Epigenetic Alterations and their Impact on Assisted Reproduction Technology Outcomes 2 1 569 2 It is already three decades since the birth of the first “test-tube baby” in 1978 and such methods have found their due place in most countries. In Denmark alone, more than 6% of children are born through assisted reproduction technologies (ARTs) (1). In the United States, more than 50,000 babies are born through IVF annually and there are more than 4 million people born through such methods around the world (2). In the beginning, there were much opposition and criticism surrounding the use of these methods, which mostly consisted of criticisms about gamete manipulation and the induction of genetic and chromsomal disorders in embryos (2). By extensive use of these methods and the birth, growth and puberty of these children, criticisms and concerns abated and the marriage and successful pregnancy of Louise Brown dampened disputes about the fertility of these children. Recently, following the rise in the number of babies born through ART and statistical evaluation on the health of IVF children, there have been reports on the increasing number of abortion, intrauterine fetal death, congenital anomalies and abnormal epigenetic modifications in these children (3). Although most of the causes of these problems are not understood exactly, but epigenetic alterations in the gametes and embryos are believed to be the potential causes (4). Studies have shown the evidences of gametes epigenetic alterations during gametogenesis and these changes seem to be essential for the natural fertilization, quality and development of embryos, fate of pregnancy and health of the offspring during his life. These changes include covalent changes such as methylation, phosphorylation, acetylation, etc at DNA level or in proteins (histones and protamines) attached to it without any changes in the sequence of DNA nucleotides affecting the developmental process and function of every cell, tissue, organ and organism (4). Patterns of epigenetic modification could be influenced by environmental conditions and health status of organisms, therefore, assisted reproductive procedures including ovulation induction protocols, fertilization, embryo culture conditions and physico-chemical environment of IVF labs could influence the epigenetic modifications of gametes, embryos and even the fetus. In addition, one of the hot topics in ART and management of IVF centers are intensive care through quality control (QC) and quality assurance (QA) for all the materials, methods, instruments and procedures (5). Taking these points into account, it seems that most of the indices used for the assessment of ART methods including the rate of fertilization and the number and quality of embryos need to be revised in the near future. Although certain changes in therapeutic protocols may not have obvious effects on ART indexes (due to genomic inactivate of early embryos), but due to epigenetic changes and subsequently gene expression patterns, it may drastically affect the implantation rate, pregnancy outcomes and health status of IVF children. This will not be possible unless we design studies to identify the conditions with minimal alterations from normal epigenetic patterns and subsequently maintain these conditions in IVF centers through rigorous implementation of QC and QA. Presently, these topics are hot subjects of papers and lectures in the field of reproduction and infertility and future studies would emphasize on the detection of normal epigenetic modifications and their abnormal aberrations throughout ART procedures and their potential effects on embryos and IVF children. In conclusion, this scenario will lead to obvious changes in the infertility treatment protocols to provide more safety and success rate with lower costs. 01 2 Mohammad Reza MR Sadeghi محمدرضا صادقی Editor-in-chief Editor-in-chief Iran No Keyword 569.pdf Dobson R. Proportion of babies born in Europe after IVF varies 20-fold. BMJ. 2006;332(7542):626.##Pauli SA, Berga SL, Shang W, Session DR. Current status of the approach to assisted reproduction. Pediatr Clin North Am. 2009;56(3):467-88.##Rimm AA, Katayama AC, Diaz M, Katayama KP. A meta-analysis of controlled studies comparing major malforma-tion rates in IVF and ICSI infants with naturally conceived children. J Assist Reprod Genet. 2004;21(12):437-43.##Carrell DT, Hammoud SS. The human sperm epigenome and its potential role in embryonic development. Mol Hum Reprod. 2010;16(1):37-47.##Lane M, Mitchell M, Cashman KS, Feil D, Wakefield S, Zander-Fox DL. To QC or not to QC: the key to a consist-ent laboratory? Reprod Fertil Dev. 2008;20(1):23-32.##

Effects of Kamdhenu Ark and Active Immunization by Gonadotropin Releasing Hormone Conjugate (GnRH-BSA) on Gonadosomatic Indices (GSI) and Sperm Parameters in Male Mus musculus 2 1 2 Background: Active immunization against GnRH decreases the secretion of gonadotropins and causes cessation of gonadal function, thereby, inducing infertility. Based on the immunoenhancing activity of Kamdhenu ark (distilled cow urine), this study was performed to evaluate its effects on the gonadosomatic indices (GSI) and sperm parameters in male mice receiving a GnRH contraceptive vaccine. Methods: Sixty adult male mice of Parke’s strain were divided into three groups of twenty. Group I served as the controls, while group II was immunized by GnRH-BSA conjugate (50/0.2/35 µg/ml/g BW) by four intraperitoneal injections at different intervals on days 1, 30, 60 and 90. However, group III was supplemented daily by oral Kamdhenu ark (100 ppm) along with GnRH-BSA immunizations. The animals were sacrificed after 30, 60, 90 and 120 days and their testis and epididymis were dissected out weighed and semen analysis was performed. Results: GSI values, sperm motility, sperm count and sperm morphology in male Mus musculus were decreased significantly in all the experimental groups as compared to the control group (p<0.01). Kamdhenu ark significantly enhanced the effect of GnRH vaccine on the aforesaid parameters especially in 90 and 120 days treated groups (p<0.05). Conclusion: The changes witnessed in sperm parameters suggested that the GnRH-BSA immunization suppressed the activities of gonadotropins and testosterone directly through hypothalamo-hypophysial-gonadal axis and indirectly by acting on the testes which may modulate the sperm morphology, sperm count and motility. However, Kamdhenu ark seems to have enhanced these effects because of its immunemodulatory properties too. 03 8 Javid J Ahmad Ganaie جاوید احمد گانعی Endocrinology Laboratory, Department of Biosciences, Barkatullah University Endocrinology Laboratory, Department of Biosciences, Barkatullah University India Varsha V Gautam وارشا گواتام Endocrinology Laboratory, Department of Biosciences, Barkatullah University Endocrinology Laboratory, Department of Biosciences, Barkatullah University India Vinoy V Kumar Shrivastava وینوی کومار شریواستاوا Endocrinology Laboratory, Department of Biosciences, Barkatullah University Endocrinology Laboratory, Department of Biosciences, Barkatullah University India vinoyks2001@yahoo.com GnRH-BSAGonadosomatic indices (GSI)ImmunizationKamdhenu arkMus musculusSperm Parameters 460.pdf Garner LL, Campbell GT, Blake CA. Luteinizing hormone (LH)-releasing hormone: chronic effects on LH and follicle-stimulating hormone cells and secretion in adult male rats. Endocrinology. 1990; 126(2):992-100.##Hoskinson RM, Rigby RD, Mattner PE, Huynh VL, D'Occhio M, Neish A, et al. Vaxstrate: an anti reproductive vaccine for cattle. Aust J Biotechnol. 1990;4(3):166-70, 176.##Prendiville DJ, Enright WJ, Crowe MA, Vaughan L, Roche JF. Immunization of prepubertal beef heifers against gonadotropin-releasing hormone: immune, estrus, ovarian, and growth responses. J Anim Sci. 1995;73(10):3030-7.##Bonneau M, Enright WJ. Immunocastration in cattle and pigs. Livest Prod Sci. 1995;42(2-3):193-200.##Robbins SC, Jelinski MD, Stotish RL. Assessment of the immunological and biological efficacy of two different doses of a recombinant GnRH vaccine in domestic male and female cats (Felis catus). J Reprod Immunol. 2004;64(1-2):107-19.##Miller LA, Johns BE, Killian GJ. Immunocontraception of white-tailed deer with GnRH vaccine. Am J Reprod Immunol. 2000;44(5):266-74.##Simms MS, Scholfield DP, Jacobs E, Michaeli D, Broome P, Humphreys JE, et al. Anti-GnRH antibodies can induce castrate levels of testosterone in patients with advanced prostate cancer. Br J Cancer. 2000;83(4):443-6.##Kumar N, Savage T, DeJesus W, Tsong YY, Didolkar A, Sundaram K. Chronic toxicity and reversibility of antifertility effect of immunization against gonadotropin-releasing hormone in male rats and rabbits. Toxicol Sci. 2000;53(1):92-9.##Bhadauria H. Gomutra-Ek Chamatkari Aushadhi (Cow urine- A Magical therapy). Vish Ayur Patrika. 2002;5:71-4.##Chauhan RS, Singh BP, Singhal LK. Immunomodulation with kamdhenu ark in mice. J Immunol Immunopathol. 2001;3(1):74-7.##Garg N, Chauhan RS, Kumar A. Assessing the effect of cow urine on immunity of white leghorn layers. International Society for Animal Hugges (ISAH). 2005;2:81-3.##Mathivanan R, Kalaiarasi K. Panchagavya and Andrographis paniculata as alternatives to antibiotic growth promoters on haematological, serum biochemical parameters and immune status of broilers. J Poult Sci. 2007;44(2):198-204.##Prasad MR, Chinoy NJ, Kadam KM. Changes in succinic dehydrogenase levels in the rat epididymis under normal and altered physiologic conditions. Fertil Steril. 1972;23(3):186-90.##Highland HN, Rao MV, Chinoy NJ, Shah VC. Analysis of the functional and nuclear integrity of human spermatozoa. Int J Fertil. 1991;36(1):43-7.##Esbenshade KL, Britt JH. Active immunization of gilts against gonadotropin-releasing hormone: effects on secretion of gonadotropins, reproductive function, and responses to agonists of gonadotropin-releasing hormone. Biol Reprod. 1985;33 (3):569-77.##Johnson HE, DeAvila DM, Chang CF, Reeves JJ. Active immunization of heifers against luteinizing hormone-releasing hormone, human chorionic gonadotropin and bovine luteinizing hormone. J Anim Sci. 1988;66(3):719-26.##Meloen RH, Turkstra JA, Lankhof H, Puijk WC, Schaaper WM, Dijkstra G, et al. Efficient immunecastration of male piglets by immunoneutralization of GnRH using a new GnRH-like peptide. Vaccine. 1994;12(8):741-6.##Grizzle TB, Esbenshade KL, Johnson BH. Active immunization of boars against gonadotrop in releasing hormone. I. Effects on reproductive parameters. Theriogenology. 1987;27(4):571-80.##Dowsett KF, Pattie WA, Knott LM, Jackson AE, Hoskinson RM, Rigby RP, et al. A preliminary study of immunological castration in colts. J Reprod Fertil Suppl. 1991;44:183-90.##McLachlan RI, Wreford NG, Tsonis C, De Kretser DM, Robertson DM. Testosterone effects on spermatogenesis in the gonadotropin-releasing hormone-immunized rat. Biol Reprod. 1994;50(2): 271-80.##Cook RB, Popp JD, Kastelic JP, Robbins S, Harland R. The effects of active immunization against gnRH on testicular development, feedlot performance, and carcass characteristics of beef bulls. J Anim Sci. 2000;78(11):2778-83.##Janett F, Stump R, Burger D, Thun R. Suppression of testicular function and sexual behavior by vaccination against GnRH (Equity) in the adult stallion. Anim Reprod Sci. 2009;115(1-4):88-102.##Levy JK, Miller LA, Cynda Crawford P, Ritchey JW, Ross MK, Fagerstone KA. GnRH immunecontraception of male cats. Theriogenology. 2004; 62(6):1116-30.##Ross MK, Miller LA, Crawford PC, Ritchey JW, Fagerstone KA. GnRH immunocontraception in cats. In: Baker H, Boyle S, Griffin B, editors. Proceedings of the 2004 ACCD International Symposium on Non-surgical Methods for Pet Population Control; 2004 June 24-27; Denver, Colorado, Belle Court (Portland): ACCD Press; 2005. p.113-5.##Prabhakar K, Singh GK, Chauhan RS, Singh DD. Effect of cow urine on lymphocyte proliferation in developing stages of chicks. Indian Cow. 2004;1 (2):3-5.##Kumar R, Chauhan RS, Singhal LK, Singh AK, Singh DD. A comparative study on immunostimulatory effects of Kamdhenu Ark and Vasant Kusumakar in mice. J Immunol Immunopathol. 2002;4 (1-2):104-6.##Garg N, Chauhan RS. Kamdhenu ark changes humoral immunity in rat. In: National Symposium on Molecular Biology in India- A Post Graduate Update; 2003 Jan 18, Gwalior, India, Gwalior (Madhya Pradesh): Cancer Hospital and Research Institute; 2003. p. 98.##Chauhan RS, Singh DD, Singhal LK, Kumar R. Effect of cow urine on interleukin-1 and 2. J Immunol Immunopathol. 2004;6(1):38-9.##Ganaie JA, Shrivastava VK. Effects of gonadotropin releasing hormone conjugate immunization and bioenhancing role of Kamdhenu ark on estrous cycle serum estradiol and progesterone levels in female Mus musculus. Iran J Reprod Med. 2010;8 (2):70-5.##

PCR-Based Evaluation of Tuberculous Endometritis in Infertile Women of North India 2 1 2 Background: Tuberculous endometritis is often a diagnostic dilemma for gynecologists in the evaluation of infertility. This study evaluated and compared different diagnostic methods in tuberculous endometritis. Methods: 100 infertile women were investigated for tuberculous endometritis. The endometrial samples obtained by curettage were evaluated by Ziehl-Neelsen (ZN) staining of the smears for acid-fast bacilli (AFB), the samples were also cultured on Lowenstein-Jensen medium, and histopathological examination and nested PCR targeting ‘hupB’ gene (Rv 2986c in M. tuberculosis and Mb3010c in M. bovis) which can differentiate between M. tuberculosis and M. bovis were performed. Antibodies against 38-kDa and 16-kDa mycobacterial antigens were detected in serum using ELISA. Statistical analysis was done by online GraphPad Prism software, version 5.0. McNemar’s test was applied and Kappa agreement coefficient was calculated for agreement between various methods. A p-value < 0.05 was considered significant. Results: Among the 100 evaluated endometrial samples, one was AFB smear positive, none was positive by culture, four were positive by histopathology and 13 were positive by PCR. Of the 13 PCR-positive cases, 38.4% were positive for M. tuberculosis, 23.07% for M. bovis, and 38.4% showed co-infection with both species. 40% of the patients had raised IgG against M. tuberculosis 38-kDa antigen. McNemar’s test was applied to PCR and the conventional methods of TB diagnosis (AFB, Culture and histopathology) and the p-value was < 0.001 (highly significant) for PCR. Detection by PCR showed a fair agreement with detection by Mantoux test and ELISA. Conclusion: In paucibacillary endometrial tuberculosis, the positive detection rate was found to be significantly higher for PCR compared to other methods. The ‘in-house’ nested PCR assay targeting the hupB gene and used in this study, can serve as a rapid diagnostic aid for tubercular endometritis. It can also differentiate between members of the Mycobacterium tuberculosis complex, namely M. tuberculosis and M. bovis. 09 15 Monika M D. Kohli مونیکا کوهلی Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital India Bimota B Nambam بیموتا نام بام Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital India Sagar S. Trivedi ساگر ترای ودی Department of Obstetrics and Gynecology, Lady Hardinge Medical College and S.S.K. Hospital Department of Obstetrics and Gynecology, Lady Hardinge Medical College and S.S.K. Hospital India Banke B L. Sherwal بنک شروال Department of Microbiology, Lady Hardinge Medical College and S.S. K. Hospital Department of Microbiology, Lady Hardinge Medical College and S.S. K. Hospital India Sarika S Arora ساریکا آرورا Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital India Anju A Jain آنجو جین Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital Department of Biochemistry, Lady Hardinge Medical College and S.S.K. Hospital India dranjujain@rediffmail.com InfertilityM. bovisM. tuberculosisPCRTuberculous endometritis 461.pdf Scrimin F, Limone A, Wiesenfeld U, Guaschino S. Tubercular endometritis visualized as endometrial micropolyps during hysteroscopic procedure. Arch Gynecol Obstet. 2010;281(6):1079-80.##Tripathy SN, Tripathy SN. Gynaecological tuberculosis -an update. Indian J Tuberc. 1998;45:193-7.##Aka N, Vural EZ. Evaluation of patients with active pulmonary tuberculosis for genital involvement. J Obstet Gynaecol Res. 1997;23(4):337-40.##Nagpal M, Pal D. Genital Tuberculosis – A diagnostic dilemma in OPD patients. J Obstet Gynecol India. 2001;51(6):127-31.##Mani R, Nayak S, Kagal A, Deshpande S, Dandge N, Bharadwaj R. Tuberculous endometritis in infertility: A bacteriological and histopathological study. Indian J Tuberc. 2003;50:161.##Singh N, Sumana G, Mittal S. Genital tuberculosis: a leading cause for infertility in women seeking assisted conception in North India. Arch Gynecol Obstet. 2008;278(4):325-7.##Shah NP, Singhal A, Jain A, Kumar P, Uppal SS, Srivatsava MV, et al. Occurrence of overlooked zoonotic tuberculosis: detection of Mycobacterium bovis in human cerebrospinal fluid. J Clin Microbiol. 2006;44(4):1352-8.##Prasad HK, Singhal A, Mishra A, Shah NP, Katoch VM, Thakral SS, et al. Bovine tuberculosis in India: potential basis for zoonosis. Tuberculosis (Edinb). 2005;85(5-6):421-8.##Grange JM. Mycobacterium bovis infection in human beings. Tuberculosis (Edinb). 2001;81(1-2):71-7.##Ananthnarayan R, Paniker CKJ. Mycobacterium-1 Tuberculosis. In: Ananthnarayan R, Paniker CKJ, editors. Textbook of Microbiology. Chennai: Orient Longman; 1996. p. 324-37.##Prabhakar S, Annapurna PS, Jain NK, Dey AB, Tyagi JS, Prasad HK. Identification of an immunegenic histone-like protein (HLPMt) of Mycobacterium tuberculosis. Tuber Lung Dis. 1998;79(1): 43-53.##Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D, et al. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998;393(6685):537-44.##Prabhakar S, Mishra A, Singhal A, Katoch VM, Thakral SS, Tyagi JS, et al. Use of the hupB gene encoding a histone-like protein of Mycobacterium tuberculosis as a target for detection and differentiation of M. tuberculosis and M. bovis. J Clin Microbiol. 2004;42(6):2724-32.##Mishra A, Singhal A, Chauhan DS, Katoch VM, Srivastava K, Thakral SS, et al. Direct detection and identification of Mycobacterium tuberculosis and Mycobacterium bovis in bovine samples by a novel nested PCR assay: correlation with conventional techniques. J Clin Microbiol. 2005;43(11): 5670-8.##Nambam B, Prasad HTK, Sherwal BL, Aneja S, Jain A. Comparative analysis of a hup B gene based diagnostic PCR with other conventional techniques for pediatric tuberculous meningitis. J Pediatr Infect Dis. 2006;1(3):143-8.##Chakravorty S, Tyagi JS. Novel use of guanidinium isothiocyanate in the isolation of Mycobacterium tuberculosis DNA from clinical material. FEMS Microbiol Lett. 2001;205(1):113-7.##Srivastava N, Manaktala U, Baveja CP. Role of ELISA (enzyme-linked immunosorbent assay) in genital tuberculosis. Int J Gynaecol Obstet. 1997; 57(2):205-6.##Biswas D, Agrawal M, Ali A, Bhargava R. Anti ES-31/41 antibodies: A potential tool in the serodiagnosis of extra-pulmonary Tuberculosis. Indian J Tuberc. 2003;50:203-7.##Senol G, Erer OF, Yalcin YA, Coskun M, Gündüz AT, Biçmen C, et al. Humoral immune response against 38-kDa and 16-kDa mycobacterial antigens in tuberculosis. Eur Respir J. 2007;29(1):143-8.##Demkow U, Zielonka TM, Nowak-Misiak M, Filewska M, Bialas B, Strzalkowski J, et al. Humoral immune response against 38-kDa and 16-kDamycobacterial antigens in bone and joint tuberculosis. Int J Tuberc Lung Dis. 2002;6(11):1023-8.##Abebe M, Lakew M, Kidane D, Lakew Z, Kiros K, Harboe M. Female genital tuberculosis in Ethiopia. Int J Gynaecol Obstet. 2004;84(3):241-6.##Manjunath N, Shankar P, Rajan L, Bhargava A, Saluja S, Shriniwas. Evaluation of a polymerase chain reaction for the diagnosis of tuberculosis. Tubercle. 1991;72(1):21-7.##Vishnevskaia EB, Marttila NJ, Semenovskiĭ AV, Oleĭnik AN, Vil'ianen MK, Vishnevskiĭ. [Polymerase chain reaction and ligase chain reaction in the diagnosis of gynecological tuberculosis]. Probl Tuberk. 2002;(1):49-51. Russian.##Mirlina ED, Lantsov VA, Semenovskiĭ AV, Oleĭnik AN, Popova SS, Manicheva OA, et al.[Diagnostic values of polymerase chain reaction test in females with genital tuberculosis]. Probl Tuberk. 1998;(1):46-8. Russian.##

Effects of Maternal Caffeine Consumption on Ovarian Follicle Development in Wistar Rats Offspring 2 1 2 Background: In recent years concerns have been raised about human reproductive disorders, specially the effects of environmental factors on human fertility and pregnancy outcome. Therefore, the present study was designed to assess the effects of maternal caffeine consumption on ovarian follicles development in rat offspring. Methods: 60 pregnant female rats were randomly divided into a control and two experimental groups. The rats in the two experimental groups received caffeine via drinking water during gestation (26 and 45 mg/kg) and lactation (25 and 35 mg/kg). The ovaries of the offspring were removed at 7, 14, 28, 60, 90 and 120 days after birth, and fixed in Bouin’s solution. By preparing serial tissue sections, structural changes in ovarian follicles and corpora lutea were studied during postnatal development. Results: The weight of ovaries decreased significantly (p<0.05) in the high dose caffeine-treated group at all stages of postnatal development. Significant (p<0.05) decreases were seen in the number of primordial follicles from day 7 to 120 after birth in the high dose caffeine-treated group. Moreover, the number of primary and secondary follicles decreased significantly on days 7, 14 and 28 as did the number of antral follicles on days 14 and 28 after birth (p<0.05) in the high dose caffeine-treated group. The diameter of secondary and antral follicles decreased significantly (p<0.05) in high dose caffeine-treated group on the early days of postnatal development. No statistically significant differences were seen in the number of corpora lutea between the groups. Conclusion: The present study shows that caffeine consumption during gestation and lactation affects the early stages of ovarian follicle development and reduces reproductive efficiency in the offspring of Wistar rats. 15 23 Mehran M Dorostghoal مهران درست قول Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz Iran mdorostghoal@yahoo.com Mahmood M Khaksari Mahabadi محمود خاکساری مهابادی Departmant of Anatomy, Faculty of Veterinary, Shahid Chamran University of Ahwaz Departmant of Anatomy, Faculty of Veterinary, Shahid Chamran University of Ahwaz Iran Sahar S Adham سحر ادهم Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz Iran CaffeineFertilityOvarian folliclesPostnatal development 462.pdf Hruska KS, Furth PA, Seifer DB, Sharara FI, Flaws JA. Environmental factors in infertility. Clin Obstet Gynecol. 2000;43(4):821-9.##Kumar S, Kumari A, Murarka S. Lifestyle factors in deteriorating male reproductive health. Indian J Exp Biol. 2009;47(8):615-24.##Al-Shoshan AA. Predictors of caffeine consumption among young women. Pak J Nutr. 2007;6(6):597-602.##Monroe J. Caffeine’s hook. Curr Health. 1998;2:5-12.##Barone JJ, Roberts HR. Caffeine consumption. Food Chem Toxicol. 1996;34(1):119-29.##Otberg N, Teichmann A, Rasuljev U, Sinkgraven R, Sterry W, Lademann J. Follicular penetration of topically applied caffeine via a shampoo formulation. Skin Pharmacol Physiol. 2007;20(4):195-8.##Golding J. Reproduction and caffeine consumption--a literature review. Early Hum Dev. 1995;43(1):1-14.##Aranda JV, Collinge JM, Zinman R, Watters G. Maturation of caffeine elimination in infancy. Arch Dis Child. 1979;54(12):946-9.##Ramlau-Hansen CH, Thulstrup AM, Bonde JP, Olsen J, Bech BH. Semen quality according to prenatal coffee and present caffeine exposure: two decades of follow-up of a pregnancy cohort. Hum Reprod. 2008;23(12):2799-805.##Nolen GA. The effect of brewed and instant coffee on reproduction and teratogenesis in the rat. Toxicol Appl Pharmacol. 1981;58(2):171-83.##Wilcox A, Weinberg C, Baird D. Caffeinated beverages and decreased fertility. Lancet. 1988;2 (8626-8627):1453-6.##Jagiello G, Ducayen M, Lin JS. Meiosis suppression by caffeine in female mice. Mol Gen Genet. 1972;118(3):209-14.##Boyd EM, Dolman M, Knight LM, Sheppard EP. The chronic oral toxicity of caffeine. Can J Physiol Pharmacol. 1965;43(6):995-1007.##Kren R, Ogushi S, Miyano T. Effect of caffeine on meiotic maturation of porcine oocytes. Zygote. 2004;12(1):31-8.##Olsen J. Cigarette smoking, tea and coffee drinking, and subfecundity. Am J Epidemiol. 1991;133 (7):734-9.##Alderete E, Eskenazi B, Sholtz R. Effect of cigarette smoking and coffee drinking on time to conception. Epidemiology. 1995;6(4):403-8.##Caan B, Quesenberry CP Jr, Coates AO. Differences in fertility associated with caffeinated beverage consumption. Am J Public Health. 1998;88(2): 270-4.##Pollard I, Williamson S, Magre S. Influence of caffeine administered during pregnancy on the early differentiation of fetal rat ovaries and testes. J Dev Physiol. 1990;13(2):59-65.##Maalouf W, Lee JH, Campbell KH. Effects of caffeine on the developmental potential of in vitro matured, aged and denuded ovine oocytes. Hum Fertil. 2005;8(2):129-30.##Grosso LM, Bracken MB. Caffeine metabolism, genetics, and perinatal outcomes: a review of exposure assessment considerations during pregnancy. Ann Epidemiol. 2005;15(6):460-6.##Moore RW, Rudy TA, Lin TM, Ko K, Peterson RE. Abnormalities of sexual development in male rats with in utero and lactational exposure to the antiandrogenic plasticizer Di(2-ethylhexyl) phthalate. Environ Health Perspect. 2001;109(3):229-37.##Odum J, Lefevre PA, Tinwell H, Van Miller JP, Joiner RL, Chapin RE, et al. Comparison of the developmental and reproductive toxicity of diethylstilbestrol administered to rats in utero, lactationally, preweaning, or postweaning. Toxicol Sci. 2002; 68(1):147-63.##Hughes RN, Beveridge IJ. Behavioral effects of exposure to caffeine during gestation, lactation or both. Neurotoxicol Teratol. 1991;13(6):641-7.##Hughes RN, Loader VG. Effects on elevated plusmaze behavior of exposure to caffeine during both gestation and lactation. Psychobiology. 1996;24 (4):314-9.##Bolon B, Bucci TJ, Warbritton AR, Chen JJ, Mat-tison DR, Heindel JJ. Differential follicle counts as a screen for chemically induced ovarian toxicity in mice: results from continuous breeding bioassays. Fundam Appl Toxicol. 1997;39(1):1-10.##Britt KL, Drummond AE, Cox VA, Dyson M, Wreford NG, Jones ME, et al. An age-related ovarian phenotype in mice with targeted disruption of the Cyp 19 (aromatase) gene. Endocrinology. 2000;141(7):2614-23.##Myers M, Britt KL, Wreford NG, Ebling FJ, Kerr JB. Methods for quantifying follicular numbers within the mouse ovary. Reproduction. 2004;127 (5):569-80.##McGee EA, Hsueh AJ. Initial and cyclic recruitment of ovarian follicles. Endocr Rev. 2000;21(2): 200-14.##Hoyer PB. Damage to ovarian development and function. Cell Tissue Res. 2005;322(1):99-106.##Guigon CJ, Magre S. Contribution of germ cells to the differentiation and maturation of the ovary: insights from models of germ cell depletion. Biol Reprod. 2006;74(3):450-8.##Kezele P, Skinner MK. Regulation of ovarian primordial follicle assembly and development by estrogen and progesterone: endocrine model of follicle assembly. Endocrinology. 2003;144(8): 3329-37.##Hirshfield AN. Development of follicles in the mammalian ovary. Int Rev Cytol. 1991;124:43-101.##Rennels EG. Influence of hormones on the histochemistry of ovarian interstitial tissue in the immature rat. Am J Anat. 1951;88(1):63-107.##Hoyer PB, Sipes IG. Assessment of follicle destruction in chemical-induced ovarian toxicity. Annu Rev Pharmacol Toxicol. 1996;36:307-31.##Hooser SB, Douds DP, DeMerell DG, Hoyer PB, Sipes IG. Long-term ovarian and gonadotropin changes in mice exposed to 4-vinylcyclohexene. Reprod Toxicol. 1994;8(4):315-23.##Generoso WM, Stout SK, Huff SW. Effects of alkylating chemicals on reproductive capacity of adult female mice. Mutat Res. 1971;13(2):172-84.##Axelrod J, Reichenthal J. The fate of caffeine in man and a method for its estimation in biological material. J Pharmacol Exp Ther. 1953;107(4):519-23.##Müller WU, Bauch T, Wojcik A, Böcker W, Streffer C. Comet assay studies indicate that caffeinemediated increase in radiation risk of embryos is due to inhibition of DNA repair. Mutagenesis. 1996;11(1):57-60.##

Reproductive Health Rights of Women in the Rural Areas of Meherpur District in Bangladesh 2 1 2 Background: This study evaluated the reproductive health rights, women empowerment and gender equity in a rural area of Bangladesh. Methods: Three hundred married women of reproductive age (15-49 years) in Meherpur District, Bangladesh were interviewed using a structured questionnaire and purposing sampling techniques. The logistic regression analysis was used to determine the dominating factors affecting reproductive health rights. To fulfill the objectives of the study the two main factors, age at marriage and family planning acceptance of the respondents, were regarded as the determinants. Results: The study results revealed that almost all the respondents were housewives (82.3%), one-third (31.0%) did not avail any modern facility, and their yearly income was very low. Moreover, about half of the women (52.7%) were very young ( 30 years), most of them (79.0%) had married early (<18 years) and about half of them (53.3%) had taken contraceptives based on their husbands’ choice. Finally, multivariate analysis identified the relationship between the profession of the respondents, yearly income, number of family members, and the availability of modern facilities with age at marriage (Model 1). The study also identified the relationship between the age of respondents, education, occupation, yearly income, and the total number of family members with family planning acceptance (Model 2). Conclusion: Regarding the results of this study, women’s reproductive health rights, marriage after the age of 18 and family planning acceptance among couples needs to be enhanced in Mehrpur District in Bangladesh. 23 33 Md Kamal K Hossain کمال حسین محمد Department of Population Science and Human Resource Development, University of Rajshahi Department of Population Science and Human Resource Development, University of Rajshahi Bangladesh Md Nazrul Islam N Mondal نظرالاسلام موندال محمد Department of Population Science and Human Resource Development, University of Rajshahi Department of Population Science and Human Resource Development, University of Rajshahi Bangladesh nazrul_ru@yahoo.com Mst Nazniz N Akter نازنین اختر سمت Department of Population Science and Human Resource Development, University of Rajshahi Department of Population Science and Human Resource Development, University of Rajshahi Bangladesh EmpowermentGenderReproductive rightsWomen 457.pdf Amin R, Becker S, Bayes A. NGO-Promoted microcredit programs and women's empowerment in rural Bangladesh: quantitative and qualitative evidence. J Dev Areas. 1998;32(2):221-36.##Naved RT. Empowerment of women: listening to the voices of women. Bangladesh Dev Stud. 1994;2 (3):176-8.##Akhter F. The eugenic and racist premise of reproductive rights and population control. Issues Reprod Genet Eng. 1992;5(1):1-8.##United Nations; Report of the International Conference on Population and Development, Cairo, 5-13 September 1994. 1st ed. New York: UN Population Fund; 1995. 193 p.##Freedman LP, Isaacs SL. Human Rights and Reproductive Choice. Stud Fam Plann. 1993;24(1):18-30.##Ashford LS. New Population Policies: Advancing Women’s Health and Rights. Population Bulletin. 2001;56(1).##Save the Children [Internet]. USA: Save the Child-ren Federation; 2011. A Study on the Impact of the workshop on family planning conducted with the traditional and religious leaders in Nasirnagar Thana. 1993 Sept 19 [cited 2011 Mar 18]; [about 63 pages]. Available from: http://pdf.usaid.gov/pdf_ docs/PNABU648.pdf##Bangladesh Bureau of Statistics. 2001 Statistical yearbook of Bangladesh. 22nd ed. Bangladesh: Bangladesh Bureau of Statistics, Planning Division, Government of the People's Republic of Bangladesh; 2003.##NCBP. Gender equality, development and peace for the twenty-first century. Dhaka: Women for Women; 2000.##Balk D. Change comes slowly for women in rural Bangladesh. Asia Pac Pop Policy. 1997;(41):1-4.##Microfinance Opportunities [Internet]. Washington DC: Assessing the Impact of Microenterprise Services (AIMS); Microfinance, Risk Management, and Poverty. 2000 Mar [cited 2011 Mar 18]; [about 115 page]. Available from: http://www.microfinanceopportunities.org/docs/Microfinance_Risk_Management_and_Poverty.pdf##Obaid TA. State of world population 2005. New York: United Nations Fund for Population Activities (UNFPA); 2005. 128 p.##UNICEF. Bangladesh Country Program Document. United Nations: United Nations International Children's Emergency Fund (UNICEF); 2005.##James C, Trail FT. Empowerment of rural women in India through literacy education. J Agric Ext Educ. 1995;2(2):18-27.##DFID; Poverty elimination and the empowerment of women. UK: Department of International Development; 2000. 39 p.##Dixon RB. Explaining cross-cultural variation in age at marriage and proportion never marriage. Popul stud. 1971;25(2):215-7.##

Iranian Oncologists’ Attitude towards Fertility Preservation in a Sample Group 2 1 2 Background: Recent advances in cancer treatment have resulted in an increased number of cancer survivors. Fertility might be impaired by cancer itself or through gonadal damage as a consequence of radio-chemotherapy. Damages to reproductive organs are seen in minimally upto 30% of cancer patients. The aim of the present study was evaluate the attitude of Iranian oncologists toward fertility preservation. Methods: Thirty oncologists filled an author-designed questionnaire which included five multiple choice questions at Mahak hospital, Tehran in 2009. The data were collected and analyzed by SPSS 11.5 software. Results: More than half of the oncologists believed that radio-chemotherapy has destructive effects on reproductive organs. This study showed that as high as 67% of the respondants believed fertility preservation should be offered to all patients at risk, but 40% of them insisted that the patients should bring up the topic themselves. Only 46% of the oncologist knew about fertility preservation techniques. The greatest barriers for accepting fertility preservation by the parents of children undergoing cancer treatment were: lack of information (41%), hopelessness from treatment prognosis (33%), fear of delay in starting the treatment (15%) and the priority of child survival (11%). Conclusion: The specific challenges of counseling are time pressure between diagnosis and start of treatment, little knowledge about fertility preservation both by treating oncologists and parents; therefore a team consisting of oncologists, infertility specialists and social workers is recommended in these settings. 33 37 Behzad B Ghorbani بهزاد قربانی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Pouran P Madahi پوران مداحی Mahak Hospital Mahak Hospital Iran Elham E Shirazi الهام شیرازی Department of Psychiatry, Faculty of Medicine, Tehran University of Medical Sciences Department of Psychiatry, Faculty of Medicine, Tehran University of Medical Sciences Iran Hooman H Sadri-Ardekani هومن صدری‌اردکانی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Koorosh K Kamali Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran K.Kamali@avicenna.ac.ir AttitudeCancerFertility preservationOncologistRadio-chemotherapy 463.pdf Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;364(9443):1405-10.##Wallace WH, Anderson RA, Irvine DS. Fertility preservation for young patients with cancer: who is at risk and what can be offered? Lancet Oncol. 2005;6(4):209-18.##Jemal A, Clegg LX, Ward E, Ries LA, Wu X, Jamison PM, et al. Annual report to the nation on the status of cancer, 1975-2001, with a special feature regarding survival. Cancer. 2004;101(1):3-27.##Maltaris T, Boehm D, Dittrich R, Seufert R, Koelbl H. Reproduction beyond cancer: a message of hope for young women. Gynecol Oncol. 2006;103(3): 1109-21.##Blatt J. Pregnancy outcome in long-term survivors of childhood cancer. Med Pediatr Oncol. 1999;33 (1):29-33.##Tschudin S, Bitzer J. Psychological aspects of fertility preservation in men and women affected by cancer and other life-threatening diseases. Hum Reprod Update. 2009;15(5):587-97.##Davis VJ. Female gamete preservation. Cancer. 2006;107(7 Suppl):1690-4.##Practice Committee of the American Society for Reproductive Medicine. Ovarian tissue and oocyte cryopreservation. Fertil Steril. 2004;82(4):993-8.##Lee SJ, Schover LR, Partridge AH, Patrizio P, Wallace WH, Hagerty K, et al. American Society of Clinical Oncology recommendations on fertility preservation in cancer patients. J Clin Oncol. 2006; 24(18):2917-31.##Schover LR, Brey K, Lichtin A, Lipshultz LI, Jeha S. Knowledge and experience regarding cancer, infertility, and sperm banking in younger male survivors. J Clin Oncol. 2002;20(7):1880-9.##Zapzalka DM, Redmon JB, Pryor JL. A survey of oncologists regarding sperm cryopreservation and assisted reproductive techniques for male cancer patients. Cancer. 1999;86(9):1812-7.##Partridge AH, Gelber S, Peppercorn J, Sampson E, Knudsen K, Laufer M, et al. Web-based survey of fertility issues in young women with breast cancer. J Clin Oncol. 2004;22(20):4174-83.##Thewes B, Meiser B, Taylor A, Phillips KA, Pendlebury S, Capp A, et al. Fertility- and menopauserelated information needs of younger women with a diagnosis of early breast cancer. J Clin Oncol. 2005;23(22):5155-65.##Duffy CM, Allen SM, Clark MA. Discussions regarding reproductive health for young women with breast cancer undergoing chemotherapy. J Clin Oncol. 2005;23(4):766-73.##Goodwin T, Elizabeth Oosterhuis B, Kiernan M, Hudson MM, Dahl GV. Attitudes and practices of pediatric oncology providers regarding fertility issues. Pediatr Blood Cancer. 2007;48(1):80-5.##

Post-Delivery Cardiomyopathy in a Patient Admitted to Critical Care Unit; A Rare Case Report 2 1 2 Background: Peripartum cardiomyopathy (PPCM) is an uncommon disease that affects women in the last month of pregnancy or within the first five months postpartum, occurring in about 1 in 3500 live births. The disease bears potentially devastating effects both on mother and the fetus if not treated early in its course. Case Presentation: The case was a 34-year old woman with a triple pregnancy who presented to the ward immediately after cesarean section with signs of dyspnea, cyanosis and pulmonary edema. She was diagnosed with PPCM upon echocardiography. The patient improved remarkably despite the PPCM’s devastating complications. This case report aims to describe a female patient who developed PPCM after a triple delivery. Conclusion: Regarding the high risks of developing PPCM in subsequent pregnancies and avoiding multiparty, especially in older age, a reliable contraception in childbearing women would be helpful. The best prevention of PPCM is to avoid subsequent pregnancies. 37 42 Mohammadreza MR Safavi محمد رضا صفوی Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Iran Azim A Honarmand عظیم هنرمند Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Iran honarmand@med.mui.ac.ir Parviz P Kashefi پرویز کاشفی Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Iran Saied Morteza SM Heidari سعید مرتضی حیدری Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Iran Afshin A Safavi افشین صفوی Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Anesthesiology and Critical Care Research Center, Isfahan University of Medical Sciences Iran Rita R Hekmat ریتا حکمت Department of Gynecology and Obstetrics, School of Medicine, Isfahan University of Medical Sciences Department of Gynecology and Obstetrics, School of Medicine, Isfahan University of Medical Sciences Iran Caesarian sectionEchocardiographyGestational hypertensionMyocarditisPeripartum cardiomyopathyPreeclampsiaThrombophilic phenomena 459.pdf Demakis JG, Rahimtoola SH, Sutton GC, Meadows WR, Szanto PB, Tobin JR, et al. Natural course of peripartum cardiomyopathy. Circulation. 1971;44 (6):1053-61.##Satpathy HK, Frey D, Satpathy R, Satpathy C, Fleming A, Mohiuddin SM, et al. Peripartum cardiomyopathy. Postgrad Med. 2008;120(1):28-32.##Lampert MB, Lang RM. Peripartum cardiomyopathy. Am Heart J. 1995;130(4):860-70.##Sliwa K, Damasceno A, Mayosi BM. Epidemiology and etiology of cardiomyopathy in Africa. Circulation. 2005;112(23):3577-83.##Elkayam U, Akhter MW, Singh H, Khan S, Bitar F, Hameed A, et al. Pregnancy-associated cardiomyopathy: clinical characteristics and a comparison between early and late presentation. Circulation. 2005; 111(16):2050-5.##Bouabdallaoui N, de Groote P, Mouquet F. [Peripartum cardiomyopathy]. Presse Med. 2009;38(6): 995-1000. French.##Veille JC. Peripartum cardiomyopathies: a review. Am J Obstet Gynecol. 1984;148(6):805-18.##Mendelson MA, Chandler J. Postpartum cardiomyopathy associated with maternal cocaine abuse. Am J Cardiol. 1992;70(11):1092-4.##Lampert MB, Hibbard J, Weinert L, Briller J, Lindheimer M, Lang RM. Peripartum heart failure associated with prolonged tocolytic therapy. Am J Obstet Gynecol. 1993;168(2):493-5.##Hibbard JU, Lindheimer M, Lang RM. A modified definition for peripartum cardiomyopathy and prognosis based on echocardiography. Obstet Gynecol. 1999;94(2):311-6.##Pearl W. Familial occurrence of peripartum cardio-myopathy. Am Heart J. 1995;129(2):421-2.##Al-Shamiri MQ, Al-Nozha MM. Peripartum cardiomyopathy searching for a better understanding. Saudi Med J. 2003;24(10):1048-51.##Homans DC. Peripartum cardiomyopathy. N Engl J Med. 1985;312(22):1432-7.##Demakis JG, Rahimtoola SH. Peripartum cardiomyopathy. Circulation. 1971;44(5):964-8.##Melvin KR, Richardson PJ, Olsen EG, Daly K, Jackson G. Peripartum cardiomyopathy due to myocarditis. N Engl J Med. 1982;307(12):731-4.##Pearson GD, Veille JC, Rahimtoola S, Hsia J, Oakley CM, Hosenpud JD, et al. Peripartum cardiomyopathy: National Heart, Lung, and Blood Institute and Office of Rare Diseases (National Institutes of Health) workshop recommendations and review. JAMA. 2000;283(9):1183-8.##Murali S, Baldisseri MR. Peripartum cardiomyopathy. Crit Care Med. 2005;33(10 Suppl):S340-6.##Homans DC. Peripartum cardiomyopathy. N Engl J Med. 1985;312(22):1432-7.##Lee W. Clinical management of gravid women with peripartum cardiomyopathy. Obstet Gynecol Clin North Am. 1991;18(2):257-71.##Cooper LT Jr, Gersh BJ. Viral infection, inflammation, and the risk of idiopathic dilated cardiomyopathy: can the fire be extinguished? Am J Cardiol. 2002;90(7):751-4.##Lapinsky SE, Kruczynski K, Slutsky AS. Critical care in the pregnant patient. Am J Respir Crit Care Med. 1995;152(2):427-55.##Midei MG, DeMent SH, Feldman AM, Hutchins GM, Baughman KL. Peripartum myocarditis and cardiomyopathy. Circulation. 1990;81(3):922-8.##Sanson BJ, Lensing AW, Prins MH, Ginsberg JS, Barkagan ZS, Lavenne-Pardonge E, et al. Safety of low-molecular-weight heparin in pregnancy: a systematic review. Thromb Haemost. 1999;81(5): 668-72.##Aziz TM, Burgess MI, Acladious NN, Campbell CS, Rahman AN, Yonan N, et al. Heart transplantation for peripartum cardiomyopathy: a report of three cases and a literature review. Cardiovasc Surg. 1999;7(5):565-7.##Lee W, Cotton DB. Peripartum cardiomyopathy: current concepts and clinical management. Clin Obstet Gynecol. 1989;32(1):54-67.##Bozkurt B, Villaneuva FS, Holubkov R, Tokarczyk T, Alvarez RJ Jr, MacGowan GA, et al. Intravenous immune globulin in the therapy of peripartum cardiomyopathy. J Am Coll Cardiol. 1999; 34(1):177-80.##Coulson CC, Thorp JM Jr, Mayer DC, Cefalo RC. Central hemodynamic effects of recombinant human relaxin in the isolated, perfused rat heart model. Obstet Gynecol. 1996;87(4):610-2.##Packer M, Fowler MB, Roecker EB, Coats AJ, Katus HA, Krum H, et al. Effect of carvedilol on the morbidity of patients with severe chronic heart failure: results of the carvedilol prospective randomized cumulative survival (COPERNICUS) study. Circulation. 2002;106(17):2194-9.##McNamara DM, Holubkov R, Starling RC, Dec GW, Loh E, Torre-Amione G, et al. Controlled trial of intravenous immune globulin in recentonset dilated cardiomyopathy. Circulation. 2001; 103(18):2254-9.##Ginsberg JS, Hirsh J. Anticoagulants during pregnancy. Annu Rev Med. 1989;40:79-86. Review.##Packer M, O'Connor CM, Ghali JK, Pressler ML, Carson PE, Belkin RN, et al. Effect of amlodipine on morbidity and mortality in severe chronic heart failure. Prospective Randomized Amlodipine Survival Evaluation Study Group. N Engl J Med. 1996;335(15):1107-14.##Sliwa K, Skudicky D, Bergemann A, Candy G, Puren A, Sareli P. 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Publications about Sperm during the Years 1897 to 2010 2 1 2 Sperm cells are one of the most stellar and fas-cinating cells in all species. One of the most distinctive characteristic of these cells is their movement dynamics which can be observed through a light microscope. The first paper regarding sperm cell biology that appeared on PubMed was pres-ented by Dr. W.F. Whitney (1). There, he showed the staining properties of spermatozoa and it was published in the Boston Society of Medical Sciences Journal in 1897. From that time on, around 82,998 papers have been published about sperm. The objective of the present paper was to evaluate the dynamic of sperm publication in the world using PubMed indexing database between 1897 and 2010 and the percentage of papers published on sperm per year. 43 44 Maya M Walter Cardona مایا والتر کاردونا Grupo Reproducción, Universidad de Antioquia Grupo Reproducción, Universidad de Antioquia Colombia wdcmaya@medicina.udea.edu.co No Keyword 458.pdf Whitney WF. The double staining of spermatozoa. J Boston Soc Med Sci. 1897;1(7):9-10.##