In vitro Human Embryo Culture; When Questions Outweigh Answers 2 1 578 2 More than five million babies have been born by the application of assisted conception methods throughout the world, and about 1% to 2% of babies in developed countries are born by the help of assisted reproductive techniques (ART) in couples with infertility. In spite of huge developments in equipments, techniques, procedures, supplies and greater control of environmental factors which affect assisted conception processes, there is mounting concern regarding anomalies and birth defects from assisted conception in the community and, especially, among infertile couples. Although this concern is very low compared to the time of application of IVF technology three decades ago, but results of more comprehensive longitudinal studies have revealed increased risk of congenital anomalies, malformation and early pregnancy loss associated with ART(1). What really increases the defects, or whether these defects are transferred through parental gametes to the offspring or whether suboptimal in vitro conditions cause permanent changes in gametes or embryos that will influence the future life of IVF babies are not clearly known yet. Comparison of babies conceived through ART with babies of infertile/subfertile couples conceived without these techniques and those of fertile couples emphasize the role of optimization of assisted conception processes on their success rate and on the present and future health of these babies. Several important factors influencing the quality and health of gametes and embryos in ART are the candidates themselves and the conditions, suboptimal for instance, under which ART processes are run. The factors also include conditions influencing controlled ovarian stimulation, gametes retrieval and processing, fertilization techniques and in vitro embryo cultures. Duration and condition of in vitro embryo culture have critical roles in IVF success rate and susceptibility of babies to different diseases in future. A large number of variables can affect development and quality of embryos during in vitro culture, including culture media, quantity of embryos, volume of each droplet, temperature, gas phase composition and quality, IVF laboratory air quality, culture ware/contact supplies, overlay oil, number and capacity of incubators, equipment validation and many other factors that need precise control during in vitro embryonic development (2). Regarding the large number of variables influencing in vitro culture of embryos, a question is raised whether which of the aforesaid factors could be of more importance and need greater attention and whether we really could control all these variables precisely similar to the controlling systems in the fallopian tubes and the uterus. As an example among hundreds of factors influencing ART outcomes, is the culture medium. For the time being, two commercial sets of media are available in the market, one set is sequential culture media with philosophy of "back to the nature" and another set is one-step culture media with philosophy of "let the embryo select". There are large numbers of documents and papers on the benefits of one set over the other in the development of embryos and many studies have not even confirmed the results of previous ones (3). The urge to design and perform studies in reputable institutes and publish the findings in high impact peer-review journals and the advertising push by companies to present their products, make it difficult for IVF clinics to select the best culture medium for in vitro development of embryos. On the other side already, most embryo development assessments are based on morphological criteria. Most changes in embryo microenvironment are not reflected on its morphology; therefore, it is time to change the assessment procedures and open the door for sub-cellular level of data integration methods such as genome, proteome, trans-criptome, glycome and metabolome. In fact, we should consider the compensatory mechanisms and whole embryonic functions at different levels of changes in each parameter (4). Hence, our knowledge on embryo culture against the unknown is too negligible. Recently, several studies have focused on the effects of physicochemical changes on the epigenetic status of cultured embryos, but their findings are preliminary and in some instances contradictory to one another (5). Therefore, to protect embryos and prevent harmful errors to their future health, we need to focus on the details of changes made to embryonic cultures and the compensatory functions of embryos towards those changes. But until the time we obtain these vital pieces of information, we should provide micro-environ-mental conditions as closely as possible to the womb conditions during in vitro embryonic cultures and this would not be possible except by implementing strict quality control/quality assurance programs. 48 50 Mohammad Reza MR Sadeghi محمدرضا صادقی Editor-in-chief Editor-in-chief Iran No Keyword 578.pdf Pinborg A, Henningsen AK, Malchau SS, Loft A. Congenital anomalies after assisted reproductive technology. Fertil Steril. 2013;99(2):327-32.##Lane M, Gardner DK. Embryo culture medium: which is the best? Best Pract Res Clin Obstet Gynaecol. 2007;21 (1):83-100.##Gruber I, Klein M. Embryo culture media for human IVF: which possibilities exist? J Turkish-German Gynecol Assoc. 2011;12:110-7.##Machtinger R, Racowsky C. Morphological systems of human embryo assessment and clinical evidence. Reprod Biomed Online. 2013;26(3):210-21.##El Hajj N, Haaf T. Epigenetic disturbances in in vitro cultured gametes and embryos: implications for human assisted reproduction. Fertil Steril. 2013 Jan 25. [Epub ahead of print].##

Investigation of Apelin Expression in Endometriosis 2 1 2 Background: Apelin is a mitogenic peptide; it has functions in vessel formation and cell proliferation. In this study we aimed to evaluate the serum and tissue levels and local expression pattern of apelin in eutopic and ectopic endometrium from patients with and without endometriosis and to compare the proliferative and secretory phase differences. Methods: Thirty women with endometriosis and 15 women without endometriosis undergoing surgery for benign indications as control group were included in the study. Serum and tissue concentrations and proliferative and secretory phase expression patterns of apelin were evaluated in the ectopic and eutopic endometrium using immunoassay and immunohistochemistry methods. The results were compared with Mann-Whitney U test. The p-values smaller than 0.05 were considered as statistically significant. Results: Apelin expression was detected in eutopic and ectopic endometrium of women with endometriosis and endometrium of control group. Intense immunoreactivity of apelin was observed in glandular cells of eutopic and ectopic endometrial tissues of women with endometriosis and endometrium of control group during secretory phase (p<0.01). In both groups, tissue concentrations of apelin were higher than of the serum (p=0.03) but, there were no significant differences between the two groups for tissue and serum concentrations of apelin. Conclusion: Apelin expression showed cyclic changes in eutopic and ectopic endometrium. Its expression may be related to menstrual changes of angiogenesis in endometrium of women. 50 56 Zehra Sema ZS Ozkan Zehra Sema Ozkan Department of Obstetrics and Gynecology, Firat University School of Medicine Department of Obstetrics and Gynecology, Firat University School of Medicine Turkey zehrasema@yahoo.com Hasan H Cilgin Hasan Cilgin Department of Obstetrics and Gynecology, Firat University School of Medicine Department of Obstetrics and Gynecology, Firat University School of Medicine Turkey Mehmet M Simsek Mehmet Simsek Department of Obstetrics and Gynecology, Firat University School of Medicine Department of Obstetrics and Gynecology, Firat University School of Medicine Turkey Bengu B Cobanoglu Bengu Cobanoglu Department of Pathology, Firat University School of Medicine Department of Pathology, Firat University School of Medicine Turkey Necip N Ilhan Necip Ilhan Department of Biochemistry, Firat University School of Medicine Department of Biochemistry, Firat University School of Medicine Turkey AngiogenesisApelinEndometriosisEndometriumExpression pattern 526.pdf Lapp T. ACOG issues recommendations for the management of endometriosis. American College of Obstetricians and Gynecologists. Am Fam Physician. 2000;62(6):1431-4.##Giudice LC, Kao LC. Endometriosis. Lancet. 2004;364(9447):1789-99.##Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM. Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol. 1984;64(2):151-4.##Kruitwagen RF, Poels LG, Willemsen WN, de Ronde IJ, Jap PH, Rolland R. Endometrial epithelial cells in peritoneal fluid during the early follicular phase. Fertil Steril. 1991;55(2):297-303.##Hudelist G, Keckstein J, Czerwenka K, Lass H, Walter I, Auer M, et al. Estrogen receptor beta and matrix metalloproteinase 1 are coexpressed in uterine endometrium and endometriotic lesions of patients with endometriosis. Fertil Steril. 2005;84 (Suppl) 2:1249-56.##Castan-Laurell I, Boucher J, Dray C, Daviaud D, Guigné C, Valet P. Apelin, a novel adipokine over-produced in obesity: friend or foe? Mol Cell Endocrinol. 2005;245(1-2):7-9.##Kasai A, Shintani N, Oda M, Kakuda M, Hashimoto H, Matsuda T, et al. Apelin is a novel angiogenic factor in retinal endothelial cells. Biochem Biophys Res Commun. 2004;325(2):395-400.##Shirasuna K, Shimizu T, Sayama K, Asahi T, Sasaki M, Berisha B, et al. Expression and localization of apelin and its receptor APJ in the bovine corpus luteum during the estrous cycle and prostaglandin F2alpha-induced luteolysis. Reproduction. 2008;135(4):519-25.##Kawamata Y, Habata Y, Fukusumi S, Hosoya M, Fujii R, Hinuma S, et al. Molecular properties of apelin: tissue distribution and receptor binding. Biochim Biophys Acta. 2001;1538(2-3):162-71.##Kunduzova O, Alet N, Delesque-Touchard N, Millet L, Castan-Laurell I, Muller C, et al. Apelin/APJ signaling system: a potential link between adipose tissue and endothelial angiogenic processes. FASEB J. 2008;22(12):4146-53.##Carpéné C, Dray C, Attané C, Valet P, Portillo MP, Churruca I, et al. Expanding role for the apelin/APJ system in physiopathology. J Physiol Biochem. 2007;63(4):359-73.##Masri B, Morin N, Cornu M, Knibiehler B, Audigier Y. Apelin (65-77) activates p70 S6 kinase and is mitogenic for umbilical endothelial cells. FASEB J. 2004;18(15):1909-11.##Katugampola SD, Maguire JJ, Matthewson SR, Davenport AP. [(125)I]-(Pyr(1))Apelin-13 is a novel radioligand for localizing the APJ orphan receptor in human and rat tissues with evidence for a vasoconstrictor role in man. Br J Pharmacol. 2001;132(6):1255-60.##Kleinz MJ, Davenport AP. Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells. Regul Pept. 2004;118(3):119-25.##Kleinz MJ, Skepper JN, Davenport AP. Immunocytochemical localisation of the apelin receptor, APJ, to human cardiomyocytes, vascular smooth muscle and endothelial cells. Regul Pept. 2005;126(3):233-40.##Hashimoto T, Kihara M, Ishida J, Imai N, Yoshida S, Toya Y, et al. Apelin stimulates myosin light chain phosphorylation in vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 2006;26(6):1267-72.##Takakura N, Kidoya H. Maturation of blood vessels by haematopoietic stem cells and progenitor cells: involvement of apelin/APJ and angiopoietin/Tie2 interactions in vessel caliber size regulation. Thromb Haemost. 2009;101(6):999-1005.##Tiani C, Garcia-Pras E, Mejias M, de Gottardi A, Berzigotti A, Bosch J, et al. Apelin signaling modulates splanchnic angiogenesis and portosystemic collateral vessel formation in rats with portal hypertension. J Hepatol. 2009;50(2):296-305.##Heinonen MV, Laaksonen DE, Karhu T, Karhunen L, Laitinen T, Kainulainen S, et al. Effect of diet-induced weight loss on plasma apelin and cytokine levels in individuals with the metabolic syndrome. Nutr Metab Cardiovasc Dis. 2009;19(9):626-33.##Kälin RE, Kretz MP, Meyer AM, Kispert A, Heppner FL, Brändli AW. Paracrine and autocrine mechanisms of apelin signaling govern embryonic and tumor angiogenesis. Dev Biol. 2007;305(2):599-614.##Noyes RW, Hertig AT, Rock J. Dating the endometrial biopsy. Fertil Steril. 1950;1:3-25.##Braun DP, Ding J, Shaheen F, Willey JC, Rana N, Dmowski WP. Quantitative expression of apoptosis-regulating genes in endometrium from women with and without endometriosis. Fertil Steril. 2007;87(2):263-8.##Kitawaki J, Kado N, Ishihara H, Koshiba H, Kitaoka Y, Honjo H. Endometriosis: the pathophysiology as an estrogen-dependent disease. J Steroid Biochem Mol Biol. 2002;83(1-5):149-55.##Huet-Hudson YM, Chakraborty C, De SK, Suzuki Y, Andrews GK, Dey SK. Estrogen regulates the synthesis of epidermal growth factor in mouse uterine epithelial cells. Mol Endocrinol. 1990;4(3):510-23.##Casimiri V, Rath NC, Parvez H, Psychoyos A. Effect of sex steroids on rat endometrial epithelium and stroma cultured separately. J Steroid Biochem. 1980;12:293-8.##Cunha GR, Chung LW, Shannon JM, Taguchi O, Fujii H. Hormone-induced morphogenesis and growth: role of mesenchymal-epithelial interactions. Recent Prog Horm Res. 1983;39:559-98.##Habata Y, Fujii R, Hosoya M, Fukusumi S, Kawamata Y, Hinuma S, et al. Apelin, the natural ligand of the orphan receptor APJ, is abundantly secreted in the colostrum. Biochim Biophys Acta. 1999;1452(1):25-35.##De Falco M, De Luca L, Onori N, Cavallotti I, Artigiano F, Esposito V, et al. Apelin expression in normal human tissues. In Vivo. 2002;16(5):333-6.##Kleinz MJ, Davenport AP. Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells. Regul Pept. 2004;118(3):119-25.##Tatemoto K, Takayama K, Zou MX, Kumaki I, Zhang W, Kumano K, et al. The novel peptide apelin lowers blood pressure via a nitric oxide-dependent mechanism. Regul Pept. 2001;99(2-3):87-92.##Ishida J, Hashimoto T, Hashimoto Y, Nishiwaki S, Iguchi T, Harada S, et al. Regulatory roles for APJ, a seven-transmembrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo. J Biol Chem. 2004;279(25):26274-9.##Zhong JC, Yu XY, Huang Y, Yung LM, Lau CW, Lin SG. Apelin modulates aortic vascular tone via endothelial nitric oxide synthase phosphorylation pathway in diabetic mice. Cardiovasc Res. 2007;74(3):388-95.##Cox CM, D'Agostino SL, Miller MK, Heimark RL, Krieg PA. Apelin, the ligand for the endothelial G-protein-coupled receptor, APJ, is a potent angiogenic factor required for normal vascular development of the frog embryo. Dev Biol. 2006;296(1):177-89.##Schilffarth S, Antoni B, Schams D, Meyer HH, Berisha B. The expression of apelin and its receptor APJ during different physiological stages in the bovine ovary. Int J Biol Sci. 2009;5(4):344-50.##Shimizu T, Kosaka N, Murayama C, Tetsuka M, Miyamoto A. Apelin and APJ receptor expression in granulosa and theca cells during different stages of follicular development in the bovine ovary: Involvement of apoptosis and hormonal regulation. Anim Reprod Sci. 2009;116(1-2):28-37.##Kidoya H, Takakura N. Biology of the apelin-APJ axis in vascular formation. J Biochem. 2012;152(2):125-31.##

Effects of L-carnitine and Pentoxifylline on the Activity of Lactate Dehydrogenase C4 isozyme and Motility of Testicular Spermatozoa in Mice 2 1 2 Background: Extracted sperm from the testis have poor motility. Moreover, their motility changes during their journey through epidydimis. Meanwhile, they face high concentration of L-carnitin. In addition, lactate dehydrogenase C4 (LDH-C4) gene disorders has been shown to cause impaired sperm motility, leading to infertility in male mice. The aim of this study was to evaluate sperm motility and LDH-C4 ezyme activity upon L-carnitine (LC) and Pentoxifylline (PTX) administrations in mice. Methods: We extracted testicular sperm of 48 mice and divided them into three equal parts. One part was incubated with Ham's F10 medium (control), the other parts were treated with Ham's F10 containing LC and PTX with a final concentration of 1.76 mM, for 30 min at room temperature. Sperm motility was assessed according to the World Health Organization (WHO) criteria. Sperm LDH-C4 enzyme activity was measured by spectrophotometery method. Statistical analyses were performed using ANOVA and Fisher's LSD test, and a p-value less than 0.05 was considered as a statistically significant difference. Results: Sperm motility increased after 30 min of incubation in LC- and PTX-treated group (p<0.001). LC and PTX administrations showed a significant increase in the LDHC4 enzyme activity of sperm compared to that of the controls after 30 min (P=0.04 and 0.01, respectively). Conclusion: The effects of LC and PTX on motility of sperm can be explained by an increase in LDH-C4 enzyme activity that may influence male fertility status. We suggest that LC as a non-toxic antioxidant is more suitable for use in assisted reproductive technique protocols than PTX. 56 62 Elham E Aliabadi Elham Aliabadi Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences Iran Fatemeh F Karimi Fatemeh Karimi Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences Iran Mozhgan M Rasti Mozhgan Rasti Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Iran rasti31@yahoo.com Masoumeh M Akmali Masoumeh Akmali Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences Iran Tahereh T Esmaeilpour Tahereh Esmaeilpour Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences Iran L-carnitineLDH-C4Male infertilityPentoxifyllineTesticular sperm 531.pdf Isidori AM, Pozza C, Gianfrilli D, Isidori A. edical treatment to improve sperm quality. Reprod Biomed Online. 2006;12(6):704-14.##Su LM, Palermo GD, Goldstein M, Veeck LL, Rosenwaks Z, Schlegel PN. Testicular sperm extraction with intracytoplasmic sperm injection for nonobstructive azoospermia: testicular histology can predict success of sperm retrieval. J Urol. 1999;161(1):112-6.##Mann T, Lutwak-Mann C. Studies on the metabolism of semen. 4. Aerobic and anaerobic utilization of fructose by spermatozoa and seminal vesicles. Biochem J. 1948;43(2):266-70.##Storey BT. Mammalian sperm metabolism: oxygen and sugar, friend and foe. Int J Dev Biol. 2008;52(5-6):427-37.##Miki K, Qu W, Goulding EH, Willis WD, Bunch DO, Strader LF, et al. Glyceraldehyde 3-phosphate dehydrogenase-S, a sperm-specific glycolytic enzyme, is required for sperm motility and male fertility. Proc Natl Acad Sci U S A. 2004;101(47):16501-6.##Li SS. Lactate dehydrogenase isoenzymes A (muscle), B (heart) and C (testis) of mammals and the genes coding for these enzymes. Biochem Soc Trans. 1989;17(2):304-7.##Duan C, Goldberg E. Inhibition of lactate dehydrogenase C4 (LDH-C4) blocks capacitation of mouse sperm in vitro. Cytogenet Genome Res. 2003;103(3-4):352-9.##Abd-Allah AR, Helal GK, Al-Yahya AA, Aleisa AM, Al-Rejaie SS, Al-Bakheet SA. Pro-inflammatory and oxidative stress pathways which compromise sperm motility and survival may be altered by L-carnitine. Oxid Med Cell Longev. 2009;2(2):73-81.##Odet F, Duan C, Willis WD, Goulding EH, Kung A, Eddy EM, et al. Expression of the gene for mouse lactate dehydrogenase C (Ldhc) is required for male fertility. Biol Reprod. 2008;79(1):26-34.##Sikka SC. Relative impact of oxidative stress on male reproductive function. Curr Med Chem. 2001;8(7):851-62.##Chaudiere J. Some chemical and biochemical constraints of oxidative stress in living cells. In: Riee-Evans CA, Burdon RH, editors. Free radical damage and its control. Amsterdam: Elsevier; 1994. p. 25-66.##Aitken RJ, Baker MA. Oxidative stress, sperm survival and fertility control. Mol Cell Endocrinol. 2006;250(1-2):66-9.##Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ Jr, et al. Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum Reprod. 2001;16(9):1922-30.##Lenzi A, Sgrò P, Salacone P, Paoli D, Gilio B, Lombardo F, et al. A placebo-controlled double-blind randomized trial of the use of combined l-carnitine and l-acetyl-carnitine treatment in men with asthenozoospermia. Fertil Steril. 2004;81(6): 1578-84.##Aliabadi E, Soleimani-Mehranjani M, Borzoei Z, Talaei-Khozani T, Mirkhani H, Tabesh H. Effects of L-carnitine and L-acetyl-carnitine on testicular sperm motility and chromatin quality. Iran J Reprod Med. 2012;10(2):77-82.##Kovacic B, Vlaisavljevic V, Reljic M. Clinical use of pentoxifylline for activation of immotile testicular sperm before ICSI in patients with azoospermia. J Androl. 2006;27(1):45-52.##Tournaye H, Janssens R, Devroey P, van Steirteghem A. The influence of pentoxifylline on motility and viability of spermatozoa from normozoospermic semen samples. Int J Androl. 1994;17(1):1-8.##Zare Z, Eimai H, Mohammadi M, Mofid M, dashtnavard H. The effect of orally administered l-carnitine on testis tissue, sperm parameters and daily sperm productionin adult mice. Cell J. 2010;11(4):382-9.##Jenkins DL, Griffith OW. Antiketogenic and hypoglycemic effects of aminocarnitine and acylaminocarnitines. Proc Natl Acad Sci U S A. 1986;83(2):290-4.##Beshay E, Croze F, Prud'homme GJ. The phosphodiesterase inhibitors pentoxifylline and rolipram suppress macrophage activation and nitric oxide production in vitro and in vivo. Clin Immunol. 2001;98(2):272-9.##Gavella M, Lipovac V, Marotti T. Effect of pentoxifylline on superoxide anion production by human sperm. Int J Androl. 1991;14(5):320-7.##Sato M, Ishikawa A. Room temperature storage of mouse epididymal spermatozoa: exploration of factors affecting sperm survival. Theriogenology. 2004;61(7-8):1455-69.##Balaban B, Urman B, Isiklar A, Alatas C, Aksoy S, Mercan R, et al. Progression to the blastocyst stage of embryos derived from testicular round spermatids. Hum Reprod. 2000;15(6):1377-82.##World Health Organization. WHO Laboratory manual for examination of human semen and sperm-cervical mucuinteraction. 4th ed. Cambridge: Cambridge University press; 2010.##Moreira SG, Lipshultz LI. Management of male infertility. Sci World J. 2004;4:214-448.##Wang YX, Yang SW, Qu CB, Huo HX, Li W, Li JD, et al. [L-carnitine: safe and effective for asthenozoospermia]. Zhonghua Nan Ke Xue. 2010;16(5):420-2.Chinese.##Shi JZ, Zhang SS, Zhang Z, Liang Q, Shi Y, Hua JL, et al. [Expressions of sperm-specific genes in carnitine-cultured testis sperm of obstructive azoospermia patients]. Zhonghua Nan Ke Xue. 2010;16(6):504-9. Chinese.##Jeulin C, Lewin LM. Role of free L-carnitine and acetyl-L-carnitine in post-gonadal maturation of mammalian spermatozoa. Hum Repord Update. 1996;2(2):87-102.##Brooks DE. Carnitine, acetylcarnitine and the activity of carnitine acyltransferases in seminal plasma and spermatozoa of men, rams and rats. J Reprod Fertil. 1979;56(2):667-73.##Casillas ER, Chaipayungpan S. The distribution of carnitine and acetylcarnitine in the rabbit epididymis and the carnitine content of rabbit spermatozoa during maturation. J Reprod Fertil. 1979;56(2):439-44.##Burgos C, Maldonado C, Gerez de Burgos NM, Aoki A, Blanco A. Intracellular localization of the testicular and sperm-specific lactate dehydrogenase isozyme C4 in mice. Biol Reprod. 1995;53(1):84-92.##Tanphaichitr N. In vitro stimulation of human sperm motility by acetylcarnitine. Int J Fertil. 1977;22(2):85-91.##Tasdemir I, Taşdemir M, Tavukcuoglu S. Effect of pentoxifylline on immotile testicular spermatozoa. J Assist Reprod Genet. 1998;15(2):90-2.##

Evaluation of Interleukin-10 (G-1082A) Promoter Polymorphism in Preeclampsia 2 1 2 Background: Preeclampsia is a pregnancy-specific syndrome that may be life-threatening, especially to the fetus. Several causes have been reported that may have a possible role in the development of the disorder. Interleukin-10 affect maternal intravascular inflammation, as well as endothelial dysfunction. The aim of this study was to investigate the association between IL-10 G-1082A polymorphism and pre-eclampsia. Methods: A total of eighty-eight pregnant women with preeclampsia and 100 women with normal pregnancy attending the Gynecological unit of Government Maternity Hospital, Petlaburz, Hyderabad, India, were considered for the study. A standard amplification refractory mutation system (ARMS) PCR was carried out for genotyping IL-10 G-1082A promoter polymorphism in all the participants. Genotypic distribution of the control and patient groups were compared with values predicted by Hardy-Weinberg equilibrium using χ2 test. Odd ratios (OR) and their respective 95% confidence intervals were used to measure the strength of association between IL-10 gene polymorphism and preeclampsia. Results: The frequencies of IL-10 G-1082A genotypes, GG, GA and AA, were 17.8%, 41.09% and 41.09% in women with preeclampsia and 25%, 28% and 47% in the controls respectively. There was no significant difference in the distribution of genotypes and alleles of IL-10 G-1082A between the two groups (Test power=0.66). Conclusion: The present study suggests that the IL-10 G-1082A gene promoter polymorphism is not a major genetic regulator in the etiology of preeclampsia. 62 67 Sowmya S Sabnavis Sowmya Sabnavis Institute of Genetics and Hospital for Genetic Diseases, Begumpet Institute of Genetics and Hospital for Genetic Diseases, Begumpet India Aruna A Ramaiah Aruna Ramaiah Government Maternity Hospital, Petlaburz Government Maternity Hospital, Petlaburz India Tella T Sunitha تلا سونیتا Institute of Genetics and Hospital for Genetic Diseases, Begumpet Institute of Genetics and Hospital for Genetic Diseases, Begumpet India Pratibha P Nallari Pratibha Nallari Department of Genetics, Osmania University Department of Genetics, Osmania University India Akka A Jyothy Institute of Genetics and Hospital for Genetic Diseases, Begumpet Institute of Genetics and Hospital for Genetic Diseases, Begumpet India Ananthapur A Venkateshwari Institute of Genetics and Hospital for Genetic Diseases, Begumpet Institute of Genetics and Hospital for Genetic Diseases, Begumpet India venkateshwari@yahoo.com ARMS PCRCytokinesInterleukin-10PolymorphismPreeclampsia 525.pdf Schobel HP, Fischer T, Heuszer K, Geiger H, Schmieder RE. Preeclampsia -- a state of sympathetic overactivity. N Engl J Med. 1996;335(20):1480-5.##Rochat RW, Koonin LM, Atrash HK, Jewett JF. Maternal mortality in the United States: report from the Maternal Mortality Collaborative. Obstet Gynecol. 1988;72(1):91-7.##Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science. 2005;308(5728):1592-4.##Roberts JM, Gammill HS. Preeclampsia: recent insights. Hypertension. 2005;46(6):1243-9.##Redman CW, Sacks GP, Sargent IL. Preeclampsia: an excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol. 1990;180:499-506.##Xue H, Lin B, An J, Zhu Y, Huang G. Interleukin-10-819 promoter polymorphism in association with gastric cancer risk. BMC Cancer. 2012;12:102.##de Jong BA, Westendorp RG, Eskdale J, Uitdehaag BM, Huizinga TW. Frequency of functional interleukin-10 promoter polymorphism is different between relapse-onset and primary progressive multiple sclerosis. Hum Immunol. 2002;63(4):281-5.##Lahiri DK, Nurnberger JI Jr. A rapid non-enzymatic method for the preparation of HMW DNA from blood for RFLP studies. Nucleic Acids Res. 1991;19(19):5444.##Perrey C, Turner SJ, Pravica V, Howell WM, Hutchinson IV. ARMS-PCR methodologies to determine IL-10, TNF-alpha, TNF-beta and TGF-beta 1 gene polymorphisms. Transpl Immunol. 1999;7(2):127-8.##Celik H, Avi B, Alper T. Comparison of maternal serum levels of interleukin-10, interleukin-12, and interleukin-2 in normal and preeclamptic pregnancies. Pregnancy Hypertens Int J Women Cardiovasc Health. 2012;2:39-42.##Roberts JM, Hubel CA. Is oxidative stress the link in the two-stage model of pre-eclampsia? Lancet. 1999;354(9181):788-9.##Orange S, Horvath J, Hennessy A. Preeclampsia is associated with a reduced interleukin-10 production from peripheral blood mononuclear cells. Hypertens Pregnancy. 2003;22(1):1-8.##Wegmann TG, Lin H, Guilbert L, Mosmann TR. Bidirectional cytokine interactions in the maternal-fetal relationship: is successful pregnancy a TH2 phenomenon? Immunol Today. 1993;14(7):353-6.##Darmochwal-Kolarz D, Leszczynska-Gorzelak B, Rolinski J, Oleszczuk J. T helper 1- and T helper 2-type cytokine imbalance in pregnant women with pre-eclampsia. Eur J Obstet Gynecol Reprod Biol. 1999;86(2):165-70.##Saito S, Umekage H, Sakamoto Y, Sakai M, Tanebe K, Sasaki Y, et al. Increased T-helper-1-type immunity and decreased T-helper-2-type immunity in patients with preeclampsia. Am J Reprod Immunol. 1999;41(5):297-306.##Roth I, Fisher SJ. IL-10 is an autocrine inhibitor of human placental cytotrophoblast MMP-9 production and invasion. Dev Biol. 1999;205(1):194-204.##Groux H, Cottrez F. The complex role of interleukin-10 in autoimmunity. J Autoimmun. 2003;20(4):281-5.##Westendorp RG, Langermans JA, Huizinga TW, Verweij CL, Sturk A. Genetic influence on cytokine production in meningococcal disease. Lancet. 1997;349(9069):1912-3.##de Craen AJ, Posthuma D, Remarque EJ, van den Biggelaar AH, Westendorp RG, Boomsma DI. Heritability estimates of innate immunity: an extended twin study. Genes Immun. 2005;6(2):167-70.##Bowen RS, Gu Y, Zhang Y, Lewis DF, Wang Y. Hypoxia promotes interleukin-6 and -8 but reduces interleukin-10 production by placental trophoblast cells from preeclamptic pregnancies. J Soc Gynecol Investig. 2005;12(6):428-32.##Rees LE, Wood NA, Gillespie KM, Lai KN, Gaston K, Mathieson PW. The interleukin-10-1082 G/A polymorphism: allele frequency in different populations and functional significance. Cell Mol Life Sci. 2002;59(3):560-9.##Vural P, Degirmencioglu S, Saral NY, Demirkan A, Akgul C, Yildirim G, et al. Tumor necrosis factor alpha, interleukin-6 and interleukin-10 polymorphisms in preeclampsia. J Obstet Gynaecol Res. 2010;36(1):64-71.##Daher S, Sass N, Oliveira LG, Mattar R. Cytokine genotyping in preeclampsia. Am J Reprod Immunol. 2006;55(2):130-5.##Stonek F, Hafner E, Metzenbauer M, Katharina S, Stümpflen I, Schneeberger C, et al. Absence of an association of tumor necrosis factor (TNF)-alpha G308A, interleukin-6 (IL-6) G174C and interleukin-10 (IL-10) G1082A polymorphism in women with preeclampsia. J Reprod Immunol. 2008;77(1):85-90.##Kamali-Sarvestani E, Kiany S, Gharesi-Fard B, Robati M. Association study of IL-10 and IFN-gamma gene polymorphisms in Iranian women with preeclampsia. J Reprod Immunol. 2006;72(1-2):118-26.##Makris A, Xu B, Yu B, Thornton C, Hennessy A. Placental deficiency of interleukin-10 (IL-10) in preeclampsia and its relationship to an IL10 promoter polymorphism. Placenta. 2006;27(4-5):445-51.##Xie C, Yao MZ, Liu JB, Xiong LK. A meta-analysis of tumor necrosis factor-alpha, interleukin-6, and interleukin-10 in preeclampsia. Cytokine. 2011;56(3):550-9.##

Effects of Chlamydia trachomatis Infection on Fertility; A Case-Control Study 2 1 2 Background: Nowadays, Chlamydia trachomatis is known as a causative agent of infertility. Because of, asymptomatic nature of infection, many may suffer from its lasting complications such as infertility. This study was performed in Tehran during April 2007 to April 2008 to compare the prevalence of Chlamydia trachomatis infection in fertile and infertile women using ELISA and PCR methods. Methods: Overall, 234 infertile and 223 pregnant women, as the fertile group, participated in this hospital-based case-control study. After completing an informed consent form and the questionnaire, first catch urine and blood sample were obtained for PCR and ELISA (IgG, IgM) tests, respectively. Logistic regression analysis was used to control possible confounding factors, and determine adjusted odds ratio of infertility due to the infection. Results: PCR results revealed that 29 (12.4%) of the infertile and 19 (8.5%) of the fertile women were positive for C. trachomatis infection (p=0.440). IgG was positive in 21 (9.0%) of the infertile and 11 (5.0%) in the fertile group (p=0.093). IgM assays identified that 2 (0.9%) of the infertile and 4 (1.8%) of the fertile women were positive for the micro-organism (p=0.375). Conclusion: We found no significant differences among fertile and infertile women for Chlamydia trachomatis infection. Nevertheless, molecular techniques which are more sensitive, more specific and non-invasive can be used to detect C. trachomatis infection. 67 73 Batool B Hossein Rashidi بتول حسین رشیدی Vali-e- Asar Reproductive Health Research Center,Tehran Medical Sciences University Vali-e- Asar Reproductive Health Research Center,Tehran Medical Sciences University Iran Leili L Chamani Tabriz لیلی چمنی تبریز Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran lchamani@avicenna.ac.ir Fedyeh F Haghollahi فدیه حق‌اللهی Vali-e- Asar Reproductive Health Research Center,Tehran Medical Sciences University Vali-e- Asar Reproductive Health Research Center,Tehran Medical Sciences University Iran Mahmood M Jeddi-Tehrani محمود جدی‌تهرانی Monoclonal Antibody Research Center, Avicenna Research Institute (ACECR) Monoclonal Antibody Research Center, Avicenna Research Institute (ACECR) Iran Mohammad Mehdi MM Naghizadeh محمدمهدی نقی زاده Department of Biostatistics and Epidemiology, Fassa University of Medical Sciences Department of Biostatistics and Epidemiology, Fassa University of Medical Sciences Iran Mamak M Shariat مامک شریعت Maternal-Fetal-Neonatal Health Research Center, Tehran University of Medical Sciences Maternal-Fetal-Neonatal Health Research Center, Tehran University of Medical Sciences Iran Mohammad Mehdi MM Akhondi محمدمهدی آخوندی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Rezvan R Bagheri Rezvan Bagheri Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Soheila S Asgari سهیلا عسگری Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Kevan K Wylie Kevan Wylie Consultant in sexual Medicine Consultant in sexual Medicine United Kingdom Case control studyChlamydia trachomatisEnzyme-linked immunosorbent assayInfertilityPolymerase chain reaction 528.pdf Weinstock H, Berman S, Cates W Jr. Sexually transmitted diseases among American youth: incidence and prevalence estimates, 2000. Perspect Sex Reprod Health. 2004;36(1):6-10.##Centers for Disease Control and Prevention [Internet]. USA: Centers for Disease Control and Prevention; 2005 Sexually transmitted diseases surveillance; 2006 Nov [cited 2008 Oct 26]; Available from: http://www.cdc.gov/std/stats05/toc2005.htm##Chen MY, Rohrsheim R, Donovan B. Chlamydia trachomatis infection in Sydney women. Aust N Z J Obstet Gynaecol. 2005;45(5):410-3.##Chen MY, Donovan B. Changes in testing methods for genital Chlamydia trachomatis in New South Wales, Australia, 1999 to 2002. Sex Health. 2005;2(4):251-3.##Stamm WE, Jones RB, Batteiger BE. Chlamydia trachomatis (trachoma, prinatal infections, lymphogranuloma venereum and other genital infections). In: Mandell GL, Dolin R, Bennette JE, editors. Principles and practice of infectious diseases. Philadelphia: Elsevier Churchill Livingstone; 2005. p. 2239-51.##Guven MA, Dilek U, Pata O, Dilek S, Ciragil P. Prevalance of Chlamydia trochomatis, Ureaplasma urealyticum and Mycoplasma hominis infections in the unexplained infertile women. Arch Gynecol Obstet. 2007;276(3):219-23.##Numazaki K, Asanuma H, Niida Y. Chlamydia trachomatis infection in early neonatal period. BMC Infect Dis. 2003;3:2.##Wagenlehner FM, Weidner W, Naber KG. Chlamydial infections in urology. World J Urol. 2006;24(1):4-12.##den Hartog JE, Morré SA, Land JA. Chlamydia trachomatis-associated tubal factor subfertility: Immunogenetic aspects and serological screening. Hum Reprod Update. 2006;12(6):719-30.##Siemer J, Theile O, Larbi Y, Fasching PA, Danso KA, Kreienberg R, et al. Chlamydia trachomatis infection as a risk factor for infertility among women in Ghana, West Africa. Am J Trop Med Hyg. 2008;78(2):323-7.##Al-Ramahi M, Mahafzah A, Saleh S, Fram K. Prevalence of Chlamydia trachomatis infection in infertile women at a university hospital in Jordan. East Mediterr Health J. 2008;14(5):1148-54.##Malik A, Jain S, Hakim S, Shukla I, Rizvi M. Chlamydia trachomatis infection & female infertility. Indian J Med Res. 2006;123(6):770-5.##Chamani-Tabriz L, Tehrani MJ, Akhondi MM, Mosavi-Jarrahi A, Zeraati H, Ghasemi J, et al. Chlamydia trachomatis prevalence in Iranian women attending obstetrics and gynaecology clinics. Pak J Biol Sci. 2007;10(24):4490-4.##Jenab A, Roghanian R, Golbang N, Golbang P, Chamani-Tabriz L. Comparison of three meth-ods of DNA extraction in endocervical specimens for Chlamydia trachomatis infection by spectrophotometry, agarose gel, and PCR. Arch Immunol Ther Exp. 2010;58(3):227-34.##Sharma M, Sethi S, Daftari S, Malhotra S. Evidence of chlamydial infection in infertile women with fallopian tube obstruction. Indian J Pathol Microbiol. 2003;46(4):680-3.##Sambrook J, Russell DW. Molecular cloning: A laboratory manual. 3rd ed. New York: Cold Spring Harbor Laboratory Press; 2001. 2344 p.##Sirmatel F, Sahin N, Sirmatel O, Telli E, Kececi S. Chlamydia trachomatis antigen positivity in women in risk groups and its relationship with the use of antibiotics. Jpn J Infect Dis. 2005;58(1):41-3.##Kucinskiene V, Sutaite I, Valiukeviciene S, Milasauskiene Z, Domeika M. Prevalence and risk factors of genital Chlamydia trachomatis infection. Medicina. 2006;42(11):885-94.##Corson SL. The role of laparoscopy in the infertility work-up. J Reprod Med. 1977;18(3):127-31.##Centers for Disease Control and Prevention [Internet]. USA: Centers for Disease Control and Prevention; 2012. Sexually transmitted disease surveillance; 2002 [cited 2008 Oct 26]; Available from: http://www.cdc.gov/std/stats02/toc2002.htm##Moaiedmohseni M, Owje M. The value of chlamydia trachomatis antibody testing in prediction of tubal factor infertility. J Family Reprod Health. 2008;2(1):29-32.##Basirat Z, Sharbatdaran M, Montazar F. Comparison of anti Chlamydia antibodies in tubal and non-tubal infertile patients. J Mazandaran Univ Med Sci. 2006;16(55)118-24.##Tukur J, Shittu SO, Abdul AM. A case control study of active genital Chlamydia trachomatis infection among patients with tubal infertility in northern Nigeria. Trop Doct. 2006;36(1):14-6.##Taylor-Robinson D. Chlamydia trachomatis and sexually transmitted disease. BMJ. 1994;308 (6922):150-1.##Olshen E, Shrier LA. Diagnostic tests for chlamydial and gonorrheal infections. Semin Pediatr Infect Dis. 2005;16(3):192-8.##Newhall WJ, Johnson RE, DeLisle S, Fine D, Hadgu A, Matsuda B, et al. Head-to-head evaluation of five chlamydia tests relative to a quality-assured culture standard. J Clin Microbiol. 1999; 37(3):681-5.##Black CM. Current methods of laboratory diagnosis of Chlamydia trachomatis infections. Clin Microbiol Rev. 1997;10(1):160-84.##Moncada J, Schachter J, Hook EW 3rd, Ferrero D, Gaydos C, Quinn TC, et al. The effect of urine testing in evaluations of the sensitivity of the Gen-Probe Aptima Combo 2 assay on endocervical swabs for Chlamydia trachomatis and neisseria gonorrhoeae: the infected patient standard reduces sensitivity of single site evaluation. Sex Transm Dis. 2004;31(5):273-7.##Shrier LA, Dean D, Klein E, Harter K, Rice PA. Limitations of screening tests for the detection of Chlamydia trachomatis in asymptomatic adolescent and young adult women. Am J Obstet Gynecol. 2004;190(3):654-62.##van Doornum GJ, Schouls LM, Pijl A, Cairo I, Buimer M, Bruisten S. Comparison between the LCx Probe system and the COBAS AMPLICOR system for detection of Chlamydia trachomatis and Neisseria gonorrhoeae infections in patients attending a clinic for treatment of sexually transmitted diseases in Amsterdam, The Netherlands. J Clin Microbiol. 2001;39(3):829-35.##

Effects of Varicocelectomy on Anti-sperm Antibody in patients with Varicocele 2 1 2 Background: Anti-sperm antibody (ASA) can decrease sperm motility and, therefore, it is a cause of male infertility. The aim of this study was to evaluate the effects of varicocelectomy on anti-sperm antibody in patients with varicocele. Methods: This observational study was conducted on 90 patients with varicocele at Sina and Imam Khomeini hospitals during 2006 to 2009. All varicocelectomy candidates were selected for ASA assessment both in semen and serum before and after surgery. ASA level was measured using a direct method for semen and an indirect method of Sperm MAR test, for serum. Paired t-test and McNemar's test were used for data analysis, and p<0.05 was considered statistically significant. Results: ASA level in semen was 13.7% before, and 15.7% after three month of varicocelectomy (p=0.881). Serum level of ASA before and after surgery were 13.6% and 21.7%, respectively (p=0.033). Three parameters including sperm count, motility and morphology showed recovery following, varicocelectomy, but only the difference in sperm motility was significant (p<0.05). Conclusion: This study showed that varicocelectomy has no effect on semen ASA. Although serum antibody has been shown to increase after varicocelectomy but sperm motility will improve. Varicocelectomy seems to have a beneficial effect on semen parameters in infertile men with varicocele. 73 79 Mohammad Reza MR Bonyadi Mohammad Reza Bonyadi Drug Applied Research Center, Faculty of Medicine, Tabriz University of Medical Sciences Drug Applied Research Center, Faculty of Medicine, Tabriz University of Medical Sciences Iran bonyadir@tbzmed.ac.ir Sayyed Kazem SK Madaen Sayyed Kazem Madaen Department of Urology, Faculty of Medicine, Tabriz University of Medical Sciences Department of Urology, Faculty of Medicine, Tabriz University of Medical Sciences Iran Maryam M Saghafi Maryam Saghafi School of Public Health and Nutrition, Tabriz University of Medical Sciences School of Public Health and Nutrition, Tabriz University of Medical Sciences Iran Anti-sperm AntibodyInfertilitySperm motilityVaricocelectomy 527.pdf Rogenio A, Lobo A, Daniel R, Richard JR, Pavlson M. Role of immunology in infertility. In: Lobo A, Mishell DR, Paulsen RJ, Shoupe D, editors. Infertility, contraception and reproductive endocrinology. Massachusetts: Blackwell Science; 1997. p. 675-9.##Bronson RA, O'Connor WJ, Wilson TA, Bronson SK, Chasalow FI, Droesch K. Correlation between puberty and the development of autoimmunity to spermatozoa in men with cystic fibrosis. Fertil Steril. 1992;58(6):1199-204.##Flickinger CJ, Baran ML, Howards SS, Herr JC. Sperm autoantigens recognized by autoantibodies in developing rats following prepubertal obstruction of the vas deferens. J Androl. 1996;17(4):433-42.##De Almeida M, Soumah A, Jouannet P. Incidence of sperm-associated immunoglobulins in infertile men with suspected autoimmunity to sperm. Int J Androl. 1986;9(5):321-30.##Heidenreich A, Bonfig R, Wilbert DM, Strohmaier WL, Engelmann UH. Risk factors for antisperm antibodies in infertile men. Am J Reprod Immunol. 1994;31(2):69-76.##Sinisi AA, Di Finizio B, Pasquali D, Scurini C, D'Apuzzo A, Bellastella A. Prevalence of antisperm antibodies by SpermMARtest in subjects undergoing a routine sperm analysis for infertility. Int J Androl. 1993;16(5):311-4.##Haas GG Jr, Lambert H, Stern JE, Manganiello P. Comparison of the direct radiolabeled antiglobulin assay and the direct immunobead binding test for detection of sperm-associated antibodies. Am J Reprod Immunol. 1990;22(3-4):130-2.##Ohl DA, Naz RK. Infertility due to antisperm antibodies. Urology. 1995;46(4):591-602.##Oshinsky GS, Rodriguez MV, Mellinger BC. Varicocele-related infertility is not associated with increased sperm-bound antibody. J Urol. 1993;150(3):871-3.##Flickinger CJ, Howards SS, Bush LA, Baker LA, Herr JC. Antisperm autoantibody responses to vasectomy and vasovasostomy in Fischer and Lewis rats. J Reprod Immunol. 1995;28(2):137-57.##Marc GO. Surgical management of male infertility and other scrotal disorders. In: Walsh PC, Retik AB, Vavghao ED, Wein AJ, editors. Campbell's Urology. Philadelphia: W.B. Saunders; 2002. p. 1571-7.##Rajah SV, Parslow JM, Howell RJ, Hendry WF. Comparison of mixed antiglobulin reaction and direct immunobead test for detection of sperm-bound antibodies in subfertile males. Fertil Steril. 1992;57(6):1300-3.##Almagor M, Margalioth EJ, Yaffe H. Density differences between spermatozoa with antisperm autoantibodies and spermatozoa covered with antisperm antibodies from serum. Hum Reprod. 1992;7(7):959-61.##Cayan S, Kadioglu TC, Tefekli A, Kadioglu A, Tellaloglu S. Comparison of results and complications of high ligation surgery and microsurgical high inguinal varicocelectomy in the treatment of varicocele. Urology. 2000;55(5):750-4.##Djaladat H, Mehrsai A, Rezazade M, Djaladat Y, Pourmand G. Varicocele and antisperm antibody: fact or fiction? South Med J. 2006;99(1):44-7.##Ozen H, Asar G, Gungor S, Peker AF. Varicocele and antisperm antibodies. Int Urol Nephrol. 1985;17(1):97-101.##Golomb J, Vardinon N, Homonnai ZT, Braf Z, Yust I. Demonstration of antispermatozoal antibodies in varicocele-related infertility with an enzyme-linked immunosorbent assay (ELISA). Fertil Steril. 1986;45(3):397-402.##Knudson G, Ross L, Stuhldreher D, Houlihan D, Bruns E, Prins G. Prevalence of sperm bound antibodies in infertile men with varicocele: the effect of varicocele ligation on antibody levels and semen response. J Urol. 1994;151(5):1260-2.##Gilbert BR, Witkin SS, Goldstein M. Correlation of sperm-bound immunoglobulins with impaired semen analysis in infertile men with varicoceles. Fertil Steril. 1989;52(3):469-73.##Gubin DA, Dmochowski R, Kutteh WH. Multivariant analysis of men from infertile couples with and without antisperm antibodies. Am J Reprod Immunol. 1998;39(2):157-60.##Liu RZ, Lu YL, Xu ZG, Zuo WJ, Xin JL, Wang ZS. [The effect of semen antisperm antibody on human sperm acrosin activity]. Zhonghua Nan Ke Xue. 2003;9(4):252-3. Chinese.##Sinisi AA, D'Apuzzo A, Pasquali D, Venditto T, Esposito D, Pisano G, et al. Antisperm antibodies in prepubertal boys treated with chemotherapy for malignant or non-malignant diseases and in boys with genital tract abnormalities. Int J Androl. 1997;20(1):23-8.##Stedronska J, Hendry WF. The value of the mixed antiglobulin reaction (MAR test) as an addition to routine seminal analysis in the evaluation of the subfertile couple. Am J Reprod Immunol. 1983;3(2):89-91.##

Using Fresh and Frozen Testicular Sperm Samples in Couples Undergoing ICSI-Micro TESE Treatment 2 1 2 Background: We performed this study to evaluate use of fresh and frozen sperm samples in non-obstructive azoospermia microdissection testicular sperm extraction (micro-TESE-ICSI) treatment. Methods: We performed a total of 82 consecutive in vitro fertilization (IVF) cycles at Fertijin IVF Center in Istanbul, Turkey from January 2010 to March 2012. In 43 participants we used fresh sperm and frozen sperm in the remaining 39 cases. We used fresh and frozen thawed micro surgical testicular sperm extraction (micro TESE) sperm for ICSI with metaphase II (MII) oocytes. Results: Frozen microTESE sperm was used in 39 cycles, while 43 ICSI cycles were performed using fresh microTESE. Neither the age of male partners (38.33±5.93 and 38.13±8.28) nor that of the female participants (33.16±6.38 and 33.33±6.97) showed significant difference between fresh versus the microTESE and frozen treatment groups, respectively. FSH concentrations were (14.66±13.93 mIU/ml) in fresh TESE group and (17.91±16.29 mIU/ml) in frozen group with no correlations or differences between the two groups. The average number of mature oocytes injected with sperm was 9.23±3.77, versus 9.26±5.26 in cycles using fresh and frozen microTESE sperm, respectively. Fertilization rate was not significantly different in the fresh microTESE (44.79%) than frozen TESE sperm group (46.76%). The average number of transferred embryos was 1.60±0.49 in fresh sperm group and 1.59±0.50 in frozen sperm group. All embryo transfers were performed on day 3. Conclusion: Cryopreservation of testicular sperm tissues is more suitable and of great benefite if carried out before ovulation induction and not after, especially in cases with non-obstructive azoospermia. 79 85 Safak S Tavukcuoglu Safak Tavukcuoglu Reproductive Medicine Unit, University of Schleswig-Holstein Reproductive Medicine Unit, University of Schleswig-Holstein Germany saftv@hotmail.com Tahani T Al-Azawi Tahani Al-Azawi Reproductive Medicine Unit, University of Schleswig-Holstein Reproductive Medicine Unit, University of Schleswig-Holstein Germany Safaa S Al-Hassani صفا الحسنی Reproductive Medicine Unit, University of Schleswig-Holstein Reproductive Medicine Unit, University of Schleswig-Holstein Germany Amir Afshin AA Khaki Amir Afshin Khaki Reproductive Medicine Unit, University of Schleswig-Holstein Reproductive Medicine Unit, University of Schleswig-Holstein Germany Arash A Khaki Arash Khaki Reproductive Medicine Unit, University of Schleswig-Holstein Reproductive Medicine Unit, University of Schleswig-Holstein Germany Seval S Tasdemir Seval Tasdemir Fertijin IVF Center Fertijin IVF Center Turkey ICSIIn Vitro fertilizationMicrosurgical testicular sperm extractionSperm retrievalSperm 524.pdf Ramasamy R, Yagan N, Schlegel PN. Structural and functional changes to the testis after conventional versus microdissection testicular sperm extraction. Urology. 2005;65(6):1190-4.##Gordon UD. Assisted conception in the azoospermic male. Hum Fertil. 2002;5(Suppl 1):S9-S14.##Anniballo R, Ubaldi F, Cobellis L, Sorrentino M, Rienzi L, Greco E, et al. Criteria predicting the absence of spermatozoa in the Sertoli cell-only syndrome can be used to improve success rates of sperm retrieval. Hum Reprod. 2000;15(11):2269-77.##Cedenho AP, Da Ros CT, Juliano RV. Treatment of non-obstructive azoospermia. Int Braz J Urol. 2004;29(Suppl 5):39-41.##Chandley AC. The chromosomal basis of human infertility. Br Med Bull. 1979;35(2):181-6.##Schoysman R, Vanderzwalmen P, Nijs M, Segal-Bertin G, van de Casseye M. Successful fertilization by testicular spermatozoa in an in-vitro fertilization programme. Hum Reprod. 1993;8(8):1339-40.##Schlegel PN. Testicular sperm extraction: microdissection improves sperm yield with minimal tissue excision. Hum Reprod. 1999;14(1):131-5.##Schoysman R, Vanderzwalmen P, Nijs M, Segal L, Segal-Bertin G, Geerts L, et al. Pregnancy after fertilisation with human testicular spermatozoa. Lancet. 1993;342(8881):1237.##Silber SJ, Van Steirteghem AC, Liu J, Nagy Z, Tournaye H, Devroey P. High fertilization and pregnancy rate after intracytoplasmic sperm injection with spermatozoa obtained from testicle biopsy. Hum Reprod. 1995;10(1):148-52.##Gianaroli L, Magli MC, Selman HA, Colpi G, Belgrano E, Trombetta C, et al. Diagnostic testicular biopsy and cryopreservation of testicular tissue as an alternative to repeated surgical openings in the treatment of azoospermic men. Hum Reprod. 1999;14(4):1034-8.##Prins GS, Dolgina R, Studney P, Kaplan B, Ross L, Niederberger C. Quality of cryopreserved testicular sperm in patients with obstructive and nonobstructive azoospermia. J Urol. 1999;161(5): 1504-8.##Habermann H, Seo R, Cieslak J, Niederberger C, Prins GS, Ross L. In vitro fertilization outcomes after intracytoplasmic sperm injection with fresh or frozen-thawed testicular spermatozoa. Fertil Steril. 2000;73(5):955-60.##Dafopoulos K, Griesinger G, Schultze-Mosgau A, Orief Y, Schöpper B, Nikolettos N, et al. Factors affecting outcome after ICSI with spermatozoa retrieved from cryopreserved testicular tissue in non-obstructive azoospermia. Reprod Biomed Online. 2005;10(4):455-60.##Dafopoulos K, Griesinger G, Schultze-Mosgau A, Orief Y, Schöpper B, Nikolettos N, et al. Cumulative pregnancy rate after ICSI with cryopreserved testicular tissue in non-obstructive azoospermia. Reprod Biomed Online. 2005;10(4): 461-6.##Amer M, Haggar SE, Moustafa T, Abd El-Naser T, Zohdy W. Testicular sperm extraction: impact of testicular histology on outcome, number of biopsies to be performed and optimal time for repetition. Hum Reprod. 1999;14(12):3030-4.##Liu J, Tsai YL, Katz E, Compton G, Garcia JE, Baramki TA. Outcome of in-vitro culture of fresh and frozen-thawed human testicular spermatozoa. Hum Reprod. 1997;12(8):1667-72.##Al Hasani S, Küpker W, Baschat AA, Sturm R, Bauer O, Diedrich C, et al. Mini-swim-up: a new technique of sperm preparation for intracytoplasmic sperm injection. J Assist Reprod Genet. 1995;12(7):428-33.##Veeck L. Preembryo grading and degree of cytoplasmic fragmentation. In: Veeck L, editor. An atlas of human gametes and conceptuses. London: Parthenon publishing; 1999. p. 46-50.##Gil-Salom M, Romero J, Minguez Y, Rubio C, De los Santos MJ, Remohi J, et al. Pregnancies after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa. Hum Reprod. 1996;11(6):1309-13.##Küpker W, Schlegel PN, Al-Hasani S, Fornara P, Johannisson R, Sandmann J, et al. Use of frozen-thawed testicular sperm for intracytoplasmic sperm injection. Fertil Steril. 2000;73(3):453-8.##Lin MH, Morshedi M, Srisombut C, Nassar A, Oehninger S. Plasma membrane integrity of cryopreserved human sperm: an investigation of the results of the hypoosmotic swelling test, the water test, and eosin-Y staining. Fertil Steril. 1998;70(6):1148-55.##Jezek D, Knuth UA, Schulze W. Successful testicular sperm extraction (TESE) in spite of high serum follicle stimulating hormone and azoospermia: correlation between testicular morphology, TESE results, semen analysis and serum hormone values in 103 infertile men. Hum Reprod. 1998;13(5):1230-4.##Tournaye H, Verheyen G, Nagy P, Ubaldi F, Goossens A, Silber S, et al. Are there any predictive factors for successful testicular sperm recovery in azoospermic patients? Hum Reprod. 1997;12(1):80-6.##Schlegel PN, Su LM. Physiological consequences of testicular sperm extraction. Hum Reprod. 1997;12(8):1688-92.##Sheikh A, Fouad S, Hussein M. Fresh and frozen testicular tissue in conjunction with intra-cytoplasmic sperm injection in the treatment of obstructive and nonobstructive azoospermia. Hum Reprod. 1998;13:117-8.##Friedler S, Raziel A, Soffer Y, Strassburger D, Komarovsky D, Ronel R. Intracytoplasmic injection of fresh and cryopreserved testicular spermatozoa in patients with nonobstructive azoospermia--a comparative study. Fertil Steril. 1997;68(5):892-7.##Oates RD, Mulhall J, Burgess C, Cunningham D, Carson R. Fertilization and pregnancy using intentionally cryopreserved testicular tissue as the sperm source for intracytoplasmic sperm injection in 10 men with non-obstructive azoospermia. Hum Reprod. 1997;12(4):734-9.##Watkins W, Nieto F, Bourne H, Wutthiphan B, Speirs A, Baker HW. Testicular and epididymal sperm in a microinjection program: methods of retrieval and results. Fertil Steril. 1997;67(3):527-35.##Devroey P, Liu J, Nagy Z, Goossens A, Tournaye H, Camus M, et al. Pregnancies after testicular sperm extraction and intracytoplasmic sperm injection in non-obstructive azoospermia. Hum Reprod. 1995;10(6):1457-60.##Van Steirteghem A, Nagy P, Joris H, Janssenswillen C, Staessen C, Verheyen G, et al. Results of intracytoplasmic sperm injection with ejaculated, fresh and frozen-thawed epididymal and testicular spermatozoa. Hum Reprod. 1998;13 Suppl 1:134-42.##

Sexual Activity of Adolescent School Girls in an Urban Secondary School in Cameroon 2 1 2 Background: The objective of this study was to describe the extent of sexual activity in adolescent school girls. Methods: This was a cross-sectional study with prolective collection of data carried out at Lycée General Leclerc, Yaounde (Cameroon), from October 1 to November 30, 2011. Heterosexual coitus was considered as sexual activity. A pretested self-administered questionnaire was proposed to all consenting girl students aged 10 to 19 years. The data were analyzed using Epi Info 3.2.1 and Microsoft Excel 2007 software. Results: Of the 2660 students who responded to the questionnaire, 21.3% (566) admitted being sexually active. Out of these, 64.3% (364) were aged between 10 and 16 years at their first heterosexual contact. The mean age at the first sexual intercourse was 15.3 years. Although 56.4% (319) of the sexually active respondents had only one sexual partner, 43.6% (247) of them had at least two partners. Sexual activity was occasional in 71.4% of those being sexually active. Meanwhile, 52.1% (295) of the sexually active adolescent girls used condoms during sexual intercourse, 41.5% (235) did so occasionally, and 6.4% (36) had regular unprotected sex. Conclusion: More than one-fifth of adolescent girls were sexually active in this study. Sexual intercourse started mostly at the age of 16 or less, and it was mostly occasional. Half of the cases had multiple sexual partners, and half were not using condoms during sexual intercourse. We, thus, recommend the implementation of interventions aimed at delaying the age of the first sexual intercourse and accessibility of condoms to students in this setting. 85 90 Pascal P Foumane Pascal Foumane Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Gynaeco-Obstetric and Pediatric Hospital Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Gynaeco-Obstetric and Pediatric Hospital Cameroon pfoumane2004@yahoo.fr Andreas A Chiabi Andreas Chiabi Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Gynaeco-Obstetric and Pediatric Hospital Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Gynaeco-Obstetric and Pediatric Hospital Cameroon Christelle C Kamdem Christelle Kamdem Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Military Hospital Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Military Hospital Cameroon Francisca F Monebenimp Francisca Monebenimp Department of Pediatrics, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, University Teaching Hospital Department of Pediatrics, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, University Teaching Hospital Cameroon Julius J Sama Dohbit Julius Sama Dohbit Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Gynaeco-Obstetric and Pediatric Hospital Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Yaoundé Gynaeco-Obstetric and Pediatric Hospital Cameroon Robinson R Enow Mbu Robinson Enow Mbu Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Central Maternity Department of Gynecology and Obstetric, Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, Central Maternity Cameroon Adolescent school girlsCameroonCondom useSexual activitySexually transmitted infectionUnwanted pregnancy 529.pdf De Muylder X. [The sexual health of adolescent girl]. Louvain Med. 2004;123(2):52-9. French.##Hamada H, Zaki A, Nejjar H, Filali A, Chraibi C, Bezad R, et al. [Pregnancy and delivery in adolescents: characteristics and profile of 311 cases]. J Gynecol Obstet Biol Reprod. 2004;33(7):607-14. French.##Bajos N, Bozon M. [Survey on sexuality in France: practice, gender and health]. 1st ed. Paris: La Decouverte; 2008. p 117-47. French.##Centers for Disease Control and Prevention [Internet]. USA: Centers for Disease Control and Prevention; Trends in the prevalence of sexual behaviors national YRBS: 1991-2007; [cited 2012 Jul 8]; [about 2 screens]. Available from: http://ppt-elect.center-chool.org/providers/304/yrbs07_us_sexual_behaviors_trend.pdf##Buga GA, Amoko DH, Ncayiyana DJ. Sexual behaviour, contraceptive practice and reproductive health among school adolescents in rural Transkei. S Afr Med J. 1996;86(5):523-7.##Courtois R, Mullet E, Malvy D. [Survey on sexual behavior by Congolese and French high-school students in an AIDS context]. Sante. 2001;11(1):49-55. French.##Anochie IC, Ikpeme EE. Prevalence of sexual activity and outcome among female secondary school students in Port Harcourt, Nigeria. Afr J Reprod Health. 2001;5(2):63-7.##Cameroon’s National Institute of Statistics [Internet]. Cameroon: National Institute of Statistics of Cameroon; The population of Cameroon in 2010; [cited 2012 Nov 21]; Available from: http://www.statistics-cameroon.org/##Cameroon’s National Institute of Statistics [Internet]. Cameroon: National Institute of Statistics of Cameroon; Third demographic and health survey, 2004; [cited 2012 May 17]; Available from: http://www.statistics-cameroon.org/##Cameroon’s National Institute of Statistics [Internet]. Cameroon: National Institute of Statistics of Cameroon; Results of the fourth demographic and health survey (DHS) combined with the multiple indicators cluster survey (MICS) of 2011; [cited 2012 May 17]; Available from: http://www.statistics-cameroon.org/##Kamtchouing P, Takougang I, Ngoh N, Yakam I. [Sexuality of adolescent students in Yaounde (Cameroon)]. Contracept Fertil Sex. 1997;25(10): 798-801. French.##Mimbila-Mayi M, Nzame Vierin Y, Biloghe AS, Moussavou A. Connaissances et comportements des adolescents en matière de santé sexuelle au Gabon. Clin Mother Child Health. 2011;8.##Makenzius M, Larsson M. Early onset of sexual intercourse is an indicator for hazardous lifestyle and problematic life situation. Scand J Caring Sci. 2013;27(1):20-6.##Westhoff WW, McDermott RJ, Holcomb DR. HIV risk behaviors: a comparison of U.S. Hispanic and Dominican Republic youth. AIDS Educ Prev. 1996;8(2):106-14.##Bumbuliene Z, Alisauskas J. Sexual behavior and high-risk human papillomavirus in 15- to 22-year-old Lithuanian women. Acta Obstet Gynecol Scand. 2012;91(4):511-3.##Kurewa NE, Mapingure MP, Munjoma MW, Chirenje MZ, Rusakaniko S, Stray-Pedersen B. The burden and risk factors of Sexually Transmitted Infections and Reproductive Tract Infections among pregnant women in Zimbabwe. BMC Infect Dis. 2010;10:127.##Valois RF, Oeltmann JE, Waller J, Hussey JR. Relationship between number of sexual intercourse partners and selected health risk behaviors among public high school adolescents. J Adolesc Health. 1999;25(5):328-35.##Braun-Courville DK, Rojas M. Exposure to sexually explicit Web sites and adolescent sexual attitudes and behaviors. J Adolesc Health. 2009;45(2):156-62.##Langille DB, Asbridge M, Flowerdew G, Allen M. Associations of sexual risk-taking with having intercourse before 15 years in adolescent females in Cape Breton, Nova Scotia, Canada. Sex Health. 2010;7(2):199-204.##Schuster MA, Bell RM, Berry SH, Kanouse DE. Impact of a high school condom availability program on sexual attitudes and behaviors. Fam Plann Perspect. 1998;30(2):67-72.##

Aspects of Psychosocial Development in Infertile Versus Fertile Men 2 1 2 Background: Infertility is one of the most difficult life experiences that a couple might encounter. Infertility as a bio-psycho-social phenomenon, could influence all aspects of life. While paying special attention to the psychological aspects of infertility in couples; many studies have investigated the non-clinical aspects of infertility, however, they rarely have evaluated the psychosocial development of infertile versus fertile men. We aimed to study the effects of infertility on psychosocial development in men. Methods: In fact, we designed the study based on "Erikson’s theory of psychosocial development". We focused on the relationship between psychosocial development and some self-conceived indices. For this purpose, we divided the participants volunteers into two groups of cases (80 infertile men) and controls (40 fertile men) and asked them to complete a 112 (questions questionnaire based on "self description"). The statistical analysis was performed by SPSS (version 13) using independent t-test, Pearson correlation coefficient and analysis of covariance. A p-value <0.05 was considered significant. Results: Data analysis showed significant inter and intra group differences. Infertile and fertile groups showed significant differences in trust, autonomy, generativity and integrity stages (p<0.05). Infertile intergroup analysis represents us to higher scores in positive than negative stages. Conclusion: Infertility as a phenomenon had its own effects on the psychosocial development of infertile men. However, good coping skills are powerful tools to manage these myriad of feelings surrounding infertile men. 90 94 Mohammad Mehdi MM Akhondi محمدمهدی آخوندی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Sima S Binaafar Sima Binaafar Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Zohreh Z Behjati Ardakani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Kourosh K Kamali Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Haleh H Kosari Haleh Kosari Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Behzad B Ghorbani بهزاد قربانی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran B.Ghorbani@avicenna.ac.ir Erikson's theoryInfertilityMenPsychosocial development 530.pdf Ramezanzadeh F, Noorbala AA, Abedinia N, Rahimi Forooshani A, Naghizadeh MM. Psychiatric intervention improved pregnancy rates in infertile couples. Malays J Med Sci. 2011;18(1):16-24.##Stouffs K, Tournaye H, Van der Elst J, Liebaers I, Lissens W. Is there a role for the nuclear export factor 2 gene in male infertility? Fertil Steril. 2008;90(5):1787-91.##Kormi Nouri R. [Psycho-Social aspects of infertility]. J Reprod Infertil. 2000;1(2):57-68. Persian.##Edelmann RJ, Connolly KJ. Psychological aspects of infertility. Br J Med Psychol. 1986;59:209-19.##Greil AL. Infertility and psychological distress: a critical review of the literature. Soc Sci Med. 1997;45(11):1679-704.##Greil AL, Slauson-Blevins K, McQuillan J. The experience of infertility: a review of recent literature. Sociol Health Illn. 2010;32(1):140-62.##Munley PH. Erik Erikson's theory of psychosocial development and vocational behavior. J Couns Psychol. 1975;22(4): 314-319.##Akhondi MA, Dadkhah A, Bagherpour A, Behjati Ardekani Z, Kamali K, Binaafar S, et al. Study of Body Image in Fertile and Infertile Men. J Reprod Infertil. 2011;12(4):295-298.##Naseri Tafti N, Pakdaman Sh, Asgari A. [The role of sport and personality traits in psychological development of students]. J Iran Psychol. 2008;5(17):53-62. Persian.##Deka PK, Sarma S. Psychological aspects of infertility. BJMP. 2010;3(3):336.##Edelmann RJ, Connolly KJ. Psychological aspects of infertility. Br J Med Psychol. 1986;59(3):209-19.##Mahlstedt PP. The psychological component of infertility. Fertil Steril. 1985;43(3):335-46.##Kedem P, Mikulincer M, Nathanson YE , Bartoov B. Psychological aspects of male infertility. Br J Med Psychol. 1990;63(1):73-80.##Dhaliwal LK, Gupta KR, Gopalan S, Kulhara P. Psychological aspects of infertility due to various causes--prospective study. Int J Fertil Womens Med. 2004;49(1):44-8.##Snarey J, Son L, Kuehne Valerie S, Hauser S, Vaillant G. The role of parenting in men's psychosocial development: A longitudinal study of early adulthood infertility and midlife generativity. Dev Psychol. 1987;23(4):593-603.##Cousineau TM, Domar AD. Psychological impact of infertility. Best Pract Res Clin Obstet Gynaecol. 2007;21(2):293-308.##Atwood J, Dobkin S. Storm clouds are coming: ways to help couples reconstruct the crisis of infertility. Contemp Fam Ther. 1992;14(5):385-403.##