Brief Summary of Minutes of Annual Meeting

The meeting was called to order at 5:00 p.m. by Secretary Curt Youngs (Chairperson Cindy Tian was attending the IETS pre-conference symposium and did not join the meeting until after it had started). Members were welcomed, and Curt expressed regrets to the group from Tom Bunch (Utah) and C.-Y. Hu (Hawaii) who were unable to attend this years annual meeting. George Seidel moved and Matt Wheeler seconded a motion to approve the minutes of last years annual meeting. The motion carried unanimously. Curt Youngs asked one of our new members, Jianbo Yao, to introduce himself to the group. Jianbo earned his PhD at McGill University in Canada and did post-doctoral training in human genetic diseases in Montreal. While at Michigan State University he worked with Paul Coussens and George Smith to develop a cDNA library from bovine oocytes. At West Virginia University, he collaborates with Paul Lewis and Keith Inskeep. Bob Godke moved and Matt Wheeler seconded a motion to elect Brett White as secretary. Motion carried unanimously. Neither C.-Y. Hu (administrative advisor) nor Mark Mirando (CSREES) were present at the meeting, so the group did not receive comments from the administrative advisor or CSREES representative. The date and location of next years annual meeting was discussed. The W-1171 group had previously decided to hold their annual meeting in conjunction with the annual conference of the International Embryo Transfer Society (IETS). The 2008 IETS conference will be held January 6-8, 2008 in Denver, Colorado, USA. Members raised concerns about the usual timing of the W-1171 meeting. Typically the W-1171 meeting is held at the same time as the IETS pre-conference symposium, and this has been a recurring conflict for the W-1171 members who are speakers and/or attendees at the IETS pre-conference symposia. After lengthy discussion, a decision was made to explore the possibility of having the W-1171 meeting on the Monday evening and Tuesday evening of the annual IETS conference. The idea of sponsoring a symposium or mini-symposium at the annual IETS conference was discussed. Potential topics that were suggested for the symposium were embryo development & manipulation, the first cell cycle, hyperactivated sperm motility, maternal/embryonic transition, and cortical granules. Curt Youngs led a discussion regarding the upcoming midterm review and evaluation of the W-1171 multistate project by RCIC. There are four main areas that will be reviewed: progress report, linkages, funding, and information & technology transfer. The administrative advisor (C.-Y. Hu) is responsible for providing a brief evaluation of the activities and success of the W-1171 multistate project to RCIC using a special form (Appendix I of the MRP manual). The initial topic of discussion pertained to section 2 of this review (linkages). The following linkages were identified: Colorado & Illinois, Colorado & Connecticut, Colorado & North Carolina, Illinois & Wisconsin, Illinois & Nebraska, Illinois & Connecticut, California & Connecticut, Louisiana & Wisconsin, Louisiana & Iowa, Iowa & Beltsville, Nebraska & Indiana, North Carolina & Beltsville, North Carolina & Clay Center, Indiana & Illinois, Connecticut & Beltsville, Indiana & Beltsville. These linkages have involved collaborative research, graduate student training, publication of joint manuscripts, and submissions of grants. Rather than taking limited time to discuss each and every section of the upcoming midterm evaluation and review, it was decided that is would be more time efficient if four subcommittees were formed. Subcommittee one is headed by Bob Godke (assisted by Curt Youngs) and will focus on the progress report. Subcommittee two is headed by Carol Keefer (assisted by Matt Wheeler) and will focus on linkages. Subcommittee three is headed by Cindy Tian (assisted by Jerry Yang) and will focus on funding. Subcommittee four is headed by Rebecca Krisher (assisted by Curt Youngs) and will focus on information and technology transfer. Curt Youngs will send out an e-mail request to the W-1171 membership no later than January 31, 2007 requesting that each station prepare concise information to provide to each subcommittee chairperson. Responses to the subcommittee chairpersons will be due February 15, 2007. Subcommittee chairpersons will have one month to compile and prepare their sections of the midterm evaluation and review, and these need to be sent to Curt Youngs no later than March 15, 2007. Curt Youngs will compile the four sections into a single, cohesive report and send the report to Gary Anderson and George Seidel for editorial review. Because C.-Y. Hu needs to have the report submitted no later than May 15, 2007, George and Gary should submit the revised report to C.-Y. no later than April 20, 2007. George requested that Rebecca, Cindy, and Curt e-mail to him the compiled W-1171 station reports for the past three years. Discussion was held regarding the establishment of a W-1171 web site. This web site could serve as a public source of scientifically-based information about embryo and reproductive technologies such as transgenic animals, nuclear transfer, sperm sexing, etc. A suggestion was made that one portion of the web site should be password protected so that only members of W-1171 could have access. This web site could be a site for chat rooms, exchange of experimental methods, reaction to research proposals, etc. Richard Fayrer-Hosken indicated that he could take the lead of developing such a web site if the group would give him some guidance and direction on what they wanted. Station reports were given, although members were asked to be very brief and to highlight only extremely novel findings and/or discuss ideas for new collaborative research projects. The members were asked to read the complied station reports on their own time after adjournment of the meeting. The W-1171 members broke out into the three main joint research groups (oocytes, stem cells, and embryo production). Before discussions began, members were reminded that the success of the multistate project hinges on productive collaborations and that the viability of W-1171 is contingent upon joint research efforts (and not on individual station research efforts). The groups were asked to share research findings, evaluate new possibilities for collaborative research efforts based on those findings, and to chart a plan of action for the coming year. Bob Godke moved and Brett White seconded a motion to adjourn the meeting. The motion carried unanimously, and the meeting was adjourned at 10:20 p.m. Respectfully submitted, Curt Youngs 2006 W-1171 Acting President and Secretary

Work during the preceding year suggests aromatase inhibition during the postnatal interval may have statistically significant effects on accessory sex gland development but suggests such effects will not be biologically significant given the previously observed positive effects on Sertoli cell proliferation. Our data suggest specific oocyte plasma membrane proteins may be affected by heat-stress during oocyte maturation and used as biomarkers of heat-stress induced reduction in oocyte quality during development. Our data provide further evidence for involvement of two specific oocyte proteins in the fertilization process. Although there were no statistical differences in the sex ratio of single- or super-ovulating beef cattle, there is compelling evidence to continue this line of research. There may indeed be a bull effect that can be exploited in future experiments to enhance the efficacy of this sorting procedure, and there may also be other mechanisms (such as trypsin treatment) that may aid the binding of the HeiferPlus® product to the sperm head. There may also be other sorting methods (electromagnetic) that are favorable to flow cytometery to sway the sex ratio without the associated low pregnancy rates and high variability and costs. Understanding the interplay among hormones and the establishment of one or more dominant follicles will provide enabling knowledge about the fertilizability of oocytes from specific follicles within a follicular wave. Further, investigation of the follicular hierarchy is the focus of at least two submitted USDA-CSREES Standard Grant Proposals in which Clemson University will serve as a collaborator. The ability to supply a follicular wave with a pure, reliable, and consistent source of FSH will decrease the considerable variation associated with embryo transfer and will eliminate potential prion disease transmission. We showed that reducing oxygen concentrations to 5%, which approximate physiological levels in the follicle and oviduct, improved the quality of resulting mouse embryos. Similar studies with maturing bovine oocytes in our laboratory show no detrimental effect of lowering oxygen during oocyte maturation. We showed that culturing embryos in the presence of phenazine ethosulfate (PES) had no detrimental effects on pregnancy rates in cattle. Embryos cultured with PES survive cryopreservation better than controls. We achieved high calving rate of sexed IVP bovine embryos and improved the bovine IVP system by improving fertilization and vitrification conditions. To study oocyte developmental competence, we have determined that we can reliably isolate adequate amounts of mRNA from 150 oocytes to hybridize to a microarray. For validation of microarray results, we can test up to 9 genes by real time PCR from a pool of 25 oocytes. We have shown that large-scale transfer of IVP Holstein heifer embryos, produced with gender-selected semen, to beef recipients is a feasible production scheme. Producers can use IVP to produce high genetic merit claves from lower genetic merit recipients. Oocytes harvested from Brahman and Angus breed donor cows were individually evaluated for lipid content. Using buoyant density gradients and Nile Red staining, the Brahman oocytes were found to have significantly more intracellular lipids than the oocytes from the Angus cows. Sexed semen was used to successfully to produce in vitro fertilized cattle embryos. These gender-selected Holstein embryos were transferred nonsurgically to mature crossbred beef cows. All these gender-selected embryos produce female calves. Mature crossbred beef cow recipient females had no difficulty giving birth with the typically large Holstein heifer calves. Our lab has identified two novel Lef1 isoforms, Lef12,3,6 and Lef16 that are expressed in bovine preimplantation embryos. These transcription factors may play a critical role in trophectoderm and placental differentiation. Fusion constructs consisting of the bovine NANOG promoter and GFP gene and the bovine Nanog protein and GFP have been produced and demonstrated to be expressed following transfection. These constructs will aid in functional studies concerning the role of Nanog in maintenance of pluripotency in early ruminant embryos. We have determined transcriptomes, functional blue prints, of bovine oocytes and early embryos. We have identified many proteins in the bovine GV stage oocytes and its surrounding cumulus cells. With respect to oocyte maturation, we: " Determined gene transcription is not required for EGF-induced oocytes maturation. " Determined that FSH-induced maturation requiring gene transcription over-rides EGF pathway when both hormones are present in maturation medium. " Identified potential candidate mRNAs associated with FSH-induced initiation of GVBD in murine and bovine cumulus-oocyte complexes matured in vitro. " Used short interfering RNA approaches to assess the functional role of NR4a1 and other mRNAs of interest during FSH-induced oocyte maturation in cattle. With respect to embryo and fetal development, we: " Determined that expression of IGF1 and IGF2R mRNAs was significantly decreased in the livers of fetuses resulting from in vitro-produced embryos at day 70 of gestation. " Determined that expression of IGF2R was significantly decreased in skeletal muscle of bovine fetuses from in vitro-produced embryos at day 70 of gestation. " Developed a new classification system for reporting of normal and abnormal outcomes following the transfer of IVF or SCNT embryos. The wide-range of potential developmental abnormalities of fetuses, placentas and calves are included in the new term, Abnormal Offspring Syndrome (AOS). " Confirmed conservation of genomic imprinting in swine placenta for over 15 known imprinted genes. We gained new information regarding the developmental biology of the early bovine embryo and factors affecting the establishment of extraembryonic endoderm. Identifying critical factors involved in this process may provide insights into aberrant mechanisms that predispose the embryo to pregnancy loss due to abnormal development and/or implantation failure. Genomic analysis of in vivo and in vitro porcine blastocysts: We constructed and characterized Serial Analysis of Gene Expression (SAGE) libraries representing comprehensive gene expression profiles of in vivo- and in vitro-derived day 6 pig blastocysts. Poultry sperm studies: a. Established methodology to differentiate distinct testicular cell populations and progenitor spermatogonia within the turkey testis. b. Developed transplantation protocol for the successful transfer of donor germ cells and repopulation of germ-cell-free recipient seminiferous tubules. c. Characterization of the turkey and chicken sperm glycocalyx composition. d. Identification of glycocalyx modulations occuring during in vitro storage of poultry semen. e. Confirmation of the ability of turkey sperm to acquire exogenous phospholipids during in vitro storage. Integrins facilitate attachment of cells to the extra-cellular matrix, often binding the arginine-glycine-aspartic acid tri-peptide motif, thus facilitating cell migration, mediating cell-cell adhesion, linking the extracellular matrix with cytoskeletal elements, and acting as signaling molecules. Adhesion activates signaling mechanisms that regulate integrin function, cytoskeletal assembly, cell behavior, and protein synthesis. Immunofluorescence was used to determine the presence of integrin a and b subunits on the surface of bovine oocytes using a panel of monoclonal antibodies specific for aL, aM, aX, aV, a2, a4, a6, b1, b2, and b3 antigens, with multiple antibodies for each subunit. Confocal microscopy indicated the presence of aV, a6, a4, a2, ß1, and ß3 integrin subunits on the plasma membrane of bovine oocytes. The presence of these subunits was verified by RT-PCR analysis using primers designed based on known gene sequences of bovine integrin subunits, or by using sequence information using bovine expressed sequence tags compared with known human and murine integrin subunit gene sequence information. Previously unpublished sequence information for bovine a6 and b3 integrins was determined. The presence of these integrin subunits on the bovine oocyte vitelline membrane supports the hypothesis that sperm-oocyte interactions in the bovine are mediated by integrins. The ability of synthetic RGD-containing peptides to induce intracellular calcium transients similar to those observed at fertilization by spermatozoa in the bovine has been reported (Campbell et al., 2000; Sessions et al., 2006). These results also indicated the ability of synthetic RGD-containing peptides to induce activation and subsequent parthenogenetic development to the blastocyst stage, although, at numbers lower than observed with control in vitro fertilization (IVF). Evidence has been provided indicating the important effect of surrounding regions on the biological activity of the RGD sequence (Zhu and Evans, 2002; Sessions et al., 2006). The current experiments were designed to use natural RGD-containing sequences (disintegrins) to understand their effects. A total of three RGD-containing snake venom peptides (Kistrin (K), Elegantin (Ele), and Echistatin (Ech)) and one non-RGD-containing venom (Erabutoxin B (EB; control) were used at three concentrations (0.1, 1 and 10 mg /ml) to induce parthenogenetic development to the blastocyst stage and in conjunction (1.0, 5.0 and 10 µg/ml) with spermatozoa to evaluate competitive inhibition of fertilization and subsequent development. A (P< 0.01) higher number of bovine oocytes developed to the blastocyst stage after incubation with K, Ele and Ech at 1.0 µg/ml and was not different (P> 0.01) from IVF control. Fertilization was significantly reduced (P<.01) at all concentrations of K, Ele and Ech as compared to IVF control. No reduction (P>.05) was observed in EB (non-RGD) treated oocytes. These results support the involvement of a disintegrin-integrin interaction at fertilization in the bovine resulting in activation and subsequent development. Integrins have been shown to be involved in the process of fertilization and many integrin-ligand interactions are mediated through the recognition of an arginine-glycine-aspartic acid (RGD) sequence. Despite the fact the RGD domain is a principal player in determining the functional characteristics of an adhesive protein, increasing evidence has accumulated implicating the amino acids flanking the RGD sequence in determining the functional properties of the RGD-containing protein. A set of linear peptides in which the amino acid sequence in and around the RGD tri-peptide was modified was synthesized to better understand the specificity of the RGD-receptor interaction. Mature oocytes were fertilized in vitro in the presence of RGD-containing and RGD-modified peptides. Both the RGD-containing and RGD-modified peptides impaired the ability of sperm to fertilize bovine oocytes, illustrated by a reduction in cleavage. The linear modified RGD containing peptides were also examined for their ability to induce parthenogenetic development with the objective of providing a linear RGD peptide with greater biological activity than the one (GRGDSPK) used previously (Campbell et al. 2000). The data demonstrate the specificity of the receptor for the RGD sequence, further implicate the involvement of integrins in the process of bovine fertilization, and illustrate the importance of the amino acids surrounding the RGD sequence in determining the binding and functional properties of RGD-containing peptides. The data support the findings that a linear RGD peptide can block fertilization and that amino acids around the RGD sequence have an impact on the biological activity of the receptor. The effects of nicotine on nuclear maturation and meiotic spindle dynamics of bovine oocytes and subsequent embryonic development were investigated. Maturation rates (85%-94%) were similar to the control (86%) at concentrations from 0.01 to 1.0 mM, but decreased significantly (P<0.01 or 0.001) at 2.0 to 6.0 mM. Haploid complements of metaphase II oocytes in 0.01 to 1.0 mM nicotine (approximately 90%) were similar to the control, while significantly lower (ranged from 63% to 76%, P<0.05 or P<0.01) haploid oocytes were observed in the 2.0 to 6.0 mM nicotine groups. The majority of diploid (2n=60) oocytes without a PB1 occurred in the 3.0 to 6.0 mM nicotine treatments. Spindle microtubules changed from characteristically being asymmetrical in the controls to being equally distributed into two separate chromosome groups in the nicotine treatments. Nicotine inhibited the movement of anaphase or telophase chromosomes to the cortical area. The inhibited two chromosome groups became two spindles that either moved close in proximity or merged entirely together resulting in diploidy within the affected oocyte. Nicotine treatment significantly reduced the rate of cleavage and blastocyst development after parthenogenetic activation. Diploidy and cell number were dramatically reduced in the resultant blastocysts. We conclude that nicotine can alter the normal process of bovine oocyte meiosis by causing a diploid state in the oocyte that subsequently interferes with embryonic development. Immunofluorescent staining and cytogenetic preparation techniques were performed to examine bovine oocyte maturation, dynamic nuclear changes, and meiotic spindle patterns after oocytes were exposed to nicotine at various concentrations and at different times. Nicotine at concentrations of 0.01 and 0.1 mM resulted in higher (P<0.05) maturation rates (92.9% and 93.6%, respectively) than the controls (84.1%); concentrations of 0.5 to 2.0 mM had maturation rates similar to the control; concentrations of 3.0 to 6.0 mM significantly decreased (P<0.01 or 0.001) maturation rates. Haploid compositions of metaphase II oocytes in 0.01 to 1.0 mM nicotine (approximately 90%) were similar to the control, while significantly lower (ranged from 63% to 75%, P<0.05 or P<0.01) haploid oocytes were obtained in the 3.0 to 6.0 mM nicotine groups. Extremely swollen and twisted spindles formed after oocytes were exposed to concentrations e 2.0 mM. Spindle microtubules were equally distributed between the separated two groups of chromosomes in nicotine groups, while in the control, microtubules were asymmetrically distributed to the separated two groups of chromosomes, and the majority of the microtubules were oriented and distributed to the first polar bodies. Nicotine exposure at 3.0, 4.0, 5.0, and 6.0 mM for 24 hr resulted in 23.1, 32.5, 41.2, and 43.1% diploid oocytes, respectively. Diploidy resulted from the inhibition of anaphase or telophase chromatid movement. The inhibited two groups of chromatids became two spindles that either moved close in proximity or merged entirely together. Nicotine exposure affected oocyte maturation in a dose-dependent manner resulting in disfiguration of meiotic spindles and causing diploidy. Examination of histological sections gave evidence of the formation of eosinophilic cells, blood islets, and contractile elements with no evidence of a forming neural system. Blood islets and eosinophilic cells were associated with the contractile elements forming an apparent vascular tissue matrix. Contractile elements were positive for the formation presence of calcium activated ATP-ase activity, and were unresponsive to acid reversal of calcium activated myo-fibriliary ATP-ase activity at pH 4.3 and pH 4.5, indicating a similarity to cardiac muscle fibers. We have demonstrated level of nuclear reprogramming of gene expression using DNA microarrays in bovine SCNT derived blastocysts. The expression patterns of key transcription factors, Nanog, Oct4 and Sox2 in ICM explants were determined. This knowledge will aid in the derivation and characterization of ruminant embryonic stem cell lines. Our study showed that culturing bovine fibroblast cells in vitro enhanced various nuclear epigenetic modifications that are likely detrimental to nuclear reprogramming during the cloning procedure in animals. In a recent study we were able to isolate and culture stem cell like cells from porcine subcutaneous fat cells (adipocytes). The enhanced lactation potential of the transgenic gilts synergizes with suckling intensity to stimulate increased milk production during early lactation, as well as stimulating piglet growth. These animals provide a method to manipulate lactation persistency in swine. Cultured bone marrow derived porcine mesenchymal stem cells have been differentiated into osteoblastic cells when cultured on chitosan/biphasic calcium phosphate porous scaffolds for up to four weeks. This allows development of engineered bone replacement tissue for humans and animals. We improved NT efficiency by using PHA; studied the reprogrammability of cells at different differentiation stages; studied gene reprogramming in porcine NT by using X-linked genes as markers. We observed that loading extra cholesterol into membranes of oocytes improved their survival after vitrification. Similar studies by others with sperm of several species show that adding cholesterol improves survival after cryopreservation. An efficacious method of vitrification of bovine and equine pre-compaction embryos was developed. This likely would also work for sheep and goat embryos. We developed a SCNT method in swine that can generate both transgenic and non-transgenic clones at high efficiencies. We have increased usefulness of Affymetrix porcine arrays. We have determined which microarray platform is best suited for swine studies. Embryo-derived cell line models: We established stable and long-lived porcine endoderm cell lines from in vivo embryos and characterized cell lines transcriptomic and proteomic of candidate genes analyses as well as by morphology and enzymatic activity. Porcine embryo stem cells: To better understand the potential importance of several well-known human and/or stem cell and differentiation associated factors in the potential maintenance or spontaneous differentiation associated with in vitro culture, transcript expression analysis of those factors was examined in undifferentiated porcine epiblasts and the effect of short-term of culture was determined. Furthermore, we demonstrated that the porcine epiblast expression profile more closely resembles that of the human embryonic stem cell. Cloning is one of several new assisted reproductive techniques being developed for clinical use in the equine industry. Potential uses of equine cloning include: (1) the preservation of genetics from individual animals that would otherwise not be able to reproduce, such as geldings; (2) the preservation of genetic material of endangered and/or exotic species, such as the Mongolian wild horse (Przewalskis horse); and (3) because of the companion animal role that horses fill for some individuals, it is likely that some horse owners will have individual animals cloned for emotional fulfillment.Although equine cloning has been successful, like other species, it remains a very inefficient process (<3% success). In most species, the inefficiency of cloning results from a high incidence of embryonic, fetal and/or placental developmental abnormalities that contribute to extremely high rates of embryonic loss, abortion and stillbirths throughout gestation and compromised neonatal health after birth. The present review describes some of the ultrasonographic, endocrinological and histopathological characteristics of successful (produced viable offspring) and unsuccessful (resulted in pregnancy failure) cloned equine (mule and horse) pregnancies we have produced.A total of 21 cloned mule pregnancies were established using fetal fibroblast cells, whereas a total of seven cloned horse pregnancies were established using adult cumulus cells. Three of the cloned mule conceptuses were carried to term, resulting in the birth of three healthy clones. We have successfully developed a course in embryo transfer and related technologies that utilizes research advancements made by members of the W-1171 multistate research project.

1. REFEREED ARTICLES/BOOK CHAPTERS Aston, K., G-P. Li, B. Hicks, B. Sessions, B. Pate, D. Hammon, T. Bunch, and K. White. 2006. Effect of the time interval between fusion and activation on nuclear state and development in vitro and in vivo of bovine somatic cell nuclear transfer embryos. Reproduction 131:1-8. Aston, K.I., G-P. Li, B.A. Hicks, B.R. Sessions, B.J. Pate, D.S. Hammon, T.D. Bunch, and K.L. White. 2006. The developmental competence of bovine nuclear transfer embryos derived from cow versus heifer cytoplasts. Anim. Reprod. Sci. 95:234-243. At-Taras, E. E., T. Berger, M. J. McCarthy, A. J. Conley, B. J. Nitta-Oda, and J. F. Roser. 2006. Reducing estrogen synthesis in developing boars increases testis size and total sperm production. J Androl 27: 552-559. At-Taras, E. E., A. J. Conley, T. Berger, and J. F. Roser. 2006. Reducing estrogen synthesis does not affect gonadotropin secretion in the developing boar. Biol Reprod 74: 58-66. At-Taras, E. E., I. Kim, T. Berger, A. J. Conley, and J. F. Roser. 2006. Reducing endogenous estrogen during development alters hormone production by porcine Leydig cells and seminiferous tubules domestic animal endocrinology Domest Anim Endocrinol doi:10.1016/j.domaniend.2006.11.003 (in press). Bakst, M., Akuffo, V., Trefil, P., and Brillard, J.P. 2006. Morphological and histochemical characterization of the seminiferous epithelial and Leydig cells of the turkey. Ani. Reprod. Sci. 97:303-313. Bakst, M.R., Long, J.A., and Kramer, M.H. 2006. Impact of fenbendazole on turkey semen quality. J. of Appl. Poultry Res. 15:307-311. Blomberg, L.A., Garrett, W.M., Guillomot, M., Miles J.R., Sonstegard, T.S., Van Tassell, C.P., and Zuelke, K.A. 2006. Transcriptome profiling of the tubular porcine conceptus identifies the differential regulation of growth and developmentally associated genes. Mol. Reprod. Dev. 73:1491-502. Bou Khalil, M., K. Chakrabandhu, H. Xu, W. Weerachatyanukul, M. Buhr, T. Berger, E. Carmona, N. Vuong, P. Kumarathasan, P. T. Wong, D. Carrier, and N. Tanphaichitr. 2006. Sperm capacitation induces an increase in lipid rafts having zona pellucida binding ability and containing sulfogalactosylglycerolipid. Dev Biol 290: 220-235. Campos-Chillon, L. F., D.J. Walker, J.F. De La Torre-Sanchez, and G.E. Seidel, Jr. 2006. In vitro assessment of a direct transfer vitrification procedure for bovine embryos. Theriogenology 65:1200-1214. Chang C-C, Nagy ZP, Abdelmassih R, Liu J-L, Yang X, Tian XC. 2006. Meiotic spindle formation and function following insertion of mitotic chromosomes into GV stage mouse oocytes. RMBonline 12:213-221. Chaubal SA, Molina JA, Ohlrichs CA, Ferre LB, Faber DC, Bols PEJ, Riesen J, Tian XC. Yang X. 2006. Different transvaginal ovum pick-up techniques in cows to optimize oocyte retrieval and embryo production over a fixed period of time. Theriogenology 66:1631-1648. De La Torre-Sanchez, J. F., K. Preis, and G.E. Seidel, Jr. 2006. Metabolic regulators of in vitro-produced bovine embryos I. Effects of metabolic regulators at different glucose concentrations with embryos produced by semen from different bulls. Reprod. Fertil. Develop. 18:585-596. De La Torre-Sanchez, J.F., D.K. Gardner, K. Preis, J. Gibbons, and G.E. Seidel, Jr. 2006. Metabolic regulation of in vitro-produced bovine embryos II. Effects of phenazine ethosulfate, sodium azide, and 2,4-dinitrophenol during post-compaction development on glucose metabolism and lipid accumulation. Reprod. Fertil. Develop. 18:597-607. Du F, Shen P, Xu J, Sung L, Jeong B-S, Nedambale TL, Riesen J, Tian XC, Cheng WTK, Lee S-N, Yang X. 2006. Phytohemagglutinin-cell agglutination agent and embryo vitrification improves the efficiency of somatic cloning in cattle (Bos Taurus). Theriogenology 65: 642-657. Ducummon, C.C. and T. Berger. 2006. Localization of the rho GTPases and some rho effector proteins in the sperm of several mammalian species. Zygote 14: 249-257. Farin CE, Rodriguez KF, Alexander JE, Hockney JE, Herrick JR, and Kennedy-Stoskopf S. 2006. The role of transcription in EGF- and FSH-mediated oocyte maturation in vitro. Anim Reprod Sci, 2006 (October 13, Epub ahead of print). Farin PW, Moore K, and Drost M. Assisted reproductive technologies in cattle. 2006. In: (RS Youngquist and WR Threlfall, Eds.) Current Therapy in Large Animal Theriogenology, 2nd Ed., Elsevier, Inc., (in press). Farin PW, Piedrahita JA, and Farin CE. 2006. Errors in development of fetuses and placentas from in vitro-produced bovine embryos. Theriogenology 65:178-191 Ford, J.A., S.G. Clark, E.M. Walters, M.B. Wheeler, W.L. Hurley 2006. Estrogenic effects of genistein on reproductive tissues of ovariectomized gilts. J. Anim. Sci. 84:834842. Giraldo, A., J. Lynn, R.A. Godke and K. Bondioli.2006. Proliferation characteristics and chromosomal stability of bovine donor cells for nuclear transfer. Mol. Reprod. Dev. 73:1230-1238. He, S., D. Pant, A. Schiffmacher, S. Bischoff, D. Melican, W. Gavin, and C. Keefer. 2006. Developmental expression of pluripotency determining factors in caprine embryos: Novel pattern of nanog protein localization in the nucleolus. Mol Reprod Dev 73: 1512-1522. Herrick, J.R., Behboodi, E., Memili, E., Blash, S., Echelard, Y., Krisher, R.L. 2006. Metabolism, protein content and in vitro embryonic development of goat cumulus-oocyte complexes matured with physiological concentrations of glucose and L-lactate. Mol. Reprod. Dev. 73:256-266. Herrick, J.R., Brad, A.M., Krisher, R.L. 2006. Chemical manipulation of glucose metabolism in porcine oocytes: Effects on nuclear and cytoplasmic maturation in vitro. Reproduction. 131: 289-298. Herrick JR, Lane M, Gardner DK, Behboodi E, Memili E, Blash S, Echelard Y, Krisher RL. 2006. Metabolism, protein content, and in vitro embryonic development of goat cumulus-oocyte complexes matured with physiological concentrations of glucose and L-lactate. Mol Reprod Dev 73:256-266. Horvath, G., and Seidel, G. E. Jr. 2006. Vitrification of bovine oocytes after treatment with cholesterol-loaded methyl-²-cyclodextrin. Theriogenology 66:1026-1033. Jiang L, Jobst P, Lai L, Samuel M, Ayares D, Prather RS, Tian XC. 2006. Expression of Growth-Regulating Imprinted Genes in Cloned Piglets. Cloning Stem Cell (in press). Jiang L, Lai L, Samuel M, Prather RS, Yang X, Tian XC. 2006. Expression of X-linked Genes in Deceased Neonates and Surviving Cloned Female Piglets. Mol Reprod Dev (in press). Keefer, C.L., Pant, D., Blomberg, L.A., and Talbot, N.C. 2006. Challenges and prospects for the establishment of embryonic stem cell lines of domesticated ungulates. Anim. Reprod. Sci. Oct 14; [Epub ahead of print]. Lazaris, A., R. Keyston, C. N. Karatzas, and C. L. Keefer. 2006. Transgenesis using nuclear transfer in goats. Methods Mol Biol 348: 213-226. Li, G-P., T.D. Bunch, K.L. White, L. Rickords, Y. Liu, and B. R. Sessions. 2006. Denuding and centrifugation of maturing oocytes alters oocyte spindle integrity and the ability of cytoplasm to support parthenogenetic and nuclear transfer embryo development. Mol. Reprod. Dev. 73:446-451. Li, G., J. Saenz, R.A. Godke and R.V. Devireddy. 2006. Effect of glycerol and cholesterolloadedcyclodextrin on freezing induced water loss in bovine spermatozoa. Reproduction 131:875-886 Lindbloom, S.L., T.A. Farmerie, C.M. Clay, G.E. Seidel, Jr., and E.M. Carnevale. 2006. Roles of EGF-like growth factors and phosphodiesterases in initiation of oocyte maturation in the equine follicle. Anim. Reprod. Sci. (in press). Liu, Y., G-P. Li, L.F. Rickords, K.L. White, B.R. Sessions, K.I. Aston, T.D. Bunch. 2006. Effect of nicotine on in vitro maturation of bovine oocytes. Reprod. Biol. Endocrinol. Online citation: http://dx.doi.org/10.1016/j.anireprosci.2006.11.013. Long, J.A. 2006. Avian semen cryopreservation: What are the biological challenges? J. Poultry Sci. 85:232-236. Long, J.A., and H.D. Guthrie. 2006. Validation of a rapid, large-scale assay to quantify ATP levels in spermatozoa. Theriogenology 65:1620-1630. Marshall, K.M., W.L. Hurley, R.D. Shanks, M.B Wheeler. 2006. Effects of suckling intensity on milk yield and piglet growth from lactation-enhanced gilts. J. Anim. Sci. 84:23462351. McCarthy, M. J., E. E. At-Taras, C. A. Pearl, B. S. Nitta-Oda, J. F. Roser, A. J. Conley, and T. Berger. 2006. Suppression of endogenous estrogen during development affects porcine epididymal sperm maturation. Mol Reprod Dev 73: 1122-1128. Memili E, Peddinti D, Shack LA, Nanduri B, McCarthy F, Sagirkaya H and Burgess SC. 2006. Global proteome of bovine germinal vesicle oocytes and cumulus cells. Reproduction. (in press). Michael, D.D., Alvarez, I.M., Ocon, O.M., Powell, A.M., Talbot, N.C., Johnson, S.E., and Ealy A.D. 2006. Fibroblast growth factor-2 is expressed by the bovine uterus and stimulates interferon-tau production in bovine trophectoderm. 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Reduced oxygen concentration improves the developmental competence of mouse oocytes following in vitro maturation. Molec. Reprod. Develop. (DOI 10.1002.mrd) (in press) Purcell, S. H., Seidel, G.E. Jr., McCue, P.M., and Squires, E.L. 2006. Aspiration of oocytes from transitional, cycling, and pregnant mares. Anim. Reprod. Sci. (in press). Ramesh, R., C. A. Pearl, E. At-Taras, J. F. Roser, and T. Berger. 2006. Ontogeny of androgen and estrogen receptor expression in porcine testis: Effect of reducing testicular estrogen synthesis. Anim Reprod Sci. doi:10.1016/j.anireprosci.2006.10.025 (in press). Rodriguez KF, Blomberg LA, Zuelke KA, Miles JR, Alexander JE, and Farin CE. 2006. Identification of candidate mRNAs associated with gonadotropin-induced maturation of murine cumulus oocyte complexes using serial analysis of gene expression. Physiol Genomics 27: 318-327. Sagirkaya H, Misirlioglu M, Kaya H, Parrish JJ, First NL and Memili E. 2006. Developmental potential of bovine oocytes cultured in defined maturation medium. Animal Reproduction Science. Oct 17; [Epub ahead of print] PMID: 17052869 [PubMed - as supplied by publisher] Sagirkaya H, Misirlioglu M, Kaya H, Parrish JJ, First NL and Memili E. 2006. Developmental and molecular correlates of bovine preimplantation embryos. Reproduction 131: 895-904 Sansinena, M. J., S.A. Taylor, P.J. Taylor, E.E. Schmidt, R.S. Denniston and R.A. Godke. 2006. In vitro production of llama (Lama glama) embryos by intracytoplasmic sperm injection: Effect of chemical activation treatments and culture conditions. Anim. Reprod. Sci. (In press). Schenk, J. L., Suh, T. K., and Seidel, G. E. Jr. 2006. Embryo production from superovulated cattle following insemination of sexed sperm. Theriogenology 65:299-307. Seidel, G. E. Jr. 2006. Modifying oocytes and embryos to improve their cryopreservation. Theriogenology 65:228-235. Seidel, G. E., Jr., and Walker, D. J. 2006. Pregnancy rates with embryos vitrified in 0.25-ml straws. J. Reprod. Develop. 52 Suppl): S71-S76. Sessions, BR, Aston, KI, Davis, AP, Pate, BJ and KL White. 2006. Effects of Amino Acid Substitutions in and Around the Arginine-Glycine-Aspartic Acid (RGD) Sequence on Fertilization and Parthenogenetic Development in Mature Bovine Oocytes. Mol. Reprod. Dev. 73:651-657. Sung L-Y, Gao S, Shen H, Yu H, Song Y, Smith SL, Chang C-C, Inoue K, Kuo K, Lian J, Li A, Tian XC, Tuck DP, Weissman SM, Cheng T, Yang X. 2006. Cloned mouse pups can be derived from adult stem cells as well as terminally differentiated somatic cells. Nature Genetics 38:1323-8. Sung LY, Shen PC, Jeong BS, Xu J, Chang CC, Cheng WT, Wu JS, Lee SN, Broek D, Faber D, Tian XC, Yang X, Du F. 2006. Premature Chromosome Condensation Is Not Essential for Nuclear Reprogramming in Bovine Somatic Cell Nuclear Transfer. Biol Reprod 2006 Nov 15; [Epub ahead of print] Suteevun T, Parnpai R, Smith SL, Chang C-C, Muenthaisong S, and XC Tian. 2006. Epigenetic characteristics of cloned and in vitro fertilized swamp buffalo (Bubalus bubalis) embryos. J Anim Sci 84:2065-2071. Suteevan T, Smith S, Muentaisong S, Yang X, Parnpai R, Tian XC. 2006. Anomalous mRNA levels of chromatin remodeling genes in swamp buffalo (Bubalus bubalis) cloned embryos. Theriogenology 65:1704-1715. Tanphaichitr, N., E. Carmona, M. Bou Khalil, H. Xu, T. Berger, and G. L. Gerton. 2007. New insights into sperm-zona pellucida interaction: Involvement of sperm lipid rafts. Front Biosci 12: 1748-1766. Trefil, P., Micakova, A., Mucksova, J., Hejnar, J., Brillard, J.P., Bakst, M. Kalina, J., and Poplstein, M. 2006. estoration of spermatogenesis and male fertility by transplantation of dispersed testicular cells in the chicken. Biol. Reprod. 75: 575  581. Tsai S, Cassady J, Freking D, Nonneman G. Rohrer G, and Piedrahita JA. 2006. Annotation of Affymetrix porcine genome microarray chip. Animal Genetics 37:423-424. Tsai S, Mir B, Martin AC, Estrada JL, Bischoff SR, Hsieh WP, Cassady JP, Freking BA, Nonneman DJ, Rohrer GA, and Piedrahita JA. 2006. Detection of transcriptional difference of porcine imprinted genes using different microarray platforms. BMC Genomics. 7:328-334. Vanderwall, DK, Woods, GL, Roser, JF, Schlafer, DH, Sellon, DC, Tester, DF, and White, KL. 2006. Equine Cloning: Applications and Outcomes. Reprod. Fertil. Devel. 18:91-98. Walker, D. J., Campos-Chillon, L. F., and Seidel, G. E. 2006. Vitrification of in vitro-produced bovine embryos by addition of ethylene glycol in one step. Reprod. Domest. Anim. 41:467-471 Wheeler, M.B., A.S. Lima, T. VanEtten, M.R.B. Mello. 2006. Transgenic Animals for Biomedicine and Agriculture. In (Rodolfo L. de la Sota and Gabriel A. Bo, Eds.) Bovine Theriogenology 23 pgs. Wheeler, M.B., M.R.B. Mello, A.S. Lima, T. VanEtten. 2006. Modification of the Domestic Animal Genome. In (Rodolfo L. de la Sota and Gabriel A. Bo, Eds.) Bovine Theriogenology 33 pgs. Wheeler, M.B., J.J Rutledge, A. Fischer-Brown, T. VanEtten, S. Malusky, D.J. Beebe. 2006. Application of sexed semen technology to in vitro embryo production in cattle. Theriogenology 65:219227. White, K.L., M. Passipieri, T.D. Bunch, K.D. Cambell and B.J. Pate. 2006. Effects of arginine-glycine-aspartic acid (RGD containing snake venom peptides on parthenogenetic development and in vitro fertilization of bovine oocytes. Mol. Reprod. Dev. 74:88-96. Woodard, J., A. Hilldore, , M. B. Wheeler, S. G. Clark, S. Lan, R. Jamison, A.J. Wagoner Johnson. 2006. The mechanical properties and osteoconductivity of hydroxyapatite bone scaffolds with multi-scale porosity. Biomaterials 28:4554. Xu J, Guo ZQ, Nedambale TL, Zhang JX, Schenk J, Tian XC, Yang X, Du F. 2006. High developmental potential of vitrified Holstein cattle embryos fertilized in Vitro with sex-sorted sperm. J Dairy Sci 89:2510-8. Youngs, C.R. and R.A. Godke. 2006. Biotechnology: Cattle Embryo Transfer Technology. In (D.R. Heldman, A. Bridges, D. Hoover, and M.B. Wheeler, Eds.) Encyclopedia of Biotechnology in Agriculture and Food. Marcel Dekker, New York, NY (in press) 2. BOOKS, NON-REFEREED BOOK CHAPTERS, PROCEEDINGS, INSTRUCTIONAL MEDIA, THESES/DISSERTATIONS Ahola, J. K., Aznarez, V. A., Seidel, G. E. Jr., and Whittier, J. C. 2006. Superimposing 14d MGA pre-feeding and(or) 7d CIDR on the SelectSynch (GnRH-PG) estrus synchronization protocol in beef cows. Proc. Western Sect. Am. Soc. Anim. Sci. 57:272-275. Curchoe C. 2006. Epigenetics of bovine clones. PhD thesis. University of Connecticut De La Torre-Sanchez, J. F., and Seidel, G. E. Jr. 2006. Evolucion de los medios para culturo de embriones bovinos. In: Memorias del XI Curso Internacional de Reproduccion Bovina, FMVZ-UNAM, Mexico, D.F., pp. 104-121. Farin PW. 2006. Estrus synchronization programs for cows and heifers. Proc 142nd Annual Convention, AVMA, July 15-19, 2006, Honolulu Farin PW. 2006. Management of embryo donor cows and recipients. Proc 142nd Annual Convention, AVMA, July 15-19, 2006, Honolulu. Farin PW. 2006. Reducing fetal loss in cattle. Proc 142nd Annual Convention, AVMA, July 15-19, 2006, Honolulu. Farin PW. 2006. Update on sexed semen, ivf and cloning in cattle. Proc 142nd Annual Convention, AVMA, July 15-19, 2006, Honolulu. Guerrerro, C. 2006. Cryopreservation and Intracytoplasmic with Bovine Epididymal Spermatozoa. PhD Dissertation, Louisiana State University. August, 2006. Jiang L. 2006. Reprogramming of epigenetically-regulated genes in cloned pigs. PhD thesis. University of Connecticut. Leed, A. 2006. Characterization of potential porcine oocyte plasma membrane receptors. M.S. thesis, University of California-Davis. Moisan, A. 2006. Vitrification and Dehydration for the Preservation of Gametes. PhD Dissertation Louisiana State University. December, 2006. Nel-Themaat, L. 2006. Gamete and Cell Technolgies for Use in Biological Resource Banking. PhD Dissertation, Louisiana State University. May, 2006. Pinisetty, D. J. Saenz, R.A. Godke and R.V. Devereddy. 2006. Theoretical predictions of optimal cooling rates for cryopreservation of caprine sperm. Proc. ASME Bioengineering Meetings. CD Publication 2006. Preis, K. 2006. Metabolic Characteristics and Gene Expression of In Vitro and In Vivo Matured Mammalian Oocytes. Ph.D. Dissertation, Colorado State University, Fort Collins, CO. Seidel, G. E., Jr. 2006. On the usefulness of an update on assisted reproductive technologies in cattle. Theriogenology 65:1-3. Seidel, G. E., Jr., and Schenk, J. L. 2006. Sex-selected semen. Proc. Applied Reproductive Strategies in Beef Cattle. Univ. Missouri, Columbia, MO, pp. 315-321. Seidel, G. E., Jr., and Wheeler, M. B.. 2006. Cloning Pets. In: Proc. Symposium on Cloning Pets, M.B. Wheeler and G.E. Seidel, Jr. (eds), pp. 1-8. Sendemir-Urkmez, A., S.G. Clark, M.S. Goldwasser, M.B. Wheeler, R.D. Jamison. 2006. Tissue Engineered Bone for Mandibular Reconstruction. American Association of Oral-Maxiofacial Surgeons Annual Meeting, October 2006, San Diego, CA pgs. 42-43. Suh, T. K., Stokes, J., Squires, E. L., and Seidel, G. E. Jr. 2006. Effect of removing acrosomes from stallion sperm before ICSI of bovine oocytes. Anim. Reprod. Sci. 303-306. Wagoner Johnson, A. J., Woodard, J., Hilldore, A., Jamison, R.D., Wheeler, M.B., Clark, S.G. 2006. Bone-Like Behavior of Brittle, porous Hydroxyapatite Implants With Microporosity. American Association of Oral-Maxiofacial Surgeons Annual Meeting, October 2006, San Diego, CA pgs. 40-41. Wheeler, M.B. 2006. Cloning and Transgenic Animals for Agriculture. 2nd International Symposium of Applied Animal Reproduction, Londrina, Parana, Brazil, October 5-7, 2006. Pgs. 1-18. White, K.L., G.L. Woods, D.K. Vanderwall, G. Li, and T.D. Bunch. 2006. Cloning the Equine. Epigenetic Risks of Cloning. CRC Press, Taylor & Francis Group, LLC. pp 59 - 69. Wilson, M. 2006. Bovine and Equine Recombinant Follicle Stimulating Hormone Produced in the Heterologous Host Trypansoma brucei. M.S. Thesis, Clemson University, Clemson, SC. Youngs, C.R. and A.E. Freeman. 2006. Breeding (Animal). In McGraw-Hill Encyclopedia of Science and Technology, 10th Ed. (in press). 3. ABSTRACTS Barcelo-Fimbres, M., Brink, Z., and Seidel, G. E. Jr. 2006. Effects of PES during in vitro culture of bovine embryos on pregnancy rates. J. Anim. Sci. 84 (Suppl. 1):432. Barcelo-Fimbres, M., and Seidel, G. E. Jr. 2006. Effects of fetal calf serum and fructose or glucose on embryonic development and lipid accumulation of bovine embryos. Reprod. Fertil. Develop. 18:186. Berger, T. and A. Leed. 2006. Putative molecules involved in porcine sperm-oocyte plasma membrane interaction 10th International Symposium on Spermatology Madrid. Burns, M., D. Lapin, A. Brown, J. Gibbons. 2006. Effects of progesterone and presence of a corpus luteum on superovulation in beef cattle. 32nd Annual Conference of the International Embryo Transfer Society, Orlando, FL. Abstract 360. Burns, M., D. Lapin, A. Brown, J. Gibbons. 2006. Effects of progesterone and presence of a corpus luteum on superovulation in beef cattle. 32nd Annual Conference of the International Embryo Transfer Society, Orlando, FL. Abstract 360. Campos-Chillon, L. F., Cox, J., Seidel, G. E. Jr., and Carnevale, E. M. 2006. Vitrification of large equine embryos. Reprod. Fertil. Develop. 18:151. Campos-Chillon, L. F., Suh, T. K., Seidel, G. E. Jr., and Carnevale, E. M. 2006. Use of bovine embryos to establish methods for vitrification of early equine embryos. Theriogenology 66:684-685. Chang CC, Nagy ZP, Elliott TA, Wright G, Kort HI, Yang X, Tian XC. The DNA methylation and histone acetylation of oocytes in juvenile and adult mice. Hum Reprod 2006; 21(S1):i14. (presented in the European Society of Human Reproduction and Embryology annual meeting, Prague). Chang CC, Nagy ZP, Kort HI, Rasmussen TP, Yang X, Tian XC. Dynamic distribution of heterochromatin protein 1b in the preimplantation embryos. Fertil Steril 2006; 86(2S):S79-80. (New Orleans, American Society for Reproductive Medicine in 2006 annual meeting). Chang CC, Nagy ZP, Shapiro DB, Mitchell-Leef D, Yang X, Tian XC. Dynamic distribution of g-tubulin during mouse oocyte maturation. Fertil Steril 2006; 86(2S):S96-97. (New Orleans, American Society for Reproductive Medicine in 2006 annual meeting). Curry, E., S. Ellis, D. Lapin, and J. Gibbons. 2006. Efficacy determination of HEIFERPlusÔ semen sexing kit in superovulated cows. Society for the Study of Reproduction Meetings, Omaha, NE. Abstract 499. Davidson, T.R., S.A. Malusky, C.E. Ferguson, S.J. Lane, M.B.Wheeler, W.L. Hurley. 2006. Isolation of multilineage stem cells from the porcine mammary fat pad. Reproduction, Fertility and Development 18:171. Du,F., J Xu, S Gao, L-Y Sung, D Stone, M Joyner, J Zhang, S Chaubal, X Tian, YE Chen, X Yang. Full-term and live rabbit clones produced by somatic cell nuclear transfer. Reprod Fert Dev 2006; 18:124 (presented at the annual meeting of the International Embryo Transfer Society, Florida, 2006). Estrada J, Lee E, and Piedrahita J. Cryopreservation of donor cells for nuclear transfer: effect of cell freezing method on the efficiency of somatic cell nuclear transfer in pigs. Reprod Fert Dev 18:125 (abst 32), 2006. Everts RE, Razzak A, Chavatte-Palmer P, Hue I, Green CA, Oliveira R, Rodriguez-Zas S, Tian XC, Yang X, Renard JP, Lewin HA. 2006. Gene expression profiling of placentomes from cloned cattle. A model for mammalian systems biology. (to be presented in "Pathways, Networks, and Systems" to be held in Mykonos, Greece, October 8-13). Everts, RE, A Razzak, P Chavatte-Palmer, I Hue, CA Green, R Oliveira, S Rodriguez-Zas, XC Tian, X Yang, J-P Renard, HA Lewin. 2006. Global gene expression profiling distinguishes placentomes derived by artificial insemination from those pregnancies derived by either in vitro fertilization or nuclear transfer. PAG abstract guide: p282. Presented at the Plant & Animal Genome XIV Conference, January 14 - 18, 2006, San Diego, California Everts RE, Sommers A, Green CA, Oliveira R, Rodriguez-Zas S, Sung LY, Du F, Evans ACO, Boland M, Fair T, Lonergan P, Renard JP, Yang X, Lewin HA, XC Tian. Major differences in Gene expression profiles revealed in Day-25 placental tissues collected from Cows carrying cloned fetuses (to be presented in the Plant & Animal Genome XV Conference, January 13-17, 2007, San Diego, USA). Farin CE, Sommer JR, Rodriguez KF, Petters RM, and Alexander JE. Functional analysis using short-interfering (si)RNA of TRAM-6, a novel transcript associated with oocyte maturation in cattle Reprod Fert Dev 18: 269-270 (abst 324), 2006. Giraldo, A.M., J.A. Jenkins, J.W. Lynn, R.A. Godke and K.R. Bondioli. 2006. Histone phosphorylation patterns and chromosomal stability of cultured bovine fibroblasts. Reprod. Fertil. Develop. 18:292. Guerrero, C.A., S.P. Leibo, K.R. Bondioli and R.A. Godke. 2006. Effect of cryoprotective agent addition and removal at 4°C on motility and plasma membrane integrity of bovine caudal epididymal spermatozoa. Reprod. Fertil. Develop. 18:156 Hartke J.L., J.M. Drnevich, M.B. Wheeler, S.M. Donovan. 2006. Transgenic Overexpression of Insulin-Like Growth Factor-I (IGF-I) in Milk Influences Intestinal Development and Gene Expression in Piglets. 13th Int.Conference Int. Soc. Research Human Milk and Lactation, Niagara-on-the-Lake, Ontario, Canada, September 22-26, 2006. Jiang L, Prather RS, Jobst P, Ayares D, Yang X, Tian XC. Expression of growth-related imprinted genes in decreased newborn and surviving cloned piglets. Reprod Fert Dev 2006; 18:232 (presented at the annual meeting of the International Embryo Transfer Society, Florida, 2006). Kaya A, Schindler J, Memili E and Parrish JJ. (2006) Melatonin improves bovine embryo production and protects embryos from heat stress in vitro. Abstract # 398. Biol Reprod Special Issue. Kim C, Ma Y, Chang C-C, Yang X, Rasmussen T, Tian XC. The Typical Pattern of Histone MacroH2A1 in Preimplantation Bovine Embryos Following Activation and In Vitro Fertilization. Reprod Fert Dev 2006; 18:232-233 (presented at the annual meeting of the International Embryo Transfer Society, Florida, 2006). Krisher, R.L. (2005) Inhibition of the pentose phosphate pathway results in meiotic arrest in porcine oocytes that can be overcome by the addition of pathway cofactors and end products. Reprod. Fertil. Dev. 17:293 (abstr. 287). 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