Challenges of Genetic Screening of In Vitro Fertilized Human Embryos Using Current Technologies 2 1 688 2 Human is usually sub-fertile in comparison to other mammals; in other words, reproduction process of other mammals is more effective than human. Chromosomal anomalies of early embryos are the major reason for the low fecundity of human per cycle. Most of embryos with major genetic defects may be implanted and it can be followed with stopping of early development and early embryo loss with no tangible evidence of pregnancy. In addition, several studies have shown that more than 70% of embryos from assisted reproductive technologies have genetic anomalies. These anomalies lead to implantation failure or early embryo loss following implantation in IVF cycles (1). According to the above evidence on in vitro produced embryos, different methods such as PGD and PGS were developed for choosing the best embryos without genetic defects. These methods are based on molecular techniques of Q-PCR, FISH, CGH and SNP microarray, NGS and many other advanced technologies. While several studies approved the efficiency of these techniques, they have different problems that put their effectiveness in doubt over time. For example, the FISH method on blastomers of day three embryos was the choice in the two past decades, but recent evidence revealed that it failed to provide reliable result of PGS on cleavage embryos. Therefore, it has been replaced by the newer techniques such as CGH microarray and SNP microarray over time. However, though the newer techniques have higher degree of accuracy, they always have their own limitations and deficiencies as well (2). Alternatively, the poor results of PDG and PGS on day three embryos lead to performing the embryo biopsy on blastocyst on day five after fertilization. Although the quantity and quality of biopsied cells are better than day three embryos, this area already raised issues such as suspicion about its limitations and deficiencies like epigenetic changes due to prolonged in vitro culture, self-correction potency for chromosomal anomalies of early embryo, different origins of biopsied cells from trophoectoderm against ICM and several other limitations. Furthermore, today in the scientific community and the media, application of the expensive technique of next generation of sequencing (NGS) is recommended for genetic screening of IVF embryos. Preliminary data using this technology reported the increase of ART outcomes up to %70-80. Based on these results, this technique may have high accuracy and reliability. However, its wide application in large portion of infertile couples, especially those with repeated IVF failure (RIF), repeated pregnancy loss (RPL) and the patients older than 40 years requires further investigation (3). Currently, application of this technique will impose huge cost on couples and if its accuracy and precision is in doubt, it may lead to loss of a large number of embryos that have the potential of implantation and live birth. In addition, the interpretation of the large volume of data from NGS is not very simple. Many of these findings may indicate normal variations or corrective potential of embryos may ameliorate most of them. So, as long as validation of the effectiveness of this technique or innovation of newer techniques with maximum sensitivity and specificity is required, care should be taken in prescribing these techniques to infertile couples. 137 138 Mohammad Reza MR Sadeghi محمدرضا صادقی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran sadeghi@avicenna.ac.ir No Keyword 688.pdf Munné S, Cohen J. Chromosome abnormalities in human embryos. Hum Reprod Update. 1998;4 (6):842-55.##Brezina PR, Kutteh WH, Bailey AP, Ke RW. Preimplantation genetic screening (PGS) is an excellent tool, but not perfect: a guide to counseling patients considering PGS. Fertil Steril. 2016;105(1):49-50.##Ly KD, Agarwal A, Nagy ZP. Preimplantation genetic screening: does it help or hinder IVF treatment and what is the role of the embryo? J Assist Reprod Genet. 2011 Sep;28(9):833-49.##

Survival Assessment of Mouse Preimplantation Embryos After Exposure to Cell Phone Radiation 2 1 672 2 Background: Using cellular phone has rapidly increased all over the world. Also, the concern on the possible health hazards of electromagnetic fields (EMF) induced from cell phones to reproduction has been growing in many countries. The aim of this study was to assess the consequences and effects of exposure to the cell phone radiation on the quality and survival rates of preimplantation embryos in mice. Methods: A total of 40 mice (20 females and 20 males), 6 weeks old and sexually mature BALB/c, were used for control and experimental groups. The ovary burses were removed and the zygotes were dissected in the morning after mating. Next, 2-cell embryos were divided into two groups of control (n=150) and experimental (n=150). EMF (900-1800 MHz) was used for four days in experimental group for 30 min/day in culture at 37°C in a CO2 incubator. The quality of embryos was recorded daily and the fluorescent staining was used for identification of viable blastocysts. All data were compared by Student’s t-test and Mann-Whitney test (p<0.05). Results: The rate of embryo survival to the blastocysts stage was similar in both groups. However, the percentage of dead embryos at the 2-cell stage was significantly higher in EMF-exposed group compared with controls (p=0.03). Also, the loss of cell viability significantly increased in experimental blastocysts (p=0.002). Conclusion: The normal embryonic development up to the blastocyst stage indicates that EMF-exposure commonly did not have adverse effect on embryo development in mice. But, it caused loss of blastocysts cell viability. 138 144 Fereshteh F Safian Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Iran Mohammad Ali MA Khalili محمد علی خلیلی Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Iran khalili59@hotmail.com Arezoo A Khoradmehr Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Iran Fatemeh F Anbari Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Iran Saeedeh S Soltani Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Iran Iman I Halvaei Iman Halvaei Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences Iran Cell phoneIn VitroMicePreimplantation embryoRF-EMFViability 672.pdf Panagopoulos DJ, Karabarbounis A, Margaritis LH. Effect of GSM 900-MHz Mobile phone radiation on the reproductive capacity of Drosophila melanogaster. Electromagn Biol Med. 2004;23(1):29-43.##Lee KS, Choi JS, Hong SY, Son TH, Yu K. Mobile phone electromagnetic radiation activates MAPK signaling and regulates viability in Drosophila. Bioelectromagnetics. 2008;29(5):371-9.##Panagopoulos DJ, Chavdoula ED, Karabarbounis A, Margaritis LH. Comparison of bioactivity between GSM 900 MHz and DCS 1800 MHz mobile telephony radiation. Electromagn Biol Med. 2007;26(1):33-44.##Challis LJ. Mechanisms for interaction between RF fields and biological tissue. Bioelectromagnetics. 2005;Suppl 7:S98-S106.##Hyland GJ. Physics and biology of mobile telephony. Lancet. 2000;356(9244):1833-6.##Zareen N, Khan MY, Minhas LA. Dose related shifts in the developmental progress of chick embryos exposed to mobile phone induced electromagnetic fields. J Ayub Med Coll Abbottabad. 2009;21(1):130-4.##Panagopoulos DJ, Margaritis LH. The effect of exposure duration on the biological activity of mobile telephony radiation. Mutat Res. 2010;699(1-2):17-22.##Pilger A, Ivancsits S, Diem E, Steffens M, Kolb HA, Rüdiger HW. No effects of intermittent 50 Hz EMF on cytoplasmic free calcium and on the mitochondrial membrane potential in human diploid fibroblasts. Radiat Environ Biophys. 2004;43(3):203-7.##Falzone N, Huyser C, Franken DR, Leszczynski D. Mobile phone radiation does not induce pro-apoptosis effects in human spermatozoa. Radiat Res. 2010;174(2):169-76.##Beraldi R, Sciamanna I, Mangiacasale R, Lorenzini R, Spadafora C. Mouse early embryos obtained by natural breeding or in vitro fertilization display a differential sensitivity to extremely low-frequency electromagnetic fields. Mutat Res. 2003;538(1-2):163-70.##Wdowiak A, Wdowiak L, Wiktor H. Evaluation of the effect of using mobile phones on male fertility. Ann Agric Environ Med. 2007;14(1):169-72.##El-Sayed A, Badr HS, Yahia R, Salem SM, Kandil AM. Effects of thirty minute mobile phone irradiation on morphological and physiological parameters and gene expression in pregnant rats and their fetuses. Afr J Biotechnol. 2011;10(84):19670-80.##D'Silva, MH, Swer RT, Anbalagan J, Bhargavan R. Effect of ultrahigh frequency radiation emitted from 2G cell phone on developing lens of chick embryo: a histological study. Advances in Anatomy. 2014;2014:1-9.##Halvaei I, Khalili MA, Soleimani M, Razi MH. Evaluating the Role of First Polar Body Morphology on Rates of Fertilization and Embryo Development in ICSI Cycles. Int J Fertil Steril. 2011;5(2):110-5.##Farin PW, Crosier AE, Farin CE. Influence of in vitro systems on embryo survival and fetal development in cattle. Theriogenology. 2001;55(1):151-70.##Luo Q, Yang J, Zeng QL, Zhu XM, Qian YL, Huang HF. 50-Hertz electromagnetic fields induce gammaH2AX foci formation in mouse preimplantation embryos in vitro. Biol Reprod. 2006;75(5):673-80.##Elledge SJ. Cell cycle checkpoints: preventing an identity crisis. Science. 1996;274(5293):1664-72.##Murray A. Cell cycle checkpoints. Curr Opin Cell Biol. 1994;6(6):872-6.##Wells PG, Bhuller Y, Chen CS, Jeng W, Kasapinovic S, Kennedy JC, et al. Molecular and biochemical mechanisms in teratogenesis involving reactive oxygen species. Toxicol Appl Pharmacol. 2005;207(2 Suppl):354-66.##Hardy K. Cell death in the mammalian blastocyst. Mol Hum Reprod. 1997;3(10):919-25.##Friedman J, Kraus S, Hauptman Y, Schiff Y, Seger R. Mechanism of short-term ERK activation by electromagnetic fields at mobile phone frequencies. Biochem J. 2007;405(3):559-68.##Henshaw DL, Reiter RJ. Do magnetic fields cause increased risk of childhood leukemia via melatonin disruption? Bioelectromagnetics. 2005;Suppl 7:S86-97.##Panagopoulos DJ, Chavdoula ED, Nezis IP, Margaritis LH. Cell death induced by GSM 900-MHz and DCS 1800-MHz mobile telephony radiation. Mutat Res. 2007;626(1-2):69-78.##Pasternak CA. Membrane transport and disease. Mol Cell Biochem. 1989;91(1-2):3-11.##Weisbrot D, Lin H, Ye L, Blank M, Goodman R. Effects of mobile phone radiation on reproduction and development in Drosophila melanogaster. J Cell Biochem. 2003;89(1):48-55.##de Pomerai D, Daniells C, David H, Allan J, Duce I, Mutwakil M, et al. Non-thermal heat-shock response to microwaves. Nature. 2000;405(6785):417-8.##Kwee S, Raskmark P. Changes in cell proliferation due to environmental non-ionizing radiation: 2. Microwave radiation. Bioelectrochem Bioenerg. 1998;44(2):251-5.##Kwee S, Raskmark P, Velizarov S. Changes in cellular proteins due to environmental non-ionizing radiation. I. Heat-shock proteins. Electromagn Biol Med. 2001;20(2):141-52.##Shallom JM, Di Carlo AL, Ko D, Penafiel LM, Nakai A, Litovitz TA. Microwave exposure induces Hsp70 and confers protection against hypoxia in chick embryos. J Cell Biochem. 2002;86(3):490-6.##Mezhevikina LM, Khramov RN, Lepikhov KA. [The simulation of the cooperative effect of development in a culture of early mouse embryos after irradiation with electromagnetic waves in the millimeter range]. Ontogenez. 2000;31(1):27-31. Russian.##Grigor'ev IuG. [Biological effects of mobile phone electromagnetic field on chick embryo (risk assessment using the mortality rate)]. Radiats Biol Radioecol. 2003;43(5):541-3. Russian.##Poole C. Invited commentary: evolution of epidemiologic evidence on magnetic fields and childhood cancers. Am J Epidemiol. 1996;143(2):129-32.##Boerjan ML, den Daas JH, Dieleman SJ. Embryonic origins of health: long-term effects of IVF in human and livestock. Theriogenology. 2000;53(2):537-47.##

The Effect of Ammonium Chloride Concentration in In Vitro Maturation Culture on Ovine Embryo Development 2 1 684 2 Background: Ammonium is produced in culture medium due to amino acids degradation and has adverse effect on in vitro culture of embryo. In the current study, the purpose was to evaluate the effects of ammuniom chloride (AC) on in vitro oocyte maturation rate and early embryo development in the sheep and its effect on the expression of Bcl-2. Methods: In vitro maturation (IVM) was performed in the presence of various concentrations (0, 29, 88,132,176 μM/ml) of ammonium chloride (NH4CL) (AC). Meiotic maturation, embryonic development and expression of Bcl2 gene in Blastocyst cells were determined. The data were analyzed by one-way ANOVA and Tukey post HOC test, and values with p<0.05 were considered statistically significant. Results: The highest concentration (176 µM) of AC significantly decreased the rate of fully expanded cumulus cells 24 hr after IVM compared with the control group (p<0.05). Moreover, significantly lower rates of MII oocytes were found in the 176 µM AC group compared with the 29 µM AC group. The percentage of zygotes developing to blastocysts in 176 µM AC was lower than the other group. Also, supplementation of the oocyte maturation media with 176 µM AC decreased Bcl2 expression. Conclusion: Our results suggested that significant increase in IVM rate could be obtained with supplementation maturation medium with AC in a dose dependent manner. Increased AC concentration led to lower blastocyst rate under normal condition. However, regulation of pro–apoptotic (Bcl-2) gene did not change with different concentrations of AC supplementing. 144 151 Ali A Golchin Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz Iran Reza R Asadpour Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz Iran rasadpour4@gmail.com Leila L Roshangar لیلا روشنگر Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences Iran Raziallah R Jafari-Jozani Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz Iran Ammonium chlorideGene expressionOvine embryo 684.pdf Alexiou M, Leese HJ. Purine utilisation, de novo synthesis and degradation in mouse preimplantation embryos. Development. 1992;114(1):185-92.##Butler WR, Calaman JJ, Beam SW. Plasma and milk urea nitrogen in relation to pregnancy rate in lactating dairy cattle. J Anim Sci. 1996;74(4):858-65.##Cory S, Adams JM. Matters of life and death: programmed cell death at Cold Spring Harbor. Biochim Biophys Acta. 1998;1377(2):R25-44.##Edwards LJ, Williams DA, Gardner DK. Intracellular pH of the mouse preimplantation embryo: amino acids act as buffers of intracellular pH. Hum Reprod. 1998;13(12):3441-8.##Ellis RE, Yuan JY, Horvitz HR. Mechanisms and functions of cell death. Annu Rev Cell Biol. 1991;7:663-98.##Elrod CC, Butler WR. Reduction of fertility and alteration of uterine pH in heifers fed excess ruminally degradable protein. J Anim Sci. 1993;71 (3):694-701.##Epstein CJ, Smith SA. Amino acid uptake and protein synthesis in preimplanatation mouse embryos. Dev Biol. 1973;33(1):171-84.##Ferguson JD, Blanchard T, Galligan DT, Hoshall DC, Chalupa W. Infertility in dairy cattle fed a high percentage of protein degradable in the rumen. J Am Vet Med Assoc. 1988;192(5):659-62.##Ferguson JD, Galligan DT, Blanchard T, Reeves M. Serum urea nitrogen and conception rate: the usefulness of test information. J Dairy Sci. 1993;76(12):3742-6.##Gardner DK. Changes in requirements and utilization of nutrients during mammalian preimplantation embryo development and their significance in embryo culture. Theriogenology. 1998;49(1):83-102.##Gardner DK, Lane M. Amino acids and ammonium regulate mouse embryo development in culture. Biol Reprod. 1993;48(2):377-85.##Gross A, McDonnell JM, Korsmeyer SJ. BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 1999;13(15):1899-911.##Hafez B, Hafez ESE. Reproduction in farm animals. 7th. ed. New York: Wiley; 2000. p. 1-13.##Hammon DS, Wang S, Holyoak GR, Knight-Sherod J, Evans RC. Effects of high ammonia concentrations during IVM on oocyte maturation and in vitro development of bovine embryos. Theriogenology. 1999;51(1):375.##Marei WF, Wathes DC, Fouladi-Nashta AA. The effect of linolenic Acid on bovine oocyte maturation and development. Biol Reprod. 2009;81(6):1064-72.##Cognié Y, Baril G, Poulin N, Mermillod P. Current status of embryo technologies in sheep and goat. Theriogenology. 2003;59(1):171-88.##Lane M. Mechanisms for managing cellular and homeostatic stress in vitro. Theriogenology. 2001;55(1):225-36.##Lindenbaum A. A survey of naturally occurring chelating ligands. Adv Exp Med Biol. 1973;40:67-77.##McEvoy TG, Robinson JJ, Aitken RP, Findlay PA, Robertson IS. Dietary excesses of urea influence the viability and metabolism of preimplantation sheep embryos and may affect fetal growth among survivors. Anim Reprod Sci. 1997;47(1-2):71-90.##Tareq K, Miah AG, Salma U, Yoshida M, Tsujii H. Effect of amino acids and dipeptides on accumulation of ammonia in the medium during in vitro maturation and fertilization of porcine oocytes. Reprod Med Biol. 2007;6(3):165-70.##Papadopoulos S, Lonergan P, Gath V, Quinn KM, Evans AC, O'Callaghan D, et al. Effect of diet quantity and urea supplementation on oocyte and embryo quality in sheep. Theriogenology. 2001;55(5):1059-69.##Schneider M, Marison IW, von Stockar U. The importance of ammonia in mammalian cell culture. J Biotechnol. 1996;46(3):161-85.##Hammon DS, Wang S, Holyoak GR. Effects of ammonia during different stages of culture on development of in vitro produced bovine embryos. Anim Reprod Sci. 2000;59(1-2):23-30.##Yuan Y, Krisher RL. Effect of ammonium during in vitro maturation on oocyte nuclear maturation and subsequent embryonic development in pigs. Anim Reprod Sci. 2010;117(3-4):302-7.##Orsi NM, Leese HJ. Ammonium exposure and pyruvate affect the amino acid metabolism of bovine blastocysts in vitro. 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Association of the +49 A/G Polymorphism of CTLA4 Gene with Idiopathic Recurrent Spontaneous Abortion in Women in Southwest of Iran 2 1 667 2 Background: Survival of the semi-allograft fetus during pregnancy opens a new area for the immunological based causes of recurrent spontaneous abortion (RSA). Cytotoxic T lymphocyte-associated antigen 4 (CTLA4) is a negative regulator of the T-cell activation, which may modulate peripheral self-tolerance of the allogeneic fetus. The present study aimed to investigate the +49 A/G CTLA4 genetic polymorphism and predisposition to RSA. Methods: The total participants were 120 women with at least two miscarriages and 120 healthy post-menopausal women as the control group. The +49 A/G polymorphism was genotyped using PCR-RFLP method. Required demographic information was collected through filling out a questionnaire. The obtained data were fed into SPSS software version 16. Results: The results showed a significant association between the minor alleles (G) with the decreased risk of the RSA. The frequency of the G allele in controls and patients was 25% and 12%, respectively. A GG genotype in the co-dominance model (OR: 0.25, 95%CI: 0.09-0.66) and in the dominant model for allele G (GG+AG vs. AA) (OR: 0.84, 95%CI: 0.8-0.87) showed significant association with RSA by imposing the protective role. The frequency of miscarriage is significantly (p=0.04) higher among the relatives of RSA women (33.3%) in comparison with the women in the control group (21.7%). Conclusion: It can be concluded that +49G allele may act as a dominant allele and reduce the risk of RSA. Family history of miscarriage increased the risk of RSA among women. 151 157 Zarnegar Z Rasti Department of Biology, Islamic Azad University, Arsanjan branch Department of Biology, Islamic Azad University, Arsanjan branch Iran Mahboobeh M Nasiri Department of Biology, Islamic Azad University, Arsanjan branch Department of Biology, Islamic Azad University, Arsanjan branch Iran nasiri@iaua.ac.ir CTLA-4PCR-RFLPPolymorphismRecurrent spontaneous abortion 667.pdf Chaithra PT, Malini Suttur S, Sharath Kumar C. An overview of genetic and molecular factors responsible for recurrent pregnancy loss. Int J Hum Genet. 2011;11(4):217-25.##Rai R, Regan L. Recurrent miscarriage. Lancet. 2006;368(9535):601-11.##Veenstra van Nieuwenhoven AL, Heineman MJ, Faas MM. The immunology of successful pregnancy. Hum Reprod Update. 2003;9(4):347-57.##Medawar PB. Some immunological and endocrinological problems raised by the evolution of viviparity in vertebrates. Symp Soc Exp Biol. 1953:320-8.##Frauwirth KA, Thompson CB. Activation and inhibition of lymphocytes by costimulation. J Clin Invest. 2002;109(3):295-9.##Pedicord VA, Montalvo W, Leiner IM, Allison JP. Single dose of anti-CTLA-4 enhances CD8 T-cell memory formation, function, and maintenance. Proc Natl Acad Sci USA. 2011;108(1):266-71.##Gibson HM, Hedgcock CJ, Aufiero BM, Wilson AJ, Hafner MS, Tsokos GC, et al. Induction of the CTLA-4 gene in human lymphocytes is dependent on NFAT binding the proximal promoter. J Immunol. 2007;179(6):3831-40.##Anjos SM, Polychronakos C. Functional evaluation of the autoimmunity-associated CTLA4 gene: the effect of the (AT) repeat in the 3'untranslated region (UTR). J Autoimmun. 2006;27(2):105-9.##Kaufman KA, Bowen JA, Tsai AF, Bluestone JA, Hunt JS, Ober C. The CTLA-4 gene is expressed in placental fibroblasts. Mol Hum Reprod. 1999;5(1):84-7.##Vaidya B, Pearce S. The emerging role of the CTLA-4 gene in autoimmune endocrinopathies. Eur J Endocrinol. 2004;150(5):619-26.##Chistiakov DA, Turakulov RI. CTLA-4 and its role in autoimmune thyroid disease. J Mol Endocrinol. 2003;31(1):21-36.##Ueda H, Howson JM, Esposito L, Heward J, Snook H, Chamberlain G, et al. Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease. Nature. 2003;423(6939):506-11.##Unfried G, Bocskor S, Endler G, Nagele F, Huber JC, Tempfer CB. A polymorphism of the interleukin-6 gene promoter and idiopathic recurrent miscarriage. Hum Reprod. 2003;18(2):267-70.##Hogge WA, Byrnes AL, Lanasa MC, Surti U. The clinical use of karyotyping spontaneous abortions. Am J Obstet Gynecol. 2003;189(2):397-400.##Li TC, Makris M, Tomsu M, Tuckerman E, Laird S. Recurrent miscarriage: aetiology, management and prognosis. Hum Reprod Update. 2002;8(5):463-81.##Laird SM, Tuckerman EM, Cork BA, Linjawi S, Blakemore AI, Li TC. A review of immune cells and molecules in women with recurrent miscarriage. Hum Reprod Update. 2003;9(2):163-74.##Mor G, Cardenas I. The immune system in pregnancy: a unique complexity. Am J Reprod Immunol. 2010;63(6):425-33.##Miskovic S, Culic V, Konjevoda P, Pavelic J. Positive reproductive family history for spontaneous abortion: predictor for recurrent miscarriage in young couples. Eur J Obstet Gynecol Reprod Biol. 2012;161(2):182-6.##Bashyam H. CTLA-4: From conflict to clinic. J Exp Med. 2007;204(6):1243.##Chan DV, Gibson HM, Aufiero BM, Wilson AJ, Hafner MS, Mi QS, et al. Differential CTLA-4 expression in human CD4 versus CD8 T cells is associated with increased NFAT1 and inhibition of CD4 proliferation. Genes Immun. 2014;15(1):25-32.##Kolar P, Knieke K, Hegel JK, Quandt D, Burmester GR, Hoff H, et al. CTLA-4 (CD152) controls homeostasis and suppressive capacity of regulatory T cells in mice. Arthritis Rheum. 2009;60(1):123-32.##Wang C, Jiang T, Wei L, Li F, Sun X, Fan D, et al. Association of CTLA4 gene polymorphisms with susceptibility and pathology correlation to pulmonary tuberculosis in Southern Han Chinese. Int J Biol Sci. 2012;8(7):945-52.##Kavvoura FK, Ioannidis JP. CTLA-4 gene polymorphisms and susceptibility to type 1 diabetes mellitus: a HuGE Review and meta-analysis. Am J Epidemiol. 2005;162(1):3-16.##Zhao SX, Pan CM, Cao HM, Han B, Shi JY, Liang J, et al. Association of the CTLA4 gene with Graves' disease in the Chinese Han population. PLoS One. 2010;5(3):e9821.##Lee YH, Harley JB, Nath SK. CTLA-4 polymorphisms and systemic lupus erythematosus (SLE): a meta-analysis. Hum Genet. 2005;116(5):361-7.##Balic I, Angel B, Codner E, Carrasco E, Perez-Bravo F. Association of CTLA-4 polymorphisms and clinical-immunologic characteristics at onset of type 1 diabetes mellitus in children. Hum Immunol. 2009;70(2):116-20.##Hou R, Cao B, Chen Z, Li Y, Ning T, Li C, et al. Association of cytotoxic T lymphocyte-associated antigen-4 gene haplotype with the susceptibility to gastric cancer. Mol Biol Rep. 2010;37(1):515-20.##Yang M, Sun T, Zhou Y, Wang L, Liu L, Zhang X, et al. The functional cytotoxic T lymphocyte-associated Protein 4 49G-to-A genetic variant and risk of pancreatic cancer. Cancer. 2012;118(19):4681-6.##Wang X, Lin Q, Ma Z, Hong Y, Zhao A, Di W, et al. Association of the A/G polymorphism at position 49 in exon 1 of CTLA-4 with the susceptibility to unexplained recurrent spontaneous abortion in the Chinese population. Am J Reprod Immunol. 2005;53(2):100-5.##Messaoudi S, Houas I, Yaseen Kh, Dandana M, Mahjoub T. CTLA-4 gene polymorphisms and risk of idiopathic recurrent pregnancy loss in a Tunisian population. BMC Genomics. 2014;15(Suppl 2):P11.##Gupta R, Prakash S, Parveen F, Agrawal S. Association of CTLA-4 and TNF-α polymorphism with recurrent miscarriage among North Indian women. 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Effect of Acetylcholinesterase and Butyrylcholinesterase on Intrauterine Insemination, Contribution to Inflammations, Oxidative Stress and Antioxidant Status; A Preliminary Report 2 1 669 2 Background: Oxidative stress affects women fertility and influences on the sperm quality by alterating activities of cholinesterases, a molecular marker of stress-related infertility. The aim of the present study was to investigate the role of acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) activities and phenotypes in patients with unexplained infertility (idiopathic). It’s possible association with inflammation marker C-reactive protein (CRP) and other oxidative stress markers, i.e. before and after intra uterine insemination (IUI). Methods: In this study, blood samples of 60 patients with unexplained infertility were collected the day before and 24 hr after IUI (between 8 AM and 9 AM after the overnight fasting) and activities of BuChE, AChE, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GpX) and serum levels of thiol proteins (TP), C-reactive protein (CRP), total antioxidant capacity (TAC) were measured. Statistical significance was assumed at p<0.05. Results: Before IUI, there was a significant (p=0.048) positive correlation between BuChE activity and plasma TAC and a significant difference in the CAT activity between various BuChE (UU and non-UU) phenotypes. However, after IUI, a significant negative correlation between the AChE activity and BuChE activity was found (p=0.045) and the level of RBC AChE activity was significantly reduced (382.4±163.19 vs. 586.7±384 IU/grHb, p=0.025). Meanwhile, after IUI, the activities of SOD (1568±847.5 IU/grHb vs. 1126±229.3, p=0.031) and CAT (310±53.4 IU/grHb vs. 338±73, p=0.025) were increased. Conclusion: This study suggests that decline in cholinesterases activities may be responsible for stimulation of oxidative stress and inflammation and reduction in fertility rates by IUI. 157 163 Lida L Haghnazari Fertility and Infertility Research Center, Kermanshah University of Medical Sciences Fertility and Infertility Research Center, Kermanshah University of Medical Sciences Iran Asad A Vaisi-Raygani Asad Vaisi-Raygani Fertility and Infertility Research Center, Kermanshah University of Medical Sciences Fertility and Infertility Research Center, Kermanshah University of Medical Sciences Iran asadvaisiraygani@kums.ac.ir, avaisiraygani@gmail.com Farahnaz F Keshvarzi Department of Obstetrics and Gynecology, Kermanshah University of Medical Sciences Department of Obstetrics and Gynecology, Kermanshah University of Medical Sciences Iran Farivar F Ferdowsi Department of Obstetrics and Gynecology, Kermanshah University of Medical Sciences Department of Obstetrics and Gynecology, Kermanshah University of Medical Sciences Iran Massoud M Goodarzi Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Iran Massoudgoodarzi1345@yahoo.com Zohreh Z Rahimi Zohreh Rahimi Medical Biology Research Center, Kermanshah University of Medical Sciences Medical Biology Research Center, Kermanshah University of Medical Sciences Iran Hossin H Baniamerian Department of Clinical Biochemistry, Kermanshah University of Medical Sciences Department of Clinical Biochemistry, Kermanshah University of Medical Sciences Iran Haidar H Tavilani Department of Clinical Biochemistry, Hamadan University of Medical Sciences Department of Clinical Biochemistry, Hamadan University of Medical Sciences Iran Hadis H Vaisi-Raygani Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Iran Hessam H Vaisi-Raygani Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Molecular Diagnostic Research Center, Kermanshah University of Medical Sciences Iran Tayehbeh T Pourmotabbed Department of Microbiology, Immunology and Biochemistry, University of Tennessee, Health Science Center Department of Microbiology, Immunology and Biochemistry, University of Tennessee, Health Science Center USA Acetylcholinesterase (AChE)Butyrylcholinesterase (BuChE)Enzymatic and non-enzymatic antioxidantIntrauterine insemination (IUI)Oxidative stress 669.pdf Agarwal A, Saleh RA, Bedaiwy MA. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril. 2003;79(4):829-43.##Tanahatoe SJ, McDonnell J, Goverde AJ, Hompes PG, Lambalk CB. Total fertilization failure and idiopathic subfertility. Reprod Biol Endocrinol. 2009;7:3.##Lee VM, Wong JS, Loh SK, Leong NK. Sperm motility in the semen analysis affects the outcome of superovulation intrauterine insemination in the treatment of infertile Asian couples with male factor infertility. BJOG. 2002;109(2):115-20.##Mor I, Grisaru D, Titelbaum L, Evron T, Richler C, Wahrman J, et al. Modified testicular expression of stress-associated "readthrough" acetylcholinesterase predicts male infertility. FASEB J. 2001;15(11):2039-41.##Raynal P, Houdeau E. [Comparison of the uterine reflex activity during artificial insemination and mating in the ewe]. J Gynecol Obstet Biol Reprod (Paris). 2004;33(8):725-33. French.##Agarwal A, Aponte-Mellado A, Premkumar BJ, Shaman A, Gupta S. 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Hum Genomics. 2004;1(5):375-80.##La Du BN, Bartels CF, Nogueira CP, Arpagaus M, Lockridge O. Proposed nomenclature for human butyrylcholinesterase genetic variants identified by DNA sequencing. Cell Mol Neurobiol. 1991;11(1):79-89.##Bartels CF, James K, La Du BN. DNA mutations associated with the human butyrylcholinesterase J-variant. Am J Hum Genet. 1992;50(5):1104-14.##Prabhu K, Kumar P, Adiga SK, Rao A, Lanka A, Singh J. Plasma protein thiols, ceruloplasmin, C-reactive protein and red blood cell acetylcholinesterase in patients undergoing intrauterine insemination. J Hum Reprod Sci. 2009;2(1):27-9.##Vaisi-Raygani A, Tavilani H, Vaisi-Raygani H, Rahimi Z, Karimi J, Pour-motabbed T. Serum butyrylcholinesterase activity and phenotype associations with Lipid profile during various phases of menstrual cycle in young healthy women with regular menses, a preliminary report. AviJMB. 2013;1(1):23-9.##Massoulie J, Pezzementi L, Bon S, Krejci E, Vallette FM. Molecular and cellular biology of cholinesterases. Prog Neurobiol. 1993;41(1):31-91.##Vaisi-Raygani A, Rahimi Z, Entezami H, Kharrazi H, Bahrhemand F, Tavilani H, et al. Butyrylcholinesterase K variants increase the risk of coronary artery disease in the population of western Iran. Scand J Clin Lab Invest. 2008;68(2):123-9.##Vaisi-Raygani A, Rahimi Z, Kharazi H, Tavilani H, Aminiani M, Kiani A, et al. Determination of butyrylcholinesterase (BChE) phenotypes to predict the risk of prolonged apnea in persons receiving succinylcholine in the healthy population of western Iran. Clin Biochem. 2007;40(9-10):629-33.##Vaisi-Raygani A, Tavilani H, Zahrai M, Rahimi Z, Sheikh N, Aminian M, et al. Serum butyrylcholinesterase activity and phenotype associations with lipid profile in stroke patients. Clin Biochem. 2009;42(3):210-4.##Kalman J, Juhasz A, Rakonczay Z, Abraham G, Zana M, Boda K, et al. Increased serum butyrylcholinesterase activity in type IIb hyperlipidaemic patients. Life Sci. 2004;75 (10):1195-204.##Rahimi Z, Ahmadi R, Vaisi-Raygani A, Rahimi Z, Bahrehmand F, Parsian A. Butyrylcholinesterase (BChE) activity is associated with the risk of preeclampsia: influence on lipid and lipoprotein metabolism and oxidative stress. J Matern Fetal Neonatal Med. 2013;26(16):1590-4.##Iwasaki T, Yoneda M, Nakajima A, Terauchi Y. Serum butyrylcholinesterase is strongly associated with adiposity, the serum lipid profile and insulin resistance. Intern Med. 2007;46(19):1633-9.##Bahrehmand F, Vaisi-Raygani A, Rahimi Z, Ahmadi R, Kiani A, Tavilani H, et al. Synergistic effects of BuChE non-UU phenotype and paraoxonase (PON1) 55 M allele on the risk of systemic lupus erythematosus: influence on lipid and lipoprotein metabolism and oxidative stress, preliminary report. Lupus. 2014;23(3):263-72.##Shahmohamadnejad S, Vaisi-Raygani A, Shakiba Y, Kiani A, Rahimi Z, Bahrehmand F, et al. Association between butyrylcholinesterase activity and phenotypes, paraoxonase192 rs662 gene polymorphism and their enzymatic activity with severity of rheumatoid arthritis: correlation with systemic inflammatory markers and oxidative stress, preliminary report. Clin Biochem. 2015;48(1-2):63-9.##WHO. WHO laboratory manual for the examination and processing of human semen. 5th ed. Switzerland: WHO press; 2010. 286 p.##Whittaker M, editor. Cholinesterase. Vol. 11, Monograph in Human Genetics. Switzerland: Karger; 1986. 31 p.##Marcus DL, Thomas C, Rodriguez C, Simberkoff K, Tsai JS, Strafaci JA, et al. Increased peroxidation and reduced antioxidant enzyme activity in Alzheimer's disease. Exp Neurol. 1998;150(1):40-4.##Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121-6.##Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959;82(1):70-7.##de Peyster A, Willis WO, Liebhaber M. Cholinesterase activity in pregnant women and newborns. 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Microdose Flare-up Gonadotropin-releasing Hormone (GnRH) Agonist Versus GnRH Antagonist Protocols in Poor Ovarian Responders Undergoing Intracytoplasmic Sperm Injection 2 1 664 2 Background: Microdose flare-up GnRH agonist and GnRH antagonist have become more popular in the management of poor ovarian responders (POR) in recent years; however, the optimal protocol for POR patients undergoing in vitro fertilization has still been a challenge. Methods: In this observational study design, two hundred forty four poor ovarian responders were retrospectively evaluated for their response to GnRH agonist protocol (group-1, n=135) or GnRH antagonist protocol (group-2, n=109). Clinical pregnancy rate was the primary end point and was compared between the groups. Student t-test, Mann Whitney U test and x2-test were used to compare the groups. The p<0.05 was considered to show a statistically significant result. Results: The mean total gonadotropin doses were 3814±891 IU in group 1 and 3539±877 IU in group 2 (p=0.02). The number of metaphase-II oocytes (3.6±2.4 vs. 2.8±1.9, p=0.005) and implantation rates (27.8% vs. 18.8%, p=0.04) in group 1 and group 2, respectively were significantly different. The fertilization rate in group 1 and group 2 was 73% vs. 68%, respectively (p=0.5) and clinical pregnancy rate was 19.8% vs. 14.4%, respectively (p=0.13). Conclusion: The GnRH agonist microdose flare-up protocol has favorable outcomes with respect to the number of oocytes retrieved and implantation rate; nevertheless, the clinical pregnancy rate was found to be similar in comparison to GnRH antagonist protocol in poor ovarian responders. GnRH antagonist protocol appears to be promising with significantly lower gonadotropin requirement and lower treatment cost in poor ovarian responders. 163 169 Aysen A Boza Goztepe Research and Training Hospital, Kadikoy Goztepe Research and Training Hospital, Kadikoy Turkey aysenboza@hotmail.com Erbil E Cakar Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Turkey Barıs B Boza Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Turkey Murat M Api Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Turkey Semra S Kayatas Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Turkey Kenan K Sofuoglu Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Zeynep Kamil Maternity and Children Research and Training Hospital, Uskudar Turkey AgonistAntagonistFlare-upPoor responder 664.pdf Keay SD, Liversedge NH, Mathur RS, Jenkins JM. Assisted conception following poor ovarian response to gonadotrophin stimulation. Br J Obstet Gynaecol. 1997;104(5):521-7.##Scott RT, Navot D. Enhancement of ovarian responsiveness with microdoses of gonadotropin-releasing hormone ago-nist during ovulation induction for in vitro fertilization. Fertil Steril. 1994;61(5):880-5.##Surrey ES, Bower J, Hill DM, Ramsey J, Surrey MW. Clinical and endocrine effects of a microdose GnRH agonist flare regimen administered to poor responders who are undergoing in vitro fertilization. Fertil Steril. 1998;69(3):419-24.##Akman MA, Erden HF, Tosun SB, Bayazit N, Ak-soy E, Bahceci M. Comparison of agonistic flare-up-protocol and antagonistic multiple dose protocol in ovarian stimulation of poor responders: results of a prospective randomized trial. Hum Reprod. 2001; 16(5):868-70.##Fasouliotis SJ, Laufer N, Sabbagh-Ehrlich S, Lewin A, Hurwitz A, Simon A. Gonadotropin-releasing hor-mone (GnRH)-antagonist versus GnRH-agonist in ovarian stimulation of poor responders undergoing IVF. J Assist Reprod Genet. 2003;20(11):455-60.##Leondires MP, Escalpes M, Segars JH, Scott RT Jr, Miller BT. Microdose follicular phase gonadotropin-releasing hor-mone agonist (GnRH-a) compared with luteal phase GnRH-a for ovarian stimulation at in vitro fertilization. Fertil Steril. 1999;72(6):1018-23.##Akman MA, Erden HF, Tosun SB, Bayazit N, Ak-soy E, Bahceci M. Addition of GnRH antagonist in cycles of poor responders undergoing IVF. Hum Reprod. 2000;15(10):2145-7.##Demirol A, Gurgan T. Comparison of microdose flare-up and antagonist multiple-dose protocols for poor-responder pa-tients: a randomized study. Fertil Steril. 2009;92(2):481-5.##Kahraman K, Berker B, Atabekoglu CS, Sonmezer M, Cetinkaya E, Aytac R, et al. Microdose gonadotropin-releasing hormone agonist flare-up protocol versus multiple dose gonadotropin-releasing hormone antagonist protocol in poor responders undergoing intracytoplasmic sperm injection-embryo transfer cycle. Fertil Steril. 2009;91(6):2437-44.##Zakia Mahdy Ibrahim, Heba Younes Mohamed Youssef, Magda Moustafa Elbialy, Mohamed Mou-selhy Farrag. Micro-dose flare-up gonadotrophin-releasing hormone (GnRH) agonist vs. flexible go-nadotrophin-releasing hormone (GnRH) antagonist protocol in patient with poor ovarian reserve. Middle East Fertil Soc J. 2011;16(4):272-7.##Patrizio P, Vaiarelli A, Levi Setti PE, Tobler KJ, Shoham G, Leong M, et al. How to define, diagnose and treat poor responders? Responses from a worldwide survey of IVF clinics. Reprod Biomed Online. 2015;30(6):581-92.##Toth TL, Awwad JT, Veeck LL, Jones HW Jr, Mua-sher SJ. Suppression and flare regimens of gonadotropin-releasing hormone agonist. Use in women with different basal gonadotropin values in an in vitro fertilization program. J Reprod Med. 1996;41 (5):321-6.##Al-Inany H, Aboulghar M. GnRH antagonist in as-sisted reproduction: a Cochrane review. Hum Re-prod. 2002;17(4):874-85.##Chang PL, Zeitoun KM, Chan LK, Thornton MH 2nd, Sauer MV. GnRH antagonist in older IVF patients. Retrieval rates and clinical outcome. J Re-prod Med. 2002;47(4):253-8.##Craft I, Gorgy A, Hill J, Menon D, Podsiadly B. Will GnRH antagonists provide new hope for patients considered 'difficult responders' to GnRH agonist protocols? Hum Reprod. 1999;14(12):2959-62.##Marci R, Caserta D, Dolo V, Tatone C, Pavan A, Moscarini M. GnRH antagonist in IVF poor-re-sponder patients: re-sults of a randomized trial. Reprod Biomed Online. 2005;11(2):189-93.##Nikolettos N, Al-Hasani S, Felberbaum R, Demirel LC, Kupker W, Montzka P, et al. Gonadotropin-releasing hormone antagonist protocol: a novel method of ovarian stimulation in poor responders. Eur J Obstet Gynecol Reprod Biol. 2001;97(2):202-7.##Franco JG Jr, Baruffi RL, Mauri AL, Petersen CG, Felipe V, Cornicelli J, et al. GnRH agonist versus GnRH antagonist in poor ovarian responders: a meta-analysis. Reprod Biomed Online. 2006;13(5):618-27.##Pandian Z, McTavish AR, Aucott L, Hamilton MP, Bhattacharya S. Interventions for 'poor responders' to controlled ovarian hyper stimulation (COH) in in-vitro fertilisation (IVF). Cochrane Database Syst Rev. 2010;(1):CD004379.##Pu D, Wu J, Liu J. Comparisons of GnRH antagonist versus GnRH agonist protocol in poor ovarian responders under-going IVF. Hum Reprod. 2011;26 (10):2742-9.##Malmusi S, La Marca A, Giulini S, Xella S, Tag-liasacchi D, Marsella T, et al. Comparison of a go-nadotropin-releasing hormone (GnRH) antagonist and GnRH agonist flare-up regimen in poor responders undergoing ovarian stimulation. Fertil Steril. 2005;84(2):402-6.##

Sex-Reassignment Rules in Shiite Jurisprudence 2 1 637 2 Background: The "Sex-Reassignment Surgery" is a solution that besides behavioral therapy has been suggested to people suffering from gender identity disorders in recent years. In Iran, this trend has become more popular over the past years due to the inclination to reach to the goal rapidly with less effort and also the surgery has attracted many patients with the problem. Religious clerics have tried to determine the religious doctrines for this practice and as a result a group of them favor an absolute permission while others choose prohibition and some of them favor a middle path. The aim of this study was to determine the religious doctrines for Sex-Reassignment and legitimate treatment for GID. Methods: The research method was a library research based on which an investigation was done by analyzing the relevant books, articles and dissertations. Primary documents of Islamic sources (Quran and tradition) along with scientific, medical and psychological materials were used in this research. Results: In this study, the survey shows that none of the reasons have the power to deliver a definitive and religious ruling on this issue because the validity of its reasons is related to the reality of "Sex-Reassignment". Conclusion: The results demonstrate that Sex-Reassignment is prohibited and it is not authorized. In case of urgency for doing the surgery, the gender of the person should not be changed. 169 177 Fahimeh F Kalbasi-Isfahani Payame Noor University Payame Noor University Iran fah_kalbasiisfahani@ yahoo.com Mohsen M Deleer Payame Noor University Payame Noor University Iran Gender identity disordersGenderSex-Reassignment SurgerySexTranssexualism 637.pdf Mo'mene Qomi M. [Juristical-legal study of sex-reassignment and its consequences]. Islam Sci Q. 2006;1(3):6-22. Persian.##Hatami MR, Mazhabi S. 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Can a Short Term of Repeated Ejaculations Affect Seminal Parameters? 2 1 674 2 Background: The aim of the study was to assess the effect of four repeated ejaculations on the same day at two-hour intervals on conventional and functional semen parameters. Methods: Three healthy men (32±3.6 years) donated the first semen samples after 3-4 days of sexual abstinence followed by three subsequent samples on the same day at two-hour interval each. Semen samples were processed and analyzed according to the World Health Organization (WHO) 2010 guidelines. Furthermore, intracellular reactive oxygen (ROS) production, sperm DNA fragmentation and mitochondrial function were evaluated by flow cytometry. Results: An overall decreasing trend was noted in the conventional semen parameters at second, third and fourth evaluations after two hours of abstinence in comparison to first evaluation after 3-4 days of abstinence. The statistical comparison of the conventional semen parameters at fourth evaluation after 2 hr of abstinence revealed significant reduction (p<0.05) in the parameters of concentration, total sperm count and total motile sperm count at fourth evaluation. The functional parameter of intracellular ROS production showed a decreasing trend with each subsequent evaluation, the difference being significant (p<0.05) at fourth evaluation in comparison to first evaluation. An increasing trend was noted for DNA fragmentation index (DFI), although it remained within acceptable levels (<29%). The ∆Ψm high spermatozoa and the integrity of the plasma membrane remained stable throughout the evaluations. Conclusion: The findings of the present study indicate the potential use of additional semen samples with repeated ejaculations at short abstinence times in assisted reproduction procedures particularly from severe oligospermic men. 177 184 Jose Manuel JM Mayorga-Torres Reproduction Group, Department of Microbiology and Parasitology, Medical School, University of Antioquia Reproduction Group, Department of Microbiology and Parasitology, Medical School, University of Antioquia Colombia Ashok A Agarwal Center for Reproductive Medicine, Cleveland Clinic, Cleveland Center for Reproductive Medicine, Cleveland Clinic, Cleveland USA Shubhadeep Sh Roychoudhury Center for Reproductive Medicine, Cleveland Clinic, Cleveland Center for Reproductive Medicine, Cleveland Clinic, Cleveland USA Angela A Cadavid Reproduction Group, Department of Microbiology and Parasitology, Medical School, University of Antioquia Reproduction Group, Department of Microbiology and Parasitology, Medical School, University of Antioquia Colombia Walter Dario WD Cardona-Maya Reproduction Group, Department of Microbiology and Parasitology, Medical School, University of Antioquia Reproduction Group, Department of Microbiology and Parasitology, Medical School, University of Antioquia Colombia wdario.cardona@udea.edu.co DNA fragmentationRepeated ejaculationROSSemen parametersSperm quality 674.pdf Amann RP. Considerations in evaluating human spermatogenesis on the basis of total sperm per ejaculate. J Androl. 2009;30(6):626-41.##Gomendio M, Roldan ER. Sperm competition influences sperm size in mammals. Proc Biol Sci. 1991;243(1308):181-5.##delBarco-Trillo J, Tourmente M, Roldan ER. Metabolic rate limits the effect of sperm competition on mammalian spermatogenesis. PLoS One. 2013;8(9):e76510.##Joseph PN, Sharma RK, Agarwal A, Sirot LK. Men ejaculate larger volumes of semen, more motile sperm, and more quickly when exposed to images of novel women. Evol Psychol Sci. 2015;1(4):195-200.##Johnson L, Varner DD. Effect of daily spermatozoan production but not age on transit time of spermatozoa through the human epididymis. Biol Reprod. 1988;39(4):812-7.##Amann RP. A critical review of methods for evaluation of spermatogenesis from seminal characteristics. J Androl. 1981;2(1):37-58.##Turner TT. De Graaf's thread: the human epididymis. J Androl. 2008;29(3):237-50.##World Health Organization. WHO laboratory manual for the examination and processing of human semen. 5th ed. Geneva: WHO Press; 2010. 286 p.##Andersen AG, Jensen TK, Carlsen E, Jorgensen N, Andersson AM, Krarup T, et al. High frequency of sub-optimal semen quality in an unselected population of young men. Hum Reprod. 2000;15(2):366-72.##Handelsman DJ, Conway AJ, Boylan LM, Turtle JR. Testicular function in potential sperm donors: normal ranges and the effects of smoking and varicocele. Int J Androl. 1984;7(5):369-82.##MacLeod J, Wang Y. Male fertility potential in terms of semen quality: a review of the past, a study of the present. Fertil Steril. 1979;31(2):103-16.##De Jonge C, LaFromboise M, Bosmans E, Ombelet W, Cox A, Nijs M. Influence of the abstinence period on human sperm quality. Fertil Steril. 2004;82(1):57-65.##Magnus O, Tollefsrud A, Abyholm T, Purvis K. Effects of varying the abstinence period in the same individuals on sperm quality. Arch Androl. 1991;26(3):199-203.##Pellestor F, Girardet A, Andreo B. Effect of long abstinence periods on human sperm quality. Int J Fertil Menopausal Stud. 1994;39(5):278-82.##Nnatu SN, Giwa-Osagie OF, Essien EE. Effect of repeated semen ejaculation on sperm quality. Clin Exp Obstet Gynecol. 1991;18(1):39-42.##Mayorga-Torres BJ, Camargo M, Agarwal A, du Plessis SS, Cadavid AP, Cardona Maya WD. Influence of ejaculation frequency on seminal parameters. Reprod Biol Endocrinol. 2015;13:47.##Sanchez-Martin P, Sanchez-Martin F, Gonzalez-Martinez M, Gosalvez J. Increased pregnancy after reduced male abstinence. Syst Biol Reprod Med. 2013;59(5):256-60.##Marshburn PB, Giddings A, Causby S, Matthews ML, Usadi RS, Steuerwald N, et al. Influence of ejaculatory abstinence on seminal total antioxidant capacity and sperm membrane lipid peroxidation. Fertil Steril. 2014;102(3):705-10.##Kucuk T, Sozen E, Buluc B. Intrauterine insemination with double ejaculate compared with single ejaculate in male factor infertility: a pilot study. J Androl. 2008;29(4):404-7.##Marshburn PB, Alanis M, Matthews ML, Usadi R, Papadakis MH, Kullstam S, et al. A short period of ejaculatory abstinence before intrauterine insemination is associated with higher pregnancy rates. Fertil Steril. 2010;93(1):286-8.##Gosalvez J, Gonzalez-Martinez M, Lopez-Fernandez C, Fernandez JL, Sanchez-Martin P. Shorter abstinence decreases sperm deoxyribonucleic acid fragmentation in ejaculate. Fertil Steril. 2011;96(5):1083-6.##Bar-Hava I, Perri T, Ashkenazi J, Shelef M, Ben-Rafael Z, Orvieto R. The rationale for requesting a second consecutive sperm ejaculate for assisted reproductive technology. Gynecol Endocrinol. 2000;14(6):433-6.##Cardona Maya WD, Berdugo Gutierrez JA, de los Rios J, Cadavid Jaramillo AP. Functional evaluation of sperm in Colombian fertile men. Arch Esp Urol. 2007;60(7):827-31.##Mayorga-Torres BJ, Cardona-Maya W, Cadavid A, Camargo M. [Evaluation of sperm functional parameters in normozoospermic infertile individuals]. Actas Urol Esp. 2013;37(4):221-7. Spanish.##Cardona-Maya W, Berdugo J, Cadavid A. [Comparing the sperm concentration determined by the Makler and the Neubauer chambers]. Actas Urol Esp. 2008;32(4):443-5. Spanish.##Evenson DP. Sperm chromatin structure assay (SCSA®). Methods Mol Biol. 2013;927:147-64.##Evenson DP, Jost LK, Marshall D, Zinaman MJ, Clegg E, Purvis K, et al. Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod. 1999;14(4):1039-49.##Gil-Villa AM, Cardona-Maya W, Agarwal A, Sharma R, Cadavid A. Role of male factor in early recurrent embryo loss: do antioxidants have any effect? Fertil Steril. 2009;92(2):565-71.##Gil-Villa AM, Cardona-Maya W, Agarwal A, Sharma R, Cadavid A. 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Simultaneous measurement by flow cytometry of sperm cell viability and mitochondrial membrane potential related to cell motility. J Histochem Cytochem. 1982;30(3):279-80.##Garner DL, Thomas CA, Joerg HW, DeJarnette JM, Marshall CE. Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa. Biol Reprod. 1997;57(6):1401-6.##Chen LB. Mitochondrial membrane potential in living cells. Annu Rev Cell Biol. 1988;4:155-81.##Frank SA, Hurst LD. Mitochondria and male disease. Nature. 1996;383(6597):224.##Ollero M, Muino-Blanco T, Lopez-Perez MJ, Cebrian-Perez JA. Viability of ram spermatozoa in relation to the abstinence period and successive ejaculations. Int J Androl. 1996;19(5):287-92.##

Homozygosity for a Robertsonian Translocation (13q;14q) in a Phenotypically Normal 44, XX Female with a History of Recurrent Abortion and a Normal Pregnancy Outcome 2 1 677 2 Background: Robertsonian translocations are structural chromosomal abnormalities caused by fusion of two acrocentric chromosomes. In carriers of such translocations, different modes of segregations would result in the formation of either balanced (alternate segregation mode) or unbalanced (adjacent 1, adjacent 2, and 3:1 segregation modes) gametes. In addition, there is an increased risk for imprinting disorders in their offspring. Although it has been estimated that 1/1000 healthy persons carry a Robertsonian translocation, homozygosity for this type of structural chromosomal abnormality has been reported rarely. Most of reported cases are phenotypically normal but experience adverse pregnancy outcomes. Case Presentation: In this paper, a report was made on a normal female with a history of 4 consecutive first trimester fetal losses and a normal son referred to Center for Comprehensive Genetics Services,Tehran, Iran, in summer 2015. Cytogenetic analyses of proband and her infant showed 44,XX, der(13;14)(q10;q10)x2 and 45, XY, der(13;14)(q10;q10), respectively. Parents of proband have been shown to have 45,XY,der(13q;14q) and 45,XX,der(13q;14q) karyotypes, respectively. Conclusion: The present report was in agreement with the few reports of homozygosity for Robertsonian translocation which demonstrated normal phenotypes for such persons and possibility of giving birth to phenotypically normal heterozygote carriers of Robertsonian translocations. 184 188 Mohammad M Miryounesi Genomic Research Center, Shahid Beheshti University of Medical Sciences Genomic Research Center, Shahid Beheshti University of Medical Sciences Iran Mehdi M Dianatpour Department of Medical Genetics, Shiraz University of Medical Sciences Department of Medical Genetics, Shiraz University of Medical Sciences Iran Elahe E Motevaseli Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences Iran Soudeh S Ghafouri-Fard Department of Medical Genetics, Shahid Beheshti University of Medical sciences Department of Medical Genetics, Shahid Beheshti University of Medical sciences Iran s.ghafourifard@sbmu.ac.ir Genetic counselingHabitual abortionTranslocation 677.pdf Keymolen K, Van Berkel K, Vorsselmans A, Staessen C, Liebaers I. Pregnancy outcome in carriers of Robertsonian translocations. Am J Med Genet A. 2011;155A(10):2381-5.##Abdalla EM, Kholeif SF, Elshaffie RM. Homozygosity for a Robertsonian Translocation (13q; 14q) in an Otherwise Healthy 44, XY Man With a History of Repeated Fetal Losses. Lab Medicine. 2013;44(3):254-7.##Xu SQ, Tang DL, Fang K, Xia YZ, Song JP, Wang WP, et al. Analysis of meiotic segregation patterns and interchromosomal effects in sperm from a Robertsonian translocation family. Biomed Res. 2014;25(2):233-9.##Yip MY. Uniparental disomy in Robertsonian translocations: strategies for uniparental disomy testing. Transl Pediatr. 2014;3(2):98-107.##Engels H, Eggermann T, Caliebe A, Jelska A, Schubert R, Schüler HM, et al. Genetic counseling in Robertsonian translocations der(13;14): frequencies of reproductive outcomes and infertility in 101 pedigrees. Am J Med Genet A. 2008;146A(20):2611-6.##Martinez-Castro P, Ramos MC, Rey JA, Benitez J, Sanchez Cascos A. Homozygosity for a Robertsonian translocation (13q14q) in three offspring of heterozygous parents. Cytogenet Cell Genet. 1984;38(4):310-2.##Eklund A, Simola KO, Ryynänen M. Translocation t(13;14) in nine generations with a case of translocation homozygosity. Clin Genet. 1988;33(2):83-6.##Omrani MD, Gargari SS. Uniparental disomy resulting from heterozygous Robertsonian translocation (13q14q) in both parents. J Res Med Sci. 2007;12(2):100-3.##Sensi A, Cavani S, Villa N, Pomponi MG, Fogli A, Gualandi F, et al. Nonhomologous Robertsonian translocations (NHRTs) and uniparental disomy (UPD) risk: an Italian multicentric prenatal survey. Prenat Diagn. 2004;24(8):647-52.##Rockman-Greenberg C, Ray M, Evans JA, Canning N, Hamerton JL. Homozygous Robertsonian translocations in a fetus with 44 chromosomes. Hum Genet. 1982;61(3):181-4.##Rajangam S, Michaelis RC, Velagaleti GV, Lincoln S, Hegde S, Lewin S, et al. Down syndrome with biparental inheritance of der(14q21q) and maternally derived trisomy 21: confirmation by fluorescent in situ hybridization and microsatellite polymorphism analysis. Am J Med Genet. 1997;70(1):43-7.##Neu RL, Valentine FA, Gardner LI. Segregation of a t(14q22q) chromosome in a large kindred. Clin Genet. 1975;8(1):30-6.##Wang B, Xia Y, Song J, Wang W, Tang Y. Case Report: Potential speciation in humans involving robertsonian translocations. Biomed Res. 2013;24(1):171-4.##Morgan R, Bixenman H, Hecht F. Human chromosome variation with two Robertsonian translocations. Hum Genet. 1985;69(2):178-80.##Simons A, Shaffer LG, Hastings RJ. Cytogenetic Nomenclature: Changes in the ISCN 2013 Compared to the 2009 Edition. Cytogenet Genome Res. 2013;141(1):1-6.##O'Neill ID. Homozygosity for constitutional chromosomal rearrangements: a systematic review with reference to origin, ascertainment and phenotype. J Hum Genet. 2010;55(9):559-64.##Franssen MT, Musters AM, van der Veen F, Repping S, Leschot NJ, Bossuyt PM, et al. Reproductive outcome after PGD in couples with recurrent miscarriage carrying a structural chromosome abnormality: a systematic review. Hum Reprod Update. 2011;17(4):467-75.##

A Case of Primary Hypogonadism with Features of Albright’s Syndrome 2 1 671 2 Background: McCune Albright syndrome is rare with an estimated prevalence of 1 in 100,000 to 1 in 1,000,000 persons. The classical clinical triad consists of fibrous dysplasia of the bone, café-au-lait skin spots and precocious puberty. However, in rare cases, there may be primary hypogonadism and amenorrhea. Case Presentation: An eighteen-year-old female presented with amenorrhea. She had a short stature, round face, thick neck, and short fourth metacarpals and metatarsals. The secondary sexual characters were absent. Serum calcium, phosphorus and parathyroid concentrations were normal, but gonadotropin hormones were very low. X-ray examination revealed short fourth and fifth metacarpals, short left metatarsal, and short fibula. Conclusion: These local bony abnormalities along with the biochemical findings helped us to diagnose this case as an unusual presentation of primary hypogonadism with features of McCune Albright’s syndrome where there was amenorrhea rather than preocious puberty. 188 191 Moushumi M Lodh Department of Biochemistry, IQ City Medical College and Narayana Multispeciality Hospital, Durgapur Department of Biochemistry, IQ City Medical College and Narayana Multispeciality Hospital, Durgapur India drmoushumilodh@gmail.com Rajarshi R Mukhopadhyay Department of Endocrinology, Mission Hospital, Durgapur Department of Endocrinology, Mission Hospital, Durgapur India Absent secondary sexual charactersBrachydactylyFibrous dysplasiaHypogonadismMcCune Albright syndromePseudohypoparathyroidism 671.pdf Pereda A, Garin I, Garcia-Barcina M, Gener B, Beristain E, Ibanez AM, et al. Brachydactyly E: isolated or as a feature of a syndrome. Orphanet J Rare Dis. 2013;8:141.##Simon A, Koppeschaar HP, Roijers JF, Hoppener JW, Lips CJ. Pseudohypoparathyroidism type Ia. Albright hereditary osteodystrophy: a model for research on G protein-coupled receptors and genomic imprinting. Neth J Med. 2000;56(3):100-9.##Cho SY, Yoon YA, Ki CS, Huh HJ, Yoo HW, Lee BH, et al. Clinical characterization and molecular classification of 12 Korean patients with pseudohypoparathyroidism and pseudopseudohypoparathyroidism. Exp Clin Endocrinol Diabe-tes. 2013;121(9):539-45.##Archibald RM, Finby N, De Vito F. Endocrine significance of short metacarpals. J Clin Endocrinol Metab. 1959;19:1312-22.##Kirkos JM. Idiopathic symmetrical shortening of the fourth and fifth metacarpal and metatarsal bilaterally. A case report. Acta Orthop Belg. 1999;65(4):532-5.##Gandhi P, Gupta RC, Chaudhary HR, Jain R, Gupta RK. Brachydactyly. J Indian Acad Clin Med. 2002;3(1):89-90.##Valizadeh N, Mehdizadeh A, Nazarbaghi S. Short fourth and fifth metacarpals in a case of idiopathic primary hypoparathyroidism. Indian J Endocrinol Metab. 2013;17(5):924-6.##Tzaveas A, Paraskevas G, Gekas C, Vrettakos A, Antoniou K, Spyridakis I. Anatomical variation of co-existence of 4th and 5th short metacarpal bones, sesamoid ossicles and exostoses of ulna and radius in the same hand: a case report. Cases J. 2008;1(1):281.##Wu YL, Hwang DY, Hsiao HP, Ting WH, Huang CY, Tsai WY, et al. Mutations in pseudohypoparathyroidism 1a and pseudopseudohypoparathyroidism in ethnic Chinese. PLoS One. 2014;9(3):e90640.##Kottler ML. [Paternal GNAS mutations: Which phenotypes? What genetic counseling?]. Ann Endocrinol (Paris). 2015;76(2):105-9. French.##Turan S, Thiele S, Tafaj O, Brix B, Atay Z, Abali S, et al. Evidence of hormone resistance in a pseudo-pseudohypoparathyroidism patient with a novel paternal mutation in GNAS. Bone. 2015;71:53-7.##