Surrogacy, an Excellent Opportunity for Women with More Threats 2 1 2 Surrogacy is one of the assisted reproduction methods which have a very long history. The first traditional surrogacy was performed in about 2000 years before the birth of Christ as mentioned in the Holy Qur'an of Muslims and the Christians Bible; Sarah was the wife of Abraham prophet and she was sterile which made her unable to conceive. Hajar was a female servant of Sarah who asked to carry a child for Abraham prophet. Afterward, Hajar delivered a son for Sarah named Ishmael. This is the traditional, genetic or partial form of surrogacy; it has already been performed by artificial insemination of the surrogate with sperm of intended father and the usage of this form of surrogacy has been greatly reduced. This surrogate mother not only carries the embryo till full term delivery but also provides the eggs that make her a genetic parent. Moreover, the most popular form is gestational, or IVF surrogacy in which an embryo from the intended parents, or from a donated oocyte or sperm, is transferred to the surrogate’s uterus. In gestational surrogacy, the mother who carries and delivers the child (the gestational carrier) has no genetic relation with the child (1). Surrogacy is used for approximately 2% of embryo transfer cycles in USA. The indications for surrogacy include absence of the uterus, recurrent pregnancy loss, repeated IVF failure, poor obstetric history, contraindication of pregnancy, excessive maternal risk, etc. Surrogacy improves pregnancy and live birth rate compared to other ART cycles without surrogacy. The best results are observed in surrogate candidate due to uterine factor infertility (2). Most of the studies reported better prenatal outcomes in surrogacy including hypertension, preeclampsia, gestational diabetes, placenta previa, preterm labor and lower birth weight in comparison to IVF cycles of their own uterus. However, when these adverse outcomes in surrogate mothers were compared with the same surrogate mothers who became pregnant through spontaneous conception, IVF manifested most of the above adverse outcomes due to laboratory manipulation of gametes and embryos and uterine milieu changes in stimulation cycles (3). The surrogacy needs IVF facilities without considering infertility etiology, so gestational carriers experienced embryo transfer, pregnancy, successful ongoing pregnancy and delivery. Therefore, precise medical, psychological and social assessments are necessary to succeed in gestation and delivery. Health status of carriers during gestation could affect future health and wellbeing of the child. The history profile of surrogate should contain at least one uncomplicated pregnancy, although not more than 5 deliveries or 3 caesarean sections (4). In spite of most benefits of surrogacy and also considering the fact that it may be the only option for many couples to have a child from their own gametes, surrogacy is prohibited in many European countries such as Germany, Sweden, Norway, and Italy. Due to financial payments and maintaining the dignity and rights of individuals, any payment to surrogate is officially forbidden in many other countries and only compensation of pregnancy-related costs is accepted in Australia, UK, and in many states of USA. Commercial surrogacy is accepted in India, Ukraine, USA and Middle East countries. One of the remarkable cases is accompaniment of surrogacy with oocyte donation cycles, so that 46% of surrogacy cycles in the USA involve donor eggs. Oocyte donation by carrier for intended parents is prohibited in almost all countries (4). All of these strict regulations have been set up to protect the rights of the parties involved in this sensitive and complex process, including child, surrogate and intended parents. Inappropriate practice of surrogacy in cases without medical indication such as celebrities, businesswomen and female politicians for the birth of their child in developed countries is another concern of the field. Cross-border commercial surrogacy in poor developing countries such as India, Nepal, Thailand, and Mexico is another concern. Therefore, in spite of the importance and critical role of surrogacy in assisted reproduction technologies and its existence as the only option for many couples in childbearing age, it requires more consideration and more strict regulation at global level compared to current status due to misuse of the practice in unnecessary cases. 063 64 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 50053.pdf Söderström-Anttila V, Wennerholm UB, Loft A, Pinborg A, Aittomäki K, Romundstad LB, et al. Surrogacy: outcomes for surrogate mothers, children and the resulting families-a systematic review. Hum Reprod Update. 2016;22(2):260-76.##Murugappan G, Farland LV, Missmer SA, Correia KF, Anchan RM, Ginsburg ES. Gestational carrier in assisted reproductive technology. Fertil Steril. 2018;109(3):420-8.##Woo I, Hindoyan R, Landay M, Ho J, Ingles SA, McGinnis LK, et al. Perinatal outcomes after natural conception versus in vitro fertilization (IVF) in gestational surrogates: a model to evaluate IVF treatment versus maternal effects. Fertil Steril. 2017;108(6):993-8.##Simopoulou M, Sfakianoudis K, Tsioulou P, Rapani A, Anifandis G, Pantou A, et al. Risks in surrogacy considering the embryo: from the preimplantation to the gestational and neonatal period. 2018;2018:6287507.##

Metabolomics of Male Infertility: A New Tool for Diagnostic Tests 2 1 2 Infertility is a major health issue worldwide. Males and females contribute equally to this problem. Diagnostic semen analysis fails to identify 50% of male infertility disorders. In this regard, metabolomics as a new field of omics has been suggested to have the potential of solving and diagnosis of the male infertility problems. Metabolome has a history of around 20 years. However, there are only limited metabolomics studies carried out regarding male infertility. In this review, the current metabolomics researches that have been done in infertile men were reviewed. Based on our own results, using human seminal plasma for metabolomics studies is highly recommended to find potential biomarkers and developing diagnosis tests for detection of main deficiencies in infertile men. 064 70 Bahareh B Mehrparavar Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Arash A Minai-Tehrani Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Babak B Arjmand Cell Therapy and Regenerative Medicine Research Center, Endocrinology and Metabolism Molecular Cellular Sciences Institute, Tehran University of Medical Sciences Cell Therapy and Regenerative Medicine Research Center, Endocrinology and Metabolism Molecular Cellular Sciences Institute, Tehran University of Medical Sciences Iran Kambiz K Gilany کامبیز گیلانی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran k.gilany@avicenna.ac.ir BiomarkersHuman seminal plasmaMale infertilityMetabolomics 50050.pdf Hamada AJ, Montgomery B, Agarwal A. Male infertility: a critical review of pharmacologic management. Expert Opin Pharmacother. 2012;13(17):2511-31.##Kovac JR, Pastuszak AW, Lamb DJ. The use of genomics, proteomics, and metabolomics in identifying biomarkers of male infertility. Fertil Steril. 2013;99(4):998-1007.##Minai‐Tehrani A, Jafarzadeh N, Gilany K. Metabolomics: a state‐of‐the‐art technology for better understanding of male infertility. Andrologia. 2016;48(6):609-16.##WHO. WHO laboratory manual for the examination and processing of human semen. 5th ed. Switzerland: World Health Organization; 2010. 287 p.##Strimbu K, Tavel JA. What are biomarkers? Curr Opin HIV AIDS. 2010;5(6):463-6.##Fiehn O. Metabolomics--the link between genotypes and phenotypes. Plant Mol Biol. 2002;48(1-2):155-71.##Goodacre R. Metabolomics of a superorganism. J Nutr. 2007;137(1 Suppl):259S-66S.##Sue T, Obolonkin V, Griffiths H, Villas-Boas SG. An exometabolomics approach to monitoring microbial contamination in microalgal fermentation processes by using metabolic footprint analysis. Appl Environ Microbiol. 2011;77(21):7605-10.##Hamamah S, Seguin F, Barthelemy C, Akoka S, Le Pape A, Lansac J, et al. 1H nuclear magnetic resonance studies of seminal plasma from fertile and infertile men. J Reprod Fertil. 1993;97(1):51-5.##Gilany K, Pouracil RS, Sadeghi MR. Fourier transform infrared spectroscopy: a potential technique for noninvasive detection of spermatogenesis. Avicenna J Med Biotechnol. 2014;6(1):47-52.##da Silva BF, Del Giudice PT, Spaine DM, Gozzo FC, Turco EL, Bertolla RP. Metabolomics of male infertility: characterization of seminal plasma lipid fingerprints in men with spinal cord injury. Fertil Steril. 2011;96(3):S233.##Jayaraman V, Ghosh S, Sengupta A, Srivastava S, Sonawat HM, Narayan PK. Identification of biochemical differences between different forms of male infertility by nuclear magnetic resonance (NMR) spectroscopy. J Assist Reprod Genet. 2014;31(9):1195-204.##Gilany K, Moazeni-Pourasil RS, Jafarzadeh N, Savadi-Shiraz E. Metabolomics fingerprinting of the human seminal plasma of asthenozoospermic patients. Mol Reprod Dev. 2014;81(1):84-6.##Zhang X, Diao R, Zhu X, Li Z, Cai Z. Metabolic characterization of asthenozoospermia using nontargeted seminal plasma metabolomics. Clin Chim Acta. 2015;450:254-61.##Chen X, Hu C, Dai J, Chen L. Metabolomics analysis of seminal plasma in infertile males with kidney-yang deficiency: a preliminary study. Evid Based Complement Alternat Med. 2015;2015:892930.##Gilany K, Mani-Varnosfaderani A, Minai-Tehrani A, Mirzajani F, Ghassempour A, Sadeghi MR, et al. Untargeted metabolomic profiling of seminal plasma of non-obstructive azoospermia men: a non-invasive detection of spermatogenesis. Biomed Chromatogr. 2017;31(8):e3931.##Gilany K, Jafarzadeh N, Mani-Varnosfaderani A, Minai-Tehrani A, Sadeghi MR, Darbandi M, et al. Metabolic fingerprinting of seminal plasma from Non-obstructive azoospermia patients: positive versus negative sperm retrieval. J Reprod Infertil. 2018;19(2):109-14.##Zhang Z, Zhang Y, Liu C, Zhao M, Yang Y, Wu H, et al. Serum metabolomic profiling identifies characterization of non-obstructive azoospermic men. Int J Mol Sci. 2017;18(2): pii: E238.##Deepinder F, Chowdary HT, Agarwal A. Role of metabolomic analysis of biomarkers in the management of male infertility. Expert Rev Mol Diagn. 2007;7(4):351-8.##Sharma RK, Agarwal A. Role of reactive oxygen species in male infertility. Urology. 1996;48(6):835-50.##Jafarzadeh N, Mani-Varnosfaderani A, Minai-Tehrani A, Savadi-Shiraz E, Sadeghi MR, Gilany K. Metabolomics fingerprinting of seminal plasma from unexplained infertile men: a need for novel diagnostic biomarkers. Mol Reprod Dev. 2015;82(3):150.##Qiao S, Wu W, Chen M, Tang Q, Xia Y, Jia W, et al. Seminal plasma metabolomics approach for the diagnosis of unexplained male infertility. PloS One. 2017;12(8):e0181115.##Zhang J, Huang Z, Chen M, Xia Y, Martin FL, Hang W, et al. Urinary metabolome identifies signatures of oligozoospermic infertile men. Fertil Steril. 2014;102(1):44-53.e12.##Zhang J, Mu X, Xia Y, Martin FL, Hang W, Liu L, et al. Metabolomic analysis reveals a unique urinary pattern in normozoospermic infertile men. J Proteome Res. 2014;13(6):3088-99.##Zhou X, Wang Y, Yun Y, Xia Z, Lu H, Luo J, et al. A potential tool for diagnosis of male infertility: Plasma metabolomics based on GC-MS. Talanta. 2016;147:82-9.##Zheng P, Wang Y, Lu H, Zhou X, Tang T, Fan R, et al. Plasma metabolomics analysis based on GC-MS in infertile males with Kidney-Yang deficiency syndrome. Evid Based Complement Alternate Med. 2017;2017:6270195.##Paiva C, Amaral A, Rodriguez M, Canyellas N, Correig X, Ballescà JL, et al. Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance (1H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS). Andrology. 2015;3(3):496-505.##Gilany K, Minai-Tehrani A, Savadi-Shiraz E, Rezadoost H, Lakpour N. Exploring the human seminal plasma proteome: an unexplored gold mine of biomarker for male infertility and male reproduction disorder. J Reprod Infertil. 2015;16(2):61-71.##Wishart DS, Jewison T, Guo AC, Wilson M, Knox C, Liu Y, et al. HMDB 3.0--the human metabolome database in 2013. Nucleic Acids Res. 2013;41(Database issue):D801-7.##

Expression Analysis of the CRISP2, CATSPER1, PATE1 and SEMG1 in the Sperm of Men with Idiopathic Asthenozoospermia 2 1 2 Background: The purpose of this study was to analyze the expression level of CRISP2, CATSPER1, PATE1 and SEMG1 genes in the sperm of men with asthenozoospermia (AZS). AZS is a cause of infertility in men in which the motility of the sperm is reduced. So far, a few genes have been associated with AZS; however, in most of the cases, its molecular etiology is unclear. Methods: A total of 35 subjects with idiopathic AZS and 35 fertile and healthy men as control were included. In study after total RNA extraction and cDNA synthesis, relative quantification was performed. B2M was used as the normalizer gene and fold change was calculated by 2−ΔΔCtmethod. Mann-Whitney test was used to compare the expression levels between the case and control groups with significance level of p<0.05. Results: Our results showed that CRISP2 (p=0.03) and SEMG1 (p=0.03) were significantly down- and up-regulated in AZS men respectively compared to the controls. But CATSPER1 and PATE1 did not show significant changes. Conclusion: Down-regulation of CRISP2 and up-regulation of SEMG1 were associated with AZS, which could be suggested as the potential candidate genes for the development of a diagnostic marker or potentially for more studies for treatment of AZS. 070 76 Zohreh Z Heidary Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Majid M Zaki-Dizaji Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Kioomars K Salimi Nejad کیومرث سلیمی‌نژاد Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Hamid Reza HR Khorram Khorshid حمیدرضا خرم‌خورشید Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran hrkk1@uswr.ac.ir, h.khorramkhorshid@avicenna.ac.ir AsthenozoospermiaCATSPER1CRISP2Gene expressionMale infertilityPATE1Real-time PCRSEMG1Sperm motility 50048.pdf Coutton C, Fissore RA, Palermo GD, Stouffs K, Toure A. Male infertility: genetics, mechanism, and therapies. Biomed Res Int. 2016;2016:7372362.##WHO Organization. WHO laboratory manual for the examination and processing of human semen. 5th ed. Geneva: World health organization. 2010. 286 p.##Ferlin A, Raicu F, Gatta V, Zuccarello D, Palka G, Foresta C. Male infertility: role of genetic background. Reprod Biomed Online. 2007;14(6):734-45.##Tüttelmann F, Röpke A. Genetics of Male Infertility. In: Simoni M, Huhtaniemi IT, editors. Endocrinology of the Testis and Male Reproduction. Cham: Springer International Publishing; 2017. p. 1029-49.##Miyamoto T, Minase G, Shin T, Ueda H, Okada H, Sengoku K. Human male infertility and its genetic causes. Reprod Med Biol. 2017;16(2):81-8.##Hargreave TB, Elliott D. Genetics and male infertility. In: Schill WB, Comhaire FH, Hargreave TB, editors. Andrology for the clinician. Berlin, Heidelberg: Springer; 2006. p. 462-80.##Neto FT, Bach PV, Najari BB, Li PS, Goldstein M. Genetics of male infertility. Curr Urol Rep. 2016;17(10):70.##Walsh TJ, Pera RR, Turek PJ. The genetics of male infertility. Semin Reprod Med. 2009;27(2):124-36.##Asero P, Calogero AE, Condorelli RA, Mongioi L, Vicari E, Lanzafame F, et al. Relevance of genetic investigation in male infertility. J Endocrinol Invest. 2014;37(5):415-27.##Curi SM, Ariagno JI, Chenlo PH, Mendeluk GR, Pugliese MN, Sardi Segovia LM, et al. Asthenozoospermia: analysis of a large population. Arch Androl. 2003;49(5):343-9.##Heidary Z, Saliminejad K, Zaki-Dizaji M, Khorram Khorshid HR. Genetic aspects of idiopathic asthenozoospermia as a cause of male infertility. Hum Fertil (Camb). 2018:1-10.##Luo S, Jia J, Hu H, Ma W, Jiao Y, Dong J. A Chinese herbal decoction can increase the intracellular Ca2 concentration and CatSper1 expression in mouse sperm tails. Mol Med Rep. 2013;7(1):195-200.##Zhou JH, Zhou QZ, Lyu XM, Zhu T, Chen ZJ, Chen MK, et al. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia. Biol Reprod. 2015;92(1):28.##Liu FJ, Liu X, Han JL, Wang YW, Jin SH, Liu XX, et al. Aged men share the sperm protein PATE1 defect with young asthenozoospermia patients. Hum Reprod. 2015;30(4):861-9.##Yu Q, Zhou Q, Wei Q, Li J, Feng C, Mao X. SEMG1 may be the candidate gene for idiopathic asthenozoospermia. Andrologia. 2014;46(2):158-66.##Mizuki N, Sarapata DE, Garcia-Sanz JA, Kasahara M. The mouse male germ cell-specific gene Tpx-1: molecular structure, mode of expression in spermatogenesis, and sequence similarity to two non-mammalian genes. Mamm Genome. 1992;3(5):274-80.##Kratzschmar J, Haendler B, Eberspaecher U, Roosterman D, Donner P, Schleuning WD. The human cysteine-rich secretory protein (CRISP) family. Primary structure and tissue distribution of CRISP-1, CRISP-2 and CRISP-3. Eur J Biochem. 1996;236(3):827-36.##Haendler B, Habenicht UF, Schwidetzky U, Schuttke I, Schleuning WD. Differential androgen regulation of the murine genes for cysteine-rich secretory proteins (CRISP). Eur J Biochem. 1997;250(2):440-6.##O'Bryan MK, Sebire K, Meinhardt A, Edgar K, Keah HH, Hearn MT, et al. Tpx-1 is a component of the outer dense fibers and acrosome of rat spermatozoa. Mol Reprod Dev. 2001;58(1):116-25.##Busso D, Cohen DJ, Hayashi M, Kasahara M, Cuasnicu PS. Human testicular protein TPX1/CRISP-2: localization in spermatozoa, fate after capacitation and relevance for gamete interaction. Mol Hum Reprod. 2005;11(4):299-305.##Gibbs GM, Bianco DM, Jamsai D, Herlihy A, Ristevski S, Aitken RJ, et al. Cysteine-rich secretory protein 2 binds to mitogen-activated protein kinase kinase kinase 11 in mouse sperm. Biol Reprod. 2007;77(1):108-14.##Jamsai D, Bianco DM, Smith SJ, Merriner DJ, Ly-Huynh JD, Herlihy A, et al. Characterization of gametogenetin 1 (GGN1) and its potential role in male fertility through the interaction with the ion channel regulator, cysteine-rich secretory protein 2 (CRISP2) in the sperm tail. Reproduction. 2008;135(6):751-9.##Jamsai D, Rijal S, Bianco DM, O'Connor AE, Merriner DJ, Smith SJ, et al. A novel protein, sperm head and tail associated protein (SHTAP), interacts with cysteine-rich secretory protein 2 (CRISP2) during spermatogenesis in the mouse. Biol Cell. 2009;102(2):93-106.##Nimlamool W, Bean BS, Lowe-Krentz LJ. Human sperm CRISP2 is released from the acrosome during the acrosome reaction and re-associates at the equatorial segment. Mol Reprod Dev. 2013;80(6):488-502.##Harper CV, Barratt CL, Publicover SJ. Stimulation of human spermatozoa with progesterone gradients to simulate approach to the oocyte. Induction of [Ca(2 )](i) oscillations and cyclical transitions in flagellar beating. J Biol Chem. 2004;279(44):46315-25.##Da Ros VG, Munoz MW, Battistone MA, Brukman NG, Carvajal G, Curci L, et al. From the epididymis to the egg: participation of CRISP proteins in mammalian fertilization. Asian J Androl. 2015;17(5):711-5.##Ren D, Navarro B, Perez G, Jackson AC, Hsu S, Shi Q, et al. A sperm ion channel required for sperm motility and male fertility. Nature. 2001;413(6856):603-9.##Sanchez-Cardenas C, Montoya F, Navarrete FA, Hernandez-Cruz A, Corkidi G, Visconti PE, et al. Intracellular Ca2 threshold reversibly switches flagellar beat off and on. Biol Reprod. 2018;99(5):1010-21.##Ueda Y, Yamaguchi R, Ikawa M, Okabe M, Morii E, Maeda Y, et al. PGAP1 knock-out mice show otocephaly and male infertility. J Biol Chem. 2007;282(42):30373-80.##Jensen-Seaman MI, Li WH. Evolution of the hominoid semenogelin genes, the major proteins of ejaculated semen. J Mol Evol. 2003;57(3):261-70.##Lundwall A. A locus on chromosome 20 encompassing genes that are highly expressed in the epididymis. Asian J Androl. 2007;9(4):540-4.##Zhou QZ, Guo XB, Zhang WS, Zhou JH, Yang C, Bian J, et al. Expressions of miR-525-3p and its target gene SEMG1 in the spermatozoa of patients with asthenozoospermia. Andrology. 2018.##Moreno LI, Tate CM, Knott EL, McDaniel JE, Rogers SS, Koons BW, et al. Determination of an effective housekeeping gene for the quantification of mRNA for forensic applications. J Forensic Sci. 2012;57(4):1051-8.##Cao X, Cui Y, Zhang X, Lou J, Zhou J, Bei H, et al. Proteomic profile of human spermatozoa in healthy and asthenozoospermic individuals. Reprod Biol Endocrinol. 2018;16(1):16.##Zhou JH, Zhou QZ, Yang JK, Lyu XM, Bian J, Guo WB, et al. MicroRNA-27a-mediated repression of cysteine-rich secretory protein 2 translation in asthenoteratozoospermic patients. Asian J Androl. 2016;19(5):591-5.##Li Y, Li M, Liu Y, Song G, Liu N. A microarray for microRNA profiling in spermatozoa from adult men living in an environmentally polluted site. Bull Environ Contam Toxicol. 2012;89(6):1111-4.##Wang H, Zhou Z, Xu M, Li J, Xiao J, Xu ZY, et al. A spermatogenesis-related gene expression profile in human spermatozoa and its potential clinical applications. J Mol Med (Berl). 2004;82(5):317-24.##Zhou J, Xue K, Chen M, Zhou Q, Yang J, Bian J, et al. [Expression of cysteine-rich secretory protein 2 in patients with asthenozoospermia and its clinical significance]. Nan Fang Yi Ke Da Xue Xue Bao. 2014;34(10):1528-33. Chinese.##Jing XW, Xing RW, Zhou QZ, Yu QF, Guo WB, Chen SM, et al. [Expressions of cysteine-rich secretory protein 2 in asthenospermia]. Zhonghua Nan Ke Xue. 2011;17(3):203-7. Chinese.##Du Y, Huang X, Li J, Hu Y, Zhou Z, Sha J. Human testis specific protein 1 expression in human spermatogenesis and involvement in the pathogenesis of male infertility. Fertil Steril. 2006;85(6):1852-4.##Jamsai D, Reilly A, Smith SJ, Gibbs GM, Baker HW, McLachlan RI, et al. Polymorphisms in the human cysteine-rich secretory protein 2 (CRISP2) gene in Australian men. Hum Reprod. 2008;23(9):2151-9.##Agarwal A, Sharma R, Durairajanayagam D, Ayaz A, Cui Z, Willard B, et al. Major protein alterations in spermatozoa from infertile men with unilateral varicocele. Reprod Biol Endocrinol. 2015;13:8.##Martinez-Heredia J, de Mateo S, Vidal-Taboada JM, Ballesca JL, Oliva R. Identification of proteomic differences in asthenozoospermic sperm samples. Hum Reprod. 2008;23(4):783-91.##Terai K, Yoshida K, Yoshiike M, Fujime M, Iwamoto T. Association of seminal plasma motility inhibitors/semenogelins with sperm in asthenozoospermia-infertile men. Urol Int. 2010;85(2):209-15.##Legare C, Droit A, Fournier F, Bourassa S, Force A, Cloutier F, et al. Investigation of male infertility using quantitative comparative proteomics. J Proteome Res. 2014;13(12):5403-14.##Robert M, Gagnon C. Semenogelin I: a coagulum forming, multifunctional seminal vesicle protein. Cell Mol Life Sci. 1999;55(6-7):944-60.##Marques PI, Fonseca F, Carvalho AS, Puente DA, Damião I, Almeida V, et al. Sequence variation at KLK and WFDC clusters and its association to semen hyperviscosity and other male infertility phenotypes. Hum Reprod. 2016;31(12):2881-91.##Tamburrino L, Marchiani S, Vicini E, Muciaccia B, Cambi M, Pellegrini S, et al. Quantification of CatSper1 expression in human spermatozoa and relation to functional parameters. Hum Reprod. 2015;30(7):1532-44.##Shu F, Zhou X, Li F, Lu D, Lei B, Li Q, et al. Analysis of the correlation of CATSPER single nucleotide polymorphisms (SNPs) with idiopathic asthenospermia. J Assist Reprod Genet. 2015;32(11):1643-9.##Wang YN, Wang B, Liang M, Han CY, Zhang B, Cai J, et al. Down-regulation of CatSper1 channel in epididymal spermatozoa contributes to the pathogenesis of asthenozoospermia, whereas up-regulation of the channel by Sheng-Jing-San treatment improves the sperm motility of asthenozoospermia in rats. Fertil Steril. 2013;99(2):579-87.##Visser L, Westerveld GH, Xie F, van Daalen SK, van der Veen F, Lombardi MP, et al. A comprehensive gene mutation screen in men with asthenozoospermia. Fertil Steril. 2011;95(3):1020-4.e1-9.##Avenarius MR, Hildebrand MS, Zhang Y, Meyer NC, Smith LL, Kahrizi K, et al. Human male infertility caused by mutations in the CATSPER1 channel protein. Am J Hum Genet. 2009;84(4):505-10.##Zhang S, Wang QM, Ding XP, Wang T, Mu XM, Chen ZY. Association of polymorphisms in PATE1 gene with idiopathic asthenozoospermia in Sichuan, China. J Reprod Immunol. 2016;118:54-60.##

Recurrent Miscarriage and Implantation Failure of Unknown Cause Studied by a Panel of Thrombophilia Conditions: Increased Frequency of FXIII Val34Leu Polymorphism 2 1 2 Background: The role of acquired thrombophilia has been accepted as an etiology of recurrent miscarriage (RM); however, the contribution of specific inherited thrombophilic genes to this disorder has remained controversial. An increased incidence of RM has been suggested in women with inherited thrombophilia. Methods: In this prospective study, assisted women with RM or repeated implant failure (RIF) were subjected to Thromboincode analysis, in order to identify 12 genetic variants for Factor V Leiden, Factor V Hong Kong, Factor V Cambridge, FII, FXIII, FXII, and A1 carriers. Patients included in this study were separated in RM cases (n=43), RIF cases (n=36) and RIF+RM (n=76). As a control group, patients undergoing IVF treatment (n=34) were used and a previously described 249 cases population as a representative sample of Spanish population were selected. Level of statistical significance was p<0.05 and groups were compared by Fisher test, except for age that was compared by t-test. Results: Regarding FXIII, higher values were observed in RM (69.76%), RIF (70%) and in RM+RIF (68.42%) group when compared with our control group (52.94%) and general Spanish population (56.5%), but these differences were statistically significant only in RIF group (p=0.043, p=0.01). Conclusion: Concerning our findings, both RM and RIF patients had a very similar panel of thrombophilia polymorphisms, suggesting that, in both, thrombophilia might have an important contribution. High frequency of Val34Leu polymorphism in RM/RIF presumably speaks in favor of a multifactorial RM genesis, wherean altered thrombophilia status plays a role. 076 83 Maria M Diaz-Nuñez Human Reproduction Unit, Cruces University Hospital, Biocruces Human Reproduction Unit, Cruces University Hospital, Biocruces Spain maria.diaznunez@osakidetza.eus Aintzane A Rabanal Human Reproduction Unit, Cruces University Hospital, Biocruces Human Reproduction Unit, Cruces University Hospital, Biocruces Spain Antonia A Expósito Human Reproduction Unit, Cruces University Hospital, Biocruces Human Reproduction Unit, Cruces University Hospital, Biocruces Spain Marcos M Ferrando IVI Bilbao IVI Bilbao Spain Fernando F Quintana IVI Bilbao IVI Bilbao Spain Jose JM Soria Unit of Genomics of Complex Diseases, Sant Pau Institute of Biomedical Research (IIB-Sant Pau) Unit of Genomics of Complex Diseases, Sant Pau Institute of Biomedical Research (IIB-Sant Pau) Spain Roberto R Matorras Human Reproduction Unit, Cruces University Hospital, Biocruces Human Reproduction Unit, Cruces University Hospital, Biocruces Spain Factor XIIImplant failureIVFMiscarriageThrombophilia 50052.pdf Jauniaux E, Farquharson RG, Christiansen OB, Exalto N. Evidence-based guidelines for the investigation and medical treatment of recurrent miscarriage. Hum Reprod. 2006;21:2216-22.##Rai R, Regan L. Recurrent miscarriage. Lancet. 2006;368(9535):601-11.##Rubio C, Pehlivan T, Rodrigo L, Simón C, Remohí J, Pellicer A. Embryo aneuploidy screening for unexplained recurrent miscarriage: a minireview. Am J Reprod Immunol. 2005;53(4):159-65.##Bradley LA, Palomaki GR, Bienstock J, Varga E, Scott JA. Can factor V leiden and prothrombin G20210A testing in women with recurrent pregnancy loss result in impoved pregnancy outcomes): Results from a targeted evidence-based review. Genet Med. 2012;14(1):39-50.##Coulam CB, Jeyendran RS, Fishel LA, Roussev R. Multiple thrombophilic gene mutations rather than specific gene mutations are risk factors for recurrent miscarriage. Am J Reprod Immunol. 2006;55(5):360-8.##Dawood F, Farquharson R, Quenby S, Toh CH. Acquired activated protein C resistance may be a risk factor for recurrent fetal loss. Fertil. Steril. 2003;80(3):649-50.##Rey E, Kahn SR, David M, Shrier I. Thrombophilic disorders and fetal loss: a meta-analysis. Lancet. 2003;361:901-8.##McNamee K, Dawood F, Farquharson R. Recurrent miscarriage and thrombophilia: an update. Curr Opin Obstet Gynecol. 2012;24(4):229-34.##Li J, Wu H, Chen Y, Wu H, Xu H, Li L. Genetic association between FXIII and β-fibrinogen genes and women with recurrent spontaneous abortion: a meta- analysis. J Assist Reprod Genet. 2015;32(5):817-25.##Christiansen OB, Nielsen HS, Kolte AM. Future directions of failed implantation and recurrent miscarriage research. Reprod Biomed Online. 2006;13(1):71-83.##Bellver J, Soares SR, Alvarez C, Muñoz E, Ramírez A, Rubio C, et al. The role of thrombophilia and thyroid autoimmunity in unexplained infertility, implantation failure and recurrent spontaneous abortion. Hum. Reprod. 2008;23(2):278-84.##Timeva T, Shterev A, Kyurkchiev S. Recurrent implantation failure: the role of the endometrium. J Reprod Infertil. 2014;15(4):173-83.##Azem F, Many A, Ben Ami I, Yovel I, Amit A, Lessing JB, et al. Increased rates of thrombophilia in women with repeated IVF failures. Hum Reprod. 2004;19(2):368-70.##Grandone E, Colaizzo D, Lo Bue A, Checola MG, Cittadini E, Margaglione M. Inherited thrombophilia and in vitro fertilization implantation failure. Fertil Steril. 2001;76(1):201-2.##Qublan HS, Eid SS, Ababneh HA, Amarin ZO, Smadi AZ, Al-Khafaji FF, et al. Acquired and inherited thrombophilia: implication in recurrent IVF and embryo transfer failure. Hum Reprod. 2006;21(10):2694-8.##Geva E, Amit A, Lerner-Geva L, Azem F, Yovel I, Lessing JB. Autoimmune disorders: another possible cause for in-vitro fertilization and embryo transfer failure. Hum Reprod. 1995;10(10):2560-3.##Nelson SM, Greer IA. The potential role of heparin in assisted conception. Hum Reprod Update. 2008;14(6):623-45.##Martinelli I, Battaglioli T, Mannucci PM. Screening of thrombophilia in women with failure of embryo implantation: far from being recommended. Haematologica. 2003;88(12):ELT36.##Simur A, Ozdemir S, Acar H, Colakoğlu MC, Görkemli H, Balci O, et al. Repeated in vitro fertilization failure and its relation with thrombophilia. Gynecol Obstet Invest. 2009;67(2):109-12.##Soria JM, Morange PE, Vila J, Souto JC, Moyano M, Trégouët DA, et al. Multilocus genetic risk scores for venous thromboembolism risk assessment. J Am Heart Assoc. 2014;3(5):e00106.##Goodman CS, Coulam CB, Jeyendran RS, Acosta VA, Roussev R. Which thrombophilic gene mutations are risk factors for recurrent pregnancy loss? Am J Reprod Immunol. 2006;56(4):230-6.##Altman DG. Practical statistics for medical research. 1st ed. London: Chapman and Hall; 1991. 624 p.##Pagano M, Gauvreau K. Principles of biostatistics. 2nd ed. Belmont, CA: Chapman and Hall; 2000. 584 p.##Deeks, J.J., Higgins, J.P.T., 2010. Statistical algorithms in review manager 5. Statistical methods group of the cochrane collaboration. August 2010. Available from https://training.cochrane.org/hand-book/statistical-methods-revman5.##de Haan HG, Bezemer ID, Doggen CJ, Le Cessie S, Reitsma PH, Arellano AR, et al. Multiple SNP testing improves risk prediction of first venous thrombosis. Blood. 2012;120(3):656-63.##Corral J, Hernández-Espinosa D, Soria JM, Gonzalez-Conejero R, Ordonez A, Gonzalez-Porras JR, et al. Antithrombin Cambridge II (A384S): an underestimated genetic risk factor for venous thrombosis. Blood. 2007;109(10):4258-63.##Corral J, Gonzalez-Conejero R, Soria JM, González-Porras JR, Pérez-Ceballos E, Lecumberri R, et al. A nonsensepolymorphism in the protein Z-dependent proteaseinhibitor increases the risk for venous thrombosis. Blood. 2006;108(1):177-83.##Dennis J, Johnson CY, Adediran AS, de Andrade M, Heit JA, Morange PE, et al. The endothelialprotein C receptor (PROCR) Ser219Gly variant andrisk of common thrombotic disorders: a HuGE review and meta-analysis of evidence from observational studies. Blood. 2012;119(10):2392-400.##Walch K, Riener EK, Tempfer CB, Endler G, Huber JC, Unfried G. The C46T polymorphism of the coagulation factor XII gene and idiopathic recurrent miscarriage. BJOG. 2005;112(10):1434-6.##Bagoly Z, Koncz Z, Hársfalvi J, Muszbek L. Factor XIII, clot structure, thrombosis. Thromb Res. 2012;129(3):382-7.##Karimi M, Bereczky Z, Cohan N, Muszbek L. Factor XIII deficiency. semin. Thromb Hemost. 2009;35(4):426-38.##Muszbek L, Bagoly Z, Cairo A, Peyvandi F. Novel aspects of factor XIII deficiency. Curr Opin Hematol. 2011;18(5):366-72.##Ariëns RA, Philippou H, Nagaswami C, Weisel JW, Lane DA, Grant PJ. The factor XIII V34L polymorphism accelerates thrombin activation of factor XIII and affects cross-linked fibrin structure. Blood. 2000;96(3):988-95.##Wartiovaara U, Mikkola H, Szôke G, Haramura G, Kárpáti L, Balogh I, et al. Effect of Val34Leu polymorphism on the activation of the coagulation factor XIII-A. Thromb Haemost. 2000;84(4):595-600.##Kohler HP, Ariëns RA, Whitaker P, Grant PJ. A common coding polymorphism in the FXIII A-subunit gene (FXIIIVal34Leu) affects cross-linking activity. Thromb Haemost. 1998;80(4):704.##Suzuki K, Henke J, Iwata M, Henke L, Tsuji H, Fukunaga T, et al. Novel polymorphisms and haplotypes in the human coagulation factor XIII A-subunit gene. Hum Genet. 1996;98(4):393-5.##Lim BC, Ariëns RA, Carter AM, Weisel JW, Grant PJ. Genetic regulation of fibrin structure and function: complex gene-environment interactions may modulate vascular risk. Lancet. 2003;361(9367):1424-31.##Vokó Z, Bereczky Z, Katona E, Adány R, Muszbek L. Factor XIII Val34Leu variant protects against coronary artery disease. a meta-analysis. 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Ovarian Sensitivity Index (OSI): Validating the Use of a Marker for Ovarian Responsiveness in IVF 2 1 2 Background: In this study, an attempt was made to validate the use of OSI as a measure of ovarian response during IVF treatment and to correlate OSI with age and BMI and other measures of ovarian response such as AMH, antral follicle count (AFC), total dose of administered gonadotrophins, and duration of stimulation. Methods: This study was a retrospective comparative cohort one. The study included a total of 2150 women who underwent the first IVF cycle between January 2008 and December 2017 at our center using long-agonist protocol. Patients were divided into four subgroups according to the circulating AMH level: below the 25th percentile (AMH 0.25-1.1 ng/ml, subgroup A), between 25th and 50th percentiles (AMH 1.2-1.6 ng/ml, subgroup B), between the 50th and 75th percentiles (AMH1.7-2.6 ng/ml, subgroup C), and above the 75th percentile (AMH 2.7-8.5 ng/ml, subgroup D). Qualitative data were analyzed by Chi-square or Fisher’s exact test. The p<0.05 was considered statistically significant. Results: The four subgroups formed on the basis of the AMH level did not significantly differ for age, BMI and infertility duration. OSI was significantly correlated to age (r=0.167; p=0.001), and has negative correlation with AFC (r=-0.236, p=0.001) and AMH levels (r=-0.123, p=0.001). Multiple linear regression analysis was done on OSI with other independent variables such as age, BMI, AFC, AMH. Analysis showed that approximately 8% variation in the value of OSI can be attributed to these variables with the highest correlation with antral follicle count. Conclusion: The present study showed that OSI appears to be a highly reliable index of ovarian responsiveness to recombinant FSH and can be useful to estimate the FSH dose. 083 89 Vikas V Yadav Department of Obstetrics and Gynecology, AIIMS Department of Obstetrics and Gynecology, AIIMS India Neena N Malhotra Department of Obstetrics and Gynecology, AIIMS Department of Obstetrics and Gynecology, AIIMS India malhotraneena@yahoo.com Reeta R Mahey Department of Obstetrics and Gynecology, AIIMS Department of Obstetrics and Gynecology, AIIMS India Neeta N Singh Department of Obstetrics and Gynecology, AIIMS Department of Obstetrics and Gynecology, AIIMS India Alka A Kriplani Department of Obstetrics and Gynecology, AIIMS Department of Obstetrics and Gynecology, AIIMS India Agonist protocolAMH (Anti mullerian hormone)IVF (In vitro fertilization)Ovarian sensitivity index (OSI) 50051.pdf Fauser BC, Devroey P, Macklon NS. Multiple birth resulting from ovarian stimulation for subfertility treatment. Lancet. 2005;365(9473):1807-16.##Briggs R, Kovacs G, Mac Lachlan V, Motteram C, Baker HW. Can you ever collect too many oocytes? Hum Reprod. 2015;30(1):81-7.##Sunkara SK, Rittenberg V, Raine-Fenning N, Bhattacharya S, Zamora J, Coomarasamy A. Association between the number of eggs and live birth in IVF treatment: an analysis of 400 135 treatment cycles. Hum Reprod. 2011;26(7):1768-74.##Drakopoulos P, Blockeel C, Stoop D, Camus M, de Vos M, Tournaye H, et al. Conventional ovarian stimulation and single embryo transfer for IVF/ICSI. How many oocytes do we need to maximize cumulative live birth rates after utilization of all fresh and frozen embryos? Hum Reprod. 2016;31(2):370-6.##Broekmans FJ, Kwee J, Hendriks DJ, Mol BW, Lambalk CB. A systematic review of tests predicting ovarian reserve and IVF outcome. Hum Reprod Update. 2006;12(6):685-718.##de Carvalho BR, Rosa e Silva AC, Rosa e Silva JC, dos Reis RM, Ferriani RA, Silva de Sa MF. Ovarian reserve evaluation: state of the art. J Assist Reprod Genet. 2008;25(7):311-22.##Polyzos NP, Devroey P. A systematic review of randomized trials for the treatment of poor ovarian responders: is there any light at the end of the tunnel? Fertil Steril. 2011;96(5):1058-61.e7.##Poseidon Group (Patient-oriented strategies encompassing individualized oocyte number), Alviggi C, Andersen CY, Buehler K, Conforti A, De Placido G, et al. A new more detailed stratification of low responders to ovarian stimulation: from a poor ovarian response to a low prognosis concept. Fertil Steril. 2016;105(6):1452-3.##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.##Biasoni V, Patriarca A, Dalmasso P, Bertagna A, Manieri C, Benedetto C, et al. Ovarian sensitivity index is strongly related to circulating AMH and may be used to predict ovarian response to exogenous gonadotropins in IVF. Reprod Biol Endocrinol. 2011;9:112.##Ledger WL. Clinical utility of measurement of anti-mullerian hormone in reproductive endocrinology. J Clin Endocrinol Metab. 2010;95(12):5144-54.##La Marca A, Nelson SM, Sighinolfi G, Manno M, Baraldi E, Roli L, et al. Anti-Müllerian hormone based prediction model for a live birth in assisted reproduction. Reprod Biomed Online. 2011;22(4):341-9.##Weenen C, Laven JS, Von Bergh AR, Cranfield M, Groome NP, Visser JA, et al. Anti-Müllerian hormone expression pattern in the human ovary: potential implications for initial and cyclic follicle recruitment. Mol Hum Reprod 2004;10(2):77-83.##Hehenkamp WJ, Looman CW, Themmen AP, de Jong FH, Te Velde ER, Broekmans FJ. Anti-Müllerian hormone levels in the spontaneous menstrual cycle do not show substantial fluctuation. J Clin Endocrinol Metab. 2006;91(10):4057-63.##Broer SL, Dolleman M, Opmeer BC, Fauser BC, Mol BW, Broekmans FJ. AMH and AFC as predictors of excessive response in controlled ovarian hyperstimulation: a meta-analysis. Hum Reprod Update. 2011;17(1):46-54.##Anckaert E, Smitz J, Schiettecatte J, Klein BM, Arce JC. The value of anti-Mullerian hormone measurement in the long GnRH agonist protocol: association with ovarian response and gonadotrophin-dose adjustments. Hum Reprod. 2012;27(6):1829-39.##Li HWR, Lee VCY, Ho PC, Ng EHY. Ovarian sensitivity index is a better measure of ovarian responsiveness to gonadotrophin stimulation than the number of oocytes during in-vitro fertilization treatment. J Assist Reprod Genet. 2014;31(2):199-203.##

Can Amlodipine Improve the Pre-ovulatory Follicle Blood Flow in Women with Polycystic Ovarian Syndrome? 2 1 2 Background: A reduction in intra-ovarian vascular resistance is necessary to achieve pregnancy in a natural cycle. The aim of this RCT was to detect whether a vasodilator calcium channel blocker, amlodipine, could increase the pre-ovulatory follicular blood flow, enhance follicular maturation in women with PCOS and improve ovulatory outcome. Methods: Sixty women received induction by clomiphene citrate (CC); thirty were given amlodipine (Amlodipine group) and the other 30 women were given placebo (Placebo group). The pattern of pre-ovulatory follicle blood flow was studied by color and power Doppler ultrasonography pre and post drug administration. Independent t-test was used to compare mean values of the 2 groups. The p<0.05 is considered statistically significant. Results: When comparing the Doppler effect of amlodipine versus placebo in the treatment cycle, it was found that mean value of ovarian arteries (OA) pulsatility index was lower in amlodipine group but it didn't reach statistical significance (p=0.063); however, the mean value of OA resistance index reached statistical significance (p=0.028) in amlodipine group. Moreover, in the second cycle, endometrial thickness was significantly higher (p=0.006) in women of the amlodipine group when compared to those of the placebo group. At least one sonographically detectable mature follicle (≥18 mm) was observed in 54.5% (36/66) during the first cycle. At the second cycle, this proportion significantly rose to 86.7% (26/30) in the amlodipine group, but marginally and non-significantly to 56.7% (17/30) in the placebo group. Conclusion: Amlodipine as calcium channel blocker was proved to have a role in improving ovarian blood flow at the time of ovulation and enhancing follicular maturation and thus, it may increase the chances of conception. 089 95 Doaa DE Faham Reproductive Health Research Department, National Research Centre Reproductive Health Research Department, National Research Centre Egypt drdody_as@hotmail.com Khaled Kh Ali Obstetrics and Gynecology Department, Ain Shams University Maternity Hospital Obstetrics and Gynecology Department, Ain Shams University Maternity Hospital Egypt Adel AE El Din Obstetrics and Gynecology Department, Ain Shams University Maternity Hospital Obstetrics and Gynecology Department, Ain Shams University Maternity Hospital Egypt Mamdouh M Bibars Reproductive Health Research Department, National Research Centre Reproductive Health Research Department, National Research Centre Egypt Osama O Azmy Reproductive Health Research Department, National Research Centre Reproductive Health Research Department, National Research Centre Egypt AmlodipineClomiphene citrateDoppler ultrasonographyInfertilityPreovulatory follicle 50049.pdf Adams J, Polson DW, Franks S. Prevalence of polycystic ovaries in women with anovulation and idiopathic hirsutism. Br Med J (Clin Res Ed). 1986;293(6543):355-9.##Kriplani A, Kachhawa G. Polycystic ovary syndrome. Novel insights into causesand therapy. Indian J Med Res. 2014;139(40):653.##Yildir IC, Kutluturk F, Tasliyurt T, Yelken BM, Acu B, Beyhan M, et al. Insulin resistance and cardiovascular risk factors in women with PCOS who have normal glucose tolerance test. Gynecol Endocrinol. 2013;29(2):148-51.##Christian RC, Dumesic DA, Behrenbeck T, Oberg AL, Sheedy PF 2nd, Fitzpatrick LA. Prevalence and predictors of coronary artery calcification in women with polycystic ovary syndrome. J Clin Endocrinol Metab. 2003;88(6):2562-8.##Dahlgren E, Janson PO, Johansson S, Lapidus L, Oden A. Polycystic ovary syndrome and risk for myocardial infarction. Evaluated from a risk factor model based on a prospective population study of women. Acta Obstet Gynecol Scand. 1992;71(8):599-604.##Ferreira A, Pires CR, Moron AF, Araujo Junior E, Traina E, Mattar R. Doppler assessment of uterine blood ﬂow in recurrent pregnancy loss. Int J Gynaecol Obstet. 2007;98(2):115-9.##Lam P, Johnson I, Raine-Fenning N. Endometrial blood ﬂow is impaired in women with polycystic ovarian syndrome who are clinically hyperandrogenic. Ultrasound Obstet Gynecol. 2009;34(3):326-34.##Cocksedge KA, Saravelos SH, Wang Q, Tuckerman E, Laird SM, Li TC. Does free androgen index predict subsequent pregnancy outcome in women with recurrent miscarriage? Hum Reprod. 2008;23(4):797-802.##Cameron IT, Campbell S. Nitric oxide in the endometrium. Hum Reprod. Update. 1998;4(5):565-9.##Ota H, Igarashi S, Hatazawa J, Tanaka T. Endothelial nitric oxide synthase in the endometrium during the menstrual cycle in patients with endometriosis an adenomyosis. Fertil Steril. 1998;69(2):303-8.##Takasaki A, Tamura H, Miwa I, Taketani T, Shimamura K, Sugino N. Endometrial growth and uterine blood flow: a pilot study for improving endometrial thickness in the patients with a thin endometrium. Fertil Steril. 2010;93(6):1851-8.##Rotterdam ESHRE/ASRM-Sponsored PCOS consensus workshop group. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome (PCOS). Hum Reprod. 2004;19(1):41-7.##Pfizer Canada Inc. Norvasc Product Monograph [Internet]. Canada: Pfizer Products Inc. 2017 Sept [cited 2018 Sept 18]. 33 p. Available from: https://www.pfizer.ca/sites/g/files/g10050796/f/201710/NORVASC_PM.pdf.##BC Drug and Poison Information Centre (DPIC). Calcium channel blocking drugs [Internet]. Vancouver, Canada: DPIC. 2017 [cited 2018 Sept 18]. Available from: http://dpic.org/sites/default/files/pdf/DIR_CCBs.pdf.##Official FDA information [Internet]. International Labs Inc. 2012 [cited 2018 Sept 18]. Available from: www.drugs.com.##Isaac S, Michael WB. Handbook in research and evaluation for education and the behavioral sci-ences. 3rd ed. San Diego: Edits Pub; 1995. 262 p##Sher G, Fisch JD. Vaginal sildenafil (Viagra): a preliminary report of a novel method to improve uterine artery blood flow and endometrial development in patients undergoing IVF. Hum Reprod. 2000;15(4):806-9.##Moretto M, Lopez FJ, Negro-Vilar A. Nitric oxide regulates luteinizing hormone-releasing hormone secretion. Endocrinology. 1993;133(5):2399-402.##Moncada S, Higgs A. The L-arginine-nitric oxide pathway. N Engl J Med. 1993;329(27):2002-12.##Palmer RM, Ferrige AG, Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 1987;327(122):524-6.##Ledee-Bataille N, Olivennes F, Lefaix JL, Chaouat G, Frydman R, Delanian S. Combined treatment by pentoxifylline and tocopherol for recipient women with a thin endometrium enrolled in an oocyte donation programme. Hum Reprod. 2002;17(5):1249-53.##

Evaluation of Sperm Chromatin Integrity Using Aniline Blue and Toluidine Blue Staining in Infertile and Normozoospermic Men 2 1 2 Background: Male infertility is defined as a man lost his ability to fertilize a fertile female naturally. Diagnosis of male infertility cannot be made just according to basic semen analysis. It is necessity to have specific tests for evaluation of chromatin integrity. In this study, an attempt was made to evaluate the sperm chromatin quality in fertile men and infertile subgroup. Methods: Among 1386 couples, 342 men were categorized into normospermia and 1044 were infertile and they were referred to Yazd Research and Clinical Center for infertility treatment. Standard semen analysis and sperm nuclear maturity tests including aniline blue (AB) and toluidine blue (TB) staining were done. Data were analyzed by SPSS software. The p≤0.05 was considered statistically significant. Results: The mean value of TB staining was significantly higher in infertile group compared to normospermic group (p=0.005). Mean of sperm normal morphology was lower in idiopathic infertile men in comparison with normozoospermic men (p=0.001). The highest negative correlation was obtained between sperm count and AB staining. Progressive motility was negatively correlated with AB and TB staining in both groups but there was no significant difference between AB staining and progressive motility in men normospermia group. Conclusion: Sperm chromatin staining using AB and TB showed a negative association between sperm chromatin condensation with sperm count, normal morphology and progressive motility. It seems that the AB and TB test may be useful for the assessment of male fertility potential. 095 102 Soheila S Pourmasumi Non-Communicable Diseases Research Center, Rafsanjan University of Medical Sciences Non-Communicable Diseases Research Center, Rafsanjan University of Medical Sciences Iran Arezoo A Khoradmehr Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences Iran Tahereh T Rahiminia Gametogenesis Research Center, Fertility, and Infertility Center, Kashan University of Medical Sciences Gametogenesis Research Center, Fertility, and Infertility Center, Kashan University of Medical Sciences Iran Parvin P Sabeti Department of Anatomy, Faculty of Medicine, Kurdistan University of Medical Sciences Department of Anatomy, Faculty of Medicine, Kurdistan University of Medical Sciences Iran Alireza A Talebi علیرضا طالبی Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences Iran prof_talebi@ssu.ac.ir; prof_talebi@hotmail.com Jalal J ghasemzadeh Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences Iran Aniline blueInfertilitySpermToluidine blue 50045.pdf Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reprod Biol Endocrinol. 2015;13:37.##Lewis SE, Agbaje I, Alvarez J. Sperm DNA tests as useful adjuncts to semen analysis. Syst Biol Reprod Med. 2008;54(3):111-25.##Evgeni E, Charalabopoulos K, Asimakopoulos B. Human sperm DNA fragmentation and its correlation with conventional semen parameters. J Reprod Infertil. 2014;15(1):2-14.##Varghese A, Bragais F, Mukhopadhyay D, Kundu S, Pal M, Bhattacharyya A, et al. Human sperm DNA integrity in normal and abnormal semen samples and its correlation with sperm characteristics. Andrologia. 2009;41(4):207-15.##Lewis S, Aitken R. DNA damage to spermatozoa has impacts on fertilization and pregnancy. Cell Tissue Res. 2005;322(1):33-41.##Zini A, Boman JM, Belzile E, Ciampi A. Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis. Hum Reprod. 2008;23(12):2663-8.##Pourmasumi S, Ghasemi N, Talebi AR, Mehrabani M, Sabeti P. The effect of vitamin E and selenium on sperm chromatin quality in couples with recurrent miscarriage. Int J Med Lab. 2018;5(1):1-10.##Sivanarayana T, Krishna CR, Prakash GJ, Krishna K, Madan K, Sudhakar G, et al. Sperm DNA fragmentation assay by sperm chromatin dispersion (SCD): correlation between DNA fragmentation and outcome of intracytoplasmic sperm injection. Reprod Med Biol. 2014;13(2):87-94.##Agarwal A, Said TM. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update. 2003;9(4):331-45.##Pourmasumi S, Sabeti P, Rahiminia T, Mangoli E, Tabibnejad N, Talebi AR. The etiologies of DNA abnormalities in male infertility: An assessment and review. Int J Reprod Biomed (Yazd). 2017;15(6):331-44.##Emelyanov AV, Fyodorov DV. Thioredoxin-dependent disulfide bond reduction is required for protamine eviction from sperm chromatin. Genes Dev. 2016;30(24):2651-6.##Virro MR, Larson-Cook KL, Evenson DP. Sperm chromatin structure assay (SCSA®) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil Steril. 2004;81(5):1289-95.##Sergerie M, Laforest G, Boulanger K, Bissonnette F, Bleau G. Longitudinal study of sperm DNA fragmentation as measured by terminal uridine nick end-labelling assay. Hum Reprod. 2005;20(7):1921-7.##Mehta A, Sigman M. Identification and preparation of sperm for ART. Urolc Clin North Am. 2014;41 (1):169-80.##Talebi A, Vahidi S, Aflatoonian A, Ghasemi N, Ghasemzadeh J, Firoozabadi R, et al. Cytochemical evaluation of sperm chromatin and DNA integrity in couples with unexplained recurrent spontaneous abortions. Andrologia. 2012;44 Suppl 1:462-70.##Talebi AR, Khalili MA, Vahidi S, Ghasemzadeh J, Tabibnejad N. Sperm chromatin condensation, DNA integrity, and apoptosis in men with spinal cord injury. J Spinal Cord Med. 2013;36(2):140-6.##Sabeti P, Amidi F, Kalantar SM, Sedighi Gilani MA, Pourmasumi S, Najafi A, et al. Evaluation of intracellular anion superoxide level, heat shock protein A2 and protamine positive spermatozoa percentages in teratoasthenozoospermia. Int J Reprod Biomed (Yazd). 2017;15(5):279-86.##Evenson DP. The sperm chromatin structure assay (SCSA®) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility. Anim Reprod Sci. 2016;169:56-75.##Practice committee of the American society for reproductive medicine. The clinical utility of sperm DNA integrity testing: a guideline. Fertil Steril. 2013;99(3):673-7.##Sellami A, Chakroun N, Ben Zarrouk S, Sellami H, Kebaili S, Rebai T, et al. Assessment of chromatin maturity in human spermatozoa: useful aniline blue assay for routine diagnosis of male infertility. Adv Urol. 2013;2013:578631.##Irez T, Sahmay S, Ocal P, Goymen A, Senol H, Erol N, et al. Investigation of the association between the outcomes of sperm chromatin condensation and decondensation tests, and assisted reproduction techniques. Andrologia. 2015;47(4):438-47.##Kim HS, Kang MJ, Kim SA, Oh SK, Kim H, Ku SY, et al. The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis. Clin Exp Reprod Med. 2013;40(1):23-8.##Hamidi J, Frainais C, Amar E, Bailly E, Clément P, Ménézo Y. A double-blinded comparison of in situ TUNEL and aniline blue versus flow cytometry acridine orange for the determination of sperm DNA fragmentation and nucleus decondensation state index. Zygote. 2015;23(4):556-62.##Varghese AC, Bragais FM, Mukhopadhyay D, Kundu S, Pal M, Bhattacharyya AK, et al. Human sperm DNA integrity in normal and abnormal semen samples and its correlation with sperm characteristics. Andrologia. 2009;41(4):207-15.##Sabeti P, Pourmasumi S, Rahiminia T, Akyash F, Talebi AR. Etiologies of sperm oxidative stress. 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Home Delivery Practices and Associated Factors in Ethiopia 2 1 2 Background: The risk of a woman in a developing country dying from a maternal-related cause is higher compared to a woman living in a developed country. Despite the fact that delivery care service utilization is essential for further improvement of mothers and newborns, the coverage of delivery service in Ethiopia is still near to the ground. This study aimed to identify factors associated with home delivery among women in Ethiopia at their last birth. Methods: The data was obtained from 2016 Ethiopia Demographic and Health Survey which is the fourth survey. The sample was selected using a stratified, two-stage cluster sampling design and the data was analyzed using mixed effect logistic regression model. Results: A total of 10,622 women were considered in this study and 67.2% of them gave birth at home. The percentage of home delivery at their last birth was high in Afar and Somali region (89.6% and 81.7%, respectively) while only 3.3% women who lived in Addis Ababa delivered at home. Living in rural areas, being uneducated, older age, not watching TV, and being poor are predictors of home delivery at 5% level of significance. Conclusion: There is a need of giving special attention to women living in rural area, women from poor families and uneducated women to decrease home delivery. 102 109 Ayele AG Chernet Statistics Department, College of Natural and Computational Sciences, Wolkite University Statistics Department, College of Natural and Computational Sciences, Wolkite University Ethiopia gebeyehu29@gmail.com Kassahun KT Dumga Department of Statistics, Wolkite University Department of Statistics, Wolkite University Ethiopia Kebadu KT Cherie Department of Statistics, Wolkite University Department of Statistics, Wolkite University Ethiopia Deliver careDeveloping countryEthiopiaHome deliveryMixed effectRandom effect 50047.pdf Wang W, Alva S, Wang S, Fort A. Levels and trends in the use of maternal health services in developing countries. Calverton, Maryland: DHS comparative reports; 2011Jun. 105 p. Report No 26; USAID Project No. GPO-C-00-08-00008-00).##World Health Organization. Global health observatory (GHO) data [Internet]. Geneva: World Health Organization; 2018. Global situation and trends; 2018 (cited 2019 Jan). [about 2 screens]. Available form: http://www. who. int/gho/tb/en.##Lozano R, Wang H, Foreman KJ, Rajaratnam JK, Naghavi M, Marcus JR, et al. Progress towards millennium development goals 4 and 5 on maternal and child mortality: an updated systematic analysis. Lancet. 2011;378(9797):1139-65.##World Health Organization. Making pregnancy safer: the critical role of the skilled attendant. 1st ed. Geneva: World Health Organization; 2004. 18 p.##Mrisho M, Schellenberg JA, Mushi AK, Obrist B, Mshinda H, Tanner M, et al. Factors affecting home delivery in rural Tanzania. Trop Med Int Health. 2007;12(7):862-72.##Bolam A, Manandhar DS, Shrestha P, Elis M, Malla K, Costello AM. Factors affecting home delivery in the Kathmandu Valley, Nepal. Health Policy Plan. 1998;13(2):152-8.##Shankwaya S. Study to explore barriers to utilization of maternal delivery services in Kazungula district [dissertation]. [Amsterdam]: Vrije Universiteit Amsterdam; 2009. 65 p.##Dhakal P, Shrestha M, Baral D, Pathak S. Factors affecting the place of delivery among mothers residing in jhorahat VDC, Morang, Nepal. Int J Community Based Nurs Midwifery. 2018;6(1):2-11.##Central statistical agency (CSA) of Ethiopia and ICF. Ethiopia demographic and health survey 2011. Addis Ababa, Ethiopia, and Rockville, Maryland, USA: CSA and ICF; 2017 Mar. 452 p.##Abeje G, Azage M, Setegn T. Factors associated with institutional delivery service utilization among mothers in Bahir Dar city administration, Amhara region: a community based cross sectional study. Reprod Health. 2014;11:22.##Central statistical agency (CSA) of Ethiopia and ICF. Ethiopia demographic and health survey 2016. Addis Ababa, Ethiopia, and Rockville, Maryland, USA: CSA and ICF; 2017 July. 551 p.##Fitzmaurice G, Davidian M, Verbeke G, Molenberghs G. Longitudinal data analysis. 1 st ed. USA: Chapman and Hall/CRC; 2008. 632 P.##Woldemicael G. Recent fertility decline in Eritrea: is it a conflict-led transition. Demogr Res. 2008;18(2):27-58.##Tesfaye B, Mathewos T, Kebede M. Skilled delivery inequality in Ethiopia: to what extent are the poorest and uneducated mothers benefiting? Int J Equity Health. 2017;16(1):82.##

Determining the Cause of Recurrent Miscarriages in a Couple: Importance of NOR in the Era of NGS 2 1 2 Background: Chromosomal abnormalities are a significant cause of human disorders. The characterization of such abnormalities helps in the identification of known/new genes. The purpose of the present study was to identify the cause of miscarriages in a couple by using combined molecular and cytogenetic techniques. Case Presentation: In this study, the clinical, cytogenetic and molecular cytogenetic evaluations were performed on a couple with recurrent miscarriages. Several methods like GTG banding, silver nitrate (NOR) staining, fluorescence in-situ hybridization (FISH) using whole chromosome paint probes (WCP) and bacterial artificial chromosome (BAC) clones were used. The chromosomal analysis on the metaphases revealed a karyotype of 46,XX in the wife and 46,XY,13p+ in her husband. To check the satellites on 13p region, NOR was performed which showed absence of satellites and presence of euchromatic material. On careful analysis, the satellites were observed on 11q terminal region. Thus, a balanced reciprocal translocation was detected which was confirmed by WCP and Acro-P-arm FISH. Fine mapping with BAC clones narrowed down the breakpoint regions. Conclusion: The application of the combined cytogenetic methods especially NOR helped in identification of the balanced reciprocal translocation with subsequent systematic characterization and the breakpoint regions were identified. The characterization of the breakpoint regions helped in identification of the carrier status which further paved the way for understanding the cause of recurrent miscarriages and proper genetic counseling. 109 115 Usha UR Dutta Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex India ushadutta@hotmail.com; usha@cdfd.org.in Venugopala V Swamy Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex India Rajitha R Ponnala Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex India Shagun S Aggarwal Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex India Ashwin A Dalal Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex Diagnostics Division, Center for DNA Fingerprinting and Diagnostics, Tuljaguda complex India BAC clonesBad obstetric historyFISHNucleolar organizing regionsSatellites 10027.pdf Royal College of Obstetricians and Gynecologists (RCOG). The management of recurrent miscarriage. (Edition???). London: Royal College of Obstetricians and Gynecologists; 1998. (Pagination??).##De Braekeleer M, Dao TN. Cytogenetic studies in couples experiencing repeated pregnancy losses. Hum Reprod. 1990;59(5):519-28.##Rutkowski S, Stene J, Gallano P. Risk estimates in balanced parental reciprocal translocations: analysis of 1120 pedigrees. 1st ed. Paris: Expansion Scien-tifique Francaise; 1988. 147 p.##Daniel A, Hook EB, Wulf G. Risk of unbalanced progeny at amniocentesis to carriers of chromosome rearrange-ments: data from United States and Canadian laboratories. Am J Med Genet. 1989;33(1):14-53.##Smeets D, van Ravenswaaij C, de Pater J, Gerssen-Schoorl K, Van Hemel J, Janssen G, et al. At least nine cases of trisomy 11 q23- qter in one generation as a result of familial t( 11;13) translocation. J Med Genet. 1997;34(1):18-23.##Dutta UR, Ponnala R, Pidugu VK, Dalal AB. Cytogenetic abnormalities in 1162 couples with recurrent miscarriages in southern region of India: report and review. J Assist Reprod Genet. 2011;28(2):145-9.##Mitelman F, Johansson B, Mertens FE. Mitelman database of chromosome aberrations in cancer. Available at: http://www.cgap.nci.nih.gov/Chromosomes/Mitelman. Accessed August 12, 2003.##Huret JL, Dessen P, Bernheim A. Atlas of genetics and cytogenetics in oncology and haematology, year 2003. Nucleic Acids Res. 2003;31(1):272-4.##Francke U, Weber F, Sparkes RS, Mattson PD, Mann J. Duplication 11(q21 to 23 leads to qter) syn-drome. Birth Defects Orig Artic Ser. 1977;13(3B):167-86.##Bader PJ, Jansch M, Hoffman D, Palmer CG, Gerber H, Taylor G. Trisomy 11q(q21 leads to qter). Birth Defects Orig Artic Ser. 1978;14(6C):383-92.##Pruitt KD, Tatusova T, Maglott DR. NCBI reference sequences (RefSeq): a curated non-redundant sequence data-base of genomes, transcripts and proteins. 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Positive and "Enriched" Environments Reverse Traumatic Stress and Reshape Epigenetic Signature of Spermatozoa and Ovulation 2 1 2 In evolutionary theories (Lamarckian and Darwinian), environment and physical changes could be transmitted to the descendants (1). Psychological factors in human beings (2–4) have a negative impact on the germ cells parameters (5). As time goes on, the quality of sperm is deteriorating (5, 6) as well as humanoocyte/egg (7). A simple change of environment and environmental preconceptional exposure (i.e., diet, physical activity, smoking, alcohol consumption, etc.) affects the functioning of the genes and the phenotype of the next generation through remodeling epigenetic blueprint of spermatozoa (8, 9). Lifestyle and environmental factors influence the sperm and egg which will affect subsequent generations (10–12). Some studies have examined the transmission of specific behavioral and structural adaptations in relation to the stimulus in the nervous system, from parents to their offspring (10, 13). Effects of psychological and environmental factors on gene expression persist even after the removal of the inducing agent passed on to subsequent generations (14). So, Genetic Memory can be contemplated in two ways (Biology and Psychology) (10, 15). Psychological factors which have the impact on fertility are treated in numerous studies on humans (5, 16–18) as psychological stress (7). The life of wars and prisons has an effect on germ cells (Sperm, menstrual irregularity and reproductive function) (7, 19, 20). Men who suffered from anxiety and depression or who have experienced high levels of stress had long-term sperm damage. Stress affects sperm quality over the long term, even slowing down the mental development of offspring. This stress is transmitted from one generation to another and has other perverse effects (21). Sperm of obese men have a distinct epigenetic signature compared to lean men. The methylome of spermatozoa is dynamically remodeled after weight loss (22). Paternal nutritional status can directly affect the health of offspring (21, 23), suggesting that an epigenetic inheritance phenomenon acquired by the environment is transmitted by gametes (22). Environmental factors as exercise and nutritional status induce acute changes in DNA methylation profiles in human skeletal muscle and adipose tissue (24–28) demonstrating that environmental factors are reshaping the epigenome of somatic tissues (22). The fertility of humans is impacted by myriad factors (Stress, obesity, alcohol and tobacco, seasonal variations, etc.) (28, 30). "Licking" (31), sense of touch and massage can be a great stress reliever, and is very important to emotional health (32), and sexual desire and performance (33). Scientists have found that the process of "Traumatic stress, etc." can be reversed if mice from traumatized lineages are put in "Enriched" environments (34). Psychotherapy could improve psychological and environmental factors and make the environment more positive and richer (35). Scientists have reached several conclusions by researching humans; the trauma would thus modify the behavior of the traumatized individual but also those of his descendants since one finds a metabolic modification until the third generation. This would mean that trauma also affects germ cells (Spermatozoa and ova) which are the only biological link between generations (34). A positive and nurturing environment help humans withstand the threat (34, 36). The decrease in anxiety (37), improvement of psychological factors (38) with reduced vulnerability to stress factor and shocks (39) are related to bgetting a pregnancy (5). Negative psychological and environmental factors, war, nutritional status, seasonal variations, physical and social environmental factors and stress have many negative consequences and effects on the germ cells. "Enriched" and positive environments can reverse these factors and have positive consequences on the germ cells in individual that could be transmitted to their off spring. Acknowledgement The author would like to thank Mr. Ahmad Alsaleh and Mrs. Sabrié Alsaleh, Dr. Ameni Ahmed, and Mrs. Régine Fabien for their participation in giving the counseling. Conflict of Interest The author declares that he has no competing interest. Author has not received the financial support for research. 115 118 Amani A Ahmed Caen University Hospital Caen University Hospital France Muaweah MA Alsaleh CesamS-CERReV, and INSERM, University of Caen Normandy CesamS-CERReV, and INSERM, University of Caen Normandy France moaouiya87@yahoo.com No Keyword 50046.pdf Lamarck J. 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Verdicts of water drop-sTRPPT on brain and life new frontiers in neuro cognitive positive psychotherapy in war and peace, normal-abnormal-medical-non-medical conditions. 1st ed. (Place of Publication????): Édi-tions universitaires européennes; 2017. 92 p.##Harlow CR, Fahy UM, Talbot WM, Wardle PG, Hull MG. Stress and stress-related hormones during in-vitro fertilization treatment. Hum Reprod. 1996;11(2):274‑9.##Sanders KA, Bruce NW. A prospective study of psychosocial stress and fertility in women. Hum Reprod. 1997;12(10):2324‑9.##Facchinetti F, Matteo ML, Artini GP, Volpe A, Genazzani AR. An increased vulnerability to stress is associated with a poor outcome of in vitro ferti-lization-embryo transfer treatment. Fertil Steril. févr 1997;67(2):309‑14.##