653?dopt=Abstract

%0 Journal Article %J Journal of agricultural and food chemistry %D 2008 %T Golfer exposure to chlorpyrifos and carbaryl following application to turfgrass. %A Putnam, Raymond A %A Doherty, Jeffery J %A Clark, J Marshall %X Exposure 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. %B Journal of agricultural and food chemistry %V 56 %P 6616-22 %8 2008 Aug 13 %G eng %N 15 %1 http://www.ncbi.nlm.nih.gov/pubmed/18598045?dopt=Abstract %0 Journal Article %J The Journal of clinical investigation %D 2008 %T Human sperm devoid of PLC, zeta 1 fail to induce Ca(2+) release and are unable to initiate the first step of embryo development. %A Yoon, Sook-Young %A Jellerette, Teru %A Salicioni, Ana Maria %A Lee, Hoi Chang %A Yoo, Myung-Sik %A Coward, Kevin %A Parrington, John %A Grow, Daniel %A Cibelli, Jose B %A Visconti, Pablo E %A Mager, Jesse %A Fissore, Rafael A %K Calcium %K Embryonic Development %K Humans %K Male %K Phosphoinositide Phospholipase C %K Spermatozoa %X

Egg 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.

%B The Journal of clinical investigation %V 118 %P 3671-81 %8 2008 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/18924610?dopt=Abstract %0 Journal Article %J The International journal of developmental biology %D 2008 %T Identification of proteins undergoing tyrosine phosphorylation during mouse sperm capacitation. %A Arcelay, Enid %A Salicioni, Ana M %A Wertheimer, Eva %A Visconti, Pablo E %K Animals %K Expressed Sequence Tags %K Fructose-Bisphosphate Aldolase %K Male %K Mice %K Models, Biological %K Phosphorylation %K Proteins %K Sperm Capacitation %K Spermatozoa %K Testis %K Tubulin %K Tyrosine %K Voltage-Dependent Anion Channels %X

Mammalian 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.

%B The International journal of developmental biology %V 52 %P 463-72 %8 2008 %G eng %N 5-6 %1 http://www.ncbi.nlm.nih.gov/pubmed/18649259?dopt=Abstract %0 Journal Article %J Developmental biology %D 2008 %T Inositol 1,4,5-trisphosphate receptor 1, a widespread Ca2+ channel, is a novel substrate of polo-like kinase 1 in eggs. %A Ito, Junya %A Yoon, Sook-Young %A Lee, Bora %A Vanderheyden, Veerle %A Vermassen, Elke %A Wojcikiewicz, Richard %A Alfandari, Dominique %A De Smedt, Humbert %A Parys, Jan B %A Fissore, Rafael A %X

To 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.

%B Developmental biology %V 320 %P 402-13 %8 2008 Aug 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/18621368?dopt=Abstract %0 Journal Article %J Human molecular genetics %D 2008 %T Manipulations of mouse embryos prior to implantation result in aberrant expression of imprinted genes on day 9.5 of development. %A Rivera, Rocío M %A Stein, Paula %A Weaver, Jamie R %A Mager, Jesse %A Schultz, Richard M %A Bartolomei, Marisa S %K Alleles %K Animals %K Basic Helix-Loop-Helix Transcription Factors %K Cyclin-Dependent Kinase Inhibitor p57 %K DNA Methylation %K Embryo Culture Techniques %K Embryo Transfer %K Embryonic Development %K Female %K Gene Expression Regulation, Developmental %K Genomic Imprinting %K Gestational Age %K Humans %K Insulin-Like Growth Factor II %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Transgenic %K Placenta %K Pregnancy %K Reproductive Techniques, Assisted %K RNA, Messenger %K RNA, Untranslated %K Yolk Sac %X In 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. %B Human molecular genetics %V 17 %P 1-14 %8 2008 Jan 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/17901045?dopt=Abstract %0 Journal Article %J Pest management science %D 2008 %T Neurotoxic implications of the agonistic action of CS-syndrome pyrethroids on the N-type Ca(v)2.2 calcium channel. %A Clark, J Marshall %A Symington, Steven B %X Cismethrin (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. %B Pest management science %V 64 %P 628-38 %8 2008 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/18383452?dopt=Abstract %0 Journal Article %J Journal of medical entomology %D 2008 %T A new ivermectin formulation topically kills permethrin-resistant human head lice (Anoplura: Pediculidae). %A Strycharz, Joseph P %A Yoon, Kyong Sup %A Clark, J Marshall %X This 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. %B Journal of medical entomology %V 45 %P 75-81 %8 2008 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/18283945?dopt=Abstract %0 Journal Article %J Blood %D 2008 %T Notch1 and TGFbeta1 cooperatively regulate Foxp3 expression and the maintenance of peripheral regulatory T cells. %A Samon, Jeremy B %A Champhekar, Ameya %A Minter, Lisa M %A Telfer, Janice C %A Miele, Lucio %A Fauq, Abdul %A Das, Pritam %A Golde, Todd E %A Osborne, Barbara A %X

Notch 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.

%B Blood %V 112 %P 1813-21 %8 2008 Sep 1 %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/18550850?dopt=Abstract %0 Journal Article %J Developmental biology %D 2008 %T PACSIN2 regulates cell adhesion during gastrulation in Xenopus laevis. %A Cousin, Hélène %A Desimone, Douglas W %A Alfandari, Dominique %X

We 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.

%B Developmental biology %V 319 %P 86-99 %8 2008 Jul 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/18495106?dopt=Abstract %0 Journal Article %J Environmental science & technology %D 2008 %T Perfluorinated compounds in human milk from Massachusetts, U.S.A. %A Tao, Lin %A Kannan, Kurunthachalam %A Wong, Chung M %A Arcaro, Kathleen F %A Butenhoff, John L %K Adult %K Environmental Monitoring %K Environmental Pollutants %K Female %K Fluorocarbons %K Humans %K Massachusetts %K Maternal Exposure %K Milk, Human %X Perfluorinated 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. %B Environmental science & technology %V 42 %P 3096-101 %8 2008 Apr 15 %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/18497172?dopt=Abstract %0 Journal Article %J J Mater Chem %D 2008 %T Protein-passivated Fe(3)O(4) nanoparticles: low toxicity and rapid heating for thermal therapy. %A Samanta, Bappaditya %A Yan, Haoheng %A Fischer, Nicholas O %A Shi, Jing %A Jerry, D Joseph %A Rotello, Vincent M %X

Thermotherapy 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.

%B J Mater Chem %V 18 %P 1204-1208 %8 2008 %G ENG %N 11 %1

http://www.ncbi.nlm.nih.gov/pubmed/19122852?dopt=Abstract

%0 Journal Article %J Breast cancer research : BCR %D 2008 %T Regulation of cancer stem cells by p53. %A Jerry, D Joseph %A Tao, Luwei %A Yan, Haoheng %X The 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. %B Breast cancer research : BCR %V 10 %P 304 %8 2008 %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/18828866?dopt=Abstract %0 Journal Article %J Breast Cancer Res %D 2008 %T Regulation of cancer stem cells by p53. %A Jerry, D Joseph %A Tao, Luwei %A Yan, Haoheng %K Animals %K Gene Expression Regulation, Neoplastic %K Genes, p53 %K Humans %K Mice %K Mice, Inbred BALB C %K Models, Biological %K Mutation %K Neoplastic Stem Cells %K Phenotype %K Tumor Suppressor Protein p53 %X

The 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.

%B Breast Cancer Res %V 10 %P 304 %8 2008 %G eng %N 4 %1

http://www.ncbi.nlm.nih.gov/pubmed/18828866?dopt=Abstract

%0 Journal Article %J Endocrinology %D 2008 %T Transcriptional responses to estrogen and progesterone in mammary gland identify networks regulating p53 activity. %A Lu, Shaolei %A Becker, Klaus A %A Hagen, Mary J %A Yan, Haoheng %A Roberts, Amy L %A Mathews, Lesley A %A Schneider, Sallie S %A Siegelmann, Hava T %A MacBeth, Kyle J %A Tirrell, Stephen M %A Blanchard, Jeffrey L %A Jerry, D Joseph %X

Estrogen 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.

%B Endocrinology %V 149 %P 4809-20 %8 2008 Oct %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/18556351?dopt=Abstract %0 Journal Article %J Endocrinology %D 2008 %T Transcriptional responses to estrogen and progesterone in mammary gland identify networks regulating p53 activity. %A Lu, Shaolei %A Becker, Klaus A %A Hagen, Mary J %A Yan, Haoheng %A Roberts, Amy L %A Mathews, Lesley A %A Schneider, Sallie S %A Siegelmann, Hava T %A MacBeth, Kyle J %A Tirrell, Stephen M %A Blanchard, Jeffrey L %A Jerry, D Joseph %K 14-3-3 Proteins %K Animals %K Breast Neoplasms %K Cell Line, Transformed %K Cell Line, Tumor %K Early Growth Response Protein 1 %K Epithelial Cells %K Epithelium %K Estradiol %K Female %K Gene Expression Profiling %K Humans %K Mammary Glands, Animal %K Mice %K Mice, Inbred BALB C %K Mice, Mutant Strains %K NF-kappa B %K Oligonucleotide Array Sequence Analysis %K Ovariectomy %K Progesterone %K Transcription, Genetic %K Tumor Suppressor Protein p53 %X

Estrogen 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.

%B Endocrinology %V 149 %P 4809-20 %8 2008 Oct %G eng %N 10 %1

http://www.ncbi.nlm.nih.gov/pubmed/18556351?dopt=Abstract

%0 Journal Article %J International journal for parasitology %D 2007 %T Anti-Trypanosoma brucei activity in Cape buffalo serum during the cryptic phase of parasitemia is mediated by antibodies. %A Guirnalda, Patrick %A Murphy, Noel B %A Nolan, Derek %A Black, Samuel J %X Cape 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. %B International journal for parasitology %V 37 %P 1391-9 %8 2007 Oct %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/17583714?dopt=Abstract %0 Journal Article %J Microbes and infection / Institut Pasteur %D 2007 %T Brucella abortus bacA mutant induces greater pro-inflammatory cytokines than the wild-type parent strain. %A Parent, Michelle A %A Goenka, Radhika %A Murphy, Erin %A Levier, Kristen %A Carreiro, Nuno %A Golding, Basil %A Ferguson, Gail %A Roop, R Martin %A Walker, Graham C %A Baldwin, Cynthia L %X The 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. %B Microbes and infection / Institut Pasteur %V 9 %P 55-62 %8 2007 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/17196866?dopt=Abstract %0 Journal Article %J Molecular immunology %D 2007 %T Comparison of gene expression by co-cultured WC1+ gammadelta and CD4+ alphabeta T cells exhibiting a recall response to bacterial antigen. %A Blumerman, Seth L %A Herzig, Carolyn T A %A Wang, Fei %A Coussens, Paul M %A Baldwin, Cynthia L %X Immunization 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. %B Molecular immunology %V 44 %P 2023-35 %8 2007 Mar %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/17081609?dopt=Abstract %0 Journal Article %J Immunity %D 2007 %T Deletion of CD4 and CD8 coreceptors permits generation of alphabetaT cells that recognize antigens independently of the MHC. %A Van Laethem, Francois %A Sarafova, Sophia D %A Park, Jung-Hyun %A Tai, Xuguang %A Pobezinsky, Leonid %A Guinter, Terry I %A Adoro, Stanley %A Adams, Anthony %A Sharrow, Susan O %A Feigenbaum, Lionel %A Singer, Alfred %K Animals %K Antigens, CD4 %K Antigens, CD8 %K Blotting, Northern %K Cell Differentiation %K Flow Cytometry %K Fluorescent Antibody Technique %K Immunoprecipitation %K Lymphocyte Specific Protein Tyrosine Kinase p56(lck) %K Major Histocompatibility Complex %K Mice %K Mice, Transgenic %K Receptors, Antigen, T-Cell, alpha-beta %K T-Lymphocytes %K Thymus Gland %X

The 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.

%B Immunity %V 27 %P 735-50 %8 2007 Nov %G eng %N 5 %1

http://www.ncbi.nlm.nih.gov/pubmed/18023370?dopt=Abstract

%0 Journal Article %J American journal of veterinary research %D 2007 %T Early laminar events involving endothelial activation in horses with black walnut- induced laminitis. %A Loftus, John P %A Black, Samuel J %A Pettigrew, Amanda %A Abrahamsen, Eric J %A Belknap, James K %X To 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. %B American journal of veterinary research %V 68 %P 1205-11 %8 2007 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/17975975?dopt=Abstract %0 Journal Article %J Am J Pathol %D 2007 %T Genetic mapping in mice identifies DMBT1 as a candidate modifier of mammary tumors and breast cancer risk. %A Blackburn, Anneke C %A Hill, Linda Z %A Roberts, Amy L %A Wang, Jun %A Aud, Dee %A Jung, Jimmy %A Nikolcheva, Tania %A Allard, John %A Peltz, Gary %A Otis, Christopher N %A Cao, Qing J %A Ricketts, Reva St J %A Naber, Stephen P %A Mollenhauer, Jan %A Poustka, Annemarie %A Malamud, Daniel %A Jerry, D Joseph %K Animals %K Breast Neoplasms %K Chromosome Mapping %K Female %K Gene Expression Profiling %K Genetic Predisposition to Disease %K Genotype %K Humans %K Intestine, Small %K Mammary Glands, Animal %K Mammary Glands, Human %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Mucins %K Oligonucleotide Array Sequence Analysis %K Risk Factors %K Survival Rate %K Tumor Suppressor Protein p53 %X

Low-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.

%B Am J Pathol %V 170 %P 2030-41 %8 2007 Jun %G eng %N 6 %1

http://www.ncbi.nlm.nih.gov/pubmed/17525270?dopt=Abstract

%0 Journal Article %J The American journal of pathology %D 2007 %T Genetic mapping in mice identifies DMBT1 as a candidate modifier of mammary tumors and breast cancer risk. %A Blackburn, Anneke C %A Hill, Linda Z %A Roberts, Amy L %A Wang, Jun %A Aud, Dee %A Jung, Jimmy %A Nikolcheva, Tania %A Allard, John %A Peltz, Gary %A Otis, Christopher N %A Cao, Qing J %A Ricketts, Reva St J %A Naber, Stephen P %A Mollenhauer, Jan %A Poustka, Annemarie %A Malamud, Daniel %A Jerry, D Joseph %X Low-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. %B The American journal of pathology %V 170 %P 2030-41 %8 2007 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/17525270?dopt=Abstract %0 Journal Article %J Equine veterinary journal %D 2007 %T Laminar xanthine oxidase, superoxide dismutase and catalase activities in the prodromal stage of black-walnut induced equine laminitis. %A Loftus, J P %A Belknap, J K %A Stankiewicz, K M %A Black, S J %X REASONS 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. %B Equine veterinary journal %V 39 %P 48-53 %8 2007 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/17228595?dopt=Abstract %0 Journal Article %J Mamm Genome %D 2007 %T Mammary tumor modifiers in BALB/cJ mice heterozygous for p53. %A Koch, Joanna G %A Gu, Xiangjun %A Han, Younghun %A El-Naggar, Adel K %A Olson, Melissa V %A Medina, Daniel %A Jerry, D Joseph %A Blackburn, Anneke C %A Peltz, Gary %A Amos, Christopher I %A Lozano, Guillermina %K Animals %K Crosses, Genetic %K Female %K Genes, p53 %K Genetic Predisposition to Disease %K Heterozygote %K Male %K Mammary Neoplasms, Animal %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Rats %K Specific Pathogen-Free Organisms %X

BALB/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)).

%B Mamm Genome %V 18 %P 300-9 %8 2007 May %G eng %N 5 %1

http://www.ncbi.nlm.nih.gov/pubmed/17557176?dopt=Abstract

%0 Journal Article %J Developmental and comparative immunology %D 2007 %T Molecular cloning of bovine chemokine receptors and expression by WC1+ gammadelta T cells. %A Blumerman, Seth L %A Wang, Fei %A Herzig, Carolyn T A %A Baldwin, Cynthia L %X Chemokine 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. %B Developmental and comparative immunology %V 31 %P 87-102 %8 2007 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/16762412?dopt=Abstract %0 Journal Article %J Nature reviews. Immunology %D 2007 %T Notch signalling during peripheral T-cell activation and differentiation. %A Osborne, Barbara A %A Minter, Lisa M %X For 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. %B Nature reviews. Immunology %V 7 %P 64-75 %8 2007 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/17170755?dopt=Abstract %0 Journal Article %J Journal of environmental monitoring : JEM %D 2007 %T Polybrominated diphenyl ethers and organochlorine pesticides in human breast milk from Massachusetts, USA. %A Johnson-Restrepo, Boris %A Addink, Rudolf %A Wong, Chung %A Arcaro, Kathleen %A Kannan, Kurunthachalam %K Adult %K Environmental Exposure %K Female %K Gas Chromatography-Mass Spectrometry %K Humans %K Hydrocarbons, Chlorinated %K Infant, Newborn %K Middle Aged %K Milk, Human %K Pesticides %K Polybrominated Biphenyls %K Quality Control %K United States %K United States Environmental Protection Agency %X Concentrations 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. %B Journal of environmental monitoring : JEM %V 9 %P 1205-12 %8 2007 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/17968447?dopt=Abstract %0 Journal Article %J Developmental biology %D 2007 %T Proteolytic processing of phospholipase Czeta and [Ca2+]i oscillations during mammalian fertilization. %A Kurokawa, Manabu %A Yoon, Sook Young %A Alfandari, Dominique %A Fukami, Kiyoko %A Sato, Ken-ichi %A Fissore, Rafael A %X

Phospholipase 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.

%B Developmental biology %V 312 %P 407-18 %8 2007 Dec 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/18028898?dopt=Abstract %0 Journal Article %J Invertebrate neuroscience : IN %D 2007 %T Pyrethroid action on calcium channels: neurotoxicological implications. %A Clark, J Marshall %A Symington, Steven B %X Actions 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. %B Invertebrate neuroscience : IN %V 7 %P 3-16 %8 2007 Mar %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/17294162?dopt=Abstract %0 Journal Article %J Breast Cancer Res %D 2007 %T Roles for estrogen and progesterone in breast cancer prevention. %A Jerry, D Joseph %K Animals %K Antineoplastic Agents %K Breast Neoplasms %K Cell Transformation, Neoplastic %K Disease Models, Animal %K Estrogens %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K Mammary Neoplasms, Animal %K Mice %K Progesterone %K Rats %X

Prevention 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.

%B Breast Cancer Res %V 9 %P 102 %8 2007 %G eng %N 2 %1

http://www.ncbi.nlm.nih.gov/pubmed/17381827?dopt=Abstract

%0 Journal Article %J Society of Reproduction and Fertility supplement %D 2007 %T Signalling pathways involved in sperm capacitation. %A Salicioni, Ana M %A Platt, Mark D %A Wertheimer, Eva V %A Arcelay, Enid %A Allaire, Alicia %A Sosnik, Julian %A Visconti, Pablo E %K Acrosome Reaction %K Animals %K Cell Membrane %K Female %K Male %K Mammals %K Phosphorylation %K Seminal Plasma Proteins %K Signal Transduction %K Sperm Capacitation %K Spermatozoa %X

After 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.

%B Society of Reproduction and Fertility supplement %V 65 %P 245-59 %8 2007 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/17644966?dopt=Abstract %0 Journal Article %J Environmental science & technology %D 2007 %T Synthetic musk fragrances in human milk from the United States. %A Reiner, Jessica L %A Wong, Chung M %A Arcaro, Kathleen F %A Kannan, Kurunthachalam %K Adult %K Benzopyrans %K Environmental Exposure %K Female %K Humans %K Infant %K Massachusetts %K Milk, Human %K Perfume %K Polycyclic Compounds %K Tetrahydronaphthalenes %K United States %K Xylenes %X Synthetic 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. %B Environmental science & technology %V 41 %P 3815-20 %8 2007 Jun 1 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/17612154?dopt=Abstract %0 Journal Article %J European journal of immunology %D 2007 %T WC1+ gammadelta T cell memory population is induced by killed bacterial vaccine. %A Blumerman, Seth L %A Herzig, Carolyn T A %A Baldwin, Cynthia L %X Limited 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. %B European journal of immunology %V 37 %P 1204-16 %8 2007 May %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/17429840?dopt=Abstract %0 Journal Article %J Cellular immunology %D 2006 %T Characterization of WC1 co-receptors on functionally distinct subpopulations of ruminant gamma delta T cells. %A Rogers, Aric N %A Vanburen, Denille G %A Zou, Baixiang %A Lahmers, Kevin K %A Herzig, Carolyn T A %A Brown, Wendy C %A Telfer, Janice C %A Baldwin, Cynthia L %X

WC1 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.

%B Cellular immunology %V 239 %P 151-61 %8 2006 Feb %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/16828467?dopt=Abstract %0 Journal Article %J Immunogenetics %D 2006 %T Differential TCR gene usage between WC1- and WC1+ ruminant gammadelta T cell subpopulations including those responding to bacterial antigen. %A Blumerman, Seth L %A Herzig, Carolyn T A %A Rogers, Aric N %A Telfer, Janice C %A Baldwin, Cynthia L %X

Ruminant 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.

%B Immunogenetics %V 58 %P 680-92 %8 2006 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/16799810?dopt=Abstract %0 Journal Article %J Molecular human reproduction %D 2006 %T Evidence for the involvement of proline-directed serine/threonine phosphorylation in sperm capacitation. %A Jha, K N %A Salicioni, A M %A Arcelay, E %A Chertihin, O %A Kumari, S %A Herr, J C %A Visconti, P E %K 1-Methyl-3-isobutylxanthine %K Amino Acid Motifs %K Animals %K Antibodies, Monoclonal %K beta-Cyclodextrins %K Blotting, Western %K Bucladesine %K Butadienes %K Cattle %K Cholesterol %K Cyclic AMP %K Flavonoids %K Humans %K Male %K Mice %K Mitogen-Activated Protein Kinase 1 %K Mitogen-Activated Protein Kinase 3 %K Mitosis %K Nitriles %K Phosphoproteins %K Phosphorylation %K Proline %K Protein Processing, Post-Translational %K Protein-Serine-Threonine Kinases %K Serum Albumin, Bovine %K Sodium Bicarbonate %K Sperm Capacitation %K Spermatozoa %X

To 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.

%B Molecular human reproduction %V 12 %P 781-9 %8 2006 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/17050774?dopt=Abstract %0 Journal Article %J Developmental cell %D 2006 %T An FGF response pathway that mediates hepatic gene induction in embryonic endoderm cells. %A Calmont, Amélie %A Wandzioch, Ewa %A Tremblay, Kimberly D %A Minowada, George %A Kaestner, Klaus H %A Martin, Gail R %A Zaret, Kenneth S %K Animals %K Body Patterning %K Cell Proliferation %K Embryo Culture Techniques %K Embryonic Induction %K Endoderm %K Fibroblast Growth Factors %K Integrases %K Liver %K MAP Kinase Signaling System %K Membrane Proteins %K Mesoderm %K Mice %K Mice, Inbred C57BL %K Mice, Transgenic %K Organogenesis %K Phosphatidylinositol 3-Kinases %K Phosphorylation %K Proteins %K Signal Transduction %X While 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. %B Developmental cell %V 11 %P 339-48 %8 2006 Sep %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/16950125?dopt=Abstract %0 Journal Article %J Developmental biology %D 2006 %T Hex homeobox gene controls the transition of the endoderm to a pseudostratified, cell emergent epithelium for liver bud development. %A Bort, Roque %A Signore, Massimo %A Tremblay, Kimberly %A Martinez Barbera, Juan Pedro %A Zaret, Kenneth S %K Animals %K Body Patterning %K Cell Differentiation %K Cell Proliferation %K Endoderm %K Epithelium %K Female %K Gene Expression Regulation, Developmental %K Hedgehog Proteins %K Homeodomain Proteins %K Liver %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Mutant Strains %K Mutation %K Organogenesis %K Stromal Cells %K Trans-Activators %K Transcription Factors %X Little 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. %B Developmental biology %V 290 %P 44-56 %8 2006 Feb 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/16364283?dopt=Abstract %0 Journal Article %J Critical reviews in immunology %D 2006 %T Host immune responses to the intracellular bacteria Brucella: does the bacteria instruct the host to facilitate chronic infection? %A Baldwin, Cynthia L %A Goenka, Radhika %X Brucella 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. %B Critical reviews in immunology %V 26 %P 407-42 %8 2006 %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/17341186?dopt=Abstract %0 Journal Article %J Mammalian genome : official journal of the International Mammalian Genome Society %D 2006 %T Identification of candidate maternal-effect genes through comparison of multiple microarray data sets. %A Mager, Jesse %A Schultz, Richard M %A Brunk, Brian P %A Bartolomei, Marisa S %K Animals %K Blastocyst %K Female %K Gene Expression Profiling %K Mice %K Models, Biological %K Oligonucleotide Array Sequence Analysis %K Reverse Transcriptase Polymerase Chain Reaction %K RNA, Messenger, Stored %X Transcriptional 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. %B Mammalian genome : official journal of the International Mammalian Genome Society %V 17 %P 941-9 %8 2006 Sep %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/16964442?dopt=Abstract %0 Journal Article %J Immunogenetics %D 2006 %T Identification of three new bovine T-cell receptor delta variable gene subgroups expressed by peripheral blood T cells. %A Herzig, Carolyn T A %A Blumerman, Seth L %A Baldwin, Cynthia L %X To 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. %B Immunogenetics %V 58 %P 746-57 %8 2006 Sep %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/16896832?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2006 %T Leukocyte emigration in the early stages of laminitis. %A Black, Samuel J %A Lunn, D Paul %A Yin, Cailing %A Hwang, Misako %A Lenz, Stephen D %A Belknap, James K %X The 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. %B Veterinary immunology and immunopathology %V 109 %P 161-6 %8 2006 Jan 15 %G eng %N 1-2 %1 http://www.ncbi.nlm.nih.gov/pubmed/16169600?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2006 %T Matrix metalloproteinase-9 in laminae of black walnut extract treated horses correlates with neutrophil abundance. %A Loftus, John P %A Belknap, James K %A Black, Samuel J %X We 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. %B Veterinary immunology and immunopathology %V 113 %P 267-76 %8 2006 Oct 15 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/16822550?dopt=Abstract %0 Journal Article %J The EMBO journal %D 2006 %T Notch1 augments NF-kappaB activity by facilitating its nuclear retention. %A Shin, Hyun Mu %A Minter, Lisa M %A Cho, Ok Hyun %A Gottipati, Sridevi %A Fauq, Abdul H %A Golde, Todd E %A Sonenshein, Gail E %A Osborne, Barbara A %X Notch1 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. %B The EMBO journal %V 25 %P 129-38 %8 2006 Jan 11 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/16319921?dopt=Abstract %0 Journal Article %J Development (Cambridge, England) %D 2006 %T Phosphorylation of IP3R1 and the regulation of [Ca2+]i responses at fertilization: a role for the MAP kinase pathway. %A Lee, Bora %A Vermassen, Elke %A Yoon, Sook-Young %A Vanderheyden, Veerle %A Ito, Junya %A Alfandari, Dominique %A De Smedt, Humbert %A Parys, Jan B %A Fissore, Rafael A %K Amino Acid Sequence %K Animals %K Antibodies, Phospho-Specific %K Calcium %K Cell Cycle %K Enzyme Activation %K Female %K Fertilization %K Humans %K Inositol 1,4,5-Trisphosphate Receptors %K MAP Kinase Signaling System %K Mice %K Mitogen-Activated Protein Kinases %K Molecular Sequence Data %K Oocytes %K Phosphorylation %K Sequence Alignment %K Xenopus laevis %K Zygote %X

A 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.

%B Development (Cambridge, England) %V 133 %P 4355-65 %8 2006 Nov %G eng %N 21 %1 http://www.ncbi.nlm.nih.gov/pubmed/17038520?dopt=Abstract %0 Journal Article %J Development (Cambridge, England) %D 2006 %T Phosphorylation of IP3R1 and the regulation of [Ca2+]i responses at fertilization: a role for the MAP kinase pathway. %A Lee, Bora %A Vermassen, Elke %A Yoon, Sook-Young %A Vanderheyden, Veerle %A Ito, Junya %A Alfandari, Dominique %A De Smedt, Humbert %A Parys, Jan B %A Fissore, Rafael A %X A 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. %B Development (Cambridge, England) %V 133 %P 4355-65 %8 2006 Nov %G eng %N 21 %1 http://www.ncbi.nlm.nih.gov/pubmed/17038520?dopt=Abstract %0 Journal Article %J The Journal of biological chemistry %D 2006 %T Regulation of the composition of the extracellular matrix by low density lipoprotein receptor-related protein-1: activities based on regulation of mRNA expression. %A Gaultier, Alban %A Salicioni, Ana Maria %A Arandjelovic, Sanja %A Gonias, Steven L %K Animals %K Biotin %K Cell Line %K Cell Membrane %K Cloning, Molecular %K Collagen %K Culture Media, Conditioned %K Electrophoresis, Gel, Two-Dimensional %K Electrophoresis, Polyacrylamide Gel %K Endocytosis %K Extracellular Matrix %K Fibrinogen %K Gene Expression Regulation %K Humans %K LDL-Receptor Related Proteins %K Low Density Lipoprotein Receptor-Related Protein-1 %K Mass Spectrometry %K Mice %K Microscopy, Fluorescence %K Neovascularization, Pathologic %K Phosphorylation %K Proteomics %K Receptors, LDL %K Reverse Transcriptase Polymerase Chain Reaction %K RNA, Messenger %K Signal Transduction %K Surface Properties %K Tumor Suppressor Proteins %X Low 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. %B The Journal of biological chemistry %V 281 %P 7332-40 %8 2006 Mar 17 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/16407289?dopt=Abstract %0 Journal Article %J The Journal of biological chemistry %D 2006 %T Sodium and epithelial sodium channels participate in the regulation of the capacitation-associated hyperpolarization in mouse sperm. %A Hernández-González, Enrique O %A Sosnik, Julian %A Edwards, Jennifer %A Acevedo, Juan José %A Mendoza-Lujambio, Irene %A López-González, Ignacio %A Demarco, Ignacio %A Wertheimer, Eva %A Darszon, Alberto %A Visconti, Pablo E %X In 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. %B The Journal of biological chemistry %V 281 %P 5623-33 %8 2006 Mar 3 %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/16407190?dopt=Abstract %0 Journal Article %J Cellular immunology %D 2006 %T Treatment of Brucella-susceptible mice with IL-12 increases primary and secondary immunity. %A Sathiyaseelan, Janaki %A Goenka, Radhika %A Parent, Michelle %A Benson, Rita M %A Murphy, Erin A %A Fernandes, Dancella M %A Foulkes, Andrea S %A Baldwin, Cynthia L %X Brucella 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. %B Cellular immunology %V 243 %P 1-9 %8 2006 Sep %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/17184756?dopt=Abstract %0 Journal Article %J Molecular cancer research : MCR %D 2006 %T Use of an aggressive MCF-7 cell line variant, TMX2-28, to study cell invasion in breast cancer. %A Gozgit, Joseph M %A Pentecost, Brian T %A Marconi, Sharon A %A Otis, Christopher N %A Wu, Chuanyue %A Arcaro, Kathleen F %X An 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. %B Molecular cancer research : MCR %V 4 %P 905-13 %8 2006 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/17189381?dopt=Abstract %0 Journal Article %J Molecular cancer research : MCR %D 2006 %T Use of an aggressive MCF-7 cell line variant, TMX2-28, to study cell invasion in breast cancer. %A Gozgit, Joseph M %A Pentecost, Brian T %A Marconi, Sharon A %A Otis, Christopher N %A Wu, Chuanyue %A Arcaro, Kathleen F %K Breast Neoplasms %K Cell Adhesion Molecules %K Cell Line, Tumor %K Cytoskeletal Proteins %K Female %K Humans %K Neoplasm Invasiveness %K Polymerase Chain Reaction %K Receptors, Estrogen %X An 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. %B Molecular cancer research : MCR %V 4 %P 905-13 %8 2006 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/17189381?dopt=Abstract %0 Journal Article %J Developmental biology %D 2005 %T Distinct populations of endoderm cells converge to generate the embryonic liver bud and ventral foregut tissues. %A Tremblay, Kimberly D %A Zaret, Kenneth S %K Animals %K Body Patterning %K Cell Lineage %K Cell Movement %K Cell Proliferation %K Digestive System %K Embryo, Mammalian %K Embryonic Structures %K Endoderm %K Liver %K Mice %K Mice, Inbred C3H %K Morphogenesis %K Stem Cells %X The 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. %B Developmental biology %V 280 %P 87-99 %8 2005 Apr 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/15766750?dopt=Abstract %0 Journal Article %J Oncogene %D 2005 %T Estrogen and progesterone regulate radiation-induced p53 activity in mammary epithelium through TGF-beta-dependent pathways. %A Becker, Klaus A %A Lu, Shaolei %A Dickinson, Ellen S %A Dunphy, Karen A %A Mathews, Lesley %A Schneider, Sallie Smith %A Jerry, D Joseph %K Animals %K Blotting, Northern %K Blotting, Western %K Epithelium %K Estrogens %K Immunohistochemistry %K Mammary Glands, Animal %K Mice %K Mice, Inbred BALB C %K Mice, Knockout %K Organ Culture Techniques %K Progesterone %K Transforming Growth Factor beta %K Tumor Suppressor Protein p53 %X

DNA 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.

%B Oncogene %V 24 %P 6345-53 %8 2005 Sep 22 %G eng %N 42 %1

http://www.ncbi.nlm.nih.gov/pubmed/15940247?dopt=Abstract

%0 Journal Article %J Oncogene %D 2005 %T Estrogen and progesterone regulate radiation-induced p53 activity in mammary epithelium through TGF-beta-dependent pathways. %A Becker, Klaus A %A Lu, Shaolei %A Dickinson, Ellen S %A Dunphy, Karen A %A Mathews, Lesley %A Schneider, Sallie Smith %A Jerry, D Joseph %X

DNA 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.

%B Oncogene %V 24 %P 6345-53 %8 2005 Sep 22 %G eng %N 42 %1 http://www.ncbi.nlm.nih.gov/pubmed/15940247?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2005 %T Function of ruminant gammadelta T cells is defined by WC1.1 or WC1.2 isoform expression. %A Rogers, Aric N %A Vanburen, Denille G %A Hedblom, Emmett %A Tilahun, Mulualem E %A Telfer, Janice C %A Baldwin, Cynthia L %X

WC1 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.

%B Veterinary immunology and immunopathology %V 108 %P 211-7 %8 2005 Oct 18 %G eng %N 1-2 %1 http://www.ncbi.nlm.nih.gov/pubmed/16144715?dopt=Abstract %0 Journal Article %J Developmental biology %D 2005 %T Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm. %A Kurokawa, Manabu %A Sato, Ken-ichi %A Wu, Hua %A He, Changli %A Malcuit, Christopher %A Black, Samuel J %A Fukami, Kiyoko %A Fissore, Rafael A %X

A 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.

%B Developmental biology %V 285 %P 376-92 %8 2005 Sep 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/16098961?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 2005 %T Gammadelta T cell function varies with the expressed WC1 coreceptor. %A Rogers, Aric N %A Vanburen, Denille G %A Hedblom, Emmett E %A Tilahun, Mulualem E %A Telfer, Janice C %A Baldwin, Cynthia L %X

WC1 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.

%B Journal of immunology (Baltimore, Md. : 1950) %V 174 %P 3386-93 %8 2005 Mar 15 %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/15749871?dopt=Abstract %0 Journal Article %J Nature immunology %D 2005 %T Inhibitors of gamma-secretase block in vivo and in vitro T helper type 1 polarization by preventing Notch upregulation of Tbx21. %A Minter, Lisa M %A Turley, Danielle M %A Das, Pritam %A Shin, Hyun Mu %A Joshi, Ila %A Lawlor, Rebecca G %A Cho, Ok Hyun %A Palaga, Tanapat %A Gottipati, Sridevi %A Telfer, Janice C %A Kostura, Lisa %A Fauq, Abdul H %A Simpson, Katherine %A Such, Kimberly A %A Miele, Lucio %A Golde, Todd E %A Miller, Stephen D %A Osborne, Barbara A %X

Notch 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.

%B Nature immunology %V 6 %P 680-8 %8 2005 Jul %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/15991363?dopt=Abstract %0 Journal Article %J J Exp Med %D 2005 %T Restricted MHC-peptide repertoire predisposes to autoimmunity. %A Logunova, Nadezda N %A Viret, Christophe %A Pobezinsky, Leonid A %A Miller, Sara A %A Kazansky, Dmitri B %A Sundberg, John P %A Chervonsky, Alexander V %K Animals %K Autoimmune Diseases %K Autoimmunity %K CD4-Positive T-Lymphocytes %K CD8-Positive T-Lymphocytes %K Dermatitis %K Histocompatibility Antigens Class I %K Histocompatibility Antigens Class II %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C3H %K Mice, Inbred C57BL %K Mice, Knockout %K Mice, Transgenic %K Peptides %K Receptors, Antigen, T-Cell %X

MHC 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.

%B J Exp Med %V 202 %P 73-84 %8 2005 Jul 4 %G eng %N 1 %1

http://www.ncbi.nlm.nih.gov/pubmed/15998789?dopt=Abstract

%0 Journal Article %J Molecular cancer research : MCR %D 2005 %T Sensitivity to DNA damage is a common component of hormone-based strategies for protection of the mammary gland. %A Tu, Yifan %A Jerry, D Joseph %A Pazik, Brooke %A Schneider, Sallie Smith %X

An 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.

%B Molecular cancer research : MCR %V 3 %P 435-42 %8 2005 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/16123139?dopt=Abstract %0 Journal Article %J Mol Cancer Res %D 2005 %T Sensitivity to DNA damage is a common component of hormone-based strategies for protection of the mammary gland. %A Tu, Yifan %A Jerry, D Joseph %A Pazik, Brooke %A Schneider, Sallie Smith %K Animals %K Apoptosis %K Cell Death %K DNA Damage %K Dose-Response Relationship, Drug %K Estrogens %K Female %K Fenretinide %K Gamma Rays %K Genes, p53 %K Immunohistochemistry %K In Situ Nick-End Labeling %K Mammary Glands, Animal %K Mice %K Mice, Inbred BALB C %K Ovary %K Progesterone %K Retinoid X Receptors %K Risk Factors %K Sensitivity and Specificity %K Transforming Growth Factor beta %K Tretinoin %K Tumor Suppressor Protein p53 %K Up-Regulation %X

An 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.

%B Mol Cancer Res %V 3 %P 435-42 %8 2005 Aug %G eng %N 8 %1

http://www.ncbi.nlm.nih.gov/pubmed/16123139?dopt=Abstract

%0 Journal Article %J Archives of insect biochemistry and physiology %D 2005 %T Sequencing and characterization of a cDNA encoding a ferritin subunit of Colorado potato beetle, Leptinotarsa decemlineata. %A Qiu, Lihong %A Gao, Jian-Rong %A Clark, J Marshall %X A 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). %B Archives of insect biochemistry and physiology %V 60 %P 140-50 %8 2005 Nov %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/16235258?dopt=Abstract %0 Journal Article %J Nature genetics %D 2005 %T Strategies for dissecting epigenetic mechanisms in the mouse. %A Mager, Jesse %A Bartolomei, Marisa S %K Animals %K DNA Methylation %K Epigenesis, Genetic %K Gene Expression Regulation %K Genes, Tumor Suppressor %K Genome %K Genomics %K Mice %K Models, Genetic %K Mutagenesis %K Phenotype %K Proteomics %X Epigenetics 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. %B Nature genetics %V 37 %P 1194-200 %8 2005 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/16254566?dopt=Abstract %0 Journal Article %J Anticancer research %D 2004 %T Beta-galactosidase and alpha-mannosidase inhibit formation of multicellular nodules in breast cancer cell cultures. %A Arcaro, Kathleen F %A Wang, Joann %A Otis, Christopher N %A Zuckerman, Bert M %X In 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. %B Anticancer research %V 24 %P 139-44 %8 2004 Jan-Feb %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/15015588?dopt=Abstract %0 Journal Article %J Toxicology and applied pharmacology %D 2004 %T Differential action of polycyclic aromatic hydrocarbons on endogenous estrogen-responsive genes and on a transfected estrogen-responsive reporter in MCF-7 cells. %A Gozgit, Joseph M %A Nestor, Kathleen M %A Fasco, Michael J %A Pentecost, Brian T %A Arcaro, Kathleen F %X Polycyclic 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. %B Toxicology and applied pharmacology %V 196 %P 58-67 %8 2004 Apr 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/15050408?dopt=Abstract %0 Journal Article %J Journal of medical entomology %D 2004 %T Field validation of Aedes aegypti (Diptera: Culicidae) age estimation by analysis of cuticular hydrocarbons. %A Gerade, B B %A Lee, S H %A Scott, T W %A Edman, J D %A Harrington, L C %A Kitthawee, S %A Jones, J W %A Clark, J M %X In 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. %B Journal of medical entomology %V 41 %P 231-8 %8 2004 Mar %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/15061283?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 2004 %T Localization of the domains in Runx transcription factors required for the repression of CD4 in thymocytes. %A Telfer, Janice C %A Hedblom, Emmett E %A Anderson, Michele K %A Laurent, Micheline N %A Rothenberg, Ellen V %K Amino Acid Sequence %K Animals %K Antigens, CD4 %K CD4-Positive T-Lymphocytes %K CD8-Positive T-Lymphocytes %K Cell Differentiation %K Core Binding Factor Alpha 2 Subunit %K Core Binding Factor Alpha 3 Subunit %K DNA-Binding Proteins %K Down-Regulation %K Fetus %K Growth Inhibitors %K Mice %K Mice, Inbred C57BL %K Mice, Inbred DBA %K Mice, Transgenic %K Molecular Sequence Data %K Nuclear Matrix %K Organ Culture Techniques %K Peptide Fragments %K Protein Isoforms %K Protein Structure, Tertiary %K Proto-Oncogene Proteins %K Repressor Proteins %K Sequence Deletion %K Spleen %K Thymus Gland %K Transcription Factors %X The 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. %B Journal of immunology (Baltimore, Md. : 1950) %V 172 %P 4359-70 %8 2004 Apr 1 %G eng %N 7 %1 http://www.ncbi.nlm.nih.gov/pubmed/15034051?dopt=Abstract %0 Journal Article %J Cancer Res %D 2004 %T Loss of heterozygosity occurs via mitotic recombination in Trp53+/- mice and associates with mammary tumor susceptibility of the BALB/c strain. %A Blackburn, Anneke C %A McLary, S Christine %A Naeem, Rizwan %A Luszcz, Jason %A Stockton, David W %A Donehower, Lawrence A %A Mohammed, Mansoor %A Mailhes, John B %A Soferr, Tamar %A Naber, Stephen P %A Otis, Christopher N %A Jerry, D Joseph %K Animals %K Blotting, Southern %K Chromosomes %K Crosses, Genetic %K Disease Susceptibility %K DNA Damage %K DNA Repair %K Female %K Heterozygote %K Karyotyping %K Loss of Heterozygosity %K Male %K Mammary Neoplasms, Experimental %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Mice, Knockout %K Mitosis %K Recombination, Genetic %K Tumor Suppressor Protein p53 %X

Loss 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.

%B Cancer Res %V 64 %P 5140-7 %8 2004 Aug 1 %G eng %N 15 %1

http://www.ncbi.nlm.nih.gov/pubmed/15289317?dopt=Abstract

%0 Journal Article %J Cancer research %D 2004 %T Loss of heterozygosity occurs via mitotic recombination in Trp53+/- mice and associates with mammary tumor susceptibility of the BALB/c strain. %A Blackburn, Anneke C %A McLary, S Christine %A Naeem, Rizwan %A Luszcz, Jason %A Stockton, David W %A Donehower, Lawrence A %A Mohammed, Mansoor %A Mailhes, John B %A Soferr, Tamar %A Naber, Stephen P %A Otis, Christopher N %A Jerry, D Joseph %X Loss 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. %B Cancer research %V 64 %P 5140-7 %8 2004 Aug 1 %G eng %N 15 %1 http://www.ncbi.nlm.nih.gov/pubmed/15289317?dopt=Abstract %0 Journal Article %J Thrombosis and haemostasis %D 2004 %T Low density lipoprotein receptor-related protein: regulation of the plasma membrane proteome. %A Gonias, Steven L %A Wu, Lihua %A Salicioni, Ana Maria %K Animals %K Cell Membrane %K Extracellular Matrix Proteins %K Humans %K Low Density Lipoprotein Receptor-Related Protein-1 %K Membrane Proteins %K Proteome %X Proteins 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. %B Thrombosis and haemostasis %V 91 %P 1056-64 %8 2004 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/15175790?dopt=Abstract %0 Journal Article %J The Journal of biological chemistry %D 2004 %T Low density lipoprotein receptor-related protein-1 promotes beta1 integrin maturation and transport to the cell surface. %A Salicioni, Ana Maria %A Gaultier, Alban %A Brownlee, Cristina %A Cheezum, Michael K %A Gonias, Steven L %K Animals %K Antigens, CD29 %K Biological Transport %K Cell Adhesion %K Cell Line %K Cell Membrane %K Cells, Cultured %K CHO Cells %K Cricetinae %K Densitometry %K Dose-Response Relationship, Drug %K Down-Regulation %K Electrophoresis, Polyacrylamide Gel %K Endocytosis %K Endoplasmic Reticulum %K Fibronectins %K Glycoside Hydrolases %K Humans %K Immunoblotting %K Low Density Lipoprotein Receptor-Related Protein-1 %K Mice %K Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase %K Time Factors %X Low 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. %B The Journal of biological chemistry %V 279 %P 10005-12 %8 2004 Mar 12 %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/14699139?dopt=Abstract %0 Journal Article %J Developmental biology %D 2004 %T A PTP-PEST-like protein affects alpha5beta1-integrin-dependent matrix assembly, cell adhesion, and migration in Xenopus gastrula. %A Cousin, Hélène %A Alfandari, Dominique %X

During 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.

%B Developmental biology %V 265 %P 416-32 %8 2004 Jan 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/14732402?dopt=Abstract %0 Journal Article %J Cancer Res %D 2003 %T BALB/c alleles for Prkdc and Cdkn2a interact to modify tumor susceptibility in Trp53+/- mice. %A Blackburn, Anneke C %A Brown, Jennifer S %A Naber, Stephen P %A Otis, Christopher N %A Wood, Jeff T %A Jerry, D Joseph %K Alleles %K Animals %K Cyclin-Dependent Kinase Inhibitor p16 %K DNA-Activated Protein Kinase %K DNA-Binding Proteins %K Female %K Genes, p16 %K Genes, p53 %K Genetic Predisposition to Disease %K Inbreeding %K Male %K Mammary Neoplasms, Experimental %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Nuclear Proteins %K Polymorphism, Genetic %K Protein-Serine-Threonine Kinases %X

In 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.

%B Cancer Res %V 63 %P 2364-8 %8 2003 May 15 %G eng %N 10 %1

http://www.ncbi.nlm.nih.gov/pubmed/12750252?dopt=Abstract

%0 Journal Article %J Cancer research %D 2003 %T BALB/c alleles for Prkdc and Cdkn2a interact to modify tumor susceptibility in Trp53+/- mice. %A Blackburn, Anneke C %A Brown, Jennifer S %A Naber, Stephen P %A Otis, Christopher N %A Wood, Jeff T %A Jerry, D Joseph %X In 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. %B Cancer research %V 63 %P 2364-8 %8 2003 May 15 %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/12750252?dopt=Abstract %0 Journal Article %J Seminars in immunology %D 2003 %T Cell death in the thymus--it' s all a matter of contacts. %A Minter, Lisa M %A Osborne, Barbara A %X Apoptosis, 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. %B Seminars in immunology %V 15 %P 135-44 %8 2003 Jun %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/14563112?dopt=Abstract %0 Journal Article %J Vaccine %D 2003 %T Comparison of three different leptospiral vaccines for induction of a type 1 immune response to Leptospira borgpetersenii serovar Hardjo. %A Brown, Rachel A %A Blumerman, Seth %A Gay, Cyril %A Bolin, Carole %A Duby, Robert %A Baldwin, Cynthia L %X Leptospira 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. %B Vaccine %V 21 %P 4448-58 %8 2003 Oct 1 %G eng %N 27-30 %1 http://www.ncbi.nlm.nih.gov/pubmed/14505928?dopt=Abstract %0 Journal Article %J Developmental biology %D 2003 %T Dynamic morphogenetic events characterize the mouse visceral endoderm. %A Rivera-Pérez, Jaime A %A Mager, Jesse %A Magnuson, Terry %K Animals %K Blastocyst %K Endoderm %K Gastrula %K Homeodomain Proteins %K Mice %K Transcription Factors %X Several 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. %B Developmental biology %V 261 %P 470-87 %8 2003 Sep 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/14499654?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 2003 %T Glucocorticoid-induced apoptosis of thymocytes: requirement of proteasome-dependent mitochondrial activity. %A Tonomura, Noriko %A McLaughlin, Kelly %A Grimm, Lisa %A Goldsby, Richard A %A Osborne, Barbara A %X Thymocytes 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. %B Journal of immunology (Baltimore, Md. : 1950) %V 170 %P 2469-78 %8 2003 Mar 1 %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/12594272?dopt=Abstract %0 Journal Article %J Journal of neuroendocrinology %D 2003 %T Oestradiol-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. %A Curran-Rauhut, M A %A Petersen, S L %X Oestradiol (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. %B Journal of neuroendocrinology %V 15 %P 296-303 %8 2003 Mar %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/12588519?dopt=Abstract %0 Journal Article %J Archives of dermatology %D 2003 %T Permethrin-resistant human head lice, Pediculus capitis, and their treatment. %A Yoon, Kyong Sup %A Gao, Jian-Rong %A Lee, Si Hyeock %A Clark, J Marshall %A Brown, Leon %A Taplin, David %X To 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. %B Archives of dermatology %V 139 %P 994-1000 %8 2003 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/12925385?dopt=Abstract %0 Journal Article %J Journal of agricultural and food chemistry %D 2003 %T The persistence and degradation of chlorothalonil and chlorpyrifos in a cranberry bog. %A Putnam, Raymond A %A Nelson, Judd O %A Clark, J Marshall %X The 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. %B Journal of agricultural and food chemistry %V 51 %P 170-6 %8 2003 Jan 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/12502403?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2002 %T Activation of bovine peripheral blood gammadelta T cells for cell division and IFN-gamma production. %A Baldwin, C L %A Sathiyaseelan, T %A Naiman, B %A White, A M %A Brown, R %A Blumerman, S %A Rogers, A %A Black, S J %X

Bovine 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.

%B Veterinary immunology and immunopathology %V 87 %P 251-9 %8 2002 Sep 10 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/12072243?dopt=Abstract %0 Journal Article %J Microbial pathogenesis %D 2002 %T Brucella abortus siderophore 2,3-dihydroxybenzoic acid (DHBA) facilitates intracellular survival of the bacteria. %A Parent, Michelle A %A Bellaire, Bryan H %A Murphy, Erin A %A Roop, R Martin %A Elzer, Phillip H %A Baldwin, Cynthia L %X Siderophores 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. %B Microbial pathogenesis %V 32 %P 239-48 %8 2002 May %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/12071680?dopt=Abstract %0 Journal Article %J Development %D 2002 %T Epithelial cell cycling predicts p53 responsiveness to gamma-irradiation during post-natal mammary gland development. %A Minter, Lisa M %A Dickinson, Ellen S %A Naber, Stephen P %A Jerry, D Joseph %K Animals %K Animals, Newborn %K Cell Differentiation %K Cell Division %K Cell Nucleus %K Cyclin-Dependent Kinase Inhibitor p21 %K Cyclins %K Epithelium %K Estrogens %K Female %K Gamma Rays %K Gene Expression Regulation %K Mammary Glands, Animal %K Mice %K Mice, Mutant Strains %K Organ Culture Techniques %K Pregnancy %K Progesterone %K Tumor Suppressor Protein p53 %X

The 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.

%B Development %V 129 %P 2997-3008 %8 2002 Jun %G eng %N 12 %1

http://www.ncbi.nlm.nih.gov/pubmed/12050146?dopt=Abstract

%0 Journal Article %J Development (Cambridge, England) %D 2002 %T Epithelial cell cycling predicts p53 responsiveness to gamma-irradiation during post-natal mammary gland development. %A Minter, Lisa M %A Dickinson, Ellen S %A Naber, Stephen P %A Jerry, D Joseph %X

The 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.

%B Development (Cambridge, England) %V 129 %P 2997-3008 %8 2002 Jun %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/12050146?dopt=Abstract %0 Journal Article %J Infection and immunity %D 2002 %T Evaluation 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. %A Naiman, Brian M %A Blumerman, Seth %A Alt, David %A Bolin, Carole A %A Brown, Rachel %A Zuerner, Richard %A Baldwin, Cynthia L %X Organisms 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. %B Infection and immunity %V 70 %P 6147-57 %8 2002 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/12379692?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2002 %T Expression of the bovine high affinity IL-12 receptor beta2. %A White, Ann Marie %A Blumerman, Seth %A Naiman, Brian %A Baldwin, Cynthia L %X Four 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. %B Veterinary immunology and immunopathology %V 84 %P 127-42 %8 2002 Jan 15 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/11777529?dopt=Abstract %0 Journal Article %J Veterinary microbiology %D 2002 %T Fundamentals of host immune response against Brucella abortus: what the mouse model has revealed about control of infection. %A Baldwin, Cynthia L %A Parent, Michelle %X The 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. %B Veterinary microbiology %V 90 %P 367-82 %8 2002 Dec 20 %G eng %N 1-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/12414157?dopt=Abstract %0 Journal Article %J Breast Cancer Res %D 2002 %T Hormonal control of p53 and chemoprevention. %A Jerry, D Joseph %A Minter, Lisa M %A Becker, Klaus A %A Blackburn, Anneke C %K Breast Neoplasms %K Chemoprevention %K Female %K Genes, Tumor Suppressor %K Genetic Predisposition to Disease %K Hormones %K Humans %K Tumor Suppressor Protein p53 %X

Improvements 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.

%B Breast Cancer Res %V 4 %P 91-4 %8 2002 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/12052250?dopt=Abstract

%0 Journal Article %J Breast cancer research : BCR %D 2002 %T Hormonal control of p53 and chemoprevention. %A Jerry, D Joseph %A Minter, Lisa M %A Becker, Klaus A %A Blackburn, Anneke C %X

Improvements 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.

%B Breast cancer research : BCR %V 4 %P 91-4 %8 2002 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/12052250?dopt=Abstract %0 Journal Article %J Apoptosis : an international journal on programmed cell death %D 2002 %T Hyperbaric oxygen enhances apoptosis in hematopoietic cells. %A Ganguly, B J %A Tonomura, N %A Benson, R M %A Osborne, B A %A Granowitz, E V %X Hyperbaric 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). %B Apoptosis : an international journal on programmed cell death %V 7 %P 499-510 %8 2002 Dec %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/12370492?dopt=Abstract %0 Journal Article %J Veterinary microbiology %D 2002 %T Immune response overview. %A Baldwin, Cynthia L %X A 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. %B Veterinary microbiology %V 90 %P 365-6 %8 2002 Dec 20 %G eng %N 1-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/12414156?dopt=Abstract %0 Journal Article %J Immunology %D 2002 %T Immunological characterization of a gammadelta T-cell stimulatory ligand on autologous monocytes. %A Sathiyaseelan, Thillainayagam %A Naiman, Brian %A Welte, Stefan %A Machugh, Niall %A Black, Samuel J %A Baldwin, Cynthia L %X

Bovine 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.

%B Immunology %V 105 %P 181-9 %8 2002 Feb %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/11872093?dopt=Abstract %0 Journal Article %J Breast cancer research : BCR %D 2002 %T Knockout and transgenic mice of Trp53: what have we learned about p53 in breast cancer? %A Blackburn, Anneke C %A Jerry, D Joseph %X The 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. %B Breast cancer research : BCR %V 4 %P 101-11 %8 2002 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/12052252?dopt=Abstract %0 Journal Article %J Breast Cancer Res %D 2002 %T Knockout and transgenic mice of Trp53: what have we learned about p53 in breast cancer? %A Blackburn, Anneke C %A Jerry, D Joseph %K Animals %K Breast Neoplasms %K Disease Models, Animal %K Female %K Genes, p53 %K Mice %K Mice, Knockout %K Mice, Transgenic %X

The 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.

%B Breast Cancer Res %V 4 %P 101-11 %8 2002 %G eng %N 3 %1

http://www.ncbi.nlm.nih.gov/pubmed/12052252?dopt=Abstract

%0 Journal Article %J The Journal of biological chemistry %D 2002 %T The low density lipoprotein receptor-related protein mediates fibronectin catabolism and inhibits fibronectin accumulation on cell surfaces. %A Salicioni, Ana M %A Mizelle, Kellie S %A Loukinova, Elena %A Mikhailenko, Irina %A Strickland, Dudley K %A Gonias, Steven L %K Animals %K Cell Line %K Cell Membrane %K CHO Cells %K Cricetinae %K Embryo, Mammalian %K Fibroblasts %K Fibronectins %K Humans %K Kinetics %K Low Density Lipoprotein Receptor-Related Protein-1 %K Methionine %K Mice %K Recombinant Proteins %K Sulfur Radioisotopes %K Transfection %X Low 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. %B The Journal of biological chemistry %V 277 %P 16160-6 %8 2002 May 3 %G eng %N 18 %1 http://www.ncbi.nlm.nih.gov/pubmed/11867643?dopt=Abstract %0 Journal Article %J FEMS immunology and medical microbiology %D 2002 %T Major 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. %A Murphy, E %A Robertson, G T %A Parent, M %A Hagius, S D %A Roop, R M %A Elzer, P H %A Baldwin, C L %X Immune 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. %B FEMS immunology and medical microbiology %V 33 %P 191-200 %8 2002 Jul 12 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/12110481?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2002 %T Partial cDNA sequences of bovine CD72 and CD166/ALCAM, ligands for SRCR-family accessory molecules CD5 and CD6. %A Rogers, Aric N %A Welte, Stefan %A Black, Samuel J %A Baldwin, Cynthia L %X

Accessory/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.

%B Veterinary immunology and immunopathology %V 85 %P 233-9 %8 2002 Mar %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/11943324?dopt=Abstract %0 Journal Article %J Journal of neuroendocrinology %D 2002 %T Regulation 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. %A Curran-Rauhut, M A %A Petersen, S L %X GABA 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. %B Journal of neuroendocrinology %V 14 %P 310-7 %8 2002 Apr %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/11963828?dopt=Abstract %0 Journal Article %J The Journal of cell biology %D 2002 %T Regulation of Rac1 activation by the low density lipoprotein receptor-related protein. %A Ma, Zhong %A Thomas, Keena S %A Webb, Donna J %A Moravec, Radim %A Salicioni, Ana Maria %A Mars, Wendy M %A Gonias, Steven L %K Animals %K Cell Line %K Cell Movement %K Culture Media, Serum-Free %K Enzyme Activation %K Fibroblasts %K Fibronectins %K Gene Expression Regulation, Enzymologic %K Ligands %K Low Density Lipoprotein Receptor-Related Protein-1 %K Mice %K Mice, Knockout %K Microscopy, Fluorescence %K Mitogen-Activated Protein Kinases %K Protein Binding %K rac1 GTP-Binding Protein %K Receptors, Cell Surface %K Receptors, Urokinase Plasminogen Activator %K Transfection %K Vitronectin %X The 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. %B The Journal of cell biology %V 159 %P 1061-70 %8 2002 Dec 23 %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/12499359?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 2002 %T Response of bovine gammadelta T cells to activation through CD3. %A Sathiyaseelan, T %A Rogers, Aric %A Baldwin, Cynthia L %X Since 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. %B Veterinary immunology and immunopathology %V 90 %P 155-68 %8 2002 Dec %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/12459163?dopt=Abstract %0 Journal Article %J Antioxidants & redox signaling %D 2002 %T Serum xanthine oxidase: origin, regulation, and contribution to control of trypanosome parasitemia. %A Wang, Jun %A Van Praagh, Andrew %A Hamilton, Erika %A Wang, Qin %A Zou, Baixiang %A Muranjan, Madhavi %A Murphy, Noel B %A Black, Samuel J %X African 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. %B Antioxidants & redox signaling %V 4 %P 161-78 %8 2002 Feb %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11970851?dopt=Abstract %0 Journal Article %J Pest management science %D 2001 %T DNA-based genotyping techniques for the detection of point mutations associated with insecticide resistance in Colorado potato beetle Leptinotarsa decemlineata. %A Clark, J M %A Lee, S H %A Kim, H J %A Yoon, K S %A Zhang, A %X Three 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. %B Pest management science %V 57 %P 968-74 %8 2001 Oct %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/11695191?dopt=Abstract %0 Journal Article %J Immunology %D 2001 %T Early Vlambda diversification in sheep. %A Jeong, Y %A Osborne, B A %A Goldsby, R A %X

This 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.

%B Immunology %V 103 %P 26-34 %8 2001 May %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11380689?dopt=Abstract %0 Journal Article %J FEMS immunology and medical microbiology %D 2001 %T Immune control of Brucella abortus 2308 infections in BALB/c mice. %A Murphy, E A %A Parent, M %A Sathiyaseelan, J %A Jiang, X %A Baldwin, C L %X BALB/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. %B FEMS immunology and medical microbiology %V 32 %P 85-8 %8 2001 Dec %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11750226?dopt=Abstract %0 Journal Article %J International journal for parasitology %D 2001 %T Innate and acquired control of trypanosome parasitaemia in Cape buffalo. %A Black, S J %A Sicard, E L %A Murphy, N %A Nolan, D %X The 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. %B International journal for parasitology %V 31 %P 562-5 %8 2001 May 1 %G eng %N 5-6 %1 http://www.ncbi.nlm.nih.gov/pubmed/11334943?dopt=Abstract %0 Journal Article %J The Journal of parasitology %D 2001 %T Innate and acquired resistance to African trypanosomiasis. %A Black, S J %A Seed, J R %A Murphy, N B %X The 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. %B The Journal of parasitology %V 87 %P 1-9 %8 2001 Feb %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11227870?dopt=Abstract %0 Journal Article %J Immunology %D 2001 %T Interferon-gamma is crucial for surviving a Brucella abortus infection in both resistant C57BL/6 and susceptible BALB/c mice. %A Murphy, E A %A Sathiyaseelan, J %A Parent, M A %A Zou, B %A Baldwin, C L %X Brucella 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. %B Immunology %V 103 %P 511-8 %8 2001 Aug %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/11529943?dopt=Abstract %0 Journal Article %J Methods in cell biology %D 2001 %T Model cell lines for the study of apoptosis in vitro. %A Valavanis, C %A Hu, Y %A Yang, Y %A Osborne, B A %A Chouaib, S %A Greene, L %A Ashwell, J D %A Schwartz, L M %B Methods in cell biology %V 66 %P 417-36 %8 2001 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/11396014?dopt=Abstract %0 Journal Article %J Development (Cambridge, England) %D 2001 %T Mouse embryos lacking Smad1 signals display defects in extra-embryonic tissues and germ cell formation. %A Tremblay, K D %A Dunn, N R %A Robertson, E J %K Animals %K Bone Morphogenetic Proteins %K Cell Differentiation %K Cell Lineage %K Crosses, Genetic %K DNA-Binding Proteins %K Ectoderm %K Embryonic and Fetal Development %K Extraembryonic Membranes %K Gastrula %K Genes, Lethal %K Genotype %K Germ Cells %K Heterozygote %K Mesoderm %K Mice %K Mice, Mutant Strains %K Phenotype %K Phosphoproteins %K Smad Proteins %K Smad1 Protein %K Smad5 Protein %K Smad8 Protein %K Stem Cells %K Tissue Distribution %K Trans-Activators %X The 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. %B Development (Cambridge, England) %V 128 %P 3609-21 %8 2001 Sep %G eng %N 18 %1 http://www.ncbi.nlm.nih.gov/pubmed/11566864?dopt=Abstract %0 Journal Article %J Infection and immunity %D 2001 %T Protective killed Leptospira borgpetersenii vaccine induces potent Th1 immunity comprising responses by CD4 and gammadelta T lymphocytes. %A Naiman, B M %A Alt, D %A Bolin, C A %A Zuerner, R %A Baldwin, C L %X Leptospira 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. %B Infection and immunity %V 69 %P 7550-8 %8 2001 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/11705932?dopt=Abstract %0 Journal Article %J Endocrinology %D 2000 %T Detection of estrogen receptor-beta messenger ribonucleic acid and 125I-estrogen binding sites in luteinizing hormone-releasing hormone neurons of the rat brain. %A Hrabovszky, E %A Shughrue, P J %A Merchenthaler, I %A Hajszán, T %A Carpenter, C D %A Liposits, Z %A Petersen, S L %X Luteinizing 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. %B Endocrinology %V 141 %P 3506-9 %8 2000 Sep %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/10965924?dopt=Abstract %0 Journal Article %J The Journal of comparative neurology %D 2000 %T Distribution of mRNAs encoding the arylhydrocarbon receptor, arylhydrocarbon receptor nuclear translocator, and arylhydrocarbon receptor nuclear translocator-2 in the rat brain and brainstem. %A Petersen, S L %A Curran, M A %A Marconi, S A %A Carpenter, C D %A Lubbers, L S %A McAbee, M D %X Dioxin 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. %B The Journal of comparative neurology %V 427 %P 428-39 %8 2000 Nov 20 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/11054704?dopt=Abstract %0 Journal Article %J Research in veterinary science %D 2000 %T Evaluation of cell replication by bovine T cells in polyclonally activated cultures using carboxyfluorescein succinimidyl ester (CFSE) loading and flow cytometric analysis. %A Sathiyaseelan, T %A Baldwin, C L %X Studies 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. %B Research in veterinary science %V 69 %P 275-81 %8 2000 Dec %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/11124100?dopt=Abstract %0 Journal Article %J Development (Cambridge, England) %D 2000 %T Formation of the definitive endoderm in mouse is a Smad2-dependent process. %A Tremblay, K D %A Hoodless, P A %A Bikoff, E K %A Robertson, E J %K Animals %K beta-Galactosidase %K Cell Differentiation %K Cell Lineage %K DNA-Binding Proteins %K Embryo, Mammalian %K Endoderm %K Female %K Gene Expression Regulation, Developmental %K Homozygote %K Male %K Mesoderm %K Mice %K Mice, Inbred C57BL %K Mice, Inbred Strains %K Mice, Mutant Strains %K Nodal Protein %K Signal Transduction %K Smad2 Protein %K Smad3 Protein %K Trans-Activators %K Transforming Growth Factor beta %X TGFbeta 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. %B Development (Cambridge, England) %V 127 %P 3079-90 %8 2000 Jul %G eng %N 14 %1 http://www.ncbi.nlm.nih.gov/pubmed/10862745?dopt=Abstract %0 Journal Article %J Clinical and experimental immunology %D 2000 %T Growth of Brucella abortus in macrophages from resistant and susceptible mouse strains. %A Sathiyaseelan, J %A Jiang, X %A Baldwin, C L %X C57Bl/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. %B Clinical and experimental immunology %V 121 %P 289-94 %8 2000 Aug %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/10931144?dopt=Abstract %0 Journal Article %J Genomics %D 2000 %T Identification and structural analysis of human RBM8A and RBM8B: two highly conserved RNA-binding motif proteins that interact with OVCA1, a candidate tumor suppressor. %A Salicioni, A M %A Xi, M %A Vanderveer, L A %A Balsara, B %A Testa, J R %A Dunbrack, R L %A Godwin, A K %K Amino Acid Motifs %K Amino Acid Sequence %K Animals %K Chromosome Mapping %K Chromosomes, Human, Pair 14 %K Chromosomes, Human, Pair 5 %K Cloning, Molecular %K Conserved Sequence %K COS Cells %K DNA, Complementary %K Female %K Gene Expression %K Genes, Tumor Suppressor %K Humans %K In Situ Hybridization, Fluorescence %K Male %K Microscopy, Fluorescence %K Models, Molecular %K Molecular Sequence Data %K Protein Binding %K Protein Isoforms %K Protein Structure, Tertiary %K Proteins %K RNA, Messenger %K RNA-Binding Proteins %K Sequence Alignment %K Sequence Analysis, DNA %K Sequence Homology, Amino Acid %K Tissue Distribution %K Tumor Suppressor Proteins %X The 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. %B Genomics %V 69 %P 54-62 %8 2000 Oct 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11013075?dopt=Abstract %0 Journal Article %J Developmental biology %D 2000 %T PACSIN2 is a regulator of the metalloprotease/disintegrin ADAM13. %A Cousin, H %A Gaultier, A %A Bleux, C %A Darribère, T %A Alfandari, D %K ADAM Proteins %K Amino Acid Sequence %K Animals %K Disintegrins %K Gene Expression Regulation, Developmental %K Immunohistochemistry %K In Situ Hybridization %K Membrane Proteins %K Metalloendopeptidases %K Microinjections %K Models, Biological %K Molecular Sequence Data %K Neural Crest %K Phenotype %K Precipitin Tests %K Protein Binding %K Proteins %K Recombinant Fusion Proteins %K RNA, Messenger %K Sequence Alignment %K Sequence Homology, Amino Acid %K src Homology Domains %K Substrate Specificity %K Xenopus laevis %K Xenopus Proteins %X ADAM13 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. %B Developmental biology %V 227 %P 197-210 %8 2000 Nov 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11076687?dopt=Abstract %0 Journal Article %J Comparative biochemistry and physiology. Toxicology & pharmacology : CBP %D 2000 %T Purine requirements for the expression of Cape buffalo serum trypanocidal activity. %A Wang, Q %A Hamilton, E %A Black, S J %X Cape 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. %B Comparative biochemistry and physiology. Toxicology & pharmacology : CBP %V 125 %P 25-32 %8 2000 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/11790327?dopt=Abstract %0 Journal Article %J Research in veterinary science %D 2000 %T Rapid changes occur in the percentage of circulating bovine WC1(+)gamma delta Th1 cells. %A Baldwin, C L %A Sathiyaseelan, T %A Rocchi, M %A McKeever, D %X gamma 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. %B Research in veterinary science %V 69 %P 175-80 %8 2000 Oct %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/11020371?dopt=Abstract %0 Journal Article %J International journal of oncology %D 2000 %T S100P calcium-binding protein overexpression is associated with immortalization of human breast epithelial cells in vitro and early stages of breast cancer development in vivo. %A Guerreiro Da Silva, I D %A Hu, Y F %A Russo, I H %A Ao, X %A Salicioni, A M %A Yang, X %A Russo, J %K Base Sequence %K Breast %K Breast Neoplasms %K Calcium-Binding Proteins %K Carcinoma in Situ %K Carcinoma, Ductal, Breast %K Cell Transformation, Neoplastic %K Epithelial Cells %K Female %K Humans %K Molecular Sequence Data %K Neoplasm Proteins %K RNA %K Tumor Cells, Cultured %X The 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. %B International journal of oncology %V 16 %P 231-40 %8 2000 Feb %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/10639564?dopt=Abstract %0 Journal Article %J Neuroendocrinology %D 2000 %T Serotonin inhibits luteinizing hormone release via 5-HT1A receptors in the zona incerta of ovariectomised, anaesthetised rats primed with steroids. %A Siddiqui, A %A Kotecha, K %A Salicioni, A M %A Kalia, V %A Murray, J F %A Wilson, C A %K 8-Hydroxy-2-(di-n-propylamino)tetralin %K Anesthesia %K Animals %K Estradiol %K Female %K Luteinizing Hormone %K Ovariectomy %K Piperazines %K Progesterone %K Rats %K Rats, Wistar %K Receptors, Serotonin %K Receptors, Serotonin, 5-HT1 %K Ritanserin %K Serotonin %K Serotonin Antagonists %K Serotonin Receptor Agonists %K Subthalamus %X The 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. %B Neuroendocrinology %V 72 %P 272-83 %8 2000 Nov %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/11124584?dopt=Abstract %0 Journal Article %J Journal of economic entomology %D 2000 %T Toxicity and residual effectiveness of insecticides on insecticide-treated spheres for controlling females of Rhagoletis pomonella (Diptera: Tephritidae). %A Hu, X P %A Prokopy, R J %A Clark, J M %X This 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. %B Journal of economic entomology %V 93 %P 403-11 %8 2000 Apr %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/10826192?dopt=Abstract %0 Journal Article %J Current opinion in immunology %D 2000 %T Transcriptional control of T cell development. %A Osborne, B A %X Transcriptional 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. %B Current opinion in immunology %V 12 %P 301-6 %8 2000 Jun %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/10781405?dopt=Abstract %0 Journal Article %J Journal of medical entomology %D 1999 %T Aedes aegypti (Diptera: Culicidae) age determination by cuticular hydrocarbon analysis of female legs. %A Desena, M L %A Edman, J D %A Clark, J M %A Symington, S B %A Scott, T W %X We 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. %B Journal of medical entomology %V 36 %P 824-30 %8 1999 Nov %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/10593086?dopt=Abstract %0 Journal Article %J The Journal of parasitology %D 1999 %T Anti-Trypanosoma brucei activity of nonprimate zoo sera. %A Black, S J %A Wang, Q %A Makadzange, T %A Li, Y L %A Van Praagh, A %A Loomis, M %A Seed, J R %X Constitutive 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. %B The Journal of parasitology %V 85 %P 48-53 %8 1999 Feb %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/10207362?dopt=Abstract %0 Journal Article %J Results and problems in cell differentiation %D 1999 %T Apoptosis and the proteasome. %A Grimm, L M %A Osborne, B A %X The 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. %B Results and problems in cell differentiation %V 23 %P 209-28 %8 1999 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/9950035?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 1999 %T Cutting edge: protective effects of notch-1 on TCR-induced apoptosis. %A Jehn, B M %A Bielke, W %A Pear, W S %A Osborne, B A %X The 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. %B Journal of immunology (Baltimore, Md. : 1950) %V 162 %P 635-8 %8 1999 Jan 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/9916679?dopt=Abstract %0 Journal Article %J Journal of agricultural and food chemistry %D 1999 %T Hapten design in the development of competitive enyme-linked immunosorbent assays for genotoxic metabolites of alachlor. %A Tessier, D M %A Clark, J M %X The 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). %B Journal of agricultural and food chemistry %V 47 %P 3925-33 %8 1999 Sep %G eng %N 9 %1 http://www.ncbi.nlm.nih.gov/pubmed/10552745?dopt=Abstract %0 Journal Article %J Infection and immunity %D 1999 %T Infection-associated decline of cape buffalo blood catalase augments serum trypanocidal activity. %A Wang, Q %A Murphy, N %A Black, S J %X Clearance 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. %B Infection and immunity %V 67 %P 2797-803 %8 1999 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/10338483?dopt=Abstract %0 Journal Article %J Journal of medical entomology %D 1999 %T Potential for aging female Aedes aegypti (Diptera: Culicidae) by gas chromatographic analysis of cuticular hydrocarbons, including a field evaluation. %A Desena, M L %A Clark, J M %A Edman, J D %A Symington, S B %A Scott, T W %A Clark, G G %A Peters, T M %X Gas 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). %B Journal of medical entomology %V 36 %P 811-23 %8 1999 Nov %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/10593085?dopt=Abstract %0 Journal Article %J Science %D 1998 %T Cloned transgenic calves produced from nonquiescent fetal fibroblasts. %A Cibelli, J B %A Stice, S L %A Golueke, P J %A Kane, J J %A Jerry, J %A Blackwell, C %A Ponce de León, F A %A Robl, J M %K Animals %K Animals, Genetically Modified %K Blastocyst %K Cattle %K Cell Aging %K Cell Division %K Cell Nucleus %K Cells, Cultured %K Clone Cells %K Cloning, Organism %K Embryo Transfer %K Female %K Fetus %K Fibroblasts %K G1 Phase %K Male %K Nuclear Transfer Techniques %K Oocytes %K Transfection %K Transgenes %X

An 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.

%B Science %V 280 %P 1256-8 %8 1998 May 22 %G eng %N 5367 %1

http://www.ncbi.nlm.nih.gov/pubmed/9596577?dopt=Abstract

%0 Journal Article %J International journal of oncology %D 1998 %T Correlation between cell cycle regulators and the immortalization and transformation of human breast epithelial cell lines. %A Salicioni, A M %A Russo, I H %A Russo, J %K Breast %K Breast Neoplasms %K Cell Cycle Proteins %K Cell Division %K Cell Transformation, Neoplastic %K Cells, Cultured %K Epithelial Cells %K Female %K Humans %K Phenotype %K Tumor Cells, Cultured %X Cellular 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. %B International journal of oncology %V 13 %P 65-71 %8 1998 Jul %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/9625804?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 1998 %T Multiple sites of V lambda diversification in cattle. %A Lucier, M R %A Thompson, R E %A Waire, J %A Lin, A W %A Osborne, B A %A Goldsby, R A %X

Ig 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.

%B Journal of immunology (Baltimore, Md. : 1950) %V 161 %P 5438-44 %8 1998 Nov 15 %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/9820519?dopt=Abstract %0 Journal Article %J Developmental biology %D 1998 %T Partial characterization of the calcium-releasing activity of porcine sperm cytosolic extracts. %A Wu, H %A He, C L %A Jehn, B %A Black, S J %A Fissore, R A %X Injection 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. %B Developmental biology %V 203 %P 369-81 %8 1998 Nov 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/9808787?dopt=Abstract %0 Journal Article %J Nat Biotechnol %D 1998 %T Transgenic bovine chimeric offspring produced from somatic cell-derived stem-like cells. %A Cibelli, J B %A Stice, S L %A Golueke, P J %A Kane, J J %A Jerry, J %A Blackwell, C %A Ponce de León, F A %A Robl, J M %K Animals %K Animals, Genetically Modified %K beta-Galactosidase %K Blastocyst %K Cattle %K Cell Differentiation %K Cell Fusion %K Chimera %K Ectoderm %K Embryo Transfer %K Endoderm %K Gene Transfer Techniques %K Genetic Therapy %K Mesoderm %K Stem Cells %X

We 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.

%B Nat Biotechnol %V 16 %P 642-6 %8 1998 Jul %G eng %N 7 %1

http://www.ncbi.nlm.nih.gov/pubmed/9661197?dopt=Abstract

%0 Journal Article %J Molecular carcinogenesis %D 1997 %T Differential expression of human ferritin H chain gene in immortal human breast epithelial MCF-10F cells. %A Higgy, N A %A Salicioni, A M %A Russo, I H %A Zhang, P L %A Russo, J %K Base Sequence %K Breast %K Breast Neoplasms %K Carcinoma, Ductal, Breast %K Carcinoma, Lobular %K Cells, Cultured %K Cloning, Molecular %K DNA, Complementary %K Epithelial Cells %K Female %K Ferritins %K Gene Expression Regulation %K Gene Expression Regulation, Neoplastic %K Gene Library %K Humans %K Models, Biological %K Molecular Sequence Data %K Neoplasm Invasiveness %K Transcription, Genetic %K Tumor Cells, Cultured %X Subtractive 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. %B Molecular carcinogenesis %V 20 %P 332-9 %8 1997 Dec %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/9433477?dopt=Abstract %0 Journal Article %J Critical reviews in eukaryotic gene expression %D 1997 %T Gene regulation associated with apoptosis. %A Jehn, B M %A Osborne, B A %X Apoptosis, 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. %B Critical reviews in eukaryotic gene expression %V 7 %P 179-93 %8 1997 %G eng %N 1-2 %1 http://www.ncbi.nlm.nih.gov/pubmed/9034721?dopt=Abstract %0 Journal Article %J Biochemical Society transactions %D 1997 %T Identification of the cape buffalo serum trypanocidal protein: xanthine: oxygen oxidoreductase. %A Black, S J %A Muranjan, M %A Wang, Q %B Biochemical Society transactions %V 25 %P 534S %8 1997 Aug %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/9388750?dopt=Abstract %0 Journal Article %J International reviews of immunology %D 1997 %T Immunoglobulin gene diversification in cattle. %A Meyer, A %A Parng, C L %A Hansal, S A %A Osborne, B A %A Goldsby, R A %X

Research 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.

%B International reviews of immunology %V 15 %P 165-83 %8 1997 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/9222818?dopt=Abstract %0 Journal Article %J European journal of pharmacology %D 1997 %T Participation of both adrenergic and opioidergic systems in the negative feedback of adrenal progesterone on LH secretion. %A Salicioni, A M %A Carón, R W %A Deis, R P %K Adrenal Glands %K Adrenergic alpha-1 Receptor Antagonists %K Adrenergic alpha-2 Receptor Antagonists %K Adrenergic alpha-Antagonists %K Adrenergic beta-1 Receptor Antagonists %K Adrenergic beta-2 Receptor Antagonists %K Adrenergic beta-Antagonists %K Animals %K Estradiol %K Feedback %K Female %K Hormone Antagonists %K Idazoxan %K Luteinizing Hormone %K Metoprolol %K Mifepristone %K Naloxone %K Ovariectomy %K Prazosin %K Progesterone %K Propanolamines %K Rats %K Rats, Wistar %K Receptors, Adrenergic %K Receptors, Opioid %X It 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. %B European journal of pharmacology %V 332 %P 283-7 %8 1997 Aug 13 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/9300262?dopt=Abstract %0 Journal Article %J Neuropharmacology %D 1997 %T Regulation of prolactin secretion by adrenal steroids in oestrogen-treated ovariectomized rats: participation of endogenous opioid peptides. %A Carón, R W %A Salicioni, A M %A Deis, R P %K Animals %K Anti-Inflammatory Agents %K Estradiol %K Female %K Hormone Antagonists %K Hydrocortisone %K Mifepristone %K Naloxone %K Opioid Peptides %K Ovariectomy %K Progesterone %K Prolactin %K Rats %K Rats, Wistar %X The 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. %B Neuropharmacology %V 36 %P 1433-8 %8 1997 Oct %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/9423931?dopt=Abstract %0 Journal Article %J Cancer research %D 1997 %T Tamoxifen down-regulates CD36 messenger RNA levels in normal and neoplastic human breast tissues. %A Silva, I D %A Salicioni, A M %A Russo, I H %A Higgy, N A %A Gebrim, L H %A Russo, J %K Antigens, CD36 %K Antineoplastic Agents, Hormonal %K Base Sequence %K Breast %K Breast Neoplasms %K Down-Regulation %K Female %K Gene Expression Regulation %K Humans %K Molecular Sequence Data %K RNA, Messenger %K Tamoxifen %K Transcriptional Activation %X Tamoxifen (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. %B Cancer research %V 57 %P 378-81 %8 1997 Feb 1 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/9012459?dopt=Abstract %0 Journal Article %J BioTechniques %D 1996 %T Addition of a competitive primer can dramatically improve the specificity of PCR amplification of specific alleles. %A Zhu, K Y %A Clark, J M %B BioTechniques %V 21 %P 586, 590 %8 1996 Oct %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/8891204?dopt=Abstract %0 Journal Article %J Current opinion in immunology %D 1996 %T Apoptosis and the maintenance of homoeostasis in the immune system. %A Osborne, B A %X The 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. %B Current opinion in immunology %V 8 %P 245-54 %8 1996 Apr %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/8725948?dopt=Abstract %0 Journal Article %J Journal of clinical microbiology %D 1996 %T Application of random amplified polymorphic DNA analysis to differentiate strains of Salmonella enteritidis. %A Lin, A W %A Usera, M A %A Barrett, T J %A Goldsby, R A %X A 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. %B Journal of clinical microbiology %V 34 %P 870-6 %8 1996 Apr %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/8815099?dopt=Abstract %0 Journal Article %J Trends in genetics : TIG %D 1996 %T Cell death in vertebrates: lessons from the worm. %A Osborne, B A %B Trends in genetics : TIG %V 12 %P 489-91 %8 1996 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/9257523?dopt=Abstract %0 Journal Article %J FEMS immunology and medical microbiology %D 1996 %T Comparison of T cell cytokines in resistant and susceptible mice infected with virulent Brucella abortus strain 2308. %A Fernandes, D M %A Jiang, X %A Jung, J H %A Baldwin, C L %X C57BL/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. %B FEMS immunology and medical microbiology %V 16 %P 193-203 %8 1996 Dec 31 %G eng %N 3-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/9116636?dopt=Abstract %0 Journal Article %J Journal of immunology (Baltimore, Md. : 1950) %D 1996 %T Gene conversion contributes to Ig light chain diversity in cattle. %A Parng, C L %A Hansal, S %A Goldsby, R A %A Osborne, B A %X

In 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.

%B Journal of immunology (Baltimore, Md. : 1950) %V 157 %P 5478-86 %8 1996 Dec 15 %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/8955197?dopt=Abstract %0 Journal Article %J Journal of cellular biochemistry %D 1996 %T Genes that regulate apoptosis in the mouse thymus. %A Osborne, B A %A Smith, S W %A McLaughlin, K A %A Grimm, L %A Kallinch, T %A Liu, Z %A Schwartz, L M %X Elimination 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. %B Journal of cellular biochemistry %V 60 %P 18-22 %8 1996 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/8825411?dopt=Abstract %0 Journal Article %J Advances in experimental medicine and biology %D 1996 %T Genetic regulation of apoptosis in the mouse thymus. %A Osborne, B A %A Smith, S W %A McLaughlin, K A %A Grimm, L %A Morgan, G %A Goldsby, R A %B Advances in experimental medicine and biology %V 406 %P 199-207 %8 1996 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/8910686?dopt=Abstract %0 Journal Article %J Pesticide biochemistry and physiology %D 1996 %T A Point Mutation of Acetylcholinesterase Associated with Azinphosmethyl Resistance and Reduced Fitness in Colorado Potato Beetle %A Zhu, K Y %A Lee, S H %A Clark, J M %X A 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. %B Pesticide biochemistry and physiology %V 55 %P 100-8 %8 1996 Jun %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/8980034?dopt=Abstract %0 Journal Article %J The EMBO journal %D 1996 %T Proteasomes play an essential role in thymocyte apoptosis. %A Grimm, L M %A Goldberg, A L %A Poirier, G G %A Schwartz, L M %A Osborne, B A %X Cell 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. %B The EMBO journal %V 15 %P 3835-44 %8 1996 Aug 1 %G eng %N 15 %1 http://www.ncbi.nlm.nih.gov/pubmed/8670888?dopt=Abstract %0 Journal Article %J European journal of immunology %D 1996 %T The role of oxygen in thymocyte apoptosis. %A McLaughlin, K A %A Osborne, B A %A Goldsby, R A %X

Signals 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.

%B European journal of immunology %V 26 %P 1170-4 %8 1996 May %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/8647183?dopt=Abstract %0 Journal Article %J BioEssays : news and reviews in molecular, cellular and developmental biology %D 1995 %T Cell death suffers a TKO. %A Osborne, B A %A Schwartz, L M %X The 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. %B BioEssays : news and reviews in molecular, cellular and developmental biology %V 17 %P 557-9 %8 1995 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/7575499?dopt=Abstract %0 Journal Article %J Insect biochemistry and molecular biology %D 1995 %T Cloning and sequencing of a cDNA encoding acetylcholinesterase in Colorado potato beetle, Leptinotarsa decemlineata (Say). %A Zhu, K Y %A Clark, J M %X A 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. %B Insect biochemistry and molecular biology %V 25 %P 1129-38 %8 1995 Dec %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/8580913?dopt=Abstract %0 Journal Article %J Annals of the New York Academy of Sciences %D 1995 %T Diversification of bovine lambda-light chain genes. %A Parng, C L %A Hansal, S %A Goldsby, R A %A Osborne, B A %B Annals of the New York Academy of Sciences %V 764 %P 155-7 %8 1995 Sep 29 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/7486515?dopt=Abstract %0 Journal Article %J Seminars in cancer biology %D 1995 %T Induction of genes during apoptosis: examples from the immune system. %A Osborne, B A %X The 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. %B Seminars in cancer biology %V 6 %P 27-33 %8 1995 Feb %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/7548838?dopt=Abstract %0 Journal Article %J Infection and immunity %D 1995 %T Lack of a role for natural killer cells in early control of Brucella abortus 2308 infections in mice. %A Fernandes, D M %A Benson, R %A Baldwin, C L %X Studies 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. %B Infection and immunity %V 63 %P 4029-33 %8 1995 Oct %G eng %N 10 %1 http://www.ncbi.nlm.nih.gov/pubmed/7558315?dopt=Abstract %0 Journal Article %J Current topics in microbiology and immunology %D 1995 %T Molecular events in thymocyte apoptosis. %A Smith, S W %A McLaughlin, K A %A Osborne, B A %B Current topics in microbiology and immunology %V 200 %P 147-62 %8 1995 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/7634829?dopt=Abstract %0 Journal Article %J Bulletin of environmental contamination and toxicology %D 1995 %T Preliminary study of synergism of acid rain and diflubenzuron. %A Martin, P J %A Clark, J M %A Edman, J D %B Bulletin of environmental contamination and toxicology %V 54 %P 833-6 %8 1995 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/7647497?dopt=Abstract %0 Journal Article %J Annual review of entomology %D 1995 %T Resistance to avermectins: extent, mechanisms, and management implications. %A Clark, J M %A Scott, J G %A Campos, F %A Bloomquist, J R %X The 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. %B Annual review of entomology %V 40 %P 1-30 %8 1995 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/7810984?dopt=Abstract %0 Journal Article %J Methods in cell biology %D 1995 %T Transient transfection assays to examine the requirement of putative cell death genes. %A Osborne, B A %A Smith, S W %A Liu, Z G %A McLaughlin, K A %A Schwartz, L M %X In 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. %B Methods in cell biology %V 46 %P 99-106 %8 1995 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/7541887?dopt=Abstract %0 Journal Article %J Nature %D 1994 %T Apoptotic signals delivered through the T-cell receptor of a T-cell hybrid require the immediate-early gene nur77. %A Liu, Z G %A Smith, S W %A McLaughlin, K A %A Schwartz, L M %A Osborne, B A %X Engagement 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. %B Nature %V 367 %P 281-4 %8 1994 Jan 20 %G eng %N 6460 %1 http://www.ncbi.nlm.nih.gov/pubmed/8121494?dopt=Abstract %0 Journal Article %J BioEssays : news and reviews in molecular, cellular and developmental biology %D 1994 %T Ced-3/ICE: evolutionarily conserved regulation of cell death. %A Schwartz, L M %A Osborne, B A %B BioEssays : news and reviews in molecular, cellular and developmental biology %V 16 %P 387-9 %8 1994 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/8080427?dopt=Abstract %0 Journal Article %J Trends in cell biology %D 1994 %T Essential genes that regulate apoptosis. %A Osborne, B A %A Schwartz, L M %X The 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. %B Trends in cell biology %V 4 %P 394-9 %8 1994 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/14731815?dopt=Abstract %0 Journal Article %J Immunological reviews %D 1994 %T Identification of genes induced during apoptosis in T lymphocytes. %A Osborne, B A %A Smith, S W %A Liu, Z G %A McLaughlin, K A %A Grimm, L %A Schwartz, L M %B Immunological reviews %V 142 %P 301-20 %8 1994 Dec %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/7698798?dopt=Abstract %0 Journal Article %J The Journal of steroid biochemistry and molecular biology %D 1994 %T Mifepristone treatment demonstrates the participation of adrenal glucocorticoids in the regulation of oestrogen-induced prolactin secretion in ovariectomized rats. %A Carón, R W %A Salicioni, A M %A Deis, R P %K Adrenal Cortex %K Adrenalectomy %K Animals %K Estradiol %K Estrogens %K Female %K Glucocorticoids %K Immune Sera %K Kinetics %K Mifepristone %K Ovariectomy %K Progesterone %K Prolactin %K Rats %K Rats, Wistar %X Accumulated 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. %B The Journal of steroid biochemistry and molecular biology %V 48 %P 385-9 %8 1994 Mar %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/8142316?dopt=Abstract %0 Journal Article %J Cellular immunology %D 1994 %T Quantitative analysis of the B cell repertoire by limiting dilution analysis and fluorescent in situ hybridization. %A Ravichandran, K S %A Osborne, B A %A Goldsby, R A %X

We 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.

%B Cellular immunology %V 154 %P 309-27 %8 1994 Apr 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/8131205?dopt=Abstract %0 Journal Article %J The Journal of endocrinology %D 1993 %T Adrenal progesterone facilitates the negative feedback of oestrogen on LH release in ovariectomized rats. %A Salicioni, A M %A Carón, R W %A Deis, R P %K Adrenal Glands %K Adrenalectomy %K Animals %K Estradiol %K Estrogens %K Feedback %K Female %K Immune Sera %K Luteinizing Hormone %K Mifepristone %K Ovariectomy %K Progesterone %K Radioimmunoassay %K Rats %K Rats, Wistar %X There 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) %B The Journal of endocrinology %V 139 %P 253-8 %8 1993 Nov %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/8308461?dopt=Abstract %0 Journal Article %J Proceedings of the National Academy of Sciences of the United States of America %D 1993 %T Do all programmed cell deaths occur via apoptosis? %A Schwartz, L M %A Smith, S W %A Jones, M E %A Osborne, B A %X During 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. %B Proceedings of the National Academy of Sciences of the United States of America %V 90 %P 980-4 %8 1993 Feb 1 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/8430112?dopt=Abstract %0 Journal Article %J Immunology today %D 1993 %T Programmed cell death, apoptosis and killer genes. %A Schwartz, L M %A Osborne, B A %X A 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. %B Immunology today %V 14 %P 582-90 %8 1993 Dec %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/8305131?dopt=Abstract %0 Journal Article %J Journal of immunological methods %D 1992 %T Immunoglobulin VH usage analysis by fluorescent in situ hybridization and flow cytometry. %A Ravichandran, K S %A Semproni, A R %A Goldsby, R A %A Osborne, B A %X

We 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.

%B Journal of immunological methods %V 153 %P 249-59 %8 1992 Aug 30 %G eng %N 1-2 %1 http://www.ncbi.nlm.nih.gov/pubmed/1517596?dopt=Abstract %0 Journal Article %J Avian diseases %D 1992 %T Salmonella enteritidis-specific monoclonal antibodies. %A Lin, A W %A Goldsby, R A %A Snoeyenbos, G H %X Three 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. %B Avian diseases %V 36 %P 455-8 %8 1992 Apr-Jun %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/1378262?dopt=Abstract %0 Journal Article %J Bulletin of environmental contamination and toxicology %D 1991 %T Airborne drift residues collected near apple orchard environments due to application of insecticide mixtures. %A Clark, J M %A Marion, J R %A Tessier, D M %A Brooks, M W %A Coli, W M %B Bulletin of environmental contamination and toxicology %V 46 %P 829-36 %8 1991 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/1863790?dopt=Abstract %0 Journal Article %J Medical and veterinary entomology %D 1990 %T Cuticular lipid differences between the malaria vector and non-vector forms of the Anopheles maculatus complex. %A Kittayapong, P %A Clark, J M %A Edman, J D %A Potter, T L %A Lavine, B K %A Marion, J R %A Brooks, M %X Two 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. %B Medical and veterinary entomology %V 4 %P 405-13 %8 1990 Oct %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/2133007?dopt=Abstract %0 Journal Article %J The Journal of experimental medicine %D 1989 %T Analysis of a novel VHS107 haplotype in CLA-2 and WSA mice. Evidence for gene conversion among IgVH genes in outbred populations. %A Ferguson, S E %A Cancro, M P %A Osborne, B A %X Gene 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. %B The Journal of experimental medicine %V 170 %P 1811-23 %8 1989 Dec 1 %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/2584925?dopt=Abstract %0 Journal Article %J Biochemical pharmacology %D 1989 %T Role of ion channels and intraterminal calcium homeostasis in the action of deltamethrin at presynaptic nerve terminals. %A Clark, J M %A Brooks, M W %X Using 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. %B Biochemical pharmacology %V 38 %P 2233-45 %8 1989 Jul 15 %G eng %N 14 %1 http://www.ncbi.nlm.nih.gov/pubmed/2546560?dopt=Abstract %0 Journal Article %J Genes Dev %D 1989 %T Tissue-specific expression, developmental regulation, and genetic mapping of the gene encoding CCAAT/enhancer binding protein. %A Birkenmeier, E H %A Gwynn, B %A Howard, S %A Jerry, J %A Gordon, J I %A Landschulz, W H %A McKnight, S L %K Adipose Tissue %K Animals %K CCAAT-Enhancer-Binding Proteins %K Chromosome Mapping %K DNA-Binding Proteins %K Gene Expression Regulation %K Immunohistochemistry %K Intestines %K Liver %K Mice %K Mice, Inbred BALB C %K Rats %K Rats, Inbred Strains %K RNA, Messenger %K Tissue Distribution %X

This 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.

%B Genes Dev %V 3 %P 1146-56 %8 1989 Aug %G eng %N 8 %1

http://www.ncbi.nlm.nih.gov/pubmed/2792758?dopt=Abstract

%0 Journal Article %J Developmental and comparative immunology %D 1988 %T Comparative idiotype network interactions: antigen mimicry by an anti-bovine idiotype monoclonal antibody in rats and cattle. %A Arulanandam, A R %A Goldsby, R A %X Idiotype/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. %B Developmental and comparative immunology %V 12 %P 669-83 %8 1988 Summer %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/2458977?dopt=Abstract %0 Journal Article %J Genetics %D 1988 %T Evolution of the IgA heavy chain gene in the genus Mus. %A Osborne, B A %A Golde, T E %A Schwartz, R L %A Rudikoff, S %X To 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. %B Genetics %V 119 %P 925-31 %8 1988 Aug %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/2842228?dopt=Abstract %0 Journal Article %J The Journal of experimental medicine %D 1988 %T Interaction and sequence diversity among T15 VH genes in CBA/J mice. %A Ferguson, S E %A Rudikoff, S %A Osborne, B A %X Nucleotide 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. %B The Journal of experimental medicine %V 168 %P 1339-49 %8 1988 Oct 1 %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/3139820?dopt=Abstract %0 Journal Article %J Veterinary immunology and immunopathology %D 1987 %T The application of hybridoma technology to the study of bovine immunoglobulins. %A Goldsby, R A %A Srikumaran, S %A Arulanandam, A %A Hague, B %A Ponce de León, F A %A Sevoian, M %A Guidry, A J %X Studies 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. %B Veterinary immunology and immunopathology %V 17 %P 25-35 %8 1987 Dec %G eng %N 1-4 %1 http://www.ncbi.nlm.nih.gov/pubmed/3501632?dopt=Abstract