@article{golkar-narenji_dziegiel_kempisty_petitte_mozdziak_bryja_2023, title={In vitro culture of reptile PGCS to preserve endangered species}, volume={5}, ISSN={["1095-8355"]}, DOI={10.1002/cbin.12033}, abstractNote={AbstractPrimordial germ cells (PGCs), are the source of gametes in vertebrates. There are similarities in the development of PGCs of reptiles with avian and mammalian species PGCs development. PGCs culture has been performed for avian and mammalian species but there is no report for reptilian PGCs culture. In vitro culture of PGCs is needed to produce transgenic animals, preservation of endangered animals and for studies on cell behaviour and research on fertility. Reptiles are traded as exotic pets and a source of food and they are valuable for their skin and they are useful as model for medical research. Transgenic reptile has been suggested to be useful for pet industry and medical research. In this research different aspects of PGCs development was compared in three main classes of vertebrates including mammalian, avian and reptilian species. It is proposed that a discussion on similarities between reptilian PGCs development with avian and mammalian species helps to find clues for studies of reptilian PGCs development details and finding an efficient protocol for in vitro culture of reptilian PG.}, journal={CELL BIOLOGY INTERNATIONAL}, author={Golkar-Narenji, Afsaneh and Dziegiel, Piotr and Kempisty, Bartosz and Petitte, James and Mozdziak, Paul Edward and Bryja, Artur}, year={2023}, month={May} } @misc{rzymski_kulus_jankowski_dompe_bryl_petitte_kempisty_mozdziak_2021, title={COVID-19 Pandemic Is a Call to Search for Alternative Protein Sources as Food and Feed: A Review of Possibilities}, volume={13}, ISBN={2072-6643}, url={https://doi.org/10.3390/nu13010150}, DOI={10.3390/nu13010150}, abstractNote={The coronavirus disease 2019 (COVID-19) pandemic is a global health challenge with substantial adverse effects on the world economy. It is beyond any doubt that it is, again, a call-to-action to minimize the risk of future zoonoses caused by emerging human pathogens. The primary response to contain zoonotic diseases is to call for more strict regulations on wildlife trade and hunting. This is because the origins of coronaviruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS-CoV, Middle East respiratory syndrome coronavirus (MERS-CoV), as well as other viral pathogens (e.g., Ebola, HIV) are traceable to wild animals. Although COVID-19 is not related to livestock animals, the pandemic increased general attention given to zoonotic viral infections—the risk of which can also be associated with livestock. Therefore, this paper discusses the potential transformation of industrial livestock farming and the production of animal products, particularly meat, to decrease the risks for transmission of novel human pathogens. Plant-based diets have a number of advantages, but it is unrealistic to consider them as the only solution offered to the problem. Therefore, a search for alternative protein sources in insect-based foods and cultured meat, important technologies enabling safer meat production. Although both of these strategies offer a number of potential advantages, they are also subject to the number of challenges that are discussed in this paper. Importantly, insect-based foods and cultured meat can provide additional benefits in the context of ecological footprint, an aspect important in light of predicted climate changes. Furthermore, cultured meat can be regarded as ethically superior and supports better food security. There is a need to further support the implementation and expansion of all three approaches discussed in this paper, plant-based diets, insect-based foods, and cultured meat, to decrease the epidemiological risks and ensure a sustainable future. Furthermore, cultured meat also offers a number of additional benefits in the context of environmental impact, ethical issues, and food security.}, number={1}, journal={NUTRIENTS}, publisher={MDPI AG}, author={Rzymski, Piotr and Kulus, Magdalena and Jankowski, Maurycy and Dompe, Claudia and Bryl, Rut and Petitte, James N. and Kempisty, Bartosz and Mozdziak, Paul}, year={2021}, month={Jan}, pages={150} } @misc{dompe_kulus_stefanska_kranc_chermula_bryl_pienkowski_nawrocki_petitte_stelmach_et al._2021, title={Human Granulosa Cells-Stemness Properties, Molecular Cross-Talk and Follicular Angiogenesis}, volume={10}, ISSN={["2073-4409"]}, url={https://doi.org/10.3390/cells10061396}, DOI={10.3390/cells10061396}, abstractNote={The ovarian follicle is the basic functional unit of the ovary, comprising theca cells and granulosa cells (GCs). Two different types of GCs, mural GCs and cumulus cells (CCs), serve different functions during folliculogenesis. Mural GCs produce oestrogen during the follicular phase and progesterone after ovulation, while CCs surround the oocyte tightly and form the cumulus oophurus and corona radiata inner cell layer. CCs are also engaged in bi-directional metabolite exchange with the oocyte, as they form gap-junctions, which are crucial for both the oocyte’s proper maturation and GC proliferation. However, the function of both GCs and CCs is dependent on proper follicular angiogenesis. Aside from participating in complex molecular interplay with the oocyte, the ovarian follicular cells exhibit stem-like properties, characteristic of mesenchymal stem cells (MSCs). Both GCs and CCs remain under the influence of various miRNAs, and some of them may contribute to polycystic ovary syndrome (PCOS) or premature ovarian insufficiency (POI) occurrence. Considering increasing female fertility problems worldwide, it is of interest to develop new strategies enhancing assisted reproductive techniques. Therefore, it is important to carefully consider GCs as ovarian stem cells in terms of the cellular features and molecular pathways involved in their development and interactions as well as outline their possible application in translational medicine.}, number={6}, journal={CELLS}, publisher={MDPI AG}, author={Dompe, Claudia and Kulus, Magdalena and Stefanska, Katarzyna and Kranc, Wieslawa and Chermula, Blazej and Bryl, Rut and Pienkowski, Wojciech and Nawrocki, Mariusz J. and Petitte, James N. and Stelmach, Boguslawa and et al.}, year={2021}, month={Jun} } @article{kulus_kranc_wojtanowicz-markiewicz_celichowski_swiatly-blaszkiewicz_matuszewska_sujka-kordowska_konwerska_zdun_bryl_et al._2021, title={New Gene Markers Expressed in Porcine Oviductal Epithelial Cells Cultured Primary In Vitro Are Involved in Ontological Groups Representing Physiological Processes of Porcine Oocytes}, volume={22}, ISSN={["1422-0067"]}, url={https://www.mdpi.com/1422-0067/22/4/2082}, DOI={10.3390/ijms22042082}, abstractNote={Changes that occur within oviducts after fertilization are dependent on post-ovulation events, including oocyte-oviduct interactions. Although general processes are well-defined, the molecular basis are poorly understood. Recently, new marker genes involved in ‘cell development’, ‘cell growth’, ‘cell differentiation’ and ‘cell maturation’ processes have been identified in porcine oocytes. The aim of the study was to assess the expression profile of genes in primary in vitro cultured oviductal epithelial cells (OECs), clustered in Gene Ontology groups which enveloped markers also identified in porcine oocytes. OECs (from 45 gilts) were surgically removed and cultured in vitro for ≤ 30 days, and then subjected to molecular analyses. The transcriptomic and proteomic profiles of cells cultured during 7, 15 and 30 days were investigated. Additionally, morphological/histochemical analyzes were performed. The results of genes expression profiles were validated after using RT-qPCR. The results showed a significant upregulation of UNC45B, NOX4, VLDLR, ITGB3, FMOD, SGCE, COL1A2, LOX, LIPG, THY1 and downregulation of SERPINB2, CD274, TXNIP, CELA1, DDX60, CRABP2, SLC5A1, IDO1, ANPEP, FST. Detailed knowledge of the molecular pathways occurring in the OECs and the gametes that contact them may contribute both to developments of basic science of physiology, and new possibilities in advanced biotechnology of assisted reproduction.}, number={4}, journal={INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES}, author={Kulus, Magdalena and Kranc, Wieslawa and Wojtanowicz-Markiewicz, Katarzyna and Celichowski, Piotr and Swiatly-Blaszkiewicz, Agata and Matuszewska, Eliza and Sujka-Kordowska, Patrycja and Konwerska, Aneta and Zdun, Maciej and Bryl, Rut and et al.}, year={2021}, month={Feb} } @article{wei_zhang_zhang_wu_petitte_hou_si_ahmad_guo_zhang_et al._2021, title={Targeting the TLR2 Receptor With a Novel Thymopentin-Derived Peptide Modulates Immune Responses}, volume={12}, ISSN={["1664-3224"]}, DOI={10.3389/fimmu.2021.620494}, abstractNote={The innate and adaptive immune systems act in concert to protect us from infectious agents and other harmful substances. As a state of temporary or permanent immune dysfunction, immunosuppression can make an organism more susceptible to infection, organ injury, and cancer due to damage to the immune system. It takes a long time to develop new immunomodulatory agents to prevent and treat immunosuppressive diseases, with slow progress. Toll-like receptor 2 (TLR2) agonists have been reported as potential immunomodulatory candidates due to their effective activation of immune responses. It has been demonstrated that thymopentin (TP5) could modulate immunity by binding to the TLR2 receptor. However, the fairly short half-life of TP5 greatly reduces its pharmacological potential for immunosuppression therapy. Although peptide cathelicidin 2 (CATH2) has a long half-life, it shows poor immunomodulatory activity and severe cytotoxicity, which seriously hampers its clinical development. Peptide hybridization is an effective approach for the design and engineering of novel functional peptides because hybrid peptides combine the advantages and benefits of various native peptides. In this study, to overcome all these challenges faced by the parental peptides, six hybrid peptides (CaTP, CbTP, CcTP, TPCa, TPCb, and TPCc) were designed by combining the full-length TP5 with different active fragments of CATH2. CbTP, the most potent TLR2 agonist among the six hybrid peptides, was effectively screened through in silico analysis and in vitro experiments. The CbTP peptide exhibited lower cytotoxicity than either CATH2 or TP5. Furthermore, the immunomodulatory effects of CbTP were confirmed in a CTX-immunosuppressed mouse model, which showed that CbTP has increased immunopotentiating activity and physiological stability compared to the parental peptides. CbTP successfully inhibited immunosuppression and weight loss, increased immune organ indices, and improved CD4+/CD8+ T lymphocyte subsets. In addition, CbTP significantly increased the production of the cytokine TNF-α and IL-6, and the immunoglobulins IgA, IgM, and IgG. The immunoenhancing effects of CbTP were attributed to its TLR2-binding activity, promoting the formation of the TLR2 cluster, the activation of the TLR2 receptor, and thus activation of the downstream MyD88-NF-кB signaling pathway.}, journal={FRONTIERS IN IMMUNOLOGY}, author={Wei, Xubiao and Zhang, Lulu and Zhang, Rijun and Wu, Rujuan and Petitte, James N. and Hou, Yanfei and Si, Dayong and Ahmad, Baseer and Guo, Henan and Zhang, Manyi and et al.}, year={2021}, month={May} } @article{wei_zhang_zhang_wu_si_ahmad_petitte_mozdziak_li_guo_et al._2020, title={A highly efficient hybrid peptide ameliorates intestinal inflammation and mucosal barrier damage by neutralizing lipopolysaccharides and antagonizing the lipopolysaccharide-receptor interaction}, volume={34}, ISSN={["1530-6860"]}, DOI={10.1096/fj.201903263RRR}, abstractNote={Intestinal inflammatory disorders, such as inflammatory bowel disease, are major contributors to mortality and morbidity in humans and animals worldwide. While some native peptides have great potential as therapeutic agents against intestinal inflammation, potential cytotoxicity, anti‐inciting action, and suppression of anti‐inflammatory activity may limit their development as anti‐inflammatory agents. Peptide hybridization is an effective approach for the design and engineering of novel functional peptides because hybrid peptides combine the advantages and benefits of various native peptides. In the present study, a novel hybrid anti‐inflammatory peptide that combines the active center of Cecropin A (C) and the core functional region of LL‐37 (L) was designed [C‐L peptide; C (1‐8)‐L (17‐30)] through in silico analysis to reduce cytotoxicity and improve the anti‐inflammatory activity of the parental peptides. The resulting C‐L peptide exhibited lower cytotoxicity than either C or L peptides alone. C‐L also exerted a protective effect against lipopolysaccharide (LPS)‐induced inflammatory responses in RAW264.7 macrophages and in the intestines of a mouse model. The hybrid peptide exhibited increased anti‐inflammatory activity compared to the parental peptides. C‐L plays a role in protecting intestinal tissue from damage, LPS‐induced weight loss, and leukocyte infiltration. In addition, C‐L reduces the expression levels of tumor necrosis factor‐alpha (TNF‐α), interleukin‐6 (IL‐6), IL‐1β, and interferon‐gamma (IFN‐γ), as well as reduces cell apoptosis. It also reduced mucosal barrier damage caused by LPS. The anti‐inflammatory effects of the hybrid peptide were mainly attributed to its LPS‐neutralizing activity and antagonizing the activation of LPS‐induced Toll‐like receptor 4‐myeloid differentiation factor 2 (TLR4/MD2). The peptide also affected the TLR4‐(nuclear factor κB) signaling pathway, modulating the inflammatory response upon LPS stimulation. Collectively, these findings suggest that the newly designed peptide, C‐L, could be developed into a novel anti‐inflammatory agent for animals or humans.}, number={12}, journal={FASEB JOURNAL}, author={Wei, Xubiao and Zhang, Lulu and Zhang, Rijun and Wu, Rujuan and Si, Dayong and Ahmad, Baseer and Petitte, James N. and Mozdziak, Paul E. and Li, Zhongxuan and Guo, Henan and et al.}, year={2020}, month={Dec}, pages={16049–16072} } @article{jankowski_mozdziak_petitte_kulus_kempisty_2020, title={Avian Satellite Cell Plasticity}, volume={10}, url={https://doi.org/10.3390/ani10081322}, DOI={10.3390/ani10081322}, abstractNote={Adult myogenesis is dependent on a population of precursor cells, located between the sarcolemma and the basal lamina of the muscle fiber. These satellite cells, usually present in a quiescent state, become activated in response to mechanical muscle strain, differentiating and fusing to add new nuclei to enlarging muscles. As their myogenic lineage commitment is induced on demand, muscle satellite cells exhibit a certain amount of plasticity, possibly being able to be directed to differentiate into non-myogenic fates. In this study, myosatellite cells were isolated from chicken muscle samples, characterized in vitro and introduced into developing blastoderms. They were further investigated using fluorescence microscopy, immunohistochemistry and PCR, to determine their location in embryos after three and eighteen days. The results of the in vitro analysis confirmed that the cells obtained from the Pectoralis thoracicus are highly myogenic, based on the expression of Pax7, Myogenin, MyoD, Desmin and the myotube assay. Furthermore, the investigation of satellite cells within the embryo showed their migration to the regions of Pectoralis thoracicus, heart, liver, gizzard, proventriculus, intestine and brain. Overall, the results of the study proved the high myogenicity of chicken Pectoralis thoracicus cell isolates, as well as provided new information about their migration pathways following introduction into the blastocyst. The presence of the introduced LacZ or eGFP transgenes across the embryo, even 20 days after myosatellite cell injection, further supports the notion that satellite cells exhibit significant plasticity, potentially transdifferentiating into non-muscle lineages.}, number={8}, journal={Animals}, publisher={MDPI AG}, author={Jankowski, Maurycy and Mozdziak, Paul and Petitte, James and Kulus, Magdalena and Kempisty, Bartosz}, year={2020}, month={Jul}, pages={1322} } @article{dompe_janowicz_hutchings_moncrieff_jankowski_nawrocki_józkowiak_mozdziak_petitte_shibli_et al._2020, title={Epigenetic Research in Stem Cell Bioengineering—Anti-Cancer Therapy, Regenerative and Reconstructive Medicine in Human Clinical Trials}, url={https://doi.org/10.3390/cancers12041016}, DOI={10.3390/cancers12041016}, abstractNote={The epigenome denotes all the information related to gene expression that is not contained in the DNA sequence but rather results from chemical changes to histones and DNA. Epigenetic modifications act in a cooperative way towards the regulation of gene expression, working at the transcriptional or post-transcriptional level, and play a key role in the determination of phenotypic variations in cells containing the same genotype. Epigenetic modifications are important considerations in relation to anti-cancer therapy and regenerative/reconstructive medicine. Moreover, a range of clinical trials have been performed, exploiting the potential of epigenetics in stem cell engineering towards application in disease treatments and diagnostics. Epigenetic studies will most likely be the basis of future cancer therapies, as epigenetic modifications play major roles in tumour formation, malignancy and metastasis. In fact, a large number of currently designed or tested clinical approaches, based on compounds regulating epigenetic pathways in various types of tumours, employ these mechanisms in stem cell bioengineering.}, journal={Cancers}, author={Dompe, Claudia and Janowicz, Krzysztof and Hutchings, Greg and Moncrieff, Lisa and Jankowski, Maurycy and Nawrocki, Mariusz J. and Józkowiak, Małgorzata and Mozdziak, Paul and Petitte, Jim and Shibli, Jamil A. and et al.}, year={2020}, month={Apr} } @article{chermuła_kranc_jopek_joanna_hutchings_dompe_moncrieff_janowicz_józkowiak_jeseta_et al._2020, title={Human Cumulus Cells in Long-Term In Vitro Culture Reflect Differential Expression Profile of Genes Responsible for Planned Cell Death and Aging—A Study of New Molecular Markers}, url={https://www.mdpi.com/2073-4409/9/5/1265}, DOI={10.3390/cells9051265}, abstractNote={In the ovarian follicle, maturation of the oocyte increases in the presence of somatic cells called cumulus cells (CCs). These cells form a direct barrier between the oocyte and external environment. Thanks to bidirectional communication, they have a direct impact on the oocyte, its quality and development potential. Understanding the genetic profile of CCs appears to be important in elucidating the physiology of oocytes. Long-term in vitro culture of CCs collected from patients undergoing controlled ovarian stimulation during in vitro fertilization procedure was conducted. Using microarray expression analysis, transcript levels were assessed on day 1, 7, 15, and 30 of culture. Apoptosis and aging of CCs strictly influence oocyte quality and subsequently the outcome of assisted reproductive technologies (ART). Thus, particular attention was paid to the analysis of genes involved in programmed cell death, aging, and apoptosis. Due to the detailed level of expression analysis of each of the 133 analyzed genes, three groups were selected: first with significantly decreased expression during the culture; second with the statistically lowest increase in expression; and third with the highest significant increase in expression. COL3A1, SFRP4, CTGF, HTR2B, VCAM1, TNFRSF11B genes, belonging to the third group, were identified as potential carriers of information on oocyte quality.}, journal={Cells}, author={Chermuła, Błażej and Kranc, Wiesława and Jopek, Karol and Joanna and Hutchings, Greg and Dompe, Claudia and Moncrieff, Lisa and Janowicz, Krzysztof and Józkowiak, Małgorzata and Jeseta, Michal and et al.}, year={2020}, month={May} } @article{dompe_kranc_jopek_kowalska_ciesiolka_chermuła_bryja_jankowski_perek_józkowiak_et al._2020, title={Muscle Cell Morphogenesis, Structure, Development and Differentiation Processes Are Significantly Regulated during Human Ovarian Granulosa Cells In Vitro Cultivation}, url={https://www.mdpi.com/2077-0383/9/6/2006}, DOI={10.3390/jcm9062006}, abstractNote={Granulosa cells (GCs) have many functions and are fundamental for both folliculogenesis and oogenesis, releasing hormones and communicating directly with the oocyte. Long-term in vitro cultures of GCs show significant stem-like characteristics. In the current study, RNA of human ovarian granulosa cells was collected at 1, 7, 15 and 30 days of long-term in vitro culture. Understanding the process of differentiation of GCs towards different cell lineages, as well as the molecular pathways underlying these mechanisms, is fundamental to revealing other possible stemness markers of this type of cell. Identifying new markers of GC plasticity may help to understand the aetiology and recurrence of a wide variety of diseases and health conditions and reveal possible clinical applications of the ovarian tissue cells, affecting not only the reproductive ability but also sex hormone production. Granulosa cells were the subject of this study, as they are readily available as remnant material leftover after in vitro fertilisation procedures and exhibit significant stem-like characteristics in culture. The change in gene expression was investigated through a range of molecular and bioinformatic analyses. Expression microarrays were used, allowing the identification of groups of genes typical of specific cellular pathways. This candidate gene study focused on ontological groups associated with muscle cell morphogenesis, structure, development and differentiation, namely, “muscle cell development”, “muscle cell differentiation”, “muscle contraction”, “muscle organ development”, “muscle organ morphogenesis”, “muscle structure development”, “muscle system process” and “muscle tissue development”. The results showed that the 10 most upregulated genes were keratin 19, oxytocin receptor, connective tissue growth factor, nexilin, myosin light chain kinase, cysteine and glycine-rich protein 3, caveolin 1, actin, activating transcription factor 3 and tropomyosin, while the 10 most downregulated consisted of epiregulin, prostaglandin-endoperoxide synthase 2, transforming growth factor, interleukin, collagen, 5-hydroxytryptmine, interleukin 4, phosphodiesterase, wingless-type MMTV integration site family and SRY-box 9. Moreover, ultrastructural observations showing heterogeneity of granulosa cell population are presented in the study. At least two morphologically different subpopulations were identified: large, light coloured and small, darker cells. The expression of genes belonging to the mentioned ontological groups suggest the potential ability of GCs to differentiate and proliferate toward muscle lineage, showing possible application in muscle regeneration and the treatment of different diseases.}, journal={Journal of Clinical Medicine}, author={Dompe, Claudia and Kranc, Wiesława and Jopek, Karol and Kowalska, Katarzyna and Ciesiolka, Sylwia and Chermuła, Błażej and Bryja, Artur and Jankowski, Maurycy and Perek, Joanna and Józkowiak, Małgorzata and et al.}, year={2020}, month={Jun} } @article{miramontes_kempisty_petitte_dasarathy_kulus_wieczorkiewicz_mozdziak_2020, title={Myogenic Response to Increasing Concentrations of Ammonia Differs between Mammalian, Avian, and Fish Species: Cell Differentiation and Genetic Study}, url={https://doi.org/10.3390/genes11080840}, DOI={10.3390/genes11080840}, abstractNote={Ammonia is very toxic to the body and has detrimental effects on many different organ systems. Using cultured myoblast cells, we examined ammonia’s effect on myostatin expression, a negative regulator of skeletal muscle growth, and myotube diameters. The objective of this study was to examine how murine, avian, and fish cells respond to increasing levels of ammonia up to 50 mM. The murine myoblast cell line (C2C12), primary chick, and primary tilapia myoblast cells were cultured and then exposed to 10, 25, and 50 mM ammonium acetate, sodium acetate, and an untreated control for 24 h. High levels of ammonia were detrimental to the C2C12 cells, causing increased Myostatin (MSTN) expression and decreased myotube diameters between 10 and 25 mM (p < 0.002). Ammonia at 10 mM continued the positive myogenic response in the chick, with lower MSTN expression than the C2C12 cells and larger myotube diameters, but the myotube diameter at 50 mM ammonium acetate was significantly smaller than those at 10 and 25 mM (p < 0.001). However, chick myotubes at 50 mM were still significantly larger than the sodium acetate-treated and untreated control (p < 0.001). The tilapia cells showed no significant difference in MSTN expression or myotube diameter in response to increasing the concentrations of ammonia. Overall, these results confirm that increasing concentrations of ammonia are detrimental to mammalian skeletal muscle, while chick cells responded positively at lower levels but began to exhibit a negative response at higher levels, as the tilapia experienced no detrimental effects. The differences in ammonia metabolism strategies between fish, avian, and mammalian species could potentially contribute to the differences between species in response to high levels of ammonia. Understanding how ammonia affects skeletal muscle is important for the treatment of muscle wasting observed in liver failure patients.}, journal={Genes}, author={Miramontes, Emily and Kempisty, Bartosz and Petitte, James and Dasarathy, Srinivasan and Kulus, Magdalena and Wieczorkiewicz, Maria and Mozdziak, Paul}, year={2020}, month={Jul} } @article{miramontes_mozdziak_petitte_kulus_wieczorkiewicz_kempisty_2020, title={Skeletal Muscle and the Effects of Ammonia Toxicity in Fish, Mammalian, and Avian Species: A Comparative Review Based on Molecular Research}, volume={21}, url={https://doi.org/10.3390/ijms21134641}, DOI={10.3390/ijms21134641}, abstractNote={Typically, mammalian and avian models have been used to examine the effects of ammonia on skeletal muscle. Hyperammonemia causes sarcopenia or muscle wasting, in mammals and has been linked to sarcopenia in liver disease patients. Avian models of skeletal muscle have responded positively to hyperammonemia, differing from the mammalian response. Fish skeletal muscle has not been examined as extensively as mammalian and avian muscle. Fish skeletal muscle shares similarities with avian and mammalian muscle but has notable differences in growth, fiber distribution, and response to the environment. The wide array of body sizes and locomotion needs of fish also leads to greater diversity in muscle fiber distribution and growth between different fish species. The response of fish muscle to high levels of ammonia is important for aquaculture and quality food production but has not been extensively studied to date. Understanding the differences between fish, mammalian and avian species’ myogenic response to hyperammonemia could lead to new therapies for muscle wasting due to a greater understanding of the mechanisms behind skeletal muscle regulation and how ammonia effects these mechanisms. This paper provides an overview of fish skeletal muscle and ammonia excretion and toxicity in fish, as well as a comparison to avian and mammalian species.}, number={13}, journal={International Journal of Molecular Sciences}, publisher={MDPI AG}, author={Miramontes, Emily and Mozdziak, Paul and Petitte, James N. and Kulus, Magdalena and Wieczorkiewicz, Maria and Kempisty, Bartosz}, year={2020}, month={Jun}, pages={4641} } @article{zhang_wei_zhang_si_petitte_ahmad_zhang_2019, title={A Novel Peptide Ameliorates LPS-Induced Intestinal Inflammation and Mucosal Barrier Damage via Its Antioxidant and Antiendotoxin Effects}, volume={20}, ISSN={["1422-0067"]}, url={https://doi.org/10.3390/ijms20163974}, DOI={10.3390/ijms20163974}, abstractNote={Intestinal inflammation is an inflammatory disease resulting from immune dysregulation in the gut. It can increase the risk of enteric cancer, which is a common malignancy globally. As a new class of anti-inflammatory agents, native peptides have potential for use in the treatment of several intestinal inflammation conditions; however, their potential cytotoxicity and poor anti-inflammatory activity and stability have prevented their development. Hybridization has been proposed to overcome this problem. Thus, in this study, we designed a hybrid peptide (LL-37-TP5, LTP) by combing the active centre of LL-37 (13–36) with TP5. The half-life and cytotoxicity were tested in vitro, and the hybrid peptide showed a longer half-life and lower cytotoxicity than its parental peptides. We also detected the anti-inflammatory effects and mechanisms of LTP on Lipopolysaccharide (LPS)-induced intestinal inflammation in murine model. The results showed that LTP effectively prevented LPS-induced weight loss, impairment of intestinal tissues, leukocyte infiltration, and histological evidence of inflammation. Additionally, LTP decreased the levels of tumour necrosis factor-alpha, interferon-gamma, and interleukin-6; increased the expression of zonula occludens-1 and occludin; and reduced permeability in the jejunum of LPS-treated mice. Notably, LTP appeared to be more potent than the parental peptides LL-37 and TP5. The anti-inflammatory effects of LTP may be associated with the neutralization of LPS, inhibition of oxidative stress, and inhibition of the NF-κB signalling pathway. The findings of this study suggest that LTP might be an effective therapeutic agent for treating intestinal inflammation.}, number={16}, journal={INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES}, publisher={MDPI AG}, author={Zhang, Lulu and Wei, Xubiao and Zhang, Rijun and Si, Dayong and Petitte, James N. and Ahmad, Baseer and Zhang, Manyi}, year={2019}, month={Aug} } @article{zhang_wei_zhang_petitte_si_li_cheng_du_2019, title={Design and Development of a Novel Peptide for Treating Intestinal Inflammation}, volume={10}, ISSN={["1664-3224"]}, url={http://europepmc.org/abstract/med/31447849}, DOI={10.3389/fimmu.2019.01841}, abstractNote={Intestinal inflammatory disorders, such as inflammatory bowel disease (IBD), are associated with increased pro-inflammatory cytokine secretion in the intestines. Furthermore, intestinal inflammation increases the risk of enteric cancer, which is a common malignancy globally. Native anti-inflammatory peptides are a class of anti-inflammatory agents that could be used in the treatment of several intestinal inflammation conditions. However, potential cytotoxicity, and poor anti-inflammatory activity have prevented their development as anti-inflammatory agents. Therefore, in this study, we designed and developed a novel hybrid peptide for the treatment of intestinal inflammation. Eight hybrid peptides were designed by combining the active centers of antimicrobial peptides, including LL-37 (13-36), YW12D, innate defense regulator 1, and cathelicidin 2 (1-13) with thymopentin or the active center of thymosin alpha 1 (Tα1) (17-24). The hybrid peptide, LL-37-Tα1 (LTA), had improved anti-inflammatory activity with minimal cytotoxicity. LTA was screened by molecule docking and in vitro experiments. Likewise, its anti-inflammatory effects and mechanisms were also evaluated using a lipopolysaccharide (LPS)-induced intestinal inflammation murine model. The results showed that LTA prevented LPS-induced impairment in the jejunum epithelium tissues and infiltration of leukocytes, which are both histological markers of inflammation. Additionally, LTA decreased the levels of tumor necrosis factor-alpha, interferon-gamma, interleukin-6, and interleukin-1β. LTA increased the expression of zonula occludens-1 and occludin, and reduced permeability and apoptosis in the jejunum of LPS-treated mice. Additionally, its anti-inflammatory effect is associated with neutralizing LPS, binding to the Toll-like receptor 4-myeloid differentiation factor 2 (TLR4/MD-2) complex, and modulating the nuclear factor-kappa B signal transduction pathway. The findings of this study suggest that LTA may be an effective therapeutic agent in the treatment of intestinal inflammation.}, journal={FRONTIERS IN IMMUNOLOGY}, author={Zhang, Lulu and Wei, Xubiao and Zhang, Rijun and Petitte, Jim N. and Si, Dayong and Li, Zhongxuan and Cheng, Junhao and Du, Mengsi}, year={2019}, month={Aug} } @article{golkar-narenji_petitte_mozdziak_2020, title={Transgenic chicken/poultry birds: serving us for survival}, ISBN={["978-0-12-816352-8"]}, DOI={10.1016/B978-0-12-816352-8.00009-6}, abstractNote={Avian transgenesis has been considered as a useful tool for many purposes including improvement of the poultry industry, production of recombinant proteins and developmental biology. Due to the importance of the poultry industry in human nutrition, many efforts have been made for the improvement of the poultry industry in the aspect of egg and meat production for higher quantity or quality. After the development of transgenic technology the idea of transgenic poultry was raised for the application of this technology in the poultry industry. Due to the important role of recombinant proteins and humanized antibodies for medical and diagnostic purposes, mass production of these biological macromolecules is very important in human health. Development of transgenic technology in poultry science possibly helps to improve the efficiency of poultry industry productions and it may cause the development of different branches of avian industry with novel productions.}, journal={GENOMICS AND BIOTECHNOLOGICAL ADVANCES IN VETERINARY, POULTRY, AND FISHERIES}, author={Golkar-Narenji, Afsaneh and Petitte, James N. and Mozdziak, Paul E.}, year={2020}, pages={211–221} } @article{mocka_stern_fletcher_anderson_petitte_mozdziak_2017, title={Chemoprevention of spontaneous ovarian cancer in the domestic hen}, volume={96}, ISSN={0032-5791}, url={http://dx.doi.org/10.3382/ps/pew422}, DOI={10.3382/ps/pew422}, abstractNote={&NA; The hen is an attractive animal model for in vivo testing of agents that thwart ovarian carcinogenesis because ovarian cancer in the domestic hen features clinical and molecular alterations that are similar to ovarian cancer in humans, including a high incidence of p53 mutations. The objective of the study was to test the potential ovarian cancer chemopreventive effect of the p53 stabilizing compound CP‐31398 on hens that spontaneously present the ovarian cancer phenotype. Beginning at 79 wk of age, 576 egg‐laying hens (Gallus domesticus) were randomized to diets containing different amounts of CP‐31398 for 94 wk, 5 d, comprising a control group (C) (n = 144), which was fed a diet containing 0 ppm (mg/kg) of CP‐31398; a low‐dose treatment (LDT) group (n = 144), which was fed a diet containing 100 ppm of CP‐31398; a moderate‐dose treatment (MDT) group (n = 144) which was fed a diet containing 200 ppm of CP‐31398; and a high‐dose treatment (HDT) group (n = 144), which was fed a diet containing 300 ppm of CP‐31398. Hens were killed at 174 wk of age to determine the incidence of ovarian and oviductal adenocarcinomas. Whereas the incidence of localized and metastatic ovarian cancers in the MDT and HDT groups was significantly lower (up to 77%) compared to levels in the C and LDT groups (P < 0.05), the incidence of oviductal cancer was unaffected by CP‐31398. CP‐31398 appears to be an effective tool for chemoprevention against ovarian malignancies, but does not appear to affect oviductal malignancies.}, number={6}, journal={Poultry Science}, publisher={Elsevier BV}, author={Mocka, E.H. and Stern, R.A. and Fletcher, O.J. and Anderson, K.E. and Petitte, J.N. and Mozdziak, P.E.}, year={2017}, month={Jun}, pages={1901–1909} } @article{nepomuceno_muddiman_petitte_2015, title={Global Proteomic Analysis of Functional Compartments in Immature Avian Follicles Using Laser Microdissection Coupled to LC-MS/MS}, volume={14}, ISSN={["1535-3907"]}, url={http://europepmc.org/abstract/med/26211554}, DOI={10.1021/acs.jproteome.5b00346}, abstractNote={Laser microdissection (LMD) was utilized for the separation of the yolk, follicular wall (granulosa and theca), and surrounding stromal cells of small white follicles (SWF) obtained from reproductively active domestic fowl. Herein, we provide an in situ proteomics-based approach to studying follicular development through the use of LMD and mass spectrometry. This study resulted in a total of 2889 proteins identified from the three specific isolated compartments. White yolk from the smallest avian follicles resulted in the identification of 1984 proteins, while isolated follicular wall and ovarian stroma yielded 2470 and 2456 proteins, respectively. GO annotations highlighted the functional differences between the compartments. Among the three compartments examined, the relative abundance of vitellogenins, steroidogenic enzymes, anti-Mullerian hormone, transcription factors, and proteins involved in retinoic acid receptors/retinoic acid synthesis, transcription factors, and cell surface receptors such as EGFR and their associated signaling pathways reflected known cellular function of the ovary. This study has provided a global proteome for SWF, white yolk, and ovarian stroma of the avian ovary that can be used as a baseline for future studies and verifies that the coupling of LMD with proteomic analysis can be used to evaluate proteins from small, physiologically functional compartments of complex tissue.}, number={9}, journal={JOURNAL OF PROTEOME RESEARCH}, author={Nepomuceno, Angelito I. and Muddiman, David C. and Petitte, James N.}, year={2015}, month={Sep}, pages={3912–3923} } @article{nepomuceno_shao_jing_ma_petitte_idowu_muddiman_fang_hawkridge_2015, title={In-depth LC-MS/MS analysis of the chicken ovarian cancer proteome reveals conserved and novel differentially regulated proteins in humans}, volume={407}, ISSN={["1618-2650"]}, url={http://europepmc.org/abstract/med/26159569}, DOI={10.1007/s00216-015-8862-4}, abstractNote={Ovarian cancer (OVC) remains the most lethal gynecological malignancy in the world due to the combined lack of early-stage diagnostics and effective therapeutic strategies. The development and application of advanced proteomics technology and new experimental models has created unique opportunities for translational studies. In this study, we investigated the ovarian cancer proteome of the chicken, an emerging experimental model of OVC that develops ovarian tumors spontaneously. Matched plasma, ovary, and oviduct tissue biospecimens derived from healthy, early-stage OVC, and late-stage OVC birds were quantitatively characterized by label-free proteomics. Over 2600 proteins were identified in this study, 348 of which were differentially expressed by more than twofold (p ≤ 0.05) in early- and late-stage ovarian tumor tissue specimens relative to healthy ovarian tissues. Several of the 348 proteins are known to be differentially regulated in human cancers including B2M, CLDN3, EPCAM, PIGR, S100A6, S100A9, S100A11, and TPD52. Of particular interest was ovostatin 2 (OVOS2), a novel 165-kDa protease inhibitor found to be strongly upregulated in chicken ovarian tumors (p = 0.0005) and matched plasma (p = 0.003). Indeed, RT-quantitative PCR and Western blot analysis demonstrated that OVOS2 mRNA and protein were also upregulated in multiple human OVC cell lines compared to normal ovarian epithelia (NOE) cells and immunohistochemical staining confirmed overexpression of OVOS2 in primary human ovarian cancers relative to non-cancerous tissues. Collectively, these data provide the first evidence for involvement of OVOS2 in the pathogenesis of both chicken and human ovarian cancer.}, number={22}, journal={ANALYTICAL AND BIOANALYTICAL CHEMISTRY}, author={Nepomuceno, Angelito I. and Shao, Huanjie and Jing, Kai and Ma, Yibao and Petitte, James N. and Idowu, Michael O. and Muddiman, David C. and Fang, Xianjun and Hawkridge, Adam M.}, year={2015}, month={Sep}, pages={6851–6863} } @article{harris_fletcher_anderson_petitte_kopelovich_mozdziak_2014, title={Epithelial Cell Tumors of the Hen Reproductive Tract}, volume={58}, ISSN={["1938-4351"]}, url={http://europepmc.org/abstract/med/24758120}, DOI={10.1637/10643-082313-reg.1}, abstractNote={SUMMARY There is a paucity of preclinical models that simulate the development of ovarian tumors in humans. At present, the egg-laying hen appears to be the most promising model to study the spontaneous occurrence of ovarian tumors in the clinical setting. Although gross classification and histologic grade of tumors have been used prognostically in women with ovarian tumors, there is currently no single system that is universally used to classify reproductive tumors in the hen. Four hundred and one 192-wk-old egg-laying hens were necropsied to determine the incidence of reproductive tumors using both gross pathology and histologic classification. Gross pathologic classifications were designated as follows: birds presenting with ovarian tumors only (class 1), those presenting with oviductal and ovarian tumors (class 2), those with ovarian and oviductal tumors that metastasized to the gastrointestinal tract (class 3), those with ovarian and oviductal tumors that metastasized to the gastrointestinal tract and other distant organs (class 4), those with oviductal tumors only (class 5), those with oviductal tumors that metastasized to other organs with no ovarian involvement (class 6), and those with ovarian tumors that metastasized to other organs with no oviductal involvement (class 7), including birds with gastrointestinal tumors and no reproductive involvement (GI only) and those with no tumors (normal). Histopathologic classifications range from grades 1 to 3 and are based on mitotic developments and cellular differentiation. An updated gross pathology and histologic classification systems for the hen reproductive malignancies provides a method to report the range of reproductive tumors revealed in a flock of aged laying hens. RESUMEN Tumores de células epiteliales del tracto reproductivo de la gallina. Hay una escasez de modelos preclínicos que simulen el desarrollo de los tumores de ovario en humanos. En la actualidad, la gallina de postura parece ser el modelo más prometedor para estudiar la aparición espontánea de tumores en el ovario en el ámbito clínico. Aunque la clasificación macroscópica y el grado histológico de los tumores se han utilizado para realizar el pronóstico en mujeres con tumores de ovario, no existe actualmente ningún sistema que se utilice universalmente para clasificar tumores reproductivos en la gallina. Se realizó la necropsia de 401 gallinas de postura de 192 semanas de edad para determinar la incidencia de los tumores del aparato reproductor utilizando tanto patología macroscópica y clasificación histológica. Las clasificaciones patológicas macroscópicas fueron designadas de la siguiente manera: las aves que presentaron únicamente tumores de ovario (clase 1), los que presentan tumores de ovario y oviducto (clase 2), aquellos con tumores de ovario y oviducto y con metástasis en el tracto gastrointestinal (clase 3), las aves que tienen tumores de ovario y oviducto, con metástasis en el tracto gastrointestinal y otros órganos distantes (clase 4), las aves que tienen tumores del oviducto solamente (clase 5), aquellas con tumores del oviducto con metástasis a otros órganos sin la participación de ovario (clase 6), y las aves que tienen tumores de ovario con metástasis a otros órganos sin la participación del oviducto (clase 7), incluidas las aves con tumores gastrointestinales y sin compromiso del aparato reproductivo (sólo tracto gastrointestinal) y las aves que no tienen tumores (normales). Las clasificaciones histopatológicas tuvieron un rango que va desde los grados 1 a 3, y se basaron en el desarrollo de mitosis y diferenciación celular. Una descripción patológica macroscópica actualizada y los sistemas de clasificación histológica de los tumores malignos reproductivos de la gallina proporcionan un método para reportar la gama de tumores reproductivos detectados en una parvada de gallinas ponedoras de edad.}, number={1}, journal={Avian Diseases}, author={Harris, E.A. and Fletcher, O.J. and Anderson, K.E. and Petitte, J.N. and Kopelovich, L. and Mozdziak, P.E.}, year={2014}, month={Mar}, pages={95–101} } @article{lu_west_jordan_jordan_west_yu_he_barrios_zhu_petitte_et al._2014, title={Induced Pluripotency in Chicken Embryonic Fibroblast Results in a Germ Cell Fate}, volume={23}, ISSN={["1557-8534"]}, url={http://europepmc.org/abstract/med/24720794}, DOI={10.1089/scd.2014.0080}, abstractNote={Germ cells (GCs) are critically important as the vehicle that passes genetic information from one generation to the next. Correct development of these cells is essential and perturbation in their development often leads to reproductive failure and disease. Despite the importance of GCs, little is known about the mechanisms underlying the acquisition and maintenance of the GC character. Using a reprogramming strategy, we demonstrate that overexpression of ectopic transcription factors in embryonic fibroblasts can lead to the generation of chicken induced primordial germ cells (ciPGCs). These ciPGCs express pluripotent markers POU5F1, SSEA1, and the GC defining proteins, CVH and DAZL, closely resembling in vivo sourced PGCs instead of embryonic stem cells. Moreover, CXCR4 expressing ciPGCs were capable of migrating to the embryonic gonad after injection into the vasculature of stage 15 embryos, indicating the acquisition of a GC fate in these cells. Direct availability of ciPGCs in vitro would facilitate the study of GC development as well as provide a potential strategy for the conservation of important genetics of agricultural and endangered birds using somatic cells.}, number={15}, journal={STEM CELLS AND DEVELOPMENT}, author={Lu, Yangqing and West, Franklin D. and Jordan, Brian J. and Jordan, Erin T. and West, Rachel C. and Yu, Ping and He, Ying and Barrios, Miguel A. and Zhu, Ziying and Petitte, James N. and et al.}, year={2014}, month={Aug}, pages={1755–1764} } @article{rodriguez_barnes_anderson_whitaker_berchuck_petitte_lancaster_wenham_turbov_day_et al._2013, title={Evidence of a Chemopreventive Effect of Progestin Unrelated to Ovulation on Reproductive Tract Cancers in the Egg-laying Hen}, volume={6}, ISSN={1940-6207 1940-6215}, url={http://dx.doi.org/10.1158/1940-6207.CAPR-12-0426}, DOI={10.1158/1940-6207.capr-12-0426}, abstractNote={Epidemiologic, laboratory, and animal evidence suggests that progestins and vitamin D may be potent ovarian cancer preventives. Our objectives were to evaluate progestins as reproductive tract cancer chemopreventives in the chicken, determine whether restricted ovulation affected the incidence of reproductive tract tumors, and assess whether vitamin D would confer cancer protection either alone or in addition to progestin. A total of 2,400 two-year-old Single Comb White Leghorns were randomized into six groups (400 each) with hormonal and dietary manipulation for 2 years as follows: (i) no intervention, regular feed/caloric intake, (ii) control, (iii) vitamin D, (iv) the progestin levonorgestrel, (v) vitamin D plus levonorgestrel, and (vi) the progestin Provera (medroxyprogesterone acetate). Groups 2 to 6 were caloric restricted to inhibit ovulation. Our results indicated that caloric restriction decreased egg production by more than 60%, and was associated with a greater than 70% decrease in reproductive tract cancers. Ovulatory events did not differ among the caloric-restricted groups (groups 2–6), except for the group receiving levonorgestrel, which had fewer ovulatory events than controls (P = 0.046). After correcting for egg production, birds receiving progestins had significantly fewer reproductive tract cancers [OR, 0.61; confidence interval (CI), 0.39–0.95; P = 0.03], with similar proportionate reductions in tumors arising in either the ovary or oviduct. Vitamin D did not significantly affect cancer incidence overall, or add to the cancer preventive effect of progestins. This study suggests a protective effect of progestins against ovarian and oviductal cancers. These data support the concept that progestins provide a chemopreventive effect unrelated to ovulation. Cancer Prev Res; 6(12); 1283–92. ©2013 AACR.}, number={12}, journal={Cancer Prevention Research}, publisher={American Association for Cancer Research (AACR)}, author={Rodriguez, G. C. and Barnes, H. J. and Anderson, K. E. and Whitaker, R. S. and Berchuck, A. and Petitte, J. N. and Lancaster, J. M. and Wenham, R. M. and Turbov, J. M. and Day, R. and et al.}, year={2013}, month={Oct}, pages={1283–1292} } @article{andrews kingon_petitte_muddiman_hawkridge_2013, title={Multi-peptide nLC-PC-IDMS-SRM-based Assay for the quantification of biomarkers in the chicken ovarian cancer model}, volume={61}, ISSN={1046-2023}, url={http://dx.doi.org/10.1016/J.YMETH.2013.04.004}, DOI={10.1016/J.YMETH.2013.04.004}, abstractNote={A novel form of ovomacroglobulin/ovostatin (OVOS2) predicted from EST data was previously identified in the chicken ovarian cancer model using a mass spectrometry-based shotgun label-free proteomics strategy. The quantitative label-free data from plasma showed a significant increase over time with the spontaneous onset and progression of ovarian cancer making it a potential protein biomarker for further study. Two other proteins of interest identified from this initial study included vitellogenin-1 (Vit-1), a lipid-transport protein tied to egg production, and transthyretin (TTR), a retinol binding transport protein currently used in the clinical management of ovarian cancer. A multiplexed protein cleavage isotope dilution mass spectrometry (PC-IDMS) assay was developed to quantify OVOS2, Vit-1, and TTR by selected reaction monitoring (SRM). A total of 6 stable isotope labeled (SIL) peptide standards were used in the assay with three tryptic peptides from OVOS2, one for Vit-1, and two for TTR. The assay was developed for use with un-depleted raw plasma combined with the filter assisted sample preparation (FASP) method and its use was also demonstrated for matched ovary tissue samples. The PC-IDMS data for the two TTR peptides did not correlate with each other with more than a 10-fold difference in concentration for all 5 time points measured. The PC-IDMS data from the longitudinal plasma samples correlated well for OVOS2 and Vit-1 whereas TTR was inconclusive. Interestingly, the absolute amount for one of the OVOS2 SIL peptides was 2-fold less compared with the other two SIL peptides. These data illustrate the successes and challenges of qualifying quantitative levels of proteins from an in-gel digestion sample preparation followed by LC-MS/MS (GeLC) label-free discovery-based approach to a targeted SRM-based quantitative assay in plasma and tissues.}, number={3}, journal={Methods}, publisher={Elsevier BV}, author={Andrews Kingon, Genna L. and Petitte, James N. and Muddiman, David C. and Hawkridge, Adam M.}, year={2013}, month={Jun}, pages={323–330} } @article{carver_barnes_anderson_petitte_whitaker_berchuck_rodriguez_2011, title={Reduction of Ovarian and Oviductal Cancers in Calorie-Restricted Laying Chickens}, volume={4}, ISSN={["1940-6215"]}, url={http://europepmc.org/abstract/med/21325563}, DOI={10.1158/1940-6207.capr-10-0294}, abstractNote={Abstract Epithelial ovarian cancer (OVAC) remains a highly lethal malignancy. It is a leading cause of cancer deaths among women in the United States causing more deaths than all other gynecologic malignancies combined. The pathogenesis of OVAC is not completely understood, but the process of repeated ovulation is believed to lead to genetic damage in the ovarian epithelium. As part of a prospective trial designed to evaluate OVAC chemopreventive strategies using the chicken model, caloric restriction (55% less energy) was used to inhibit ovulation in groups of hens receiving chemopreventives, thereby minimizing the impact of ovulation on the incidence of reproductive tract cancer. A separate group of chickens was maintained concurrently in the same environment, and managed similarly, except that caloric intake was not restricted. Among birds not receiving chemopreventive agents, we compared caloric versus noncaloric restricted birds to determine the relations between calorie restriction and risk of developing adenocarcinoma of the reproductive tract. Mortality in the calorie-restricted group was almost half that of those on full feed. Calorie-restricted chickens maintained body weights averaging 1.423 kg compared with the full-fed birds at 1.892 kg. Ovulation rate varied with the full-fed group producing 64% more eggs than the calorie-restricted group. Total reproductive cancers occurred in 57 (33.3%) birds for the full-fed group and 26 (10.3%) birds for the calorie-restricted group. On the basis of histopathology, 45 (26.3%) birds in the full-fed group had ovarian adenocarcinoma compared with 16 (6.3%) birds in the calorie-restricted group. Calorie restriction in laying hens resulted in a near five-fold reduction in OVAC. Cancer Prev Res; 4(4); 562–7. ©2011 AACR.}, number={4}, journal={CANCER PREVENTION RESEARCH}, author={Carver, Donna K. and Barnes, H. John and Anderson, Kenneth E. and Petitte, James N. and Whitaker, Regina and Berchuck, Andrew and Rodriguez, Gustavo C.}, year={2011}, month={Apr}, pages={562–567} } @misc{puetzer_petitte_loboa_2010, title={Comparative Review of Growth Factors for Induction of Three-Dimensional In Vitro Chondrogenesis in Human Mesenchymal Stem Cells Isolated from Bone Marrow and Adipose Tissue}, volume={16}, ISSN={["1937-3376"]}, url={http://europepmc.org/abstract/med/20196646}, DOI={10.1089/ten.teb.2009.0705}, abstractNote={The ability of bone-marrow-derived mesenchymal stem cells (MSCs) and adipose-derived stem cells (ASCs) to undergo chondrogenic differentiation has been studied extensively, and it has been suggested that the chondrogenic potential of these stem cells differ from each other. Here, we provide a comprehensive review and analysis of the various growth factor induction agents for MSC and ASC three-dimensional in vitro chondrogenic differentiation. In general, the most common growth factors for chondrogenic induction come from the transforming growth factor beta (TGFbeta) superfamily. To date, the most promising growth factors for chondrogenesis appear to be TGFbeta-3 and bone morphogenetic protein (BMP)-6. A thorough review of the literature indicates that human MSCs (hMSCs) appear to exhibit the highest chondrogenic potential in three-dimensional culture in the medium containing both dexamethasone and TGFbeta-3. Some reports indicate that the addition of BMP-6 to TFGbeta-3 and dexamethasone further increases hMSC chondrogenesis, but these results are still not consistently supported. Induction of human ASC (hASC) chondrogenesis appears most successful when dexamethasone, TGFbeta-3, and BMP-6 are used in combination. However, to date, current formulations do not always result in stable differentiation to the chondrocytic lineage by hMSCs and hASCs. Continued research must be performed to examine the expression cascades of the TFGbeta superfamily to further determine the effects of each growth factor alone and in combination on these stem cell lines.}, number={4}, journal={TISSUE ENGINEERING PART B-REVIEWS}, author={Puetzer, Jennifer L. and Petitte, John N. and Loboa, Elizabeth G.}, year={2010}, month={Aug}, pages={435–444} } @article{bosquet_peedicayil_maguire_chien_rodriguez_whitaker_petitte_anderson_barnes_shridhar_et al._2011, title={Comparison of gene expression patterns between avian and human ovarian cancers}, volume={120}, ISSN={["1095-6859"]}, url={http://europepmc.org/abstract/med/21093898}, DOI={10.1016/j.ygyno.2010.10.030}, abstractNote={Objectives A putative model of spontaneous cancer has been described in the laying hen that bears significant similarities to human ovarian cancer. Our objective was to characterize and compare the patterns of gene expression in chicken and human forms of this disease. Methods RNA from 20 localized and metastatic ovarian and oviductal chicken tumor samples was isolated, amplified using in vitro transcription, and hybridized against normal ovarian epithelium to a customized cDNA microarray constructed for these studies. Differentially expressed genes were identified for localized ovarian, metastatic ovarian, and oviductal (or tubal) cancer by class comparison using BRB-ArrayTools. Results were validated with semi-quantitative PCR. A gene list (prediction model) constructed with the class prediction tool was used in a human ovarian cancer microarray obtained from the GEO datasets (GSE6008) in order to compare these results across species. Results Class comparison analysis between localized ovarian, metastatic ovarian and oviductal cancer yielded 41 different informative probes that coded for 27 unique genes. Localized ovarian samples clustered between metastatic ovarian and oviductal cancer samples. Using our chicken data as a training set and leaving oviductal samples out of the analysis, we created a prediction model that classified early stage and advanced stage human ovarian cancer gene expression arrays with 78% overall accuracy. Conclusions Gene expression of spontaneous ovarian cancer in the chicken is comparable to gene expression patterns of human ovarian cancer.}, number={2}, journal={GYNECOLOGIC ONCOLOGY}, author={Bosquet, Jesus Gonzalez and Peedicayil, Abraham and Maguire, Jacie and Chien, Jeremy and Rodriguez, Gustavo C. and Whitaker, Regina and Petitte, James N. and Anderson, Kenneth E. and Barnes, H. John and Shridhar, Viji and et al.}, year={2011}, month={Feb}, pages={256–264} } @article{hawkridge_wysocky_petitte_anderson_mozdziak_fletcher_horowitz_muddiman_2010, title={Measuring the intra-individual variability of the plasma proteome in the chicken model of spontaneous ovarian adenocarcinoma}, volume={398}, ISSN={["1618-2650"]}, url={http://europepmc.org/abstract/med/20640409}, DOI={10.1007/s00216-010-3979-y}, abstractNote={The domestic chicken (Gallus domesticus) has emerged as a powerful experimental model for studying the onset and progression of spontaneous epithelial ovarian cancer (EOC) with a disease prevalence that can exceed 35% between 2 and 7 years of age. An experimental strategy for biomarker discovery is reported herein that combines the chicken model of EOC, longitudinal plasma sample collection with matched tissues, advanced mass spectrometry-based proteomics, and concepts derived from the index of individuality (Harris, Clin Chem 20: 1535–1542, 1974). Blood was drawn from 148 age-matched chickens starting at 2.5 years of age every 3 months for 1 year. At the conclusion of the 1 year sample collection period, the 73 birds that remained alive were euthanized, necropsied, and tissues were collected. Pathological assessment of resected tissues from these 73 birds confirmed that five birds (6.8%) developed EOC. A proteomics workflow including in-gel digestion, nanoLC coupled to high-performance mass spectrometry, and label-free (spectral counting) quantification was used to measure the biological intra-individual variability (CVW) of the chicken plasma proteome. Longitudinal plasma sample sets from two birds within the 73-bird biorepository were selected for this study; one bird was considered "healthy" and the second bird developed late-stage EOC. A total of 116 proteins from un-depleted plasma were identified with 80 proteins shared among all sample sets. Analytical variability (CVA) of the label-free proteomics workflow was measured using a single plasma sample analyzed five times and was found to be ≥CVW in both birds for 16 proteins (20%) and in either bird for 25 proteins (31%). Ovomacroglobulin (ovostatin) was found to increase (p < 0.001) over a 6 month period in the late-stage EOC bird providing an initial candidate protein for further investigation.}, number={2}, journal={ANALYTICAL AND BIOANALYTICAL CHEMISTRY}, author={Hawkridge, Adam M. and Wysocky, Rebecca B. and Petitte, James N. and Anderson, Kenneth E. and Mozdziak, Paul E. and Fletcher, Oscar J. and Horowitz, Jonathan M. and Muddiman, David C.}, year={2010}, month={Sep}, pages={737–749} } @article{hawkridge_wysocky_petitte_anderson_mozdziak_fletcher_horowitz_muddiman_2010, title={Measuring the intra-individual variability of the plasma proteome in the chicken model of spontaneous ovarian adenocarcinoma (vol 398, pg 737, 2010)}, volume={398}, ISSN={["1618-2642"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-77957848897&partnerID=MN8TOARS}, DOI={10.1007/s00216-010-4107-8}, number={4}, journal={ANALYTICAL AND BIOANALYTICAL CHEMISTRY}, author={Hawkridge, Adam M. and Wysocky, Rebecca B. and Petitte, James N. and Anderson, Kenneth E. and Mozdziak, Paul E. and Fletcher, Oscar J. and Horowitz, Jonathan M. and Muddiman, David C.}, year={2010}, month={Oct}, pages={1835–1835} } @article{dixon_bereman_petitte_hawkridge_muddiman_2011, title={One-year plasma N-linked glycome intra-individual and inter-individual variability in the chicken model of spontaneous ovarian adenocarcinoma}, volume={305}, ISSN={["1387-3806"]}, url={http://europepmc.org/abstract/med/21845070}, DOI={10.1016/j.ijms.2010.05.023}, abstractNote={Spontaneous epithelial ovarian cancer (EOC) in the chicken presents a similar pathogenesis compared with humans including CA-125 expression and genetic mutational frequencies (e.g., p53). The high prevalence of spontaneous EOC chickens also provides a unique experimental model for biomarker discovery at the genomic, proteomic, glycomic, and metabolomic level. In an effort to exploit this unique model for biomarker discovery, longitudinal plasma samples were collected from chickens at three month intervals for one year. The study described herein involved cleaving the N-glycans from these longitudinal chicken plasma samples and analyzing them via nanoLC-FTMS/MS. Glycans identified in this study were previously found in human plasma and this work provides a promising methodology to enable longitudinal studies of the N-linked plasma glycome profile during EOC progression. The structure, abundance, and intra-variability and inter-variability for 35 N-linked glycans identified in this study are reported. The full potential of the chicken model for biomarker discovery has yet to be realized, but the initial interrogation of longitudinally-procured samples provides evidence that supports the value of this strategy in the search for glycomic biomarkers.}, number={2-3}, journal={INTERNATIONAL JOURNAL OF MASS SPECTROMETRY}, author={Dixon, R. Brent and Bereman, Michael S. and Petitte, James N. and Hawkridge, Adam M. and Muddiman, David C.}, year={2011}, month={Aug}, pages={79–86} } @article{anderson_mozdziak_petitte_2010, title={The impact of scheduled cage cleaning on older hens (Gallus gallus)}, volume={39}, ISSN={["1548-4475"]}, url={http://europepmc.org/abstract/med/20567230}, DOI={10.1038/laban0710-210}, abstractNote={Researchers are increasingly using the domestic hen (Gallus gallus) as an animal model for ovarian cancer. The authors analyzed mortality rates of two large flocks of older hens that were being used for ovarian adenocarcinoma studies. All hens were fed the same maintenance diets, though some hens in each flock received experimental chemopreventive treatments. Per the request of a collaborating institution, partway through the study, the authors started to remove the hens in one of the flocks for cage changing once every 4 weeks. After the authors began cleaning some of the hens' cages, the mortality rate in this flock increased significantly. Throughout the study, within each flock, hens in the treatment and control groups had similar mortality rates. These results suggest that regularly cleaning the cages of older hens may not promote better welfare or improve flock mortality.}, number={7}, journal={LAB ANIMAL}, author={Anderson, Kenneth E. and Mozdziak, Paul E. and Petitte, James N.}, year={2010}, month={Jul}, pages={210–215} } @misc{mozdziak_petitte_2010, title={Transgenic snakes and methods of making}, volume={7,663,019}, number={2010 Feb. 16}, author={Mozdziak, P. E. and Petitte, J. N.}, year={2010} } @article{hakim_barry_barnes_anderson_petitte_whitaker_lancaster_wenham_carver_turbov_et al._2009, title={Ovarian Adenocarcinomas in the Laying Hen and Women Share Similar Alterations in p53, ras, and HER-2/neu}, volume={2}, ISSN={["1940-6215"]}, url={http://europepmc.org/abstract/med/19174584}, DOI={10.1158/1940-6207.CAPR-08-0065}, abstractNote={Abstract We examined alterations in the p53 tumor suppressor gene and the ras and HER-2/neu oncogenes in chicken ovarian cancers to determine if these tumors have genetic alterations similar to those in human ovarian adenocarcinomas. Mutations in the p53 tumor suppressor gene and the H-ras and K-ras oncogenes were assessed by direct sequencing in 172 ovarian cancers obtained from 4-year-old birds enrolled at age 2 in two separate 2-year chemoprevention trials. Birds in trial B had approximately twice as many lifetime ovulations as those in trial A. Immunohistochemical staining for the HER-2/neu oncogene was done on a subset of avian ovarian and oviductal adenocarcinomas. Alterations in p53 were detected in 48% of chicken ovarian cancers. Incidence of p53 alterations varied according to the number of lifetime ovulations, ranging from 14% in trial A to 96% in trial B (P < 0.01). No mutations were seen in H-ras, and only 2 of 172 (1.2%) tumors had K-ras mutations. Significant HER-2/neu staining was noted in 10 of 19 ovarian adenocarcinomas but in only 1 of 17 oviductal adenocarcinomas. Similar to human ovarian cancers, p53 alterations are common in chicken ovarian adenocarcinomas and correlate with the number of lifetime ovulations. Ras mutations are rare, similar to high-grade human ovarian cancers. HER-2/neu overexpression is common and may represent a marker to exclude an oviductal origin in cancers involving both the ovary and oviduct.}, number={2}, journal={CANCER PREVENTION RESEARCH}, author={Hakim, Amy A. and Barry, Catherine P. and Barnes, H. John and Anderson, Kenneth E. and Petitte, James and Whitaker, Regina and Lancaster, Jonathan M. and Wenham, Robert M. and Carver, Donna K. and Turbov, Jane and et al.}, year={2009}, month={Feb}, pages={114–121} } @article{mozdziak_hodgson_petitte_2008, title={Avian somitic cell chimeras using surrogate eggshell technology.}, volume={6}, url={http://europepmc.org/abstract/AGR/IND44063426}, journal={Asian-Australasian Journal of Animal Sciences}, author={Mozdziak, PE and Hodgson, D and Petitte, JN}, year={2008}, month={Jun} } @misc{petitte_zhang_2008, title={Culture of undifferentiated chicken cells}, volume={7,422,897}, number={2008 Sep. 9}, author={Petitte, J. N. and Zhang, Y. G.}, year={2008} } @article{ge_yu_petitte_zhang_2009, title={Epidermal Growth Factor-Induced Proliferation of Chicken Primordial Germ Cells: Involvement of Calcium/Protein Kinase C and NFKB1}, volume={80}, ISSN={["1529-7268"]}, url={http://europepmc.org/abstract/med/19005168}, DOI={10.1095/biolreprod.108.072728}, abstractNote={Abstract Epidermal growth factor (EGF) has been shown to stimulate survival in diverse cells in vitro. In the present study, the effects of EGF and the EGF-related signaling pathway on proliferation of chicken primordial germ cells (PGCs) were investigated. Results showed that EGF (10–100 ng/ml) increased the number and area of PGC colonies in a time- and dose-dependent manner. EGF also activated PKC, a process that was inhibited by AG1478 (an EGFR tyrosine kinase inhibitor) and ethyleneglycol-bis-(beta-aminoethyl ether)-N,N′-tetraacetic acid (EGTA; an intracellular Ca2+ chelator). In addition, the degradation of NFKBIA and NFKB1 (p65) translocation was observed after EGF treatment, which was significantly blocked by pretreatment with AG1478, EGTA, H7, or SN50 (NFKB1-specific inhibitor). Furthermore, we found that EGF-induced cell proliferation was significantly attenuated by AG1478, EGTA, H7, and SN50, respectively. On the other hand, inhibition of EGFR, Ca2+/PKC, or NFKB1 abolished the EGF-stimulated increase in the expression of cyclins CCND1 and CCNE1, cyclin-dependent kinase 6 (CDK6), CDK2, and BCL2, and restored the EGF-induced inhibition of BAX expression and caspase 3/9 activity, indicating that EGFR, PKC, and NFKB1 signaling cascades were involved in EGF-stimulated DNA synthesis and antiapoptosis action. In conclusion, EGF stimulated proliferation of chicken PGCs via activation of Ca2+/PKC involving NFKB1 signaling pathway. These observations suggest that EGF signaling is important in regulating germ cell proliferation in the chicken embryonic gonad.}, number={3}, journal={BIOLOGY OF REPRODUCTION}, author={Ge, Chutian and Yu, Minli and Petitte, James N. and Zhang, Caiqiao}, year={2009}, month={Mar}, pages={528–536} } @misc{shears_reynolds_petitte_2008, title={Use of a transgene encoding a vertebrate phytase to increase capacity to utilize phytic acid in livestock feed}, volume={7,351,580}, number={2008 Apr. 1}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Shears, S. and Reynolds, P. and Petitte, J.}, year={2008} } @article{petitte_mozdziak_2007, title={The incredible, edible, and therapeutic egg}, volume={104}, ISSN={["0027-8424"]}, url={http://europepmc.org/abstract/med/17272493}, DOI={10.1073/pnas.0611652104}, abstractNote={The domestic fowl has a long and unique history, serving multiple purposes in society and science. A cursory review of the Nobel Prize awards in physiology or medicine since 1901 points to the significance of birds, and the chicken in particular, as the premier nonmammalian vertebrate animal model (see www.fbresearch.org/education/nobels.htm and http://nobelprize.org/nobel_prizes/medicine/laureates). The domestic fowl aided in the discovery of essential vitamins and gave the first clue to differences between T and B cells. In fact, B cell nomenclature is based on the origin of B cells from the avian bursa of Fabricius. In addition, the chicken model provided the foundation for understanding the chemical processes for vision, insights into animal behavior, and our first introduction to tumor viruses [e.g., Rous sarcoma virus (RSV)] and the cellular origin of retroviral oncogenes. Even today, avian oncogenic viruses provide valuable models for human disease. Furthermore, for many developmental biologists, the avian embryo remains the premier animal model (1). On the practical side, the general public is protected from yearly influenza outbreaks through vaccine production in chicken eggs. In addition to its scientific and biomedical importance, poultry as an agricultural commodity has grown over the last 60 years into a global industry providing billions of people with inexpensive high-quality animal protein in the form of meat and eggs. Much of the success of the poultry industry is directly related to the application of population genetics for the selection of commercial lines for efficient protein production (2). Today, estimates of the cost of egg production in the U.S. hover around 5 cents per egg. Given that the albumin from a single egg contains ≈3.6 g of protein, the domestic laying hen is a very efficient protein bioreactor. Now, with the report of Lillico et al. (3) in this issue of …}, number={6}, journal={PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA}, author={Petitte, James N. and Mozdziak, Paul E.}, year={2007}, month={Feb}, pages={1739–1740} } @article{petitte_2006, title={Avian germplasm preservation: Embryonic stem cells or primordial germ cells?}, volume={85}, ISSN={["1525-3171"]}, url={http://europepmc.org/abstract/med/16523620}, DOI={10.1093/ps/85.2.237}, abstractNote={Presently, avian genetic resources are best maintained as living collections of birds. Unfortunately, these stocks have been under constant pressure to be destroyed because of the decline in the number of Poultry Science Departments and pressures to cut costs at land grant institutions. Cryopreservation of semen is often suggested as a means to bank avian germplasm. However, this is only applicable for single-gene traits and does not allow for full reconstitution of the genetics of the original line. Over the last 15 yr, advances in the manipulation of the early chick embryo, manipulation of primordial germ cells (PGC), and the culture of embryonic stem cells (ESC) suggests that cryopreservation of blastodermal cells, ESC, or PGC might offer a means to preserve the entire genome of highly selected, specialized stocks of poultry. Freezing each of these cell types is possible with varying degrees of efficiency. Similarly, the effectiveness of generating germ line chimeras using blastodermal cells, ESC, or PGC also varies greatly. Other factors that must be considered include the choice of the recipient lines to develop the germ line chimeras and the number of individuals needed to reconstitute the line. Finally, the low efficiency rate of reconstitution and the high cost associated with current technologies makes these approaches prohibitive. Significant challenges remain to be overcome before the entire genome of poultry stocks can be routinely cryoperserved and reconstituted.}, number={2}, journal={POULTRY SCIENCE}, author={Petitte, JN}, year={2006}, month={Feb}, pages={237–242} } @article{cho_choi_darden_reynolds_petitte_shears_2006, title={Avian multiple inositol polyphosphate phosphatase is an active phytase that can be engineered to help ameliorate the planet's "phosphate crisis"}, volume={126}, ISSN={["1873-4863"]}, url={http://europepmc.org/abstract/med/16759730}, DOI={10.1016/j.jbiotec.2006.04.028}, abstractNote={Contemporary phytase research is primarily concerned with ameliorating the problem of inadequate digestion of inositol hexakisphosphate (phytate; InsP6) in monogastric farm animal feed, so as to reduce the pollution that results from the high phosphate content of the manure. In the current study we pursue a new, safe and cost-effective solution. We demonstrate that the rate of hydrolysis of InsP6 by recombinant avian MINPP (0.7 μmol/mg protein/min) defines it as by far the most active phytase found to date in any animal cell (the corresponding activity of recombinant mammalian MINPP is only 0.006 μmol/mg protein/min). Although avian MINPP has less than 20% sequence identity with microbial phytases, we create a homology model of MINPP in which it is predicted that the structure of the phytase active site is well-conserved. This model is validated by site-directed mutagenesis and by use of a substrate analogue, scyllo-InsP6, which we demonstrate is only a weak MINPP substrate. In a model chicken cell line, we overexpressed a mutant form of MINPP that is secretion-competent. This version of the enzyme was actively secreted without affecting either cell viability or the cellular levels of any inositol phosphates. Our studies offer a genetic strategy for greatly improving dietary InsP6 digestion in poultry.}, number={2}, journal={JOURNAL OF BIOTECHNOLOGY}, author={Cho, Jaiesoon and Choi, Kuicheon and Darden, Thomas and Reynolds, Paul R. and Petitte, James N. and Shears, Stephen B.}, year={2006}, month={Nov}, pages={248–259} } @article{jackson_anderson_ashwell_petitte_mozdziak_2007, title={CA125 expression in spontaneous ovarian adenocarcinomas from laying hens}, volume={104}, ISSN={0090-8258}, url={http://dx.doi.org/10.1016/j.ygyno.2006.07.024}, DOI={10.1016/j.ygyno.2006.07.024}, abstractNote={Objective Currently, there is not a fully characterized model for human ovarian cancer; however, 2- to 4-year-old laying hens spontaneously develop ovarian tumors. CA125 expression is a hallmark of ovarian cancer in women. The major objective of this study was to characterize the in vitro growth of avian ovarian tumor cells, and CA125 expression in avian ovarian tumors. Methods Immunohistochemistry was employed to evaluate CA125 expression in avian ovarian tumor tissue. A high temperature antigen retrieval step was an essential part of the CA125 staining procedure. In vitro growth curves were constructed for avian ovarian cancer cells. Western blotting was used to estimate the size of the CA125 reactive protein and to confirm CA125 expression. Results The growth of avian tumors in culture fits a sigmoidal curve for cell growth and suggests a cell cycle time of 28 h. The tumors taken from the chicken stained positive for CA125. Approximately 90% of cells isolated from avian ovarian tumors also stained positive for CA125. Western blots show a band of approximately 25 kDa when immunodetected with CA125. Conclusions Similar to human ovarian tumors, chicken ovarian tumors express CA125. Cultured chicken ovarian cancer cells express CA125 and CA125 expression does not appear to change with time in culture.}, number={1}, journal={Gynecologic Oncology}, publisher={Elsevier BV}, author={Jackson, Emily and Anderson, Ken and Ashwell, Chris and Petitte, James and Mozdziak, Paul E.}, year={2007}, month={Jan}, pages={192–198} } @article{mozdziak_wu_bradford_pardue_borwornpinyo_giamario_petitte_2006, title={Identification of the lacZ insertion site and beta-galactosidase expression in transgenic chickens}, volume={324}, ISSN={["1432-0878"]}, url={http://europepmc.org/abstract/med/16408197}, DOI={10.1007/s00441-005-0060-9}, abstractNote={The quail:chick chimera system is a classical research model in developmental biology. An improvement over the quail:chick chimera system would be a line of transgenic chickens expressing a reporter gene. Transgenic chickens carrying lacZ and expressing bacterial beta-galactosidase have been generated, but complete characterization of the insertion event and characterization of beta-galactosidase expression have not previously been available. The genomic sequences flanking the retroviral insertion site have now been identified by using inverse polymerase chain reaction (PCR), homozygous individuals have been identified by using PCR-based genotyping, and beta-galactosidase expression has been evaluated by using Western analysis and histochemistry. Based upon the current draft of the chicken genome, the viral insertion carrying the lacZ gene has been located on chromosome 11 within the predicted gene for neurotactin/fractalkine (CX3CL1); neurotactin mRNA expression appears to be missing from the brain of homozygous individuals. When Generation 2 (G2) lacZ-positive individuals were inter-mated, they generated 361 G3 progeny; 82 were homozyous for lacZ (22.7%), 97 were wild-type non-transgenic (26.9%), and 182 (50.4%) were hemizygous for lacZ. Western analysis revealed the highest expression in the muscle and liver. With the identification of homozygous birds, the line of chickens is now designated NCSU-Blue1.}, number={1}, journal={CELL AND TISSUE RESEARCH}, author={Mozdziak, PE and Wu, Q and Bradford, JM and Pardue, SL and Borwornpinyo, S and Giamario, C and Petitte, JN}, year={2006}, month={Apr}, pages={41–53} } @article{mozdziak_wysocki_angerman-stewart_pardue_petitte_2006, title={Production of chick germline chimeras from fluorescence-activated cell-sorted gonocytes}, volume={85}, ISSN={["1525-3171"]}, url={http://europepmc.org/abstract/med/17012166}, DOI={10.1093/ps/85.10.1764}, abstractNote={Modification of the chicken germline has been difficult, because it has been challenging to fractionate sufficient numbers of primordial germ cells for manipulation and implantation into developing embryos. A technique to enrich cell suspensions for primordial germ cells, using fluorescence-activated cell sorting (FACS), has recently been developed. The objective of the current study was to demonstrate that the FACS-enriched early embryonic gonocytes could fully participate in development of the germline. Therefore, cells were disassociated from stage 27 gonads, incubated with mouse anti-stage-specific embryonic antigen-1, which was detected with goat-antimouse IgM-fluorescein isothiocyanate, and the fluorescently labeled cells were sorted from the unlabeled cells using FACS. The isolated gonocyte population was injected into the blastoderm of unincubated stage X embryos, the germinal crescent of 3-d embryos, and into the circulation of stage 17 embryos that were pretreated with busulfan. Barred Plymouth Rock gonocytes were implanted exclusively into recipient White Leghorn embryos, and White Leghorn gonocytes were implanted exclusively into Barred Plymouth Rock recipient embryos. Embryos were cultured until hatch, and male putative chimeras were reared to sexual maturity. Germline chimerism was evaluated by observing feather color of the progeny. All injection methods resulted in germline chimeras demonstrating that FACS-sorted gonocytes can fully participate in development. Moreover, it was demonstrated that gonocytes isolated from stage 27 embryonic gonads can be introduced into embryos at an earlier stage of development, and the introduced gonocytes can fully participate in germline development.}, number={10}, journal={POULTRY SCIENCE}, author={Mozdziak, P. E. and Wysocki, R. and Angerman-Stewart, J. and Pardue, S. L. and Petitte, J. N.}, year={2006}, month={Oct}, pages={1764–1768} } @article{borwompinyo_brake_mozdziak_petitte_2005, title={Culture of chicken embryos in surrogate eggshells}, volume={84}, ISSN={0032-5791}, url={http://dx.doi.org/10.1093/ps/84.9.1477}, DOI={10.1093/ps/84.9.1477}, abstractNote={The chick embryo is a classical model to study embryonic development. However, most researchers have not studied the effect of embryonic manipulation on chick hatchability. The objective of this study was to determine the effect of egg orientation and type of sealing film on the hatchability of cultured embryos. Windows were made in the small end of recipient surrogate chicken eggshells, and donor embryos were placed into the recipient eggshell for the first 3 d of incubation. Survival over the first 3 d was maximized (P < 0.05) when windowed eggs sealed with Saran Wrap were positioned with the window-end down compared with window-end up. Three-day-old cultured embryos were transferred into recipient turkey eggshells, sealed with cling film, and cultured until hatch. Water weight loss of the surrogate eggshell cultures regardless of cling film type was not significantly different from control intact eggs. The embryos cultured in turkey eggshells and sealed with Handi Wrap exhibited higher hatchability (75% +/- 10.2%) than cultures sealed with Saran Wrap (45.2% +/- 13.8%). Hatchability of control intact eggs (86.4% +/- 5.3%) was not significantly (P > 0.05) different from the hatchability of eggs sealed with Handi Wrap, which suggested that Handi Wrap was an excellent sealant for chick embryos cultured after 3 d of incubation.}, number={9}, journal={Poultry Science}, publisher={Elsevier BV}, author={Borwompinyo, S. and Brake, J. and Mozdziak, P.E. and Petitte, J.N.}, year={2005}, month={Sep}, pages={1477–1482} } @article{kloss_linscheid_johnson_lawson_edwards_linder_stocker_petitte_kern_2005, title={Effects of conjugated linoleic acid supplementation on blood lipids and adiposity of rats fed diets rich in saturated versus unsaturated fat.}, volume={6}, url={http://europepmc.org/abstract/med/15829429}, DOI={10.1016/j.phrs.2004.12.005}, abstractNote={Conjugated linoleic acid (CLA) may decrease adiposity and improve blood lipid profiles under some conditions. The goal of this study was to determine the effects of CLA supplementation on blood lipid profiles and adiposity of rats fed a diet containing a primarily saturated fat versus a diet containing a primarily unsaturated fat. Twenty-eight male Sprague–Dawley rats were randomly assigned to one of four diets containing coconut oil, coconut oil with CLA, corn oil or corn oil with CLA. After 28 days, blood was collected and serum concentrations of total cholesterol (TC), HDL-cholesterol (HDL-C), and triacylglycerols (TG) were assessed. Food intake, body weights, and epididymal fat pads were measured. No significant differences (p > 0.05) were noted among groups for amount of food consumed, weight gained, food efficiency ratio or serum TG concentrations. TC concentrations were lower (p < 0.05) in the CLA-supplemented rats that were fed coconut oil but not those consuming corn oil. Serum HDL-C was lower (p < 0.05) in rats consuming corn oil but was not significantly different (p > 0.05) for CLA supplemented groups. Epididymal fat pads weighed significantly more (p < 0.05) in the coconut oil fed group compared to the corn oil fed group, but there was no significant difference (p > 0.05) between the corn oil and coconut oil + CLA group. Overall, this study suggests that CLA is more beneficial for control of blood lipids and adiposity when supplemented to a diet rich in saturated versus unsaturated fat.}, journal={Pharmacological research}, author={Kloss, R and Linscheid, J and Johnson, A and Lawson, B and Edwards, K and Linder, T and Stocker, K and Petitte, J and Kern, M}, year={2005}, month={Jun} } @article{mozdziak_angerman-stewart_rushton_pardue_petitte_2005, title={Isolation of chicken primordial germ cells using fluorescence-activated cell sorting}, volume={84}, ISSN={["1525-3171"]}, url={http://europepmc.org/abstract/med/15844816}, DOI={10.1093/ps/84.4.594}, abstractNote={Presently, it is difficult to undertake germ line modification of the chicken with primordial germ cells (PGC) because it has been difficult to efficiently fractionate the PGC from the total somatic cell population. The objective of this study was to develop a method that allows isolation of an enriched population of viable PGC from embryonic blood and embryonic gonadal tissue. Blood was harvested from early chick embryos (stages 13 to 15), and cells were liberated from the gonads of stage 27 chick embryos. Subsequently, viable PGC were labeled with anti-stage-specific embryonic antigen-1 (SSEA-1), which was detected with goat-anti-mouse IgM-fluorescein isothiocyanate. Fluorescently labeled cells were sorted from the unlabeled cells using fluorescence-activated cell sorting (FACS), and the identities of the PGC were confirmed using periodic acid-Schiff (PAS) staining or anti-embryonic mouse antigen-1 (EMA-1) staining followed by microscopic evaluation. Finally, PGC were sorted from somatic cells of sex-identified embryos. Less than 0.1% of the blood cell population was collected as SSEA-1-positive cells. Similarly, approximately 2% of the gonadal cell population were collected as SSEA-1-positive cells. Therefore, fewer (-1,000 to 9,000) PGC were recovered from each isolate. Placing the sorted SSEA-1-positive cells on a glass slide from a microcentrifuge tube resulted in a recovery rate of 53 to 73% relative to the number detected by FACS. Furthermore, the proportions of sorted cells that stained with PAS or anti-EMA-1 following sorting were 92+/-4% PAS positive and 94+/-1% anti-EMA-1 positive. Finally, the sorted SSEA-1-positive cells were maintained in vitro to demonstrate their viability after sorting. It was demonstrated that it is possible to label blood and gonadal chicken PGC with SSEA-1 and subsequently to sort viable SSEA-1-positive PGC from somatic cells.}, number={4}, journal={POULTRY SCIENCE}, author={Mozdziak, PE and Angerman-Stewart, J and Rushton, B and Pardue, SL and Petitte, JN}, year={2005}, month={Apr}, pages={594–600} } @article{song_s d'costa_pardue_petitte_2005, title={Production of germline chimeric chickens following the administration of a busulfan emulsion}, volume={70}, ISSN={["1098-2795"]}, url={http://europepmc.org/abstract/med/15685638}, DOI={10.1002/mrd.20218}, abstractNote={Busulfan (1,4‐butanediol dimethanesulfonate) was used to deplete endogenous germ cells for the enhanced production of chicken germline chimeras. Utilizing immunohistochemical identification of primordial gem cells (PGCs) in Stage 27 chicken embryos, two delivery formulations were compared relative to the degree of endogenous PGC depletion, a busulfan suspension (BS) and a solublized busulfan emulsion (SBE). Both busulfan treatments resulted in a significant reduction in PGCs when compared to controls. However, the SBE resulted in a more consistent and extensive depletion of PGCs than that observed with the BS treatment. Repopulation of SBE‐treated embryos with exogenous PGCs resulted in a threefold increase of PGCs in Stage 27 embryos. Subsequently, germline chimeras were produced by the transfer of male gonadal PGCs from Barred Plymouth Rock embryos into untreated and SBE‐treated White Leghorn embryos. Progeny testing of the presumptive chimeras with adult Barred Plymouth Rock chickens was performed to evaluate the efficiency of germline chimera production. The frequency of germline chimerism in SBE‐treated recipients increased fivefold when compared to untreated recipients. The number of donor‐derived offspring from the germline chimeras also increased eightfold following SBE‐treatment of the recipient embryos. These results demonstrated that the administration of a busulfan emulsion into the egg yolk of unincubated eggs improved the depletion of endogenous PGCs in the embryo and enhanced the efficiency of germline chimera production. Mol. Reprod. Dev. 70: 438–444, 2005. © 2005 Wiley‐Liss, Inc.}, number={4}, journal={MOLECULAR REPRODUCTION AND DEVELOPMENT}, author={Song, YH and S D'Costa and Pardue, SL and Petitte, JN}, year={2005}, month={Apr}, pages={438–444} } @article{mozdziak_petitte_carson_2004, title={An introductory undergraduate course covering animal cell culture techniques}, volume={32}, ISSN={["1539-3429"]}, url={http://europepmc.org/abstract/med/21706746}, DOI={10.1002/bmb.2004.494032050381}, abstractNote={AbstractAnimal cell culture is a core laboratory technique in many molecular biology, developmental biology, and biotechnology laboratories. Cell culture is a relatively old technique that has been sparingly taught at the undergraduate level. The traditional methodology for acquiring cell culture training has been through trial and error, instruction when undertaking the first graduate student position, or instruction when hired for a specific industrial cell culture position. However, cell culture is an important candidate course for any biotechnology‐related training program because it is a technique that must be performed by investigators before they perform many molecular procedures, and vertebrate cell culture is becoming increasingly important for biomanufacturing of therapeutic proteins. Therefore, a cell culture techniques course is an important offering for undergraduate students who aspire to graduate training, and also undergraduate students who will seek employment with biotechnology companies immediately after graduation. Recently, a cell culture techniques course was developed and delivered to students at North Carolina State University as a component of an undergraduate Biotechnology minor curricula. Currently, the instructors at North Carolina State University are seeking to provide students with the necessary technical and critical reasoning skills to successfully perform animal cell culture.}, number={5}, journal={BIOCHEMISTRY AND MOLECULAR BIOLOGY EDUCATION}, author={Mozdziak, PE and Petitte, JN and Carson, SD}, year={2004}, pages={319–322} } @misc{petitte_liu_yang_2004, title={Avian pluripotent stem cells}, volume={121}, ISSN={["1872-6356"]}, url={http://europepmc.org/abstract/med/15296979}, DOI={10.1016/j.mod.2004.05.003}, abstractNote={Pluripotent embryonic stem cells are undifferentiated cells capable of proliferation and self-renewal and have the capacity to differentiate into all somatic cell types and the germ line. They provide an in vitro model of early embryonic differentiation and are a useful means for targeted manipulation of the genome. Pluripotent stem cells in the chick have been derived from stage X blastoderms and 5.5 day gonadal primordial germ cells (PGCs). Blastoderm-derived embryonic stem cells (ESCs) have the capacity for in vitro differentiation into embryoid bodies and derivatives of the three primary germ layers. When grafted onto the chorioallantoic membrane, the ESCs formed a variety of differentiated cell types and attempted to organize into complex structures. In addition, when injected into the unincubated stage X blastoderm, the ESCs can be found in numerous somatic tissues and the germ line. The potential give rise to somatic and germ line chimeras is highly dependent upon the culture conditions and decreases with passage. Likewise, PGC-derived embryonic germ cells (EGCs) can give rise to simple embryoid bodies and can undergo some differentiation in vitro. Interestingly, chicken EG cells contribute to somatic lineages when injected into the stage X blastoderm, but only germ line chimeras have resulted from EGCs injected into the vasculature of the stage 16 embryo. To date, no lines of transgenic chickens have been generated using ESCs or EGCs. Nevertheless, progress towards the culture of avian pluripotent stem cells has been significant. In the future, the answers to fundamental questions regarding segregation of the avian germ line and the molecular basis of pluripotency should foster the full use of avian pluripotent stem cells.}, number={9}, journal={MECHANISMS OF DEVELOPMENT}, author={Petitte, JN and Liu, G and Yang, Z}, year={2004}, month={Sep}, pages={1159–1168} } @misc{pardue_petitte_d'costa_song_2004, title={Methods for gamete production in birds}, volume={6,691,638}, number={2004 Feb. 17}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Pardue, S. and Petitte, J. and D'Costa, S. and Song, Y.-H.}, year={2004} } @misc{mozdziak_petitte_2004, title={Status of transgenic chicken models for developmental biology}, volume={229}, ISSN={["1097-0177"]}, url={http://europepmc.org/abstract/med/14991696}, DOI={10.1002/dvdy.10461}, abstractNote={AbstractThe chick embryo is a classic model that has been used to gain insight into developmental processes and cell fate within the embryo for over a century. For the most part, investigators have implanted quail cells into a chicken embryo. A more powerful tool for developmental biology research than the quail:chick chimera system would be to have lines of transgenic chickens expressing reporter genes that are readily available to the research community. However, avian transgenic technology has been fraught with technical difficulties, and transgenic chickens expressing reporter genes have only recently been developed. The goal of this review is to report the technologies that have been used to generate transgenic chickens and to discuss the challenges in generating avian transgenics for developmental biology research. Developmental Dynamics 229:414–421, 2004. © 2004 Wiley‐Liss, Inc.}, number={3}, journal={DEVELOPMENTAL DYNAMICS}, author={Mozdziak, PE and Petitte, JN}, year={2004}, month={Mar}, pages={414–421} } @article{mozdziak_borwornpinyo_mccoy_petitte_2003, title={Development of transgenic chickens expressing bacterial beta-galactosidase}, volume={226}, ISSN={["1097-0177"]}, url={http://europepmc.org/abstract/med/12619130}, DOI={10.1002/dvdy.10234}, abstractNote={AbstractReplication‐defective retroviral vectors are efficient vehicles for the delivery of exogenous genes, and they may be used in the generation of transgenic animals. The replication‐defective retroviral SNTZ vector carrying the lacZ gene with a nuclear localized signal was injected into the subgerminal cavity of freshly laid eggs. Subsequently, the eggs were allowed to hatch, and the chickens were screened for the lacZ gene by using the polymerase chain reaction. Eight of 15 male chickens that survived to sexual maturity contained the lacZ gene in their semen. Subsequently, these males were mated with wild‐type female chickens. From one of the eight lacZ‐positive G0 males, two lacZ‐positive male chickens were produced from a total of 224 G1 progeny for a germline transmission rate of 0.89%. Both G1 male chickens carrying the lacZ gene were mated with wild‐type female chickens and 46.5% of the G2 progeny contained the lacZ gene, which is consistent with the expected Mendelian 50% ratio for a heterozygous dominant allele. The product of the lacZ gene, nuclear localized β‐galactosidase, was expressed in primary myoblast cultures derived from G2 chickens, and it was also expressed in whole G2 chicken embryos. Developmental Dynamics, 2003. © 2003 Wiley‐Liss, Inc.}, number={3}, journal={DEVELOPMENTAL DYNAMICS}, author={Mozdziak, PE and Borwornpinyo, S and McCoy, DW and Petitte, JN}, year={2003}, month={Mar}, pages={439–445} } @misc{petitte_ricks_spence_2003, title={Gene transfer in poultry by introduction of embryo cells in ovo}, volume={6,515,199}, number={2003 Feb. 4}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. and Ricks, C. A. and Spence, S. E.}, year={2003} } @article{giamario_petitte_mozdziak_2003, title={Hatchability of chicken embryos following somite manipulation}, volume={34}, ISSN={["1940-9818"]}, url={http://europepmc.org/abstract/med/12813875}, DOI={10.2144/03346bm01}, abstractNote={The avian embryo has been a classical model to study early development because the embryo is easily accessible for manipulation of embryonic cells and structures. The usefulness of the chicken embryo increased with the development of quail-chick transplantation techniques (1,2). The ability to transplant Japanese quail cells into a chicken embryo provides a method to trace the developmental fate of the implanted quail cells in a chicken host embryo because the dense magentacolored heterochromatin and nucleoli of the Japanese quail nuclei in Feuglenstained histological sections distinguishes donor quail nuclei from host chicken nuclei. The quail nucleolar marker is heritable, making it possible to determine the ultimate developmental fate of quail cells transplanted into host chicken embryos. Furthermore, monoclonal antibodies have been developed that recognize quail nuclei, but they do not recognize chicken nuclei (QCPN; Developmental Studies Hybridoma Bank, Iowa City, IA, USA). Therefore, it is possible to distinguish quail cells from chicken cells in a quailchick chimera using classical histochemistry (Feuglen staining) (1–3) or immunohistochemistry (4–6). Although the quail-chick chimera has been a powerful tool for developmental biology research, it would be better to employ a chicken/chicken implantation system because it would eliminate any potentially confounding species-specific effects on any experiments. However, it has been previously impossible to employ a chicken/chicken system to study cell fate because there have not been any previously reported useful lines of transgenic chickens expressing histologically convenient reporter genes. Lines of transgenic chickens carrying the Escherichia coli lacZ reporter gene and expressing β-galactosidase have recently been developed (7), making it possible to follow the developmental fate of transgenic chicken cells implanted into wild-type embryos. Therefore, a new tool for developmental biology research that is a significant improvement over the quail-chicken system is now available. A potential difficulty in avian embryonic developmental biology research is the ability to follow embryonic cell fate through hatching and adult maturity. Few researchers have attempted to study the effect of embryonic manipulation on post-hatch chickens. In one case, a portion of the neural tube and the somites was removed from 621 chicken embryos and replaced with the same portion of the neural tube from quail embryos, but only 46 somatic chimeras hatched (7%). Seventeen of the 46 somatic chimeras exhibited various abnormalities in the limbs at hatching. Therefore, only 29 hatched somatic chimeras from the original 621 manipulated embryos were fully viable at hatch (5%) (8). The surviving quailchick chimeras appeared normal at hatch but died after a few weeks of age because of an interspecies-related demylenation of the spinal cord (9). It appears that the quail-chicken system is not applicable for studying the effects of embryonic manipulations on adult birds. Furthermore, chick-chick neural tube/somite chimeras also resulted in a very low hatchability (2 out of 40; 5%) (8). Therefore, it also appears that it is difficult to achieve a reasonable level of hatchability following embryonic manipulations to study post-hatch development. It is important to note that these experiments (8,9) included an invasive transfer of the spinal cord between embryos. However, other less invasive manipulations have also resulted in relatively low hatchability. For example, primordial germ cells were injected into the dorsal aorta of recipient stage 15 (10) embryos, and only 7%–14% of the injected embryos hatched in one study (11), and approximately 26% of the injected embryos hatched in a second study (12). Furthermore, Naito et al. (11,12) used a laborious surrogate eggshell culturing procedure for their studies, and the procedures reported in the present study employ a simple eggshell windowing technique. For the most part, hatchability data for somatic manipulations do not appear readily available in the scientific literature. Therefore, the objective of this manuscript is to report our procedures for the manipulation of embryonic chick somites and the associated level of hatchability that was achieved using the reported procedures. The rationale was to demonstrate the results of microinjection and the level of hatchability associated with the experimental manipulations. Future studies will focus on implanted cell fate. The experimental procedures are useful for studies aimed at understanding the effect of embryonic manipulations on post-hatch development. Freshly laid eggs were placed into an incubator until they reached stages 10–15 (10). Fertile eggs were stored with the blunt end up for 2–3 h. Subsequently, a hole was cut in the shell on the blunt end of the egg with surgical scissors (Figure 1). The embryos were visualized in the egg using lateral illumination though a wratten 47 blue gelatin filter (Sigma, St. Louis, MO, USA). The blue filter visualizes the somites in the embryo while it is still in the egg (Figure 2). Therefore, it is possible with blue light illumination to manipulate the chicken embryos without traditional India ink staining. All embryos were manipulated between approximately stage 10 and stage 15 (10). Figure 3 also demonstrates that it is possible to sufficiently visualize the somites and other structures for implantation studies without India ink staining because 3 μL of a 50 μg/mL solution of propidium iodide in 0.1% SDS were successfully injected into a somite. Subsequently, the injected embryo was fixed with 4% paraBenchmarks}, number={6}, journal={BIOTECHNIQUES}, author={Giamario, C and Petitte, JN and Mozdziak, PE}, year={2003}, month={Jun}, pages={1128–1130} } @article{giamario_petitte_mozdziak_2003, title={Myonuclear accrestion-a brief review}, volume={21}, number={Suppl. 1}, journal={Animal Science Papers and Reports}, author={Giamario, C. and Petitte, J. N. and Mozdziak, P. E.}, year={2003}, pages={121–131} } @article{mozdziak_pophal_borwornpinyo_petitte_2003, title={Transgenic chickens expressing beta-galactosidase hydrolyze lactose in the intestine}, volume={133}, ISSN={["1541-6100"]}, url={http://europepmc.org/abstract/med/14519787}, DOI={10.1093/jn/133.10.3076}, abstractNote={Chickens do not possess the necessary enzymes to efficiently hydrolyze lactose into glucose and galactose. The bacterial enzyme beta-galactosidase can convert lactose into glucose and galactose. Transgenic chickens that carry the E. coli lacZ gene and express beta-galactosidase could potentially utilize lactose as an energy source. The objective of this study was to determine the ability of the transgenic chicken small intestinal mucosa to hydrolyze lactose into glucose and galactose. Lactase activity was examined in the intestinal muscosa from wild-type chickens and two lines of chickens that carry the lacZ gene and express beta-galactosidase. Lactase activity was significantly higher in both transgenic lines compared with wild-type birds (P < 0.05). The presence of the beta-galactosidase enzyme was revealed by X-gal staining in the intestine of transgenic chickens, whereas it was not present in the wild-type chickens. Overall, it appears that inserting the lacZ gene, which encodes beta-galactosidase, has resulted in a chicken that can utilize lactose as an energy source. This study demonstrates that transgenic technology can be used to modify nutrient utilization in domestic poultry.}, number={10}, journal={JOURNAL OF NUTRITION}, author={Mozdziak, PE and Pophal, S and Borwornpinyo, S and Petitte, JN}, year={2003}, month={Oct}, pages={3076–3079} } @misc{s d'costa_pardue_petitte_2001, title={Comparative development of avian primordial germ cells and production of germ line chimeras}, volume={12}, ISSN={["1470-2061"]}, DOI={10.3184/147020601783698477}, number={4}, journal={AVIAN AND POULTRY BIOLOGY REVIEWS}, author={S D'Costa and Pardue, SL and Petitte, JN}, year={2001}, pages={151–168} } @article{fasenko_christensen_wineland_petitte_2001, title={Examining the effects of prestorage incubation of turkey breeder eggs on embryonic development and hatchability of eggs stored for four or fourteen days}, volume={80}, ISSN={["0032-5791"]}, url={http://europepmc.org/abstract/med/11232999}, DOI={10.1093/ps/80.2.132}, abstractNote={Thirty-six hundred British United Turkey hatching eggs were used in two separate trials to test whether prestorage incubation (PRESI) treatments of 0, 6, and 12 h (Trial 1) or 0, 7, and 14 h (Trial 2) could improve the hatchability of eggs stored (17 C) for 14 versus 4 d. The development of the embryos (n = 30) was staged before and after exposing eggs to the various PRESI treatments. Embryonic development was also established after storage to ascertain whether embryonic development was occurring during storage. The remaining eggs in each trial were split into three groups (n = 500) and incubated for 28 d to examine embryonic mortality and hatchability. No changes were observed in embryonic development due to egg storage. Embryos were significantly more developed as the number of PRESI h increased; therefore, embryos from different PRESI treatments were placed in storage at different stages of development. Early mortality (1 to 7 d of incubation), mortality at internal and external pipping, and hatchability of fertile eggs were significantly reduced in eggs stored for 14 versus 4 d. The various PRESI treatments did not significantly affect the mortality or hatchability of eggs stored for 4 d. However, the hatchability of eggs incubated prior to storage for 12 h and then stored for 14 d was restored to the levels reported for eggs subjected to the treatment that represents the industry norm (0 h of PRESI and 4 d storage). These results indicate that embryos of eggs stored for 14 d, which have developmentally advanced to the stage of complete hypoblast formation (PRESI for 12 h), have a survival advantage over eggs stored for 14 d that have not been subjected to any PRESI.}, number={2}, journal={POULTRY SCIENCE}, author={Fasenko, GM and Christensen, VL and Wineland, MJ and Petitte, JN}, year={2001}, month={Feb}, pages={132–138} } @article{nolan_killian_petitte_jirtle_2001, title={Imprint status of M6P/IGF2R and IGF2 in chickens}, volume={211}, ISSN={["0949-944X"]}, url={http://europepmc.org/abstract/med/11455432}, DOI={10.1007/s004270000132}, abstractNote={Genomic imprinting is a method of gene regulation whereby a gene is expressed in a parent-of-origin-dependent fashion; however, it is hypothesized that imprinting should not occur in oviparous taxa such as birds. Therefore, we examined the allelic expression of two genes in the chicken that are reciprocally imprinted in most mammals, mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R) and insulin-like growth factor 2 (IGF2). Single nucleotide polymorphisms were identified in these genes, and cDNA was prepared from several tissues of embryos heterozygous for these polymorphisms. Both alleles of M6P/IGF2R and IGF2 were expressed in all tissues examined by RT-PCR. Since the expression of these genes was independent of the parent from which they were inherited, we conclude that neither M6P/IGF2R nor IGF2 are imprinted in the chicken.}, number={4}, journal={DEVELOPMENT GENES AND EVOLUTION}, author={Nolan, CM and Killian, JK and Petitte, JN and Jirtle, RL}, year={2001}, month={Apr}, pages={179–183} } @misc{petitte_chang_2001, title={Method of producing an undifferentiated avian cell culture using avian primordial germ cells}, volume={6,333,192}, number={2001 Dec. 25}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. N. and Chang, I.-K.}, year={2001} } @article{s d'costa_kulik_petitte_2000, title={Expression and purification of biologically active recombinant quail stem cell factor in E-coli}, volume={24}, ISSN={["1095-8355"]}, url={http://europepmc.org/abstract/med/10805965}, DOI={10.1006/cbir.1999.0500}, abstractNote={AbstractStem cell factor (SCF) is a multifunctional cytokine involved in hematopoiesis, melanogenesis and gametogenesis. Previous studies have demonstrated that avian SCF is a requirement for the proliferation and survival of various cell types in vivo and in vitro. In the current study, recombinant quail stem cell factor was produced inEscherichia coli using a prokaryotic expression system. SCF was expressed as a fusion protein with a histidine hexamer tag at the N‐terminal end of the protein. Following expression, the protein was purified by affinity chromatography on the Ni‐NTA column. The uninduced and induced protein lysates and the purified protein were separated by SDS‐PAGE and transferred onto nitrocellulose membrane. Western blot analysis with the monoclonal antibody to the histidine tag identified SCF in the induced cell lysates and the purified sample. The recombinant SCF was approximately 22–23kD in size. This protein was generated devoid of the signal peptide, the transmembrane domain, and the intracellular domain and, hence, resembles the soluble form of SCF. Biological activity was assayed using the in vitro survival of E12 chicken dorsal root ganglion‐derived sensory neurons. The addition of recombinant quail SCF improved neuronal survival. Survival (20.6%) was the highest at the 50ng/ml concentration of SCF. The availability of quail SCF will be a valuable tool to further resolve the function of stem cell factor in birds.}, number={5}, journal={CELL BIOLOGY INTERNATIONAL}, author={S D'Costa and Kulik, MJ and Petitte, JN}, year={2000}, pages={311–317} } @misc{pardue_petitte_d'costa_2000, title={Methods for gamete production in birds}, volume={6,354,242}, number={2000 Mar 23}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Pardue, S. L. and Petitte, J. N. and D'Costa, S.}, year={2000} } @article{karagenc_petitte_2000, title={Soluble factors and the emergence of chick primordial germ cells in vitro}, volume={79}, ISSN={["0032-5791"]}, url={http://europepmc.org/abstract/med/10685893}, DOI={10.1093/ps/79.1.80}, abstractNote={Previous observations obtained from a culture of blastodermal cells on a mouse fibroblast feeder layer (STO) suggested that STO cells provide a factor or factors that facilitate development of avian primordial germ cells (PGC) from dispersed embryo cells. The purpose of the current study was to test the hypothesis that soluble factors produced by STO cells are responsible, at least in part, in supporting the development of PGC in culture and to examine the effect of stem cell factor (SCF), ciliary neurotrophic factor (CNTF), and basic fibroblast growth factor (bFGF) in the development of PGC in culture. Blastodermal cells on gelatin-coated plastic or on feeder layers of CV-1 cells yielded a small number of PGC. When blastodermal cells were cultured on STO cells, a marked increase in PGC was observed. The addition of STO cell-conditioned medium (STO-CM) to blastodermal cells cultured on gelatin-coated plastic and on feeder layers of CV-1 cells resulted in a significant increase in the number of PGC, indicating that soluble factors produced by STO cells can enhance the development of chicken PGC in culture. Supplementation of blastodermal cells with SCF (100 ng/mL) or CNTF (2 ng/mL) or with CNTF and SCF together resulted in a significant increase in the number of PGC after 48 h of culture on feeder layers of CV-1 cells. However, addition of bFGF (100 ng/mL) did not increase PGC. We concluded from these observations that the culture of blastodermal cells on feeder layers of STO and CV-1 cells can be used as a useful biological system in examining the regulatory factors that govern the ontogeny of the germ cell lineage in the avian embryo.}, number={1}, journal={POULTRY SCIENCE}, author={Karagenc, L and Petitte, JN}, year={2000}, month={Jan}, pages={80–85} } @article{d'costa_petitte_1999, title={Characterization of stage-specific embryonic antigen-1 (SSEA-1) expression during early development of the turkey embryo.}, volume={7}, url={http://europepmc.org/abstract/med/10470652}, number={4}, journal={The International journal of developmental biology}, author={D'Costa, S and Petitte, JN}, year={1999}, month={Jul}, pages={349–356} } @misc{petitte_ricks_1998, title={Apparatus for injecting avian embryo muscle tissue in ovo}, volume={5,784,992}, number={1998 July 28}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. and Ricks, C. A.}, year={1998} } @misc{petitte_ricks_phelps_williams_1998, title={Gene transfer in chickens by introduction of DNA into muscle in ovo}, volume={6,395,961}, number={1998 Jun 12}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. M. and Ricks, C. A. and Phelps, P. V. and Williams, C.}, year={1998} } @article{s d'costa_petitte_1998, title={Sex identification of turkey embryos using a multiplex polymerase chain reaction}, volume={77}, ISSN={["0032-5791"]}, url={http://europepmc.org/abstract/med/9603360}, DOI={10.1093/ps/77.5.718}, abstractNote={A considerable portion of the W chromosome in Gallinaceous birds consists of tandem repetitive DNA. In the turkey, a 0.4-kb PstI element is repeated about 10,000 times in the female diploid genome but is undetectable as such a unit in males. In this study a multiplex polymerase chain reaction was developed to identify the sex of turkeys based upon the PstI repeat. The technique utilized two pairs of primers, the first pair was designed to amplify a region of the PstI repetitive element, resulting in the production of a 177-bp fragment in females. The other pair was designed to amplify a region of the adenosine triphosphate (ATP) synthase gene, present in both males and females. The simultaneous use of all four primers in the same reaction resulted in the coamplification of a 177-bp and a 250-bp fragment in females and a 250-bp fragment in males. This technique was used to verify the sex of 45 adults of known sex and to identify the sex of 74 embryos from Day 5 to hatch. This procedure is rapid and permits the sexing of many embryos in a short time. The ability to sex early embryos can facilitate studies on avian sex determination.}, number={5}, journal={POULTRY SCIENCE}, author={S D'Costa and Petitte, JN}, year={1998}, month={May}, pages={718–721} } @misc{petitte_yang_1998, title={Veterinary pharmaceutical formulation containing avian embryonic stem cells}, volume={5,830,510}, number={1998 Nov. 3}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. N. and Yang, Z.-M.}, year={1998} } @misc{petitte_yang_1997, title={Avian embryonic stem cells}, volume={5,656,479}, number={1997 Aug. 12}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. N. and Yang, Z.-M.}, year={1997} } @article{brake_walsh_benton_petitte_meijerhof_penalva_1997, title={Egg handling and storage}, volume={76}, ISSN={0032-5791}, url={http://dx.doi.org/10.1093/ps/76.1.144}, DOI={10.1093/ps/76.1.144}, abstractNote={The temperature and relative humidity of storage, as well as the gaseous environment, interact with the fertile egg over time during storage in such a way as to affect the success of incubation either negatively or positively. This interaction occurs both above and below the "physiological zero", at which embryonic metabolism is minimal. This interaction below physiological zero implies that certain physical aspects of the egg must be affected by the environmental conditions. As the eggshell is a relatively fixed component, changes in albumen, shell membranes, cuticle, yolk, or embryo proper must account for these time- and environment-related effects. It is concluded that the major contributor is the albumen, as it is obviously the most dynamic component below physiological zero and is strategically positioned.}, number={1}, journal={Poultry Science}, publisher={Elsevier BV}, author={Brake, J. and Walsh, T.J. and Benton, C.E., Jr and Petitte, J.N. and Meijerhof, R. and Penalva, G.}, year={1997}, month={Jan}, pages={144–151} } @article{petitte_karagenc_ginsburg_1997, title={The origin of the avian germ line and transgenesis in birds}, volume={76}, ISSN={["1525-3171"]}, url={http://europepmc.org/abstract/med/9251133}, DOI={10.1093/ps/76.8.1084}, abstractNote={The origin of the germ cell lineage in vertebrates is a fundamental question that has preoccupied developmental biologists. Recent work on the origin of the avian germ line has extended and clarified our understanding of the temporal and spatial segregation of primordial germ cells (PGC) during prestreak stages of development. The germ cells first appear at Stage X (Eyal-Giladi and Kochav, 1976) in the ventral surface of the area pellucida in a scattered pattern among polyingressing cells. Subsequently, the PGC gradually translocate from the epiblast to the hypoblast. The entire process appears to be dependent upon the maintenance of an organized area pellucida. Little is known about the regulatory events governing germ cell emergence during this period; however, the culture of dispersed blastodermal cells on a mouse fibroblast feeder layer can compensate for a disorganized area pellucida and offers an in vitro system to examine the molecular basis of germ cell development. Such basic information is valuable for current approaches towards the production of transgenic poultry with targeted changes to the genome through the use of avian embryonic stem cells or primordial germ cells. Refinement of the culture of primordial germ cells or their precursors should allow academic and industrial research laboratories to answer significant biological questions and to improve the genetic potential of commercial poultry stocks. A better understanding of the biology of avian primordial germ cells during early embryo development can only enhance this process.}, number={8}, journal={POULTRY SCIENCE}, author={Petitte, JN and Karagenc, L and Ginsburg, M}, year={1997}, month={Aug}, pages={1084–1092} } @article{petitte_kulik_1996, title={Cloning and characterization of cDNAs encoding two forms of avian stem cell factor}, volume={1307}, ISSN={["0167-4781"]}, url={http://europepmc.org/abstract/med/8679698}, DOI={10.1016/0167-4781(96)00062-0}, abstractNote={Stem cell factor (SCF), also known as Steel factor, is a transmembrane cytokine involved in several developmental processes and the ligand for the receptor tyrosine kinase c-kit. In several mammalian species, two isoforms of stem cell factor have been reported, a long form in which soluble SCF is released after proteolytic cleavage of the extracellular domain and a short membrane-anchored form in which a region containing a cleavage site is deleted. Currently, only the longer, soluble form has been identified in birds. Therefore, the cDNAs encoding two forms of quail stem cell factor (qSCF) were obtained using RT-PCR with nested primers. The deduced amino acid sequence of the long form of qSCF showed a high degree of homology with chicken (98%) and relatively low homology (∼ 53%) with various mammalian SCFs. Northern blot analysis with the qSCF cDNA revealed the expression of a 5.9 and a 2.7 kb transcript in several quail tissues.}, number={2}, journal={BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION}, author={Petitte, JN and Kulik, MJ}, year={1996}, month={Jun}, pages={149–151} } @article{petitte_karagenc_1996, title={Growth factors during early events in avian embryo development}, volume={7}, number={2-3}, journal={Poultry and Avian Biology Reviews}, author={Petitte, J. N. and Karagenc, L.}, year={1996}, pages={75} } @article{karagenç_cinnamon_ginsburg_petitte_1996, title={Origin of primordial germ cells in the prestreak chick embryo.}, url={http://europepmc.org/abstract/med/9023982}, DOI={10.1002/(sici)1520-6408(1996)19:4<290::aid-dvg2>3.3.co;2-s}, abstractNote={The temporal and spatial pattern of segregation of the avian germline from the formation of the area pellucida to the beginning of primitive streak formation (stages VII–XIV, EG&K) was investigated using the culture of whole embryos and central and peripheral embryo fragments on vilelline membranes at stages VII–IX, immunohistological analysis of whole mount embryos and sections with monoclonal antibodies MC-480 against stage-specific embryonic antigen-1 (SSEA-1) and EMA-1, and with the culture of dispersed blastoderms at stages IX–XIV with and without an STO feeder layer. Whole embryos at intrauterine stages developed up to the formation of the primitive streak despite the absence of area pellucida expansion. Primordial germ cells (PGCs) appeared in the cultures of whole embryos and only in central fragments containing a partially formed area pellucida at stages VII–IX. When individual stage IX–XIV embryos were dispersed and cultured without a feeder layer, 25–45 PGCs/embryo were detected only with stage X–XIV, but not with stage IX blastoderms. However, the culture of dispersed cells from the area pellucida of stages IX–XIII on STO feeder layers yielded about 150 PGCs/embryo. The carbohydrate epitopes recognized by anti-SSEA-1 and EMA-1 first appeared at stage X on cells in association with polyingressing cells on the ventral surface of the epiblast and later on the dorsal surface of the hypoblast. The SSEA-1-positive hypoblast cells gave rise to chicken PGCs when cultured on a feeder layer of quail blastodermal cells. From these observations, we propose that the segregation and development of avian germline is a gradual, epigenetic process associated with the translocation of SSEA-1/EMA-1-positive cells from the ventral surface of the area pellucida at stage X to the dorsal side of the hypoblast at stages XI–XIV. © 1996 Wiley-Liss, Inc.}, journal={Developmental genetics}, author={Karagenç, L and Cinnamon, Y and Ginsburg, M and Petitte, JN}, year={1996}, month={Jan} } @article{petitte_kegelmeyer_1995, title={Rapid sex determination of chick embryos using the polymerase chain reaction}, volume={6}, ISSN={["1532-2378"]}, DOI={10.1080/10495399509525841}, abstractNote={Abstract The sex of early chicken embryos was determined by DNA dot‐blot hybridization and the polymerase chain reaction (PCR). An oligonucleotide probe, specific to the XhoI family of repetitive DNA of the W chromosome, was hybridized to crude and purified preparations of DNA obtained from blood or cells from embryos and detected using chemiluminescence. Complete agreement was observed in sexing 100 embryos using the dot‐blot assay and visual inspection of embryonic gonadal tissue. Although the dot‐blot system was useful for determining the sex of embryos prior to and throughout incubation, the time required for hybridization and detection did not allow for the immediate manipulation of embryos. Therefore, a PCR assay was developed using primers specific to the XhoI repetitive element and rapid thermocycling with an air thermocycler. A female‐specific amplification product was observed using DNA from as few as two cells in the PCR. Furthermore, female DNA was detected in mixtures of male and female DNA a...}, number={2}, journal={ANIMAL BIOTECHNOLOGY}, author={Petitte, JN and Kegelmeyer, AE}, year={1995}, pages={119–130} } @article{nicolasbolnet_johnston_kemper_ricks_petitte_1995, title={SYNERGISTIC ACTION OF 2 SOURCES OF AVIAN GROWTH-FACTORS ON PROLIFERATIVE DIFFERENTIATION OF CHICK EMBRYONIC HEMATOPOIETIC-CELLS}, volume={74}, ISSN={["1525-3171"]}, url={http://europepmc.org/abstract/med/7479487}, DOI={10.3382/ps.0741102}, abstractNote={During embryonic development, the components of the avian immune system undergo ontogeny in several distinct organs, including the bone marrow, spleen, thymus, and bursa of Fabricius. This process is regulated and controlled by the complex interactions of various cytokines and colony-stimulating factors (CSF). The objective was to examine the action of two different sources of hematopoietic growth factors, spleen-conditioned media (SCM) and chick embryo extract (CEE), on the proliferation of hematopoietic cells from various organs and on the differentiation of progenitor cells in semi-solid culture. Spleen and bone marrow cells obtained at Day 16 of incubation responded in a dose-dependent manner to the addition of SCM and CEE alone or in combination. No proliferative effect of SCM was observed on cells obtained from embryonic thymus or bursa. Clonal analysis of bone marrow and spleen cells suggested that CEE may contain the avian equivalents of stem cell factor, interleukin-3, granulocyte-macrophage CSF, granulocyte-CSF, and macrophage-CSF. Clonal analysis of SCM cultures suggested that in addition to myelomonocytic growth factor, which affects primarily macrophage-granulocyte lineages, a thrombocyte-CSF-like activity was also apparent. The SCM alone tended to act upon committed late progenitors. The combination of CEE and SCM amplified the size and the total number of colonies obtained and appeared to act synergistically upon progenitors with a high level of proliferative potential. This response on young progenitors was confirmed when cells were cultured in CEE and SCM prior to clonal analysis. These results document the presence of thrombocyte CSF in SCM and the effect of both CEE and SCM on the proliferative differentiation of avian embryonic hematopoietic progenitors.}, number={7}, journal={POULTRY SCIENCE}, author={NICOLASBOLNET, C and JOHNSTON, PA and KEMPER, AE and RICKS, C and PETITTE, JN}, year={1995}, month={Jul}, pages={1102–1116} } @article{petitte_kegelmeyer_kulik_1994, title={Isolation of genomic DNA from avian whole blood.}, volume={10}, url={http://europepmc.org/abstract/med/7833022}, journal={BioTechniques}, author={Petitte, JN and Kegelmeyer, AE and Kulik, MJ}, year={1994}, month={Oct} } @misc{petitte_yang_1994, title={Method of producing an avian embryonic stem cell culture and the avian embryonic stem cell culture produced by the process}, volume={5,340,740}, number={1994 Aug. 23}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Petitte, J. and Yang, Z.}, year={1994} } @article{qureshi_marsh_dietert_sung_nicolasbolnet_petitte_1994, title={PROFILES OF CHICKEN MACROPHAGE EFFECTOR FUNCTIONS}, volume={73}, ISSN={["1525-3171"]}, url={http://europepmc.org/abstract/med/7524053}, DOI={10.3382/ps.0731027}, abstractNote={In contrast to the mammalian system, avian species lack the so-called "resident" or "harvestable" macrophage population in the abdominal exudate. However, macrophages can be recruited into the chicken's abdominal cavity (presumably from the blood monocyte pool) if an inflammatory agent such as Sephadex is injected. The kinetics of inflammatory cell recruitment in terms of time, cell type, and state of activation to perform a particular effector function is currently an active area of research. This report will provide information on several chicken macrophage effector functions, including in vivo chemotaxis, phagocytosis, bacterial uptake and killing, biosynthesis of nitric oxide and various enzymes, and monokines such as interleukin-1 and granulocyte colony-stimulating factor.}, number={7}, journal={POULTRY SCIENCE}, author={QURESHI, MA and MARSH, JA and DIETERT, RR and SUNG, YJ and NICOLASBOLNET, C and PETITTE, JN}, year={1994}, month={Jul}, pages={1027–1034} } @article{petitte_brazolot_clark_liu_gibbins_etches_1993, title={Accessing the genome of the chicken using germline chimeras}, ISBN={0849342163}, journal={Manipulation of the avian genome}, publisher={Boca Raton : CRC Press}, author={Petitte, J. N. and Brazolot, C. L. and Clark, M. E. and Liu, G. D. and Gibbins, A. M. V. and Etches, R. J.}, editor={R. J. Etches and Gibbins, A. M. VerrinderEditors}, year={1993}, pages={81} } @article{qureshi_petitte_laster_dietert_1993, title={Avian macrophages: contribution to cellular microenvironment and changes in effector functions following activation.}, volume={7}, url={http://europepmc.org/abstract/med/8346153}, DOI={10.3382/ps.0721280}, abstractNote={The capacity of macrophages to generate metabolites and monokines having effector and regulatory functions can result in a major impact on their cellular microenvironment. Macrophage products synthesized in response to bacterial lipopolysaccharide stimulation include reactive oxygen and nitrogen intermediates as well as tumor necrosis factor (TNF). These secreted products of macrophages exhibit bioactivity either locally or systemically. Although the mechanism of action of avian monokines such as TNF-like factor may be similar to their mammalian counterparts, chicken TNF seems to lyse cells of chicken origin and not of mammalian origin. Furthermore, the generation and activity of products such as TNF is directly influenced by environmental stressors such as heat and toxins.}, journal={Poultry science}, author={Qureshi, MA and Petitte, JN and Laster, SM and Dietert, RR}, year={1993}, month={Jul} } @article{craig-veit_petitte_1993, title={Brain content of cGnRH I and II during embryonic development in chickens.}, volume={11}, url={http://europepmc.org/abstract/med/8282179}, DOI={10.1006/gcen.1993.1167}, abstractNote={We used specific radioimmunoassays to measure chicken gonadotropin-releasing hormones I and II (cGnRH I and II) in extracts of chicken brain as a first step in determining whether these peptides may function identically during embryonic development in birds. In three experiments chicken embryo brains were removed at various times between Day 6 of incubation and hatching. In Experiment 3, the sex of embryos was identified by means of polymerase chain reaction amplification of a W chromosome-specific DNA sequence. Brain concentrations of cGnRH I increased sharply from Day 6 of incubation to Day 8, then decreased until Day 10 to Day 12, followed by an increase beginning on approximately Day 17 and continuing until hatch. In contrast, cGnRH II concentration remained low until about Day 14 of incubation, then increased progressively until, at hatch, brain content of cGnRH II was approximately 9 to 11 times that of cGnRH I. The difference in development pattern and total content of these peptides supports the view that any physiologic function they may have might be differentiated as early as Day 7 of incubation.}, journal={General and comparative endocrinology}, author={Craig-Veit, CB and Petitte, JN}, year={1993}, month={Nov} } @article{watt_petitte_etches_1993, title={Early development of the chick embryo.}, volume={2}, url={http://europepmc.org/abstract/med/8445661}, DOI={10.1002/jmor.1052150205}, abstractNote={AbstractThe ultrastructure of the early chick embryo was investigated, using scanning (SEM) and transmission electron microscopy (TEM). Eggs were obtained from the shell gland by injecting hens intravenously with a synthetic prostaglandin or arginine vasopressin. Embryos were examined during late cleavage (stages IV–VI, Eyal‐Giladi and Kochav, '76), formation of the area pellucida (stages VII–XI), and formation of the hypoblast (stages X–XIV). SEM highlighted the reduction in cell number at the underside of the embryo during formation of the area pellucida although it became apparent that the thickness of the embryo is not reduced to a single layer of cells at stage X. In addition, blastomeres at the perimeter of embryos (stages V–VI) project filopodial extensions onto a smooth membrane that separates the sub‐embryonic cavity from the yolk. During hypoblast formation, epiblast cells generate stellate projections at their basal aspect, thus providing a meshwork for the advancing secondary hypoblast cells. By stage XII the epiblast was one cell thick and reminiscent of a columnar epithelium when viewed transversely. Cells of the deep portion of the posterior marginal zone were distinguished morphologically in the stage XII embryo by their many cell surface projections and ruffled appearance. Blastomeres at the perimeter of stage V–VI embryos projected filopodial extensions onto a smooth membrane which separates the sub‐embryonic cavity from the yolk. This membrane is presumed to be confluent with the cytolemma. Evidence is presented demonstrating the presence of intracellular membrane‐bound droplets which are hypothesised to contain sub‐embryonic fluid. © 1993 Wiley‐Liss, Inc.}, journal={Journal of morphology}, author={Watt, JM and Petitte, JN and Etches, RJ}, year={1993}, month={Feb} } @article{petitte_clark_etches_1991, title={Assessment of functional gametes in chickens after transfer of primordial germ cells.}, volume={5}, url={http://europepmc.org/abstract/med/2056493}, DOI={10.1530/jrf.0.0920225}, abstractNote={The ability of primordial germ cells (PGCs) transferred from donor to recipient embryos to form functional gametes was assessed using feather colour as a phenotypic marker. Donor primordial germ cells were obtained in blood samples taken from Dwarf White Leghorn embryos, homozygous for the dominant allele at the locus for 'dominant white' plumage (I), which had been incubated for 52 h. Blood samples containing PGCs were transferred by intravascular injection to Barred Plymouth Rock embryos (ii) incubated for 53, 72 and 96 h. Of the embryos which hatched, 28 were male and 31 were female. All chicks were raised to sexual maturity and test mated with Barred Plymouth Rock fowl. All of the 3117 offspring exhibited the typical Barred Plymouth Rock phenotype; no Barred Plymouth Rock x Dwarf White Leghorn chicks were obtained. The results of this study suggest that the frequency of transmission of the donor line genotype after PGC transfer must be improved for this technique to be useful for the routine development of transgenic poultry.}, journal={Journal of reproduction and fertility}, author={Petitte, JN and Clark, ME and Etches, RJ}, year={1991}, month={May} } @article{petitte_etches_1991, title={Daily infusion of corticosterone and reproductive function in the domestic hen (Gallus domesticus).}, volume={9}, url={http://europepmc.org/abstract/med/1936920}, DOI={10.1016/0016-6480(91)90145-v}, abstractNote={The purpose of this study was to examine the effect of the daily infusion of corticosterone on reproductive function in the laying hen and to determine the relationship between the cyclic pattern of plasma concentrations of corticosterone on the open-period for the preovulatory release of LH. An exogenous rhythm of plasma levels of corticosterone was generated using an osmotic pump. Corticosterone was infused subcutaneously into laying hens at rates of 5, 10, 15 or 30 μg/hr for a duration of 10hr beginning with the onset of darkness or at 15 μg/hr for 4hr, or continuously at 30 μg/hr. Daily infusions greater than 15 μg/hr and the continuous infusion resulted in cessation of ovulation, ovarian and oviductal regression, hyperphagia, and elevated levels of plasma corticosterone compared to that observed in control hens. The hens which were infused with 5 or 10 μg/hr of corticosterone maintained normal reproductive function with plasma concentrations of corticosterone that were approximately the same as those in the control hens. The effect of infusing 10 μg/hr of corticosterone on the open-period for the preovulatory release of LH was determined under constant light. No significant changes were observed in the frequency distribution of the times of oviposition when hens were infused with 10 μg/hr of corticosterone for 12hr from 9:00 to 21:00hr or 21:00 to 9:00hr each day. The results of this study indicate that a circadian rhythm in plasma levels of corticosterone cannot drive the open-period under constant illumination and that the effects of the infusion of corticosterone on reproductive function and feed intake are dependent upon the dose and duration of infusion.}, journal={General and comparative endocrinology}, author={Petitte, JN and Etches, RJ}, year={1991}, month={Sep} } @article{petitte_etches_anderson-langmuir_1991, title={Effect of metyrapone on the timing of oviposition and ovarian steroidogenesis in the laying hen.}, volume={9}, url={http://europepmc.org/abstract/med/1933451}, DOI={10.1080/00071669108417406}, abstractNote={1. The purpose of this study was to observe the effects of metyrapone on the time of oviposition and LH-stimulated steroidogenesis by granulosa cells and small yellow follicles. 2. In experiment 1, White Leghorn hens were injected for 11 d with 240 mg metyrapone 5 h before 'lights off'. Control hens were injected with 1 ml of vehicle (PEG-400). Metyrapone treatment resulted in a 28% decrease in the rate of lay and the modal frequency of the time of oviposition was phase-shifted by 15 h. 3. In experiment 2, hens were injected with 240 mg metyrapone 5 h before 'lights off' or at 'lights on'. While metyrapone treatment reduced the rate of lay, a clear phase-shift in the distribution of oviposition was not observed. Basal and LH-stimulated progesterone synthesis by the granulosa cells of the largest follicle and oestradiol synthesis by small yellow follicles was significantly reduced. 4. Metyrapone treatment significantly reduced basal, but not LH-stimulated output of androstenedione by whole small yellow follicles compared to that observed in control hens. 5. The addition of metyrapone in vitro to isolated granulosa cells from the three largest preovulatory follicles inhibited LH-stimulated progesterone production in a dose-specific manner. 6. The results of this study suggest that the ability of metyrapone to perturb the open-period is a pharmacological effect mediated through inhibition of ovarian and adrenal steroidogenesis.}, journal={British poultry science}, author={Petitte, JN and Etches, RJ and Anderson-Langmuir, CE}, year={1991}, month={Sep} } @article{brazolot_petitte_etches_verrinder_1991, title={Efficient transfection of chicken cells by lipofection, and introduction of transfected blastodermal cells into the embryo.}, volume={12}, url={http://europepmc.org/abstract/med/1751034}, DOI={10.1002/mrd.1080300404}, abstractNote={AbstractChicken blastodermal cells (CBCs) and primary chicken fibroblasts (PCFs) have been lipofected with a variety of lacZ constructs encoding Escherichia coli β‐galactosidase (β‐gal). A reporter construct (phspPTlacZpA) containing a mouse heat‐shock protein 68 gene (hsp 68) promoter was used to establish conditions for efficient lipofection. The construct, in circular or linear plasmid form or as reporter sequences alone, was transferred efficiently by incubating the cells for 3.5 h in a mixture of 6.2 μg LipofectinTM (a cationic liposome preparation from Bethesda Research Laboratories) and 1.55‐3.1 μg DNA per mL DMEM. These lipofection conditions were used to transfer a reporter construct (pCBcMtlacZ) containing a Zn2+‐inducible chicken metallothionein (cMt) promoter, and constructs showing constitutive expression due to Rous sarcoma virus plus chicken β‐actin (pmiwZ) or cytomegalovirus (pMaori3) promoters.Endogenous chicken β‐gal and transferred bacterial β‐gal activity could be distinguished clearly by incubating the cells with the substrate, Xgal, at pH 4.3 or 7.4, respectively. Expression of phspPTlacZpA in chicken cells did not appear to require specific induction of the mouse hsp68 promoter, whereas expression of pCBcMtlacZ required treatment of the cells for 6–12 h with 150 μM ZnCl2. Bacterial β‐gal activity was observed following lipofection of CBCs that were cultured in suspension or plated. The efficiency of lipofection was at least 1 in 25 for CBCs, judging by the proportion of cells shown to have b̃‐gal activity 16–24 h after lipofection treatment began; these events could represent transient or stable incorporation of the construct. Plated PCFs were lipofected as well, with stable incorporation of the gene construct indicated in 10% of positive events.Lipofected CBCs were injected into the subgerminal cavity of stage X (Eyal‐Giladi and Kochav, 1976) chicken embryos in a manner that has been shown to produce somatic and germline chimeras for untreated CBCs (Petitte et al., 1990). After 65 h of incubation, cells expressing β‐gal activity were observed in the prosencephalon, head ectoderm, and ventricle of the heart of a stage 11 (Hamburger and Hamilton, 1951) embryo. In other cases, bacterial β‐gal activity was detected extraembryonically, both in individual cells and in foci of expressing cells, 24, 48, and 65 h after injection. Few, if any, single expressing cells and no foci were detected following injection of the LipofectinTM:DNA mixture directly into the embryo.Refinement of these procedures could contribute to the development of transgenic poultry, without reliance on retroviral vectors for DNA transmission or incorporation.}, journal={Molecular reproduction and development}, author={Brazolot, CL and Petitte, JN and Etches, RJ and Verrinder, Gibbins AM}, year={1991}, month={Dec} } @article{etches_petitte_verrinder_brazolot_liu_1991, title={Production of chimeric chicks by blastodermal stem cell transfer and the prospects for gene manipulation.}, url={http://europepmc.org/abstract/AGR/IND92009441}, journal={Poultry Science Symposium.}, author={Etches, RJ and Petitte, JN and Verrinder, Gibbins AM and Brazolot, CL and Liu, G}, year={1991}, month={Jan} } @article{fasenko_robinson_armstrong_church_hardin_petitte_1991, title={Variability in preincubation embryo development in domestic fowl. 1. Effects of nest holding time and method of egg storage.}, volume={9}, url={http://europepmc.org/abstract/med/1780257}, DOI={10.3382/ps.0701876}, abstractNote={Embryos of eggs from Single Comb White Leghorn hens were analyzed to determine whether nest holding time and method of storage had a significant effect on postoviposition embryonic growth prior to incubation. Eggs were collected from 38- to 42-wk-old hens naturally inseminated and housed in floor pens. The experiment had a 2 x 2 factorial arrangement of treatments with two nest holding times and two storage methods. Eggs were collected within 1 h of oviposition, placed on cardboard egg flats, and stored unpacked (Treatment 1), or put on flats, and packed in 30-dozen egg cases (Treatment 2). Eggs in Treatments 3 and 4 were marked within 1 h of oviposition, but remained in the nest for 6 to 7 h. These eggs were separated into unpacked (Treatment 3), and packed (Treatment 4) groups. All eggs were stored at 13.8 C for 4 days. A total of 250 embryos were staged after storage for development using the Eyal-Giladi and Kochav classification. Least square means (LSM) for stage of development were: Treatment 1, 10.76; Treatment 2, 11.52; Treatment 3, 12.41; and Treatment 4, 12.36. For the main effects, nest holding time significantly affected stage of development (P = .0001), but storage method (P = .1140) and nest holding time by storage method interaction (P = .0730) did not. Comparison of LSM of Treatment 1 versus 3 (P = .0001), 2 versus 4 (P = .0152), and 1 versus 2 (P = .0214) were significant, but Treatment 3 versus 4 (P = .8595) was not.(ABSTRACT TRUNCATED AT 250 WORDS)}, journal={Poultry science}, author={Fasenko, GM and Robinson, FE and Armstrong, JG and Church, JS and Hardin, RT and Petitte, JN}, year={1991}, month={Sep} } @article{petitte_clark_liu_verrinder_etches_1990, title={Production of somatic and germline chimeras in the chicken by transfer of early blastodermal cells.}, volume={1}, url={http://europepmc.org/abstract/med/2351062}, journal={Development (Cambridge, England)}, author={Petitte, JN and Clark, ME and Liu, G and Verrinder, Gibbins AM and Etches, RJ}, year={1990}, month={Jan} } @article{etches_petitte_1990, title={Reptilian and avian follicular hierarchies: models for the study of ovarian development.}, url={http://europepmc.org/abstract/med/1974772}, DOI={10.1002/jez.1402560419}, abstractNote={The presence of an ovarian follicular hierarchy is a characteristic feature of reptiles and birds. The hierarchy contains follicles at all stages of maturation and therefore, varying degrees of sensitivity to the ovulation-inducing effects of the gonadotropins. In the hen, ovulability is gained as the ability of the follicle to produce androgens and estrogens declines and the ability to produce progesterone increases. In the mature follicle, the granulosa cells are the site of progesterone production whereas the theca cells produce androgens and estrogens. Small follicles that have not yet been (and may never be) recruited into the yolk-filled hierarchy are the major producers of androgens and estrogens within the ovary. In reptiles the ovarian follicular hierarchy includes non-vitellogenic follicles and in some species includes follicles destined to become atretic. These two features distinguish the reptiles from the birds and provide experimental biologists with a unique model to investigate the physiological events that regulate the most common fate of ovarian follicles, atresia.}, journal={The Journal of experimental zoology. Supplement : published under auspices of the American Society of Zoologists and the Division of Comparative Physiology and Biochemistry}, author={Etches, RJ and Petitte, JN}, year={1990}, month={Jan} } @article{petitte_etches_1989, title={The effect of corticosterone on the response of the ovary to pregnant mare's serum gonadotrophin in sexually immature pullets.}, volume={6}, url={http://europepmc.org/abstract/med/2744407}, DOI={10.1016/s0016-6480(89)80034-6}, abstractNote={Several studies have suggested that a functional relationship exists between the adrenal gland and the ovary. The purpose of this study was to determine the effect of corticosterone on the sensitivity of the ovary to exogenous gonadotrophin. Sexually immature White Lehorn pullets, 18 weeks of age, were infused with 30 μg/hr of corticosterone for 14 days. After 7 days of infusion, the pullets were injected with 3, 15, 75, or 375 IU pregnant mare's serum gonadotrophin (PMSG) for 7 days. Controls consisted of noninfused pullets injected according to the same doses of PMSG. Blood samples were obtained on Days, 3, 6, 8, 10, and 12 of infusion and were assayed for plasma concentrations of luteinizing hormone (LH), estradiol, and corticosterone by radioimmunoassay. The daily injections of 75 or 375 IU PMSG in noninfused pullets resulted in an increase in ovarian and oviductal weight and the formation of a hierarchy of yolk-filled follicles. This was accompanied by an increase in the plasma concentrations of estradiol and a decrease in the plasma concentrations of LH. The infusion of corticosterone significantly increased the plasma concentrations of this steroid over that observed in the control pullets and was not related to the dose of PMSG. This increase in plasma levels of corticosterone was associated with a significant decline in the plasma concentrations of LH and estradiol before the injection of PMSG. After 6 days of PMSG treatment, the plasma concentrations of estradiol increased to that observed in control pullets and was dependent upon the dose of PMSG injected. Ovarian and oviductal weight was, however, significantly lower in the corticosterone-infused pullets than in the control pullets. It is suggested that the mechanism of corticosterone-induced gonadosup-pression resides at the level of the hypothalamus and the ovary.}, journal={General and comparative endocrinology}, author={Petitte, JN and Etches, RJ}, year={1989}, month={Jun} } @article{petitte_etches_1988, title={The effect of corticosterone on the photoperiodic response of immature hens.}, volume={3}, url={http://europepmc.org/abstract/med/3360298}, DOI={10.1016/0016-6480(88)90034-2}, abstractNote={White Leghorn pullets, 18 weeks of age, were infused with 30 μg/hr of corticosterone for 14 days. After 7 days of continuous infusion, the pullets were photostimulated by transfer from 8L:16D to 16L:8D. Noninfused controls were either photostimulated on Day 7 or remained on an 8L:16D photoschedule. Blood samples were obtained on 3, 6, 8, 10, and 12 days of infusion and were assayed for plasma concentrations of LH, estradiol, and corticosterone by radioimmunoassay. On Day 14 all birds were weighed and sacrificed, and the ovarian and oviducal weights were recorded. Photostimulation had no effect on plasma concentrations of corticosterone. The infusion of corticosterone significantly raised the plasma concentration to 5.2 ng/ml, suppressed the photo-induced rise in plasma concentrations of LH, and resulted in significantly lower plasma concentrations of estradiol. After 7 days of photostimulation either with or without corticosterone infusion, there were no significant differences in mean ovarian weight. The oviducal weight of hens infused with corticosterone was, however, significantly lowered. It is suggested that one of the mechanisms associated with the antigonadal effect of corticosterone involves an inhibition of Gn-RH release by the hypothalamus.}, journal={General and comparative endocrinology}, author={Petitte, JN and Etches, RJ}, year={1988}, month={Mar} } @article{etches_petitte_anderson-langmuir_1984, title={Interrelationships between the hypothalamus, pituitary gland, ovary, adrenal gland, and the open period for LH release in the hen (Gallus domesticus).}, volume={12}, url={http://europepmc.org/abstract/med/6394694}, DOI={10.1002/jez.1402320317}, abstractNote={AbstractThe asynchronous ovulatory cycle of the hen is believed to be the consequence of two interacting systems, one of which is circadian and regulates the timing of the preovulatory LH surge. In suport of this proposition, the open period for LH release was shown to oscillate with the same periodicity as the photoschedule when the hens were exposed to 14 L:7 D, 14 L:10 D, and 14 L:14 D. In addition, it was demonstrated that follicular maturation is not affected by or synchronized with the photoperiod. The physiological system that transduces the light/dark cycle into an open period for LH release has not been identified although circumstantial evidence supports the idea that the adrenal gland plays a role in this function. This evidence includes the anatomical juxtaposition of the left ovary and adrenal gland, innervation of steroid‐producing cells within the follicle by nerve tracts passing through the adrenal glands, the ability of injections of metyrapone to alter the timing of preovulatory LH release, the ability of injections of corticosterone to induce ovulation when a mature follicle is present in the ovary, and the ability of dexamethasone or infusions of corticosterone to block ovulation. Recently we have also shown that infusions of corticosterone will block the gonadotropic effect of PMSG, will inhibit the photoperiodic response, and do not affect the release of LH in response to injections of GnRH. The addition of corticosterone to incubations of dispersed granulosa cells does not affect their response to LH. These data suggest that corticosterone may modulate the responsiveness of the hypothalamus to tropic stimuli and demonstrate that exposure to corticosterone can alter the responsiveness of some ovarian tissues to gonadotropins.}, journal={The Journal of experimental zoology}, author={Etches, RJ and Petitte, JN and Anderson-Langmuir, CE}, year={1984}, month={Dec} } @article{petitte_hawes_gerry_1983, title={The influence of cage versus floor pen management of broiler breeder hens on subsequent performance of cage reared broilers.}, volume={7}, url={http://europepmc.org/abstract/med/6622365}, DOI={10.3382/ps.0621241}, abstractNote={Chicks from breeder hens maintained in cages or floor pens were reared in Lohman battery cages in three separate trials. The ages of the breeder flock at the time of egg collection were 29, 36, and 54 weeks, respectively. The fertility of the artificially inseminated caged hens was significantly (P less than .05) lower than that of the naturally mated hens. The source of hatching eggs had no effect on early embryonic mortality, feed conversion, or growing period mortality in any broiler trials. Hatchability of all eggs set was significantly (P less than .05) lower for caged breeders in Trials 2 and 3. In all trials, eggs from caged hens produced significantly (P less than .05) larger day-old chicks than their floor-housed counterparts; however, these chicks were significantly (P less than .05) heavier at slaughter only in Trial 2. Carcass evaluations for breast blisters, keel malformations, and leg abnormalities revealed that the severity of each condition was associated with the sex of the broiler and that, within sexes, maternal housing management had no effect.}, journal={Poultry science}, author={Petitte, JN and Hawes, RO and Gerry, RW}, year={1983}, month={Jul} } @article{petitte_hawes_gerry_1982, title={The influence of flock uniformity on the reproductive performance of broiler breeder hens housed in cages and floor pens.}, volume={11}, url={http://europepmc.org/abstract/med/7163101}, DOI={10.3382/ps.0612166}, abstractNote={Two flocks of broiler breeder hens, differing in uniformity of body weight, were evaluated under cage and floor management systems over a 28-week period. Hens were housed singly in cages (N = 280) or in litter floor pens (N = 304). Caged hens were artificially inseminated with .05 ml of pooled semen once weekly. Floor birds were mated naturally. During the first 10 weeks of production, the more uniform flock exhibited significantly higher egg production than the less uniform flock. Uniformity did not influence cumulative egg production, egg weight, fertility, or mortality. Initial egg production was similar for caged and floor housed hens, but the caged birds attained significantly higher egg production during peak production. Caged females showed higher egg weights and body weights than did the floor females throughout the study. Naturally mated birds attained significantly higher fertility than artificially inseminated birds (94.9 vs. 90.6). Correlation coefficients for the weights of caged hens at 20 and 24 weeks with 24, 32, 40, and 50 week weights, total eggs per bird, and age at first egg indicated that body weight at 24 weeks was a better indicator of subsequent flock performance than was body weight at 20 weeks.}, journal={Poultry science}, author={Petitte, JN and Hawes, RO and Gerry, RW}, year={1982}, month={Nov} }