Low Success Rate of ART, an Illusion, a Reality or Simply a Too High Expectation? 2 1 575 2 Assisted reproductive technologies have spread worldwide to help infertile couples but access to these advanced treatments is of varying degrees in different countries. Access to infertility treatment is very limited and insurance coverage of these treatments is insufficient in developing, underdeveloped and low-income countries. From birth of the first IVF baby in 1978, procedures of diagnosis and treatment of infertility have been im-proving and they have revolutionized infertility treatment in such a way that even infertile men without mature sperm in their semen could conceive a child by retrieving a few numbers of sperm from their testicular tissue. In comparison to most other surgical and medical treatments, success rate of assisted reproductive technologies are low and consequently, successful pregnancies and take home babies require several bouts of treatment. Since these treatments are highly expensive and time-consuming, their repetition is not affordable for most infertile couples. Therefore, infertile couples’ growing discontent with low success rates of infertility treatments and their search for IVF centers with higher IVF success rates and new technologies or treatment options seem to be rea-sonable reactions. Over the past three decades of successful assisted reproductive practices, scientists and physicians have tried to improve infertility diagnosis and increase its successful treatment. An important part of these findings has been the production of new instruments to support the best environment for in vitro cultures and maturation of embryos and gametes. Another part of these efforts has focused on the preparation of infertile couples to produce adequate numbers of high quality sperm and oocytes and it has engaged in producing the best quality embryos and preparing the endometrium for their successful implantation. Overall, these efforts and progress have changed the situation so favorably that the clinical pregnancy rate of each treatment cycle has increased up to 60% (1). In fact, whether future research on ovarian stimulating drugs and protocols, fertilization and culture methods, culture media, endometrium preparation, embryo selection, embryo transfer and luteal phase support could in-crease ART outcomes more than the present ones is yet to be determined. In other words, whether the success rate of infertility treatment could be greater than the ones from natural fecundation is still a goal to be set for future investigations. The estimated natural fecundity rate is about 20% per month and the rates of pregnancy for those who try to conceive naturally will be 45%, 65% and 85% following 3, 6, and 12 cycles, respectively (2). According to the results of a study published in the New England Journal of Medicine recently, live birth rates from ART can approach those of natural fecundity in the general population, as long as the couple and embryo criteria are favorable and there are no contraindications for treatment continuation. However, financial limitations, environmental stresses and repeated implantation failures in the first two or three cycles or implication of other factors reduce live birth rates from ART dramatically. The researchers from the same study found the con-servative and optimal estimates of cumulative live-birth rates to be, respectively, 42.7% and 65.3% for transfer of cleavage embryos and 52.4% and 80.7% for transfer of blastocyst from fresh autologous oocytes in three con-secutive ART cycles. However, success rates are lower in older women than younger candidates when their own oocytes are used, but these rates could increase similar to younger women if donated oocytes are used instead (3). According to the above results, cumulative live-birth rates from ART is equal to natural fecundity rates at present but scientists believe that ART success rates could be increased by optimizing the above mentioned ART variables. Optimally, ART will be able to overcome other biological barriers of pregnancy such as pregnancy in postmenopausal women. 123 124 Mohammad Reza MR Sadeghi محمدرضا صادقی Editor-in-chief Editor-in-chief Iran No Keyword 575.pdf Zhao Y, Brezina P, Hsu CC, Garcia J, Brinsden PR, Wallach E. In vitro fertilization: four decades of reflections and promises. Biochim Biophys Acta. 2011;1810(9):843-52.##Practice Committee of American Society for Reproductive Medicine in collaboration with Society for Reproductive Endocrinology and Infertility. Optimizing natural fertility. Fertil Steril. 2008;90(5 Suppl):S1-6.##Luke B, Brown MB, Wantman E, Lederman A, Gibbons W, Schattman GL, et al. Cumulative birth rates with linked assisted reproductive technology cycles. N Engl J Med. 2012;366(26):2483-91.##

Strategies for Pituitary Down-regulation to Optimize IVF/ICSI Outcome in Poor Ovarian Responders 2 1 2 The ovarian stimulation of poor responders still remains a challenging task for clinicians. There are numerous strategies that have been suggested to improve the outcome in poor responders but there is still no one pituitary down-regulation protocol that best suits all women with such condition. Traditional GnRH agonist flare and long luteal phase protocols do not appear to be advantageous. Reduction of GnRH agonist doses, "stop" protocols, and microdose GnRH agonist flare regimes all appear to improve outcomes, although the proportional benefit of one approach over another has not been convincingly established. GnRH antagonists improve outcomes in this patient population, although, in general, pregnancy rates appear to be lower in comparison to microdose GnRH agonist flare regimes. 124 131 Ahmed A Badawy Ahmed Badawy College of Medicine, Al-Jouf University College of Medicine, Al-Jouf University Egypt ambadawy@yahoo.com Alaa A Wageah Alaa Wageah College of Medicine, Al-Jouf University College of Medicine, Al-Jouf University Egypt Mohamed M EL-Gharib College of Medicine, Al-Jouf University College of Medicine, Al-Jouf University Egypt Ezz Eldin E Osman Ezz Eldin Osman College of Medicine, Al-Jouf University College of Medicine, Al-Jouf University Egypt ICSIIVFPoor ovarian response 504.pdf Howles CM, Macnamee MC, Edwards RG. Short term use of an LHRH agonist to treat poor responders entering an in-vitro fertilization programme. Hum Reprod. 1987;2(8):655-6.##Marcus SF, Brinsden PR, Macnamee M, Rainsbury PA, Elder KT, Edwards RG. Comparative trial between an ultra-short and long protocol of luteinizing hormone-releasing hormone agonist for ovarian stimulation in in-vitro fertilization. Hum Reprod. 1993;8(2):238-43.##Scott RT, Neal GS, Illions EH, Hayslip CA, Hofmann GE. The duration of leuprolide acetate administration prior to ovulation induction does not impact ovarian responsiveness to exogenous gonadotropins. Fertil Steril. 1993;60(2):247-53.##Deaton JL, Bauguess P, Huffman CS, Miller KA. Pituitary response to early follicular-phase minidose gonadotropin releasing hormone agonist (GnRHa) therapy: evidence for a second flare. J Assist Reprod Genet. 1996;13(5):390-4.##Padilla SL, Dugan K, Maruschak V, Shalika S, Smith RD. Use of the flare-up protocol with high dose human follicle stimulating hormone and human menopausal gonadotropins for in vitro fertilization in poor responders. Fertil Steril. 1996;65(4):796-9.##Toth TL, Awwad JT, Veeck LL, Jones HW Jr, Muasher 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.##Karande V, Morris R, Rinehart J, Miller C, Rao R, Gleicher N. Limited success using the "flare" protocol in poor responders in cycles with low basal follicle-stimulating hormone levels during in vitro fertilization. Fertil Steril. 1997;67(5):900-3.##Surrey ES, Bower J, Hill DM, Ramsey J, Surrey M W. 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.##Schoolcraft W, Schlenker T, Gee M, Stevens J, Wagley L. Improved controlled ovarian hyperstimulation in poor responder in vitro fertilization patients with a microdose follicle-stimulating hormone flare, growth hormone protocol. Fertil Steril. 1997;67(1): 93-7.##Leondires MP, Escalpes M, Segars JH, Scott RT Jr, Miller BT. Microdose follicular phase gonadotropin-releasing hormone agonist (GnRH-a) compared with luteal phase GnRH-a for ovarian stimulation at in vitro fertilization. Fertil Steril. 1999;72 (6):1018-23.##Scott RT, Navot D. Enhancement of ovarian responsiveness with microdoses of gonadotropin-releasing hormone agonist during ovulation induction for in vitro fertilization. Fertil Steril. 1994;61 (5):880-5.##Kyrou D, Kolibianakis EM, Venetis CA, Papanikolaou EG, Bontis J, Tarlatzis BC. How to improve the probability of pregnancy in poor responders undergoing in vitro fertilization: a systematic review and meta-analysis. Fertil Steril. 2009;91(3): 749-66.##Weissman A, Farhi J, Royburt M, Nahum H, Glezerman M, Levran D. Prospective evaluation of two stimulation protocols for low responders who were undergoing in vitro fertilization-embryo transfer. Fertil Steril. 2003;79(4):886-92.##Latouche J, Crumeyrolle-Arias M, Jordan D, Kopp N, Augendre-Ferrante B, Cedard L, et al. GnRH receptors in human granulosa cells: anatomical localization and characterization by autoradiographic study. Endocrinology. 1989;125(3):1739-41.##Kowalik A, Barmat L, Damario M, Liu HC, Davis O, Rosenwaks Z. Ovarian estradiol production in vivo. Inhibitory effect of leuprolide acetate. J Reprod Med. 1998;43(5):413-7.##Dirnfeld M, Fruchter O, Yshai D, Lissak A, Ahdut A, Abramovici H. Cessation of gonadotropin-releasing hormone analogue (GnRH-a) upon down-regulation versus conventional long GnRH-a protocol in poor responders undergoing in vitro fertilization. Fertil Steril. 1999;72(3):406-11.##Garcia-Velasco JA, Isaza V, Requena A, Martínez-Salazar FJ, Landazábal A, Remohí J, et al. High doses of gonadotrophins combined with stop versus non-stop protocol of GnRH analogue administration in low responder IVF patients: a prospective, randomized, controlled trial. Hum Reprod. 2000;15(11):2292-6.##Faber BM, Mayer J, Cox B, Jones D, Toner JP, Oehninger S, et al. Cessation of gonadotropin-releasing hormone agonist therapy combined with high-dose gonadotropin stimulation yields favorable pregnancy results in low responders. Fertil Steril. 1998;69(5):826-30.##Wang PT, Lee RK, Su JT, Hou JW, Lin MH, Hu YM. Cessation of low-dose gonadotropin releasing hormone agonist therapy followed by high-dose gonadotropin stimulation yields a favorable ovarian response in poor responders. J Assist Reprod Genet. 2002;19(1):1-6.##Schachter M, Friedler S, Raziel A, Strassburger D, Bern O, Ron-el R. Improvement of IVF outcome in poor responders by discontinuation of GnRH analogue during the gonadotropin stimulation phase--a function of improved embryo quality. J Assist Reprod Genet. 2001;18(4):197-204##Pinkas H, Orvieto R, Avrech OM, Rufas O, Ferber A, Ben-Rafael Z, et al. Gonadotropin stimulation following GnRH-a priming for poor responders in in vitro fertilization-embryo transfer programs. Gynecol Endocrinol. 2000;14(1):11-4.##Olivennes F, Righini C, Fanchin R, Torrisi C, Hazout A, Glissant M, et al. A protocol using a low dose of gonadotrophin-releasing hormone agonist might be the best protocol for patients with high follicle-stimulating hormone concentrations on day 3. Hum Reprod. 1996;11(6):1169-72.##Feldberg D, Farhi J, Ashkenazi J, Dicker D, Shalev J, Ben-Rafael Z. Minidose gonadotropin-releasing hormone agonist is the treatment of choice in poor responders with high follicle-stimulating hormone levels. Fertil Steril. 1994;62(2):343-6.##Albuquerque LE, Saconato H, Maciel MC, Baracat EC, Freitas V. Depot versus daily administration of GnRH agonist protocols for pituitary desensitization in assisted reproduction cycles: a Cochrane Review. Hum Reprod. 2003;18(10):2008-17.##Halmos G, Schally AV. Changes in subcellular distribution of pituitary receptors for luteinizing hormone-releasing hormone (LH-RH) after treatment with the LH-RH antagonist cetrorelix. Proc Natl Acad Sci U S A. 2002;99(2):961-5.##Kovacs M, Schally AV, Csernus B, Rekasi Z. Luteinizing hormone-releasing hormone (LH-RH) antagonist Cetrorelix down-regulates the mRNA expression of pituitary receptors for LH-RH by counteracting the stimulatory effect of endogenous LH-RH. Proc Natl Acad Sci U S A. 2001;98(4): 1829-34.##Al-Inany HG, Abou-Setta AM, Aboulghar M. Gonadotrophin-releasing hormone antagonists for assisted conception. Cochrane Database Syst Rev. 2006;(3):CD001750.##Mahutte NG, Arici A. Role of gonadotropin-releasing hormone antagonists in poor responders. Fertil Steril. 2007;87(2):241-9.##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.##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.##Akman MA, Erden HF, Tosun SB, Bayazit N, Aksoy E, Bahceci M. Addition of GnRH antagonist in cycles of poor responders undergoing IVF. Hum Reprod. 2000;15(10):2145-7.##Sunkara SK, Coomarasamy A, Khalaf Y, Braude P. A three-arm randomised controlled trial comparing Gonadotrophin Releasing Hormone (GnRH) agonist long regimen versus GnRH agonist short regimen versus GnRH antagonist regimen in women with a history of poor ovarian response undergoing in vitro fertilisation (IVF) treatment: Poor responders intervention trial (PRINT). Reprod Health. 2007;4:12.##Akman MA, Erden HF, Tosun SB, Bayazit N, Aksoy E, Bahceci M. Comparison of agonistic flareup-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.##Malmusi S, La Marca A, Giulini S, Xella S, Tagliasacchi D, Marsella T, et al. Comparison of a gonadotropin-releasing hormone (GnRH) antagonist and GnRH agonist flare-up regimen in poor responders undergoing ovarian stimulation. Fertil Steril. 2005;84(2):402-6.##Demirol A, Gurgan T. Comparison of microdose flare-up and antagonist multiple-dose protocols for poor-responder patients: a randomized study. Fertil Steril. 2009;92(2):481-5.##Martinez F, Coroleu B, Marques L, Parera N, Buxaderas R, Tur R, et al. [Comparison of “Short Protocol” versus “Antagonists” with or without Clomiphene Citrate for stimulation in IVF of patients with “low response”]. Rev Iberoam Fertil Reprod Hum. 2003;20:355–60. Italian.##Schmidt DW, Bremner T, Orris JJ, Maier DB, Benadiva CA, Nulsen JC. A randomized prospective study of microdose leuprolide versus ganirelix in in vitro fertilization cycles for poor responders. Fertil Steril. 2005;83(5):1568-71.##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.##Devesa M, Martínez F, Coroleu B, Tur R, González C, Rodríguez I, et al. Poor prognosis for ovarian response to stimulation: results of a randomised trial comparing the flare-up GnRH agonist protocol vs. the antagonist protocol. Gynecol Endocrinol. 2010;26(7):509-15.##Berin I, Stein DE, Keltz MD. A comparison of gonadotropin-releasing hormone (GnRH) antagonist and GnRH agonist flare protocols for poor responders undergoing in vitro fertilization. Fertil Steril. 2010;93(2):360-3.##Lainas TG, Sfontouris IA, Papanikolaou EG, Zorzovilis JZ, Petsas GK, Lainas GT, et al. Flexible GnRH antagonist versus flare-up GnRH agonist protocol in poor responders treated by IVF: a randomized controlled trial. Hum Reprod. 2008;23(6): 1355-8.##Cheung LP, Lam PM, Lok IH, Chiu TT, Yeung S Y, Tjer CC, et al. GnRH antagonist versus long GnRH agonist protocol in poor responders undergoing IVF: a randomized controlled trial. Hum Reprod. 2005;20(3):616-21.##Marci R, Caserta D, Dolo V, Tatone C, Pavan A, Moscarini M. GnRH antagonist in IVF poor-responder patients: results of a randomized trial. Reprod Biomed Online. 2005;11(2):189-93.##Tazegül A, Görkemli H, Ozdemir S, Aktan TM. Comparison of multiple dose GnRH antagonist and minidose long agonist protocols in poor responders undergoing in vitro fertilization: a randomized controlled trial. Arch Gynecol Obstet. 2008;278(5): 467-72.##Sun Y, Zhu YM. [Gonadotropin-releasing hormone antagonist protocol in patients with risk of poor response to ovarian stimulation in IVF-ET]. Zhejiang Da Xue Xue Bao Yi Xue Ban. 2009;38(3):305-10. Chinese.##Tehraninejad ES, Fazel A, Samiei A, Rashidi B, Kiani K. Flexible multi-dose GnRH antagonist versus long GnRH agonist protocol in poor responders: A randomized controlled trial. Int J Fertil Stril. 2009;2(4):165-8.##Griesinger G, Diedrich K, Tarlatzis BC, Kolibianakis EM. GnRH-antagonists in ovarian stimulation for IVF in patients with poor response to gonadotrophins, polycystic ovary syndrome, and risk of ovarian hyperstimulation: a meta-analysis. Reprod Biomed Online. 2006;13(5):628-38.##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.##Orvieto R, Kruchkovich J, Rabinson J, Zohav E, Anteby EY, Meltcer S. Ultrashort gonadotropinreleasing hormone agonist combined with flexible multidose gonadotropin-releasing hormone antagonist for poor responders in in vitro fertilization/ embryo transfer programs. Fertil Steril. 2008;90 (1):228-30.##Humaidan P, Bungum L, Bungum M, Hald F, Agerholm I, Blaabjerg J, et al. Reproductive outcome using a GnRH antagonist (cetrorelix) for luteolysis and follicular synchronization in poor responder IVF/ICSI patients treated with a flexible GnRH antagonist protocol. Reprod Biomed Online. 2005;11(6):679-84.##Nilsson L, Andersen AN, Lindenberg S, Hausken J, Andersen CY, Kahn JA. Ganirelix for luteolysis in poor responder patients undergoing IVF treatment: a Scandinavian multicenter 'extended pilot study'. Acta Obstet Gynecol Scand. 2010;89(6): 828-31.##

RHAMM Expression in the Rat Endometrium during the Estrous Cycle and following Implantation 2 1 2 Background: Receptor for hyaluronic acid mediated motility (RHAMM) has intracellular and extracellular functions. In this study, we focus on the expression of RHAMM in the rat uterus during estrous cycle and implantation period. Methods: The female adult rats were divided into six groups following estrous cycle determination (n=36). The utreri of rats were collected according to estrous cycle phases (menstruation group). For the implantation groups, uteri were obtained on D4, D5 and D6 (day of implantation) of pregnancy. The tissue samples were fixed and cut into 5 µm thick sections. RHAMM was investigated using immunohistochemical techniques and the intensity of RHAMM was evaluated by using the Hscore technique. Comparisons between groups were performed using Kruskal-Wallis test. Results: The RHAMM immunoreactivity of uterine antimesometrial epithelium (343.00±12.81), mesometrial subepithelium (285.00±27.26) and mesometrial stroma (270.00±36.00) were more prominent (p<0.05) in the proestrus than estrus (275.00± 25.96; 220.00±14.48; 218.00±11.19) and diestrus (262.00±20.71; 192.50± 29.25; 216.00±12.97) groups, respectively. The most intense staining was seen in the epithelium on day four (275.50±30.06) and six (293.50±34.47) of pregnancy (p<0.05). Strong RHAMM expressions were in both mature and predecidual cells on D5 (256.00±18.71), (247.50±22.14) and D6 (256.00±30.72), (265.00±14.87), respectively. RHAMM expression was prominent in the nondecidual region on D5 (270.00± 13.36). Conclusion: Considering the role of RHAMM in cell proliferation, differentiation and angiogenesis, spatiotemporal expression of RHAMM in the uterus during estrous cycle and peri-implantation period is a means through which uterus becomes receptive for developing an embryo. 131 138 Kemal K Ozbilgin Kemal Ozbilgin Department of Histology and Embryology, School of Medicine, Celal Bayar University Department of Histology and Embryology, School of Medicine, Celal Bayar University Turkey kemalozbilgin@yahoo.com Banu B Boz Banu Boz Department of Histology and Embryology, School of Medicine, Celal Bayar University Department of Histology and Embryology, School of Medicine, Celal Bayar University Turkey Kazım K Tuğyan Kazım Tuğyan Department of Histology and Embryology, School of Medicine, Dokuz Eylul University Department of Histology and Embryology, School of Medicine, Dokuz Eylul University Turkey Sevinç S Inan Sevinç Inan Department of Histology and Embryology, School of Medicine, Celal Bayar University Department of Histology and Embryology, School of Medicine, Celal Bayar University Turkey Seda S Vatansever Seda Vatansever Department of Histology and Embryology, School of Medicine, Celal Bayar University Department of Histology and Embryology, School of Medicine, Celal Bayar University Turkey Estrous cycleImmunohistochemistryImplantationRHAMMUterus 505.pdf Hardwick C, Hoare K, Owens R, Hohn HP, Hook M, Moore D, et al. Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility. J Cell Biol. 1992;117(6):1343-50.##Choudhary M, Zhang X, Stojkovic P, Hyslop L, Anyfantis G, Herbert M, et al. Putative role of hyaluronan and its related genes, HAS2 and RHAMM, in human early preimplantation embryogenesis and embryonic stem cell characterization. Stem Cells. 2007;25(12):3045-57.##Savani RC, Cao G, Pooler PM, Zaman A, Zhou Z, DeLisser HM. Differential involvement of the hyaluronan (HA) receptors CD44 and receptor for HA-mediated motility in endothelial cell function and angiogenesis. J Biol Chem. 2001;276(39):36770-8.##Wang C, Thor AD, Moore DH 2nd, Zhao Y, Kerschmann R, Stern R, et al. The overexpression of RHAMM, a hyaluronan-binding protein that regulates ras signaling, correlates with overexpression of mitogen-activated protein kinase and is a significant parameter in breast cancer progression. Clin Cancer Res. 1998;4(3):567-76.##Rein DT, Roehrig K, Schöndorf T, Lazar A, Fleisch M, Niederacher D, et al. Expression of the hyaluronan receptor RHAMM in endometrial carcinomas suggests a role in tumour progression and metastasis. J Cancer Res Clin Oncol. 2003;129(3):161-4.##Lugli A, Zlobec I, Günthert U, Minoo P, Baker K, Tornillo L, et al. Overexpression of the receptor for hyaluronic acid mediated motility is an independent adverse prognostic factor in colorectal cancer. Mod Pathol. 2006;19(10):1302-9.##Li H, Guo L, Li JW, Liu N, Qi R, Liu J. Expression of hyaluronan receptors CD44 and RHAMM in stomach cancers: relevance with tumor progression. Int J Oncol. 2000;17(5):927-32.##Kornovski BS, McCoshen J, Kredentser J, Turley E. The regulation of sperm motility by a novel hyaluronan receptor. Fertil Steril. 1994;61(5):935-40.##Stojkovic M, Krebs O, Kölle S, Prelle K, Assmann V, Zakhartchenko V, et al. Developmental regulation of hyaluronan-binding protein (RHAMM/ IHABP) expression in early bovine embryos. Biol Reprod. 2003;68(1):60-6.##Li S, Davis B. Evaluating rodent vaginal and uterine histology in toxicity studies. Birth Defects Res B Dev Reprod Toxicol. 2007;80(3):246-52.##Westwood FR. The female rat reproductive cycle: a practical histological guide to staging. Toxicol Pathol. 2008;36(3):375-84.##Maruyama T, Masuda H, Ono M, Kajitani T, Yoshimura Y. Human uterine stem/progenitor cells: their possible role in uterine physiology and pathology. Reproduction. 2010;140(1):11-22.##Taylor HS. Endometrial cells derived from donor stem cells in bone marrow transplant recipients. JAMA. 2004;292(1):81-5.##Maxwell CA, Keats JJ, Crainie M, Sun X, Yen T, Shibuya E, et al. RHAMM is a centrosomal protein that interacts with dynein and maintains spindle pole stability. Mol Biol Cell. 2003;14(6):2262-76.##Assmann V, Jenkinson D, Marshall JF, Hart IR. The intracellular hyaluronan receptor RHAMM/ IHABP interacts with microtubules and actin filaments. J Cell Sci. 1999;112 ( Pt 22):3943-54.##Turley EA, Austen L, Moore D, Hoare K. Rastransformed cells express both CD44 and RHAMM hyaluronan receptors: only RHAMM is essential for hyaluronan-promoted locomotion. Exp Cell Res. 1993;207(2):277-82.##Salamonsen LA, Shuster S, Stern R. Distribution of hyaluronan in human endometrium across the menstrual cycle. Implications for implantation and menstruation. Cell Tissue Res. 2001;306(2):335-40.##Carson DD, Dutt A, Tang JP. Glycoconjugate synthesis during early pregnancy: hyaluronate synthesis and function. Dev Biol. 1987;120(1):228-35.##Teixeira Gomes RC, Verna C, Nader HB, dos Santos Simões R, Dreyfuss JL, Martins JR, et al. Concentration and distribution of hyaluronic acid in mouse uterus throughout the estrous cycle. Fertil Steril. 2009;92(2):785-92.##Gellersen B, Brosens IA, Brosens JJ. Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med. 2007;25(6):445-53.##Evanko SP, Tammi MI, Tammi RH, Wight TN. Hyaluronan-dependent pericellular matrix. Adv Drug Deliv Rev. 2007;59(13):1351-65.##Samuel SK, Hurta RA, Spearman MA, Wright JA, Turley EA, Greenberg AH. TGF-beta 1 stimulation of cell locomotion utilizes the hyaluronan receptor RHAMM and hyaluronan. J Cell Biol. 1993;123 (3):749-58.##Tolg C, Hamilton SR, Nakrieko KA, Kooshesh F, Walton P, McCarthy JB, et al. Rhamm-/- fibroblasts are defective in CD44-mediated ERK1,2 motogenic signaling, leading to defective skin wound repair. J Cell Biol. 2006;175(6):1017-28.##Chakraborty I, Das SK, Dey SK. Differential expression of vascular endothelial growth factor and its receptor mRNAs in the mouse uterus around the time of implantation. J Endocrinol. 1995;147(2): 339-52.##Douglas NC, Tang H, Gomez R, Pytowski B, Hicklin DJ, Sauer CM, et al. Vascular endothelial growth factor receptor 2 (VEGFR-2) functions to promote uterine decidual angiogenesis during early pregnancy in the mouse. Endocrinology. 2009;150 (8):3845-54.##Matou-Nasri S, Gaffney J, Kumar S, Slevin M. Oligosaccharides of hyaluronan induce angiogenesis through distinct CD44 and RHAMM-mediated signalling pathways involving Cdc2 and gamma-adducin. Int J Oncol. 2009;35(4):761-73.##Matou-Nasri S, Gaffney J, Kumar S, Slevin M. Oligosaccharides of hyaluronan induce angiogenesis through distinct CD44 and RHAMM-mediated signalling pathways involving Cdc2 and gamma-adducin. Int J Oncol. 2009;35(4):761-73.##Losa GA, Alini M. Sulfated proteoglycans in the extracellular matrix of human breast tissues with infiltrating carcinoma. Int J Cancer. 1993;54(4): 552-7.##

Antispasmodic Effects of Aqueous and Hydroalcoholic Punica granatum Flower Extracts on the Uterus of Non-pregnant Rats 2 1 2 Background: Punica granatum Linn. (PG) is native to the Mediterranean region. Its flower exhibited antioxidant activity. The present study attempt to investigate the effect of these extract on uterine contraction and its possible mechanism(s). Methods: Thirty five female Wistar rats (200-300 g) at estrous phases of cycle was examined in this study; pieces of virgin adult rat uterus (1.5 cm) were suspended in an organ bath containing 10 ml of De Jalon solution at 29 °C. Tissue contractility was isometrically recorded. KCl (60 mM), BaCl2 (4 mM) and oxytocin (10 mU/ml) were applied to the tissue in the presence and absence of aqueous and hydroalcoholic extracts of the plant (0.05, 0.1, 0.2, 0.4 and 0.8 mg/ml). Propranolol (1 µM) and naloxane (1 µM) were added in KCl induced contractions. The results were analyzed by ANOVA and p<0.05 were considered as significant. Results: Cumulative concentration of extracts reduced uterine contractions induced by KCl dose-dependently (p<0.01). Extracts in a dose dependent (p<0.05) reduced uterine contractions decreased dose-dependently after of addition oxytocin. The extracts added cumulatively to the organ bath reduced contractions but they did not affect uterine contractions induced by BaCl2 except the last dose. Spasmolytic effects of the extracts were not affected by propranolol or naloxane in KCl induced contractions. Conclusion: Extracts diminished K+-induced contraction in uterus, therefore it seems that substances that decrease K+-induced contraction can also block voltage dependent calcium channel. The extracts did not have any effect on β-adrenoceptors or potassium channels. 138 143 Akram A Ahangarpour اکرم آهنگرپور Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Iran ahang1002002@yahoo.com Razieh R Heidari Razieh Heidari Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Iran Mahsa M Abdolahzadeh Mahsa Abdolahzadeh Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Iran Ali Akbar AA Oroojan Ali Akbar Oroojan Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Department of Physiology, Physiology and Diabetes Research Centers, Jundishapur University of Medical Sciences Iran NaloxaneOxytocinPropranololPunica granatum flowerUterus 489.pdf Xie Y, Morikawa T, Ninomiya K, Imura K, Muraoka O, Yuan D, et al. Medicinal flowers. XXIII. New taraxastane-type triterpene, punicanolic acid, with tumor necrosis factor-alpha inhibitory activity from the flowers of Punica granatum. Chem Pharm Bull. 2008;56(11):1628-31.##Huang TH, Peng G, Kota BP, Li GQ, Yamahara J, Roufogalis BD, et al. Anti-diabetic action of Punica granatum flower extract: activation of PPAR-gamma and identification of an active component. Toxicol Appl Pharmacol. 2005;207(2):160-9.##Prashanth D, Asha MK, Amit A. Antibacterial activity of Punica granatum. Fitoterapia. 2001;72(2):171-3.##Das AK, Mandal SC, Banerjee SK, Sinha S, Das J, Saha BP, et al. Studies on antidiarrhoeal activity of Punica granatum seed extract in rats. J Ethnopharmacol. 1999;68(1-3):205-8.##Dutta BK, Rahman I, Das TK. Antifungal activity of Indian plant extracts. Mycoses. 1998;41(11-12):535-6.##Gharzouli K, Khennouf S, Amira S, Gharzouli A. Effects of aqueous extracts from Quercus ilex L. root bark, Punica granatum L. fruit peel and Artemisia herba-alba Asso leaves on ethanol-induced gastric damage in rats. Phytother Res. 1999;13(1): 42-5.##Gujral ML, Varma DR, Sareen KN. Oral contraceptives. Part I. Preliminary observations on the antifer-tility effect of some indigenous drugs. Indian J Med Res. 1960;48:46-51.##Hayouni EA, Miled K, Boubaker S, Bellasfar Z, Abedrabba M, Iwaski H, et al. Hydroalcoholic extract based-ointment from Punica granatum L. peels with enhanced in vivo healing potential on dermal wounds. Phytomedicine. 2011;18(11):976-84.##Lansky EP, Newman RA. Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethno-pharmacol. 2007;109(2):177-206.##Hassanpour Fard M, Ghule AE, Bodhankar SL, Dikshit M. Cardioprotective effect of whole fruit extract of pomegranate on doxorubicin-induced toxicity in rat. Pharm Biol. 2011;49(4):377-82.##Gasmi J, Sanderson JT. Growth Inhibitory, Antiandrogenic, and Pro-apoptotic Effects of Punicic Acid in LNCaP Human Prostate Cancer Cells. J Agric Food Chem. 2010 Nov 10. [Epub ahead of print]##Aviram M, Dornfeld L, Kaplan M, Coleman R, Gaitini D, Nitecki S, et al. Pomegranate juice flavornoids inhibit low-density lipoprotein oxidation and cardiovascular diseases: studies in atherosclerotic mice and in humans. Drugs Exp Clin Res. 2002;28(2-3):49-62.##Jafri MA, Aslam M, Javed K, Singh S. Effect of Punica granatum Linn. (flowers) on blood glucose level in normal and alloxan-induced diabetic rats. J Ethnopharmacol. 2000;70(3):309-14.##Zargari A. Medicinal plants. 5th ed. Tehan: Tehran University Press; 1992. p. 344.##Ahangarpour A, Oroojan AA. The effects of Cassia italica leaves aqueous extract on non-pregnant uterus contraction in rats. Iran J Reprod Med. 2010;8(4):179-84.##Naseri MK, Arabian M, Badavi M, Ahangarpour A. Vasorelaxant and hypotensive effects of Alliumcepa peel hydroalcoholic extract in rat. Pak J Biol Sci. 2008;11(12):1569-75.##Acritopoulou-Fourcroy S, Marçais-Collado H. Involvement of alpha-adrenoceptors in myometrial responses in the pro-oestral rat. Br J Pharmacol. 1988;93(1):185-91.##Gharib Naseri MK, Mohammadian M, Gharib Naseri Z. Antispasmodic effect of Physalis alkekengi fruit extract. Iran J Reprod Med. 2008;6(4):193-8.##Oropeza MV, Ponce-Monter H, Villanueva-Tello T, Palma-Aguirre JA, Campos MG. Anatomical differences in uterine sensitivity to prostaglandin F(2alpha) and serotonin in non-pregnant rats. Eur J Pharmacol. 2002;446(1-3):161-6.##Karaki H, Ozaki H, Hori M, Mitsui-Saito M, Amano K, Harada K, et al. Calcium movements, distribution, and functions in smooth muscle. Pharmacol Rev. 1997;49(2):157-230.##Zhou XB, Lutz S, Steffens F, Korth M, Wieland T. Oxytocin receptors differentially signal via Gq and Gi proteins in pregnant and nonpregnant rat uterine myocytes: implications for myometrial contractility. 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The Protective Effects of Exogenous Melatonin on Nicotine-induced Changes in Mouse Ovarian Follicles 2 1 2 Background: Nicotine exposure causes impaired fertility and ovarian dysfunction. The aim of this study was to investigate the possible protective role of melatonin, which is known as an antioxidant agent on altered ovarian functions upon nicotine exposure. Methods: A total of 32 female adult NMRI mice were divided randomly into four groups (n=8). The control group received vehicle, while group 2 received nicotine (40 μg/kg) for 15 days and group 3 melatonin (10 mg/kg) for 5 days. Group 4 received both nicotine (40μg/kg) and melatonin (10 mg/kg) for the same periods. All animals were treated intraperitoneally. After autopsy on the 16th day, histopathological and morphometrical examinations were performed and serum estradiol concentrations were measured. The data were analyzed using ANOVA and Tukey post hoc test. A value of p<0.05 was considered significant. Results: Nicotine significantly reduced the number of pre-antral and antral follicles, as well as estradiol concentration compared to the control group (p<0.05). However, the decrease in the number of primordial follicles was not significant in the nicotine treated group. A significant increase in the atretic follicles were observed in group 2 compared to the control group (p<0.05). Moreover, melatonin caused a marked normalization in the number of ovarian follicles and estradiol levels in group 4 compared to group 2. Conclusion: The results from this study suggest that melatonin may have a protective effect against nicotine-induced ovarian changes on the number of different stages of follicle growth. 143 151 Fahimeh F Mohammadghasemi فهیمه محمد قاسمی Cellular and Molecular Research Center, Guilan University of Medical Sciences Cellular and Molecular Research Center, Guilan University of Medical Sciences Iran Sina S Khajeh Jahromi سینا خواجه جهرمی Student Research Committee, Guilan University of Medical Sciences Student Research Committee, Guilan University of Medical Sciences Iran sina.khajehjahromi@gmail.com Hadi H Hajizadeh Hadi Hajizadeh Department of Pathology, Poursina Hospital, Guilan University of Medical Sciences Department of Pathology, Poursina Hospital, Guilan University of Medical Sciences Iran Mohammad Amin MA Homafar Mohammad Amin Homafar Student Research Committee, Guilan University of Medical Sciences Student Research Committee, Guilan University of Medical Sciences Iran Nazanin N Saadat Nazanin Saadat Student Research Committee, Guilan University of Medical Sciences Student Research Committee, Guilan University of Medical Sciences Iran NicotineMelatoninMouseOvaryProtectionOvarian follicles 486.pdf Sanders SR, Cuneo SP, Turzillo AM. Effects of nicotine and cotinine on bovine theca interna and granulosa cells. Reprod Toxicol. 2002;16(6):795-800.##Neal MS, Hughes EG, Holloway AC, Foster WG. Sidestream smoking is equally as damaging as mainstream smoking on IVF outcomes. Hum Reprod. 2005;20(9):2531-5.##Tuttle AM, Stämpfli M, Foster WG. Cigarette smoke causes follicle loss in mice ovaries at concentrations representative of human exposure. Hum Reprod. 2009;24(6):1452-9.##Hukkanen J, Jacob P 3rd, Benowitz NL. Metabolism and disposition kinetics of nicotine. Pharmacol Rev. 2005;57(1):79-115.##Xiao D, Huang X, Yang S, Zhang L. Direct effects of nicotine on contractility of the uterine artery in pregnancy. J Pharmacol Exp Ther. 2007;322(1): 180-5.##Patil SR, Ravindra, Patil SR, Londonkar R, Patil S B. Nicotine induced ovarian and uterine changes in albino mice. Indian J Physiol Pharmacol. 1998;42 (4):503-8.##Zeidler R, Albermann K, Lang S. Nicotine and apoptosis. Apoptosis. 2007;12(11):1927-43.##Petrik JJ, Gerstein HC, Cesta CE, Kellenberger LD, Alfaidy N, Holloway AC. Effects of rosiglitazone on ovarian function and fertility in animals with reduced fertility following fetal and neonatal exposure to nicotine. Endocrine. 2009;36(2):281-90.##Adriaens I, Jacquet P, Cortvrindt R, Janssen K, Smitz J. Melatonin has dose-dependent effects on folliculogenesis, oocyte maturation capacity and steroidogenesis. Toxicology. 2006;228(2-3):333-43.##Reiter RJ, Tan DX, Osuna C, Gitto E. Actions of melatonin in the reduction of oxidative stress. A review. J Biomed Sci. 2000;7(6):444-58.##Sanchez-Hidalgo M, de la Lastra CA, Carrascosa-Salmoral MP, Naranjo MC, Gomez-Corvera A, Caballero B, et al. Age-related changes in melatonin synthesis in rat extrapineal tissues. Exp Gerontol. 2009;44(5):328-34.##Velkov ZA, Velkov YZh, Galunska BT, Paskalev DN, Tadjer AV. Melatonin: Quantum-chemical and biochemical investigation of antioxidant activity. Eur J Med Chem. 2009;44(7):2834-9.##Ekmekcioglu C. Melatonin receptors in humans: biological role and clinical relevance. Biomed Pharmacother. 2006;60(3):97-108.##Bordel R, Laschke MW, Menger MD, Vollmar B. Nicotine does not affect vascularization but inhibits growth of freely transplanted ovarian follicles by inducing granulosa cell apoptosis. Hum Reprod. 2006;21(3):610-7.##Khajeh Jahromi S, Mohammadghasemi F, Hajizadeh Fallah H. [Evaluation of Proliferative activity of adult mouse male germ cells following administration of different doses of nicotine]. J Iran Anat Sci. 2011;9(36):229-40. Persian.##Poeggeler B, Saarela S, Reiter RJ, Tan DX, Chen L D, Manchester LC, et al. Melatonin--a highly potent endogenous radical scavenger and electron donor: new aspects of the oxidation chemistry of this indole accessed in vitro. Ann N Y Acad Sci. 1994;738:419-20.##Bandyopadhyay D, Biswas K, Bandyopadhyay U, Reiter RJ, Banerjee RK. Melatonin protects against stress-induced gastric lesions by scavenging the hydroxyl radical. J Pineal Res. 2000;29(3):143-51.##Hardeland R, Reiter RJ, Poeggeler B, Tan DX. The significance of the metabolism of the neurohormone melatonin: antioxidative protection and formation of bioactive substances. Neurosci Biobehav Rev. 1993;17(3):347-57.##Myers M, Britt KL, Wreford NG, Ebling FJ, Kerr J B. Methods for quantifying follicular numbers within the mouse ovary. Reproduction. 2004;127 (5):569-80.##Domingues SFS, Diniz LV, Furtado SHC, Ohashi OM, Rondina D, Silva LDM. Histological study of capuchin monkey (Cebus apella) ovarian follicles. Acta Amazon. 2004;34(3):495-501.##Dorostghoal M, Khaksari Mahabadi M, Adham S. Effects of maternal caffeine consumption on ovarian follicle development in wistar rats offspring. J Reprod Infertil. 2011;12(1):15-22.##Holloway AC, Kellenberger LD, Petrik JJ. Fetal and neonatal exposure to nicotine disrupts ovarian function and fertility in adult female rats. Endocrine. 2006;30(2):213-6.##Kim KH, Joo KJ, Park HJ, Kwon CH, Jang MH, Kim CJ. Nicotine induces apoptosis in TM3 mouse Leydig cells. Fertil Steril. 2005;83 Suppl 1:1093-9.##Holloway AC, Cuu DQ, Morrison KM, Gerstein HC, Tarnopolsky MA. Transgenerational effects of fetal and neonatal exposure to nicotine. Endocrine. 2007;31(3):254-9.##Holloway AC, Petrik JJ, Bruin JE, Gerstein HC. Rosiglitazone prevents diabetes by increasing betacell mass in an animal model of type 2 diabetes characterized by reduced beta-cell mass at birth. Diabetes Obes Metab. 2008;10(9):763-71.##Demiralay R, Gürsan N, Erdem H. Regulation of nicotine-induced apoptosis of pulmonary artery endothelial cells by treatment of N-acetylcysteine and vitamin E. Hum Exp Toxicol. 2007;26(7):595-602.##Machaalani R, Waters KA, Tinworth KD. Effects of postnatal nicotine exposure on apoptotic markers in the developing piglet brain. Neuroscience. 2005;132(2):325-33.##Jang MH, Shin MC, Jung SB, Lee TH, Bahn GH, Kwon YK, et al. Alcohol and nicotine reduce cell proliferation and enhance apoptosis in dentate gyrus. Neuroreport. 2002;13(12):1509-13.##Blackburn CW, Peterson CA, Hales HA, Carrell D T, Jones KP, Urry RL, et al. Nicotine, but not cotinine, has a direct toxic effect on ovarian function in the immature gonadotropin-stimulated rat. Reprod Toxicol. 1994;8(4):325-31.##Gocze PM, Szabo I, Freeman DA. Influence of nicotine, cotinine, anabasine and cigarette smoke extract on human granulosa cell progesterone and estradiol synthesis. Gynecol Endocrinol. 1999;13 (4):266-72.##Bódis J, Hanf V, Török A, Tinneberg HR, Borsay P, Szabó I. Influence of nicotine on progesterone and estradiol production of cultured human granulosa cells. Early Pregnancy. 1997;3(1):34-7.##Barbieri RL, McShane PM, Ryan KJ. Constituents of cigarette smoke inhibit human granulosa cell aromatase. Fertil Steril. 1986;46(2):232-6.##Vijayalaxmi, Reiter RJ, Tan DX, Herman TS, Thomas CR Jr. Melatonin as a radioprotective agent: a review. Int J Radiat Oncol Biol Phys. 2004;59(3): 639-53.##Woo MM, Tai CJ, Kang SK, Nathwani PS, Pang S F, Leung PC. Direct action of melatonin in human granulosa-luteal cells. J Clin Endocrinol Metab. 2001;86(10):4789-97.##Kim JK, Lee CJ. Effect of exogenous melatonin on the ovarian follicles in gamma-irradiated mouse. Mutat Res. 2000;449(1-2):33-9.##Cagnacci A, Soldani R, Yen SS. Exogenous melatonin enhances luteinizing hormone levels of women in the follicular but not in the luteal menstrual phase. Fertil Steril. 1995;63(5):996-9.##Bódis J, Koppán M, Kornya L, Tinneberg HR, Török A. Influence of melatonin on basal and gonadotropin-stimulated progesterone and estradiol secretion of cultured human granulosa cells and in the superfused granulosa cell system. Gynecol Obstet Invest. 2001;52(3):198-202.##Webley GE, Leidenberger F. The circadian pattern of melatonin and its positive relationship with progesterone in women. J Clin Endocrinol Metab. 1986;63(2):323-8.##Tamura H, Nakamura Y, Korkmaz A, Manchester LC, Tan DX, Sugino N, et al. Melatonin and the ovary: physiological and pathophysiological implications. Fertil Steril. 2009;92(1):328-43.##Mohamad Ghasemi F, Faghani M, Khajeh Jahromi S, Bahadori M, Nasiri E, Hemadi M. Effect Of Melatonin On Pproliferative Activity And Apoptosis In Spermatogenic Cells In Mouse Under Chemotherapy. J Reprod Contracept. 2010; 21(2):79–94.##Hill SM, Blask DE. Effects of the pineal hormone melatonin on the proliferation and morphological characteristics of human breast cancer cells (MCF-7) in culture. Cancer Res. 1988;48(21):6121-6.##Lissoni P, Barni S, Meregalli S, Fossati V, Cazzaniga M, Esposti D, et al. 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Derivation of Adipocytes from Human Endometrial Stem Cells (EnSCs) 2 1 2 Background: Due to increasing clinical demand for adipose tissue, a suitable cell for reconstructive adipose tissue constructs is needed. In this study, we investigated the ability of Human Endometrial-derived stem cells (EnSCs) as a new source of mesenchymal stem cells to differentiate into adipocytes. EnSCs are the abundant and easy available source with no immunological response, for cell replacement therapy. Methods: Single-cell suspensions of EnSCs were obtained from endometrial tissues from 10 women experiencing normal menstrual cycles, and were cultured at clonal density (10 cells/cm2) or limiting dilution. Endometrial mesenchymal stem cell markers were examined flow cytometry. These cells were treated with adipogenic-inducing medium for 28 days. The adipogenic differentiation of the EnSC was assessed by cellular morphology and further confirmed by Oil Red O staining and RT-PCR. The BM-MSC differentiated into adipocytes in the presence of adipogenic stimuli for 3 weeks. Results: The flow cytometric analysis showed that the cells were positive for CD90, CD105, CD146 and were negative for CD31, CD34.We showed that the key adipocytes marker PPARa was expressed in mRNA level after 28 days post treatment (PT). Conclusion: According to our finding, it can be concluded that EnSCs represent a useful in vitro model for human adipogenesis, and provide opportunities to study the stages prior to commitment to the adipocyte lineage. 151 158 Jafar Ai جعفر آی Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences Iran jafar_ai@tums.ac.ir Ahmad Reza AR Shahverdi Ahmad Reza Shahverdi Department of Pharmaceutical Biotechnology, School of Pharmacy, Tehran University of Medical Sciences Department of Pharmaceutical Biotechnology, School of Pharmacy, Tehran University of Medical Sciences Iran Somayeh S Ebrahimi Barough Somayeh Ebrahimi Barough Department of Biological Science, Faculty of Biology, University of Kharazmi Department of Biological Science, Faculty of Biology, University of Kharazmi Iran Homa H Mohseni Kouchesfehani Homa Mohseni Kouchesfehani Department of Biological Science, Faculty of Biology, University of Kharazmi Department of Biological Science, Faculty of Biology, University of Kharazmi Iran Saeed S Heidari Saeed Heidari Clinical Proteomics Research Center, Faculty of Paramedical Science, Shahid Beheshti University of Medical Sciences Clinical Proteomics Research Center, Faculty of Paramedical Science, Shahid Beheshti University of Medical Sciences Iran Reza R Roozafzoon Reza Roozafzoon Stem Cell Preparation Unit, Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences Stem Cell Preparation Unit, Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences Iran Javad J Verdi Javad Verdi Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences Iran Ahad A Khoshzaban Ahad Khoshzaban Stem Cell Preparation Unit, Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences Stem Cell Preparation Unit, Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences Iran Adipocyte cellDifferentiationEndometrial stem cell 499.pdf Langstein HN, Robb GL. Reconstructive approaches in soft tissue sarcoma. Semin Surg Oncol. 1999;17 (1):52-65.##Hedrick MH, Daniels EJ. The use of adult stem cells in regenerative medicine. Clin Plast Surg. 2003;30 (4):499-505.##Vashi AV, Keramidaris E, Abberton KM, Morrison WA, Wilson JL, O'Connor AJ, et al. Adipose differentiation of bone marrow-derived mesenchymal stem cells using Pluronic F-127 hydrogel in vitro. Biomaterials. 2008;29(5):573-9.##Conrad C, Huss R. Adult stem cell lines in regenerative medicine and reconstructive surgery. J Surg Res. 2005;124(2):201-8.##Pelled G, G T, Aslan H, Gazit Z, Gazit D. Mesenchymal stem cells for bone gene therapy and tissue engineering. Curr Pharm Des. 2002;8(21):1917-28.##Alhadlaq A, Tang M, Mao JJ. Engineered adipose tissue from human mesenchymal stem cells maintains predefined shape and dimension: implications in soft tissue augmentation and reconstruction. Tissue Eng. 2005;11(3-4):556-66.##Choi YS, Park SN, Suh H. Adipose tissue engineering using mesenchymal stem cells attached to injectable PLGA spheres. Biomaterials. 2005;26(29): 5855-63.##Neubauer M, Hacker M, Bauer-Kreisel P, Weiser B, Fischbach C, Schulz MB, et al. Adipose tissue engineering based on mesenchymal stem cells and basic fibroblast growth factor in vitro. Tissue Eng. 2005;11(11-12):1840-51.##Hong L, Peptan I, Clark P, Mao JJ. Ex vivo adipose tissue engineering by human marrow stromal cell seeded gelatin sponge. Ann Biomed Eng. 2005;33 (4):511-7.##Morganstein DL, Wu P, Mane MR, Fisk NM, White R, Parker MG. Human fetal mesenchymal stem cells differentiate into brown and white adipocytes: a role for ERRalpha in human UCP1 expression. Cell Res. 2010;20(4):434-44.##Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284(5411):143-7.##Gomillion CT, Burg KJ. Stem cells and adipose tissue engineering. Biomaterials. 2006;27(36):6052-63.##Smith AG. Embryo-derived stem cells: of mice and men. Annu Rev Cell Dev Biol. 2001;17:435-62.##Wagers AJ, Weissman IL. Plasticity of adult stem cells. Cell. 2004;116(5):639-48.##Dani C, Smith AG, Dessolin S, Leroy P, Staccini L, Villageois P, et al. Differentiation of embryonic stem cells into adipocytes in vitro. J Cell Sci. 1997; 110 ( Pt 11):1279-85.##Xiong C, Xie CQ, Zhang L, Zhang J, Xu K, Fu M, et al. Derivation of adipocytes from human embryonic stem cells. Stem Cells Dev. 2005;14(6):671-5.##De Ugarte DA, Alfonso Z, Zuk PA, Elbarbary A, Zhu M, Ashjian P, et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow. Immunol Lett. 2003;89(2-3):267-70.##Chan RW, Schwab KE, Gargett CE. Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod. 2004;70(6):1738-50.##Patel AN, Park E, Kuzman M, Benetti F, Silva FJ, Allickson JG. Multipotent menstrual blood stromal stem cells: isolation, characterization, and differentiation. Cell Transplant. 2008;17(3):303-11.##Schwab KE, Hutchinson P, Gargett CE. Identification of surface markers for prospective isolation of human endometrial stromal colony-forming cells. Hum Reprod. 2008;23(4):934-43.##Gargett CE, Chan RW, Schwab KE. Endometrial stem cells. Curr Opin Obstet Gynecol. 2007;19(4): 377-83.##Meng X, Ichim TE, Zhong J, Rogers A, Yin Z, Jackson J, et al. Endometrial regenerative cells: a novel stem cell population. J Transl Med. 2007; 5:57.##Gargett CE, Schwab KE, Zillwood RM, Nguyen H P, Wu D. Isolation and culture of epithelial progenitors and mesenchymal stem cells from human endometrium. Biol Reprod. 2009;80(6):1136-45.##Taherian MZ, Ai J, Ebrahimi Barough S, Yazdani BF, Rezayat Sorkhabadi SM, Vasei M, et al. Human endometrial stem cells as a new source for programming to neural cells. Cell Biol Int Rep. 2012;19(1):7-14.##Dimitrov R, Timeva T, Kyurkchiev D, Stamenova M, Shterev A, Kostova P, et al. Characterization of clonogenic stromal cells isolated from human endometrium. Reproduction. 2008;135(4):551-8.##Matthai C, Horvat R, Noe M, Nagele F, Radjabi A, van Trotsenburg M, et al. Oct-4 expression in human endometrium. Mol Hum Reprod. 2006;12 (1):7-10.##Gargett CE. Uterine stem cells: what is the evidence? Hum Reprod Update. 2007;13(1):87-101.##Bühring HJ, Battula VL, Treml S, Schewe B, Kanz L, Vogel W. Novel markers for the prospective isolation of human MSC. Ann N Y Acad Sci. 2007;1106:262-71.##Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, et al. Stem cell properties of human dental pulp stem cells. J Dent Res. 2002;81(8):531-5.##Crisan M, Yap S, Casteilla L, Chen CW, Corselli M, Park TS, et al. A perivascular origin for mesenchymal stem cells in multiple human organs. 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Pregnancy Predictors after Intrauterine Insemination: Analysis of 3012 Cycles in 1201 Couples 2 1 2 Background: Intrauterine insemination (IUI) is the first therapeutic step in assisted reproductive techniques and many factors, including male and female infertility and technique-dependent factors, have been reported to influence pregnancy rates after IUI. Methods: We carried out this retrospective study on 1201 couples undergoing 3012 intrauterine insemination cycles during 2002 to 2009. Pregnancy rate per cycle in terms of female infertility factors, male infertility factors, and technique-dependent factors were evaluated. The χ2, t-test, Kaplan-meier method, and multiple logistics regression model, were used for data analysis. The p<0.05 was considered statistically significant. Results: The highest pregnancy rates were obtained in cases whose infertility duration was shorter (p<0.05), Body Mass Index (BMI) was ≥25 (p<0.05), FSH<9 IU/L (p<0.05), anovulation due to polycystic ovary syndrome (p<0.05), donor sperm was used due to azoospermia (p<0.01), three IUI cycles (p<0.01), at least two follicles were recruited through controlled ovarian hyperstimulation (p<0.01), and where higher total doses of FSH were administered as necessary (p<0.05). Conclusion: This study characterizes predictors of pregnancy following IUI, for cases with shorter periods of infertility, BMI of 25 or more, FSH value below 9 IU/L, anovulation, donor sperm and performance of three intrauterine insemination cycles. 158 167 Macizo M Soria Macizo Soria Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Spain maribelmacizosoria@gmail.com Gálvez G Pradillo Gálvez Pradillo Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Spain Jorquera J García Jorquera García Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Spain Peinado P Ramón Peinado Ramón Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Spain Alvarez A Castillo Alvarez Castillo Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Spain Canteras C Jordana Canteras Jordana Department of Biostatistics, School of Medicine, University of Murcia Department of Biostatistics, School of Medicine, University of Murcia Spain Parrilla P Paricio Parrilla Paricio Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Department of Obstetrics and Gynecology, Human Reproduction Unit, Virgen de la Arrixaca University Hospital Spain GonadotropinIntra uterine inseminationOvarian hyperstimulationPregnancy rateSemen analysis 508.pdf Katzorke T, Kolodziej FB. 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Fertil Steril. 1989;51 (4):651-4.##Härkki P, Tiitinen A, Ylikorkala O. Endometriosis and assisted reproduction techniques. Ann N Y Acad Sci. 2010;1205:207-13.##Vlahos NF, Coker L, Lawler C, Zhao Y, Bankowski B, Wallach EE. Women with ovulatory dysfunction undergoing ovarian stimulation with clomiphene citrate for intrauterine insemination may benefit from administration of human chorionic gonadotropin. Fertil Steril. 2005;83(5):1510-6.##Dickey RP, Taylor SN, Lu PY, Sartor BM, Rye P H, Pyrzak R. Effect of diagnosis, age, sperm quality, and number of preovulatory follicles on the outcome of multiple cycles of clomiphene citrateintrauterine insemination. Fertil Steril. 2002;78(5): 1088-95.##Toma SK, Stovall DW, Hammond MG. The effect of laparoscopic ablation or danocrine on pregnancy rates in patients with stage I or II endometriosis undergoing donor insemination. Obstet Gynecol. 1992;80(2):253-6.##Dorjpurev U, Kuwahara A, Yano Y, Taniguchi T, Yamamoto Y, Suto A, et al. Effect of semen characteristics on pregnancy rate following intrauterine insemination. J Med Invest. 2011;58(1-2):127-33.##Sakhel K, Abozaid T, Schwark S, Ashraf M, Abuzeid M. Semen parameters as determinants of success in 1662 cycles of intrauterine insemination after controlled ovarian hyperstimulation. Fertil Steril. 2005;84 Suppl 1:S248-9.##Hughes EG. The effectiveness of ovulation induction and intrauterine insemination in the treatment of persistent infertility: a meta-analysis. Hum Reprod. 1997;12(9):1865-72.##Aboulghar M, Mansour R, Serour G, Abdrazek A, Amin Y, Rhodes C. Controlled ovarian hyperstimulation and intrauterine insemination for treatment of unexplained infertility should be limited to a maximum of three trials. Fertil Steril. 2001;75 (1):88-91.##Khalil MR, Rasmussen PE, Erb K, Laursen SB, Rex S, Westergaard LG. Intrauterine insemination with donor semen. An evaluation of prognostic factors based on a review of 1131 cycles. Acta Obstet Gynecol Scand. 2001;80(4):342-8.##Dickey RP, Olar TT, Taylor SN, Curole DN, Rye PH. Relationship of follicle number and other factors to fecundability and multiple pregnancy in clomiphene citrate-induced intrauterine insemination cycles. Fertil Steril. 1992;57(3):613-9.##Plosker SM, Jacobson W, Amato P. Predicting and optimizing success in an intrauterine insemination programme. Hum Reprod. 1994;9(11):2014-21.##Ransom MX, Blotner MB, Bohrer M, Corsan G, Kemmann E. Does increasing frequency of intrauterine insemination improve pregnancy rates significantly during superovulation cycles? Fertil Steril. 1994;61(2):303-7.##Ragni G, Maggioni P, Guermandi E, Testa A, Baroni E, Colombo M, et al. Efficacy of double intrauterine insemination in controlled ovarian hyperstimulation cycles. Fertil Steril. 1999;72(4):619-22.##Bagis T, Haydardedeoglu B, Kilicdag EB, Cok T, Simsek E, Parlakgumus AH. Single versus double intrauterine insemination in multi-follicular ovarian hyperstimulation cycles: a randomized trial. Hum Reprod. 2010;25(7):1684-90.##Cohlen BJ, Vandekerckhove P, te Velde ER, Habbema JD. Timed intercourse versus intrauterine insemination with or without ovarian hyperstimulation for subfertility in men. Cochrane Database Syst Rev. 2000;(2):CD000360.##Bry-Gauillard H, Coulondre S, Cédrin-Durnerin I, Hugues JN. [Benefits and risks of ovarian stimulition before intrauterine insemination]. Gynecol Obstet Fertil. 2000;28(11):820-31. French.##Cantineau AE, Cohlen BJ, Heineman MJ. Ovarian stimulation protocols (anti-oestrogens, gonadotrophins with and without GnRH agonists/antagonists) for intrauterine insemination (IUI) in women with subfertility. Cochrane Database Syst Rev. 2007;(2):CD005356.##Casadei L, Zamaro V, Calcagni M, Ticconi C, Dorrucci M, Piccione E. Homologous intrauterine insemination in controlled ovarian hyperstimulation cycles: a comparison among three different regimens. Eur J Obstet Gynecol Reprod Biol. 2006;129 (2):155-61.##Dankert T, Kremer JA, Cohlen BJ, Hamilton CJ, Pasker-de Jong PC, Straatman H, et al. A randomized clinical trial of clomiphene citrate versus low dose recombinant FSH for ovarian hyperstimulation in intrauterine insemination cycles for unexplained and male subfertility. Hum Reprod. 2007; 22(3):792-7.##Gerli S, Bini V, Di Renzo GC. Cost-effectiveness of recombinant follicle-stimulating hormone (FSH) versus human FSH in intrauterine insemination cycles: a statistical model-derived analysis. Gynecol Endocrinol. 2008;24(1):18-23.##Demirol A, Gurgan T. Comparison of different gonadotrophin preparations in intrauterine insemination cycles for the treatment of unexplained infertility: a prospective, randomized study. Hum Reprod. 2007;22(1):97-100.##Kocak M, Dilbaz B, Demir B, Taşci Y, Tarcan A, Dede S, et al. Lyophilised hMG versus rFSH in women with unexplained infertility undergoing a controlled ovarian stimulation with intrauterine insemination: a prospective, randomised study. Gynecol Endocrinol. 2010;26(6):429-34.##Sagnella F, Moro F, Lanzone A, Tropea A, Martinez D, Capalbo A, et al. A prospective random-ized noninferiority study comparing recombinant FSH and highly purified menotropin in intrauterine insemination cycles in couples with unexplained infertility and/or mild-moderate male factor. Fertil Steril. 2011;95(2):689-94.##Matorras R, Osuna C, Exposito A, Crisol L, Pijoan JI. Recombinant FSH versus highly purified FSH in intrauterine insemination: systematic review and metaanalysis. Fertil Steril. 2011;95(6):1937-42.##Balasch J, Fábregues F, Peñarrubia J, Creus M, Vidal R, Casamitjana R, et al. 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Contraception Use among Egyptian Women: Results from Egypt Demographic and Health Survey in 2005 2 1 2 Background: The reports of a rise in contraceptive practices have not been matched by a similar decrease in population: therefore, there is a need to look into the causes of this discrepancy. The objectives of this study were to determine the prevalence of different methods of contraception used by Egyptian women, to compare different contraception methods used among various socio-demographic groups and, finally, to identify the main decision makers of contraception use within Egyptian families. Methods: The 2005 Egypt Demographic and Health Survey (EDHS) is a nationally representative household survey of 18134 married women aged 15-49 years. The 2005 EDHS provides a wealth of information on fertility, family planning, maternal and child health and nutrition, and violence against women. The study sample was selected using a multistage sampling technique. A face-to-face structured interview was conducted with each of the selected women. The response rate was 99.5% for completing the questionnaires. Results: The prevalence of contraception was 57.5%, nearly one third of the participants (33.1%) used IUD as a method of contraception. Both male and female were responsible for decision making regarding the use of contraception among different educational levels. Most women reporting use of contraceptive methods were 30-39 years old, were employed, were rich, educated and belonged to urban governorates. Conclusion: More than half of the participants used contraception while IUDs and pills were the most commonly used methods. Whatever the level of education, the majority of women thought that family planning decisions should be made by both partners. 167 174 Hala H Ibrahim Awadalla Hala Ibrahim Awadalla Medical Science Department, Institute of Environmental Studies and Research, Ain Shams University, Abbassia Medical Science Department, Institute of Environmental Studies and Research, Ain Shams University, Abbassia Egypt Hala_Awadalla@yahoo.com ContraceptivesEgyptHusbandSociodemographic 507.pdf Yeakey MP, Muntifering CJ, Ramachandran DV, Myint Y, Creanga AA, Tsui AO. How contraceptive use affects birth intervals: results of a literature review. Stud Fam Plann. 2009;40(3):205-14.##Trussell J, Vaughan B. Contraceptive failure, method related discontinuation and resumption of use: results from the 1995 National Survey of Family Growth. Fam Plann Perspect. 1999;31(2):64-72.##Vaughan B, Trussell J, Kost K, Singh S, Jones R. Discontinuation and resumption of contraceptive use: results from the 2002 National Survey of Family Growth. Contraception. 2008;78(4):271-83.##Abasiattai AM, Bassey EA, Udoma EJ. Profile of intrauterine contraceptive device acceptors at the University of Uyo Teaching Hospital, Uyo, Nigeria. Ann Afr Med. 2008;7(1):1-5.##Eltigani EE. Toward replacement fertility in Egypt and Tunisia. Stud Fam Plann. 2009;40(3):215-26.##El-Zeini LO. The Status of Fertility Transition in Egypt and Morocco: Explaining the Differences. Paper presented at: 26th IUSSP International Population Conference; 2009 Sept 27-Oct 3; Morocco, Marrakech.##El-Zeini LO. The path to replacement fertility in Egypt: acceptance, preference, and achievement. Stud Fam Plann. 2008;39(3):161-76.##El-Zanaty F, Way A. Egypt demographic and health survey. 5th ed. Cairo: Ministry of Health and Population, National Population Council; 2005.##Leite IC, Gupta N. Assessing regional differences in contraceptive discontinuation, failure and switching in Brazil. Reprod Health. 2007;4:6.##D'Antona Ade O, Chelekis JA, D'Antona MF, Siqueira AD. Contraceptive discontinuation and nonuse in Santarém, Brazilian Amazon. Cad Saude Publica. 2009;25(9):2021-32.##MEASURE DHS [Internet]. USA: MEASURE DHS; 2012. 2009 Demographic and health surveys; [cited 2010 Oct 10]; [about 2 screens]. Available from: http://www.measuredhs.com##Curtis SL, Neitzel K. Contraceptive knowledge, use, and sources. Issue 19 of Demographic and Health Surveys comparative studies. Calverton: Macro International; 1996.##Kumar M, Meena J, Sharma S, Poddar A, Dhalliwal V, Modi-Satish Chander Modi SC, et al. Contraceptive use among low-income urban married women in India. J Sex Med. 2011;8(2):376-82.##Agha SY, Rasheed BO. Family planning and unmet need among Iraqi Kurds. East Mediterr Health J. 2007;13(6):1382-91.##United Nations, Department of Economic and Social Affairs, Population Division. World contraceptive use 2005. USA: United Nations; 2006.##El-Zanaty FH, Way AA, lil-Sukkān M, Macro International, Institute for Resource Development, Demographic and Health Surveys. Egypt interim demographic and health survey, 2003. Calverton: ORC Macro; 2004.##Adeyemi AS, Adekanle DA, Komolafe JO. Pattern of contraceptives choice among the married women attending the family planning clinic of a tertiary health institution. Niger J Med. 2008;17 (1):67-70.##Alvergne A, Gurmu E, Gibson MA, Mace R. Social transmission and the spread of modern contraception in rural Ethiopia. PLoS One. 2011;6(7): e22515.##Hong R, Montana L, Mishra V. Family planning services quality as a determinant of use of IUD in Egypt. BMC Health Serv Res. 2006;6:79.##Gubhaju B. The influence of wives' and husbands' education levels on contraceptive method choice in Nepal, 1996-2006. Int Perspect Sex Reprod Health. 2009;35(4):176-85.##Brown W, Ottney A, Nguyen S. Breaking the barrier: the Health Belief Model and patient perceptions regarding contraception. Contraception. 2011; 83(5):453-8.##al-Almaie SM. The pattern and factors associated with child spacing in eastern Saudi Arabia. J R Soc Promot Health. 2003;123(4):217-21.##Wambui T, Ek AC, Alehagen S. Perceptions of family planning among low-income men in Western Kenya. Int Nurs Rev. 2009;56(3):340-5.##Hussain S. Gender and Reproductive Behaviour: The Role of Men. Indian J Gend Stud. 2003;10:45-76.##Kabbash IA, El-Sayed NM, Al-Nawawy AN, Shady IK, Abou Zeid MS. Condom use among males (15-49 years) in Lower Egypt: knowledge, attitudes and patterns of use. East Mediterr Health J. 2007;13(6):1405-16.##DeJong J, El-Khoury G. Reproductive health of Arab young people. BMJ. 2006;333(7573):849-51.##Govindasamy P, Malhotra A. Women's position and family planning in Egypt. Stud Fam Plann. 1996;27(6):328-40.##Aryeetey R, Kotoh AM, Hindin MJ. Knowledge, perceptions and ever use of modern contraception among women in the Ga East District, Ghana. Afr J Reprod Health. 2010;14(4 Spec no.):26-31.##Nte AR, Odu N, Enyindah CE. Male involvement in family planning: women's perception. Niger J Clin Pract. 2009;12(3):306-10.##Wilson-Williams L, Stephenson R, Juvekar S, Andes K. Domestic violence and contraceptive use in a rural Indian village. Violence Against Women. 2008;14(10):1181-98.##

Detection of Partial Deletions of Y-chromosome AZFc in Infertile Men Using the Multiplex Ligation-dependent Probe Amplification Assay 2 1 2 Background: In recent studies, partial deletions of the azoospermia factor c region (AZFc) on the Y-chromosome have been detected in males with infertility problems. However, there has been a lot of debate about their significance. In order to study such deletions, a simple but accurate method for their detection was applied in this study. Methods: We present data obtained from the Multiplex Ligation-dependent Probe Amplification (MLPA) assay using a new Y-chromosome-specific MLPA probemix (P360) which allows the easy detection of partial AZFc deletions. Results: Partial AZFc deletions were detected in 8% of our cohort of previously mutation-negative infertile males (and 0% of the fertile control cohort). Conclusion: These results provide further evidence of the causality of partial AZFc deletions. None of the partial AZFc deletions were detectable by the standard multiplex PCR method, demonstrating the advantage of the MLPA method. 174 179 David J DJ Bunyan Wessex Regional Genetics Laboratory, Salisbury District Hospital Wessex Regional Genetics Laboratory, Salisbury District Hospital England Dave.Bunyan@salisbury.nhs.uk Jonathan L.A JLA Callaway Wessex Regional Genetics Laboratory, Salisbury District Hospital Wessex Regional Genetics Laboratory, Salisbury District Hospital England Nadja N Laddach Nadja Laddach MRC-Holland MRC-Holland Netherland CausalityGene dosageInfertilityMicrodeletionsMolecular geneticsY-Chromosome 506.pdf Tiepolo L, Zuffardi O. Localization of factors controlling spermatogenesis in the nonfluorescent portion of the human Y chromosome long arm. Hum Genet. 1976;34(2):119-24.##Navarro-Costa P, Gonçalves J, Plancha CE. The AZFc region of the Y chromosome: at the cross-roads between genetic diversity and male infertility. Hum Reprod Update. 2010;16(5):525-42.##Fernandes S, Huellen K, Goncalves J, Dukal H, Zeisler J, Rajpert De Meyts E, et al. High frequency of DAZ1/DAZ2 gene deletions in patients with severe oligozoospermia. Mol Hum Reprod. 2002;8 (3):286-98.##Hucklenbroich K, Gromoll J, Heinrich M, Hohoff C, Nieschlag E, Simoni M. Partial deletions in the AZFc region of the Y chromosome occur in men with impaired as well as normal spermatogenesis. Hum Reprod. 2005;20(1):191-7.##Zhang F, Li Z, Wen B, Jiang J, Shao M, Zhao Y, et al. A frequent partial AZFc deletion does not render an increased risk of spermatogenic impairment in East Asians. Ann Hum Genet. 2006;70(Pt 3):304-13.##Giachini C, Guarducci E, Longepied G, Degl'Innocenti S, Becherini L, Forti G, et al. The gr/gr deletion(s): a new genetic test in male infertility? J Med Genet. 2005;42(6):497-502.##Krausz C, Giachini C, Xue Y, O'Bryan MK, Gromoll J, Rajpert-de Meyts E, et al. Phenotypic variation within European carriers of the Y-chromosomal gr/gr deletion is independent of Y-chromosomal background. J Med Genet. 2009;46(1):21-31.##Simoni M, Bakker E, Krausz C. EAA/EMQN best practice guidelines for molecular diagnosis of y-chromosomal microdeletions. State of the art 2004. Int J Androl. 2004;27(4):240-9.##Schouten JP, McElgunn CJ, Waaijer R, Zwijnen-burg D, Diepvens F, Pals G. Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res. 2002;30(12):e57.##Noordam MJ, Westerveld GH, Hovingh SE, van Daalen SK, Korver CM, van der Veen F, et al. Gene copy number reduction in the azoospermia factor c (AZFc) region and its effect on total motile sperm count. Hum Mol Genet. 2011;20(12):2457-63.##Wu B, Lu NX, Xia YK, Gu AH, Lu CC, Wang W, et al. A frequent Y chromosome b2/b3 subdeletion shows strong association with male infertility in Han-Chinese population. 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