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Comparison of reactive oxygen species in neat and washed semen of infertile men مقایسه رادیکال های آزاد اکسیژن در نمونه سیمن خام و شسته شده مردان نابارور 2 1 مقدمه: فاکتور مردانه در 50% موارد ناباروری دخیل بوده و رادیکال­های آزاد اکسیژن (ROS) نقش مهمی را در کاهش پتانسیل باروری ایفا می­کنند. اندازه­گیری دقیق این رادیکال­ها می­تواند در ارزیابی ناباروری مردان مفید باشد. هدف: هدف از انجام این مطالعه اندازه­گیری و مقایسه رادیکال­های آزاد اکسیژن در نمونه خام و شسته شده سیمن مردان نابارور و تعیین بهترین روش اندازه­گیری ROS بود. مواد و روش­ها: در این مطالعه سطح رادیکال­های آزاد اکسیژن در 35 مرد نابارور بدون آزوسپرمی تعیین شد. مایع سیمن این بیماران به دو قسمت تقسیم، و میزان ROS و پارامترهای اسپرم در هر دو نمونه خام و شسته شده اندازه­گیری شد. همچنین وجود پیوسپرمی با استفاده از تست پراکسیداز در این نمونه­ها انجام شد. نتایج: تعداد و حرکت پیشرونده اسپرم بین دو گروه تفاوت قابل توجهی داشت و میزان ROS در گروه سیمن خام به صورت معنی­داری بالاتر از سیمن شسته شده گزارش شد (RLU 7/50 در مقابل RLU 1/20). همچنین میزان ROS بین دو گروه مردان نابارور دارای پیوسپرمی (RLU 378/67) و بدون پیوسپرمی (RLU 9/48) تفاوت معنی­دار آماری را نشان داد. نتیجه­گیری: مطالعه ما نشان داد که نمونه سیمن خام شاخص بهتری برای ارزیابی استرس­های اکسیداتیو در نمونه­های سیمن است. زیرا فاکتورهای مصنوعی که نقش مهمی در وضعیت تعادلی استرس­های اکسیداتیو دارند به آن اضافه یا کم نشده اند. 2 Background: Male are involved in near 50% of cases of infertility and reactive oxygen species (ROS) playing an important role in decreasing fertility potential. Accurate measurement of ROS seems to be important in evaluation of infertile male patients. Objective: To compare ROS measurement in neat and washed semen samples of infertile men and define the best method for evaluation of ROS in these patients. Materials and Methods: We measured the level of ROS in semen samples of thirty five non-azoospermic men with infertility. The semen samples were divided into two parts and the semen parameters and ROS levels in neat and washed samples were evaluated. We also evaluated the presence of pyospermia using peroxidase test. Results: The differences regarding sperm count and quick motility were significant in neat and washed semen samples. The mean ROS level was significantly higher in neat samples compared with washed spermatozoa (7.50 RLU vs. 1.20 RLU respectively). Difference in ROS levels was more significant in patients with pyospermia compared to whom with no pyospermia (378.67 RLU vs. 9.48 RLU respectively). Conclusion: Our study confirmed that neat or unprocessed samples are better index of normal oxidative status of semen samples. Because we do not artificially add or remove factors that may play an important role in oxidative equilibrium status. 301 0 2017/10/1 1396/7/9 2018/02/4 1396/11/15 محمدرضا معین Mohammad Reza Moein Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران moein1339@yahoo.com سراج الدین وحیدی Serajedin Vahidi Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران جلال قاسم زاده Jalal Ghasemzadeh Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران نسیم طبیب نژاد Nasim Tabibnejad Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران ROS Neat sperm Washed sperm Pyospermia رادیکال های آزاد اکسیژن سیمن خام سیمن شسته شده پیوسپرمی. Sharlip ID, Jarow JP, Belker AM, Lipshultz LI, Sigman M, Thomas AJ, et al. Best practice policies for male infertility. Fertil Steril 2002; 77: 873-882.##Alkan I, Simsek F, Haklar G, Kervancioglu E, Ozveri H, Yalcin S, et al. Reactive oxygen species production by spermatozoa of patients with idiopathic infertility: relationship to seminal plasma antioxidants. J Urol 1997; 157: 140-143.##Agarwal A, Said TM. Oxidative stress, DNA damage and apoptosis in male infertility: a clinical approach. BJU Int 2005; 95: 503-507.##Benedetti S, Tagliamonte MC, Catalani S, Primiterra M, Canestrari F, De Stefani S, et al. Differences in blood and semen oxidative status in fertile and infertile men, and their relationship with sperm quality. Reprod Biomed Online 2012; 25: 300- 306.##De Lamirande E, Gagnon C. Human sperm hyperactivation and capacitation as part of an oxidative process. Free Radic Biol Med 1993; 14: 157-166.##Agarwal A, Makker K, Sharma R. Clinical relevance of oxidative stress in male factor infertility: an update. Am J Reprod Immunol 2008; 59: 2-11.##Aitken RJ, McLaughlin EA. Molecular mechanisms of sperm capacitation: progesterone-induced secondary calcium oscillation reflect the attainment of a capacitated state. Soc Reprod Fertil Suppl 2007; 63: 273-293.##De Lamirande E, O'Flaherry C. Sperm activation: role of reactive oxygen species and kinases. Biochim Biophys Acta 2008; 1784: 106-115.##Roveri A, Ursini F, Flohe L, Maiorino M. PHGPx and spermatogenesis. Biofactors 2001; 14: 213-222.##Aitken RJ, Irvines DS, Wu FC. Prospective analysis of sperm-oocyte fusion and reactive oxygen species generation as criteria for the diagnosis of infertility. Am J Obstet Gynecol 1991; 164: 542-551.##Zorn B, Vidmar J, Meden-Vrtovec H. Seminal reactive oxygen species as predictors of fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization and intracytoplasmic sperm injection. Int J Androl 2003; 26: 279-285.##Padron OF, Brackett NL, Sharma RK, Lynne CM, Thomas AJ Jr, Agarwal A. Seminal reactive oxygen species and sperm motility and morphology in men with spinal cord injury. Fertil Steril 1997; 67: 1115-1120.##Agarwal A, Allamaneni SS, Said TM. Chemiluminescence technique for measuring reactive oxygen species. Reprod Biomed Online 2004; 9: 466-468.##Venkatesh S, Shamsi MB, Dudeja S, Kumar R, Dada R. Reactive oxygen species measurement in neat and washed semen: comparative analysis and its significance in male infertility assessment. Arch Gynecol Obstet 2011; 283: 121-126.##Agarwal A, Ikemoto I, Loughlin KR. Effect of sperm washing on levels of reactive oxygen species in semen. Arch Androl 1994; 33: 157-162.##Allamaneni SS, Agarwal A, Nallella KP, Sharma RK, Thomas AJ Jr, Sikka SC Characterization of oxidative stress status by evaluation of reactive oxygen species levels in whole semen and isolated spermatozoa. Fertil Steril 2005; 83: 800-803.##World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucus interaction, 4th Ed. Cambridge University Press, New York; 1999.##Shekarriz M, Sharma RK, Thomas AJ Jr, Agarwal A. Positive myeloperoxidase staining (Endtz test) as an indicator of excessive reactive oxygen species in semen. J Assist Reprod Genet 1995; 12: 70-74.##Endtz AW. A rapid staining method for differentiating granulocytes from "germinal cells" in Papanicolaou-stained semen. Acta Cytol 1974; 18: 2-7.##Agarwal A, Saleh RA, Bedaiwy MA. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril 2003; 79: 829-843.##Henkel R, Kierspel E, Stalf T, Mehnert C, Menkveld R, Tinneberg HR, et al. Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. Fertil Steril 2005; 83: 635-642.##Moustafa MH, Sharma RK, Thornton J, Mascha E, Abdel-Hafez MA, Thomas AJ Jr, et al. Relationship between ROS production, apoptosis and DNA denaturation in spermatozoa from patients examined for infertility. Hum Reprod 2004; 19: 129-138.##Saleh RA, Agarwal A, Kandirali E, Sharma RK, Thomas AJ, Nada EA, et al. Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa. Fertil Steril 2002; 78: 1215-1224.##Saleh RA, Agarwal A, Sharma RK, Nelson DR, Thomas AJ. Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: a prospective study. Fertil Steril 2002; 78: 491-499.##Cocuzza M, Atayde KS, Agarwal A, Pagani R, Sikka SC, Lucon AM, et al. Impact of clinical varicocele and testis size on seminal reactive oxygen species levels in a fertile population: a prospective controlled study. Fertil Steril 2008; 90: 1103-1108.##Desai N, Sharma RK, Makler K, Sabanegh E, Agarwal A. Physiologic and pathologic levels of reactive oxygen species in neat semen of infertile men. Fertil Steril 2009; 92: 1626-1631.##Iwasaki A, Gagnon C. Formation of reactive oxygen species in spermatozoa of infertile patients. Fertil Steril 1992; 57: 409-416.##Pasqualotto FF, Shrma RK, Kobayashi H, Nelson DR, Thomas AJ, Agarwal A. Oxidative stress in normospermic men undergoing infertility evaluation. J Androl 2001; 22: 316-322.## ##

Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice مصرف ساخارین میزان قطعه قطعه شدن DNA و آپوپتوز را در اسپرم موش افزایش می دهد 2 1 مقدمه: ساخارین یک شیرین­کننده مصنوعی و فاقد کالری است که برای شیرین کردن محصولاتی همچون نوشیدنی­ها، شیرینی­جات، بیسکوئیت­ها، داروها و خمیردندان استفاده می­شود. این در حالی است که بدن ما قادر به متابولیزه کردن آن نیست. سدیم ساخارین به عنوان یک فاکتور مهم در پیشبرد تومور و تحریک تومورهای مثانه در رت­های مذکر اما نه در انسان تلقی می­گردد. به هر حال بنابر دانش ما، داده­هایی مبنا بر تأثیرات ساخارین بر عملکردهای تولیدمثلی به ویژه پتانسیل باروری اسپرم وجود ندارد. هدف: از آنجایی که اسپرم دارای یک نقش حیاتی در عملکرد تولیدمثلی است، هدف از این مطالعه بررسی تأثیر مصرف ساخارین بر پارامترهای اسپرم و آپوپتوز در موش­های بالغ است. مواد و روش­ها: مجموعا 14 موش نر بالغ به 2 گروه تقسیم شدند. گروه 1 به عنوان کنترل، غذای روتین موش و گروه 2 یا حیوانات آزمایشی، آب حاوی ساخارین (W/V 0/2) را به مدت 35 روز دریافت نمودند. بعد از این مدت، از هر حیوان دم اپیدیدیم مربوط به سمت چپ جدا و در محیط Ham’s F10 قرار داده شد. اسپرم­های خارج شده برای ارزیابی تعداد، حرکت مورفولوژی (رنگ آمیزی پاپانیکولا) و حیات (رنگ آمیزی ائوزین) مورد استفاده قرار گرفتند. یکپارچگی DNA اسپرم به عنوان اندیکاتور آپوپتوز با تست­های SCD و TUNEL مورد ارزیابی قرار گرفت. نتایج: در پی مصرف ساخارین، تعداد و حرکت اسپرم­ها کاهش و میزان ناهنجاری­های مورفولوژیکی افزایش یافت. همچنین میزان آسیب DNA و آپوپتوز در گروه آزمایشی در مقایسه با گروه کنترل از لحاظ آماری به طور معنی­داری افزایش یافت. نتیجه­گیری: بنابر داده­های ما، مصرف ساخارین ممکن است دارای تأثیرات منفی بر روی پارامترهای اسپرم باشد و منجر به افزایش میزان قطعه قطعه شدن DNA و آپوپتوز در اسپرم موش گردد. 2   Background: Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans. Objective: The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice. Materials and Methods: Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin (0.2% w/v) for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham’s F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology (Pap-staining) and viability (eosin-Y staining). Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD (sperm chromatin dispersion) and terminal deoxynucleotidyl transferase (TUNEL) assay. Results: Following saccharin consumption, we had a reduction in sperm motility with respect to control animals (p=0.000). In addition, the sperm count diminished (17.70±1.11 in controls vs. 12.80±2.79 in case group, p=0.003) and the rate of sperm normal morphology decreased from 77.00±6.40 in control animals into 63.85±6.81 in saccharin-treated mice (p=0.001). Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one (p=0.001, p=0.002 respectively). Conclusion: Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice. 307 0 2017/10/12017/10/1 1396/7/9 2018/02/42018/02/4 1396/11/15 مرضیه رحیمی پور Marzieh Rahimipour Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران علیرضا طالبی Ali Reza Talebi Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران prof_talebi@ssu.ac.ir مرتضی انوری Morteza Anvari Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران ابوالقاسم عباسی سرچشمه Abolghasem Abbasi Sarcheshmeh Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران مرجان امیدی Marjan Omidi Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران Sperm Saccharin Apoptosis Mice اسپرم ساخارین آپوپتوز موش. Mitchell H. Sweeteners and sugar alternatives in food technology. 1th Ed. Blackwell; 2006.##Schardt D. Sweet nothings. Nutr Act 2004; 9: 8-11.##No DCR. Saccharin and its salts. 2th Ed. 1987.##Zurlo J, Squire RA. Is saccharin safe? Animal testing revisited. J Natl Cancer Inst 1998; 90: 2-3.##Turner SD, Tinwell H, Piegorsch W, Schmezer P, Ashby J. The male rat carcinogens limonene and sodium saccharin are not mutagenic to male Big BlueTM rats. Mutagenesis 2001; 16: 329-332.##Wolff S, Rodin B. Saccharin-induced sister chromatid exchanges in Chinese hamster and human cells. Science 1978; 200: 543-545.##Hikim AS, Swerdloff RS. Hormonal and genetic control of germ cell apoptosis in the testis. Rev Reprod 1999; 4: 38-47.##Porcelli F, Meggiolaro D, Carnevali A, Ferrandi B. Fas ligand in bull ejaculated spermatozoa: a quantitative immunocytochemical study. Acta Histochem 2006; 108: 287-292.##Nagata S. Fas ligand-induced apoptosis. Annu Rev Genet 1999; 33: 29-55.##Ozmen B, Koutlaki N, Youssry M, Diedrich K, Al-Hasani S. DNA damage of human spermatozoa in assisted reproduction: origins, diagnosis, impacts and safety. Reprod Biomed Online 2007; 14: 384-395.##Liu W, Staecker H, Stupak H, Malgrange B, Lefebvre P, Van De Water TR. Caspase inhibitors prevent cisplatin-induced apoptosis of auditory sensory cells. Neuroreport 1998; 9: 2609-2614.##Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol 1980; 68: 251.##Talebi AR. Sperm Nuclear Maturation: a basic and clinical approach. 1th Ed. New York, Nova Science Publishers; 2011.##Schulte RT, Ohl DA, Sigman M, Smith GD. Sperm DNA damage in male infertility: etiologies, assays, and outcomes. J Assist Reprod Genet 2010; 27: 3-12.##Zini A, Bielecki R, Phang D, Zenzes MT. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril 2001; 75: 674.##Makler A. The improved ten-micrometer chamber for rapid sperm count and motility evaluation. Fertil Steril 1980; 33: 337.##Kermani-Alghoraishi M, Anvari M, Talebi AR, Amini-Rad O, Ghahramani R, Miresmaili SM. The effects of acrylamide on sperm parameters and membrane integrity of epididymal spermatozoa in mice. Eur J Obstet Gynecol Reprod Biol 2010; 153: 52-55.##Kruger TF, Menkveld R, Stander F, Lombard C, Van der Merwe J, Van Zyl J, et al. Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil Steril 1986; 46: 1118.##Fernández JL, Muriel L, Rivero MT, Goyanes V, Vazquez R, Alvarez JG. The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation. J Androl 2003; 24: 59-66.##Gandini L, Lombardo F, Paoli D, Caponecchia L, Familiari G, Verlengia C, et al. Study of apoptotic DNA fragmentation in human spermatozoa. Hum Reprod 2000; 15: 830-839.##Ellwein LB, Cohen SM. The health risks of saccharin revisited. Crit Rev Toxicol 1990; 20: 311-326.##Dybing E. Development and implementation of the IPCS conceptual framework for evaluating mode of action of chemical carcinogens. Toxicology 2002; 181: 121-125.##Sarıözkan S, Bucak MN, Canturk F, Özdamar S, Yay A, Tuncer PB, et al. The effects of different sugars on motility, morphology and DNA damage during the liquid storage of rat epididymal sperm at 4°C. Cryobiology 2012; 65: 93-97.##Corcuera B, Marigorta P, Sagüés A, Saiz-Cidoncha F, Pérez-Gutiérrez J. Effect of lactose and glycerol on the motility, normal apical ridge, chromatin condensation and chromatin stability of frozen boar spermatozoa. Theriogenology 2007; 67: 1150-1157.##Sánchez R, Risopatrón J, Schulz M, Villegas J, Isachenko V, Kreinberg R, et al. Canine sperm vitrification with sucrose: effect on sperm function. Andrologia 2011; 43: 233-241.##Dasgupta J, Elliott RA, Doshani A, Tincello DG. Enhancement of rat bladder contraction by artificial sweeteners via increased extracellular Ca< sup> 2+</sup> influx. Toxicol Appl Pharmacol 2006; 217: 216-224.##Andreatta M, Mu-oz S, Lantieri M, Eynard A, Navarro A. Artificial sweetener consumption and urinary tract tumors in Cordoba, Argentina. Prev Med 2008; 47: 136-139.##Tavalaee M, Nasr-Esfahani MH, Deemeh MR. Etiology and evaluation of sperm chromatin anomalies. Int J Fertil Steril 2008; 2: 1-8.##Kensler TW, Trush MA. Role of oxygen radicals in tumor promotion. Environ Mutagen 1984; 6: 593-616.##Al-Saleh AM. Effect of Artificial Sweeteners on Insulin Secretion, ROS, and Oxygen Consumption in Pancreatic [Beta]-cells. Boston University, Boston, 2010.## ##

Expression of Glycogen synthase kinase 3-β (GSK3-β) gene in azoospermic men بیان ژن گلیکوژن سنتاز کیناز 3بتا (GSK3-β) در مردان آزواسپرم 2 1 مقدمه: مسیر پیام رسانی Wnt / β-catenin هم در دوره جنینی و هم در بلوغ در بسیاری از فرآیندهای رشد و نمو دخالت دارد. ناهنجاری­های آن می­تواند باعث اختلالاتی از جمله انواع مختلف سرطان و نقص عملکرد سلول­ها و بافت­های خاص در انسان و حیوانات شود. نقش آن در فرآیندهای تولید مثلی ثابت شده است. هدف: این مطالعه به منظور بررسی بیان GSK3-β، ژن کلیدی تنظیم کننده این مسیر سیگنالینگ و نقش احتمالی آن در آزواسپرمی طراحی شد. مواد و روش­ها: غلظت پروتئین WNT3a و بیان ژن GSK3-β در دو گروه از مردان نابارور (10 نفر مبتلا به آزوسپرمی انسدادی و 10 نفر مبتلا به آزوسپرمی غیر انسدادی) اندازه­گیری و مقایسه شد. گروه کنترل شامل 10 نفر از مردان باروری بود که پس از عمل وازکتومی مجددا مایل به داشتن فرزند بودند. نمونه گیری با بیوپسی بیضه انجام شده و پیش از ورود به تحقیق از نظر شایع­ترین جهش­های ناحیه SRY شامل (84، 86 و 255) غربالگری شدند. بیان ژن GSK3-β به روش Real time-PCR مورد بررسی کمی قرار گرفت. نتایج: غلظت پروتئین WNT3a بین دو گروه آزمون و کنترل تفاوت معنی­داری نداشت. بیان GSK3-β در گروه آزواسپرمی غیر انسدادی (0/19±3/10) در مقایسه با گروه نرمال (0/39±7/12) و آزواسپرمی انسدادی (0/42±6/32) پایین تر بود (0/001=p) نتیجه­گیری: کاهش بیان GSK-3β می­تواند باعث آزواسپرمی غیر انسدادی شود. تنظیم بیان ژن GSK-3β با استفاده از داروها می تواند به عنوان یک راه حل درمانی استفاده شود. 2 Background: The Wnt/β- The Wnt/β-catenin signaling pathway is involved in many developmental processes in both fetal and adult life; its abnormalities can lead to disorders including several types of cancers and malfunction of specific cells and tissues in both animals and humans. Its role in reproductive processes has been proven. Objective: This study was designed to evaluate the expression of the key regulator of this signaling pathway GSK3-β and its presumed role in azoospermia. Materials and Methods: WNT3a protein concentration and GSK3-β gene expression levels were measured and compared between two groups of infertile men. The test groups consisted of 10 patients with obstructive and 10 non-obstructive azoospermia. The control group was selected among healthy men after vasectomies that were willing to conceive a child using a testicular biopsy technique. Samples were obtained by testicular biopsy and screened for the most common mutations (84, 86 and 255) in the SRY region before analyzing. GSK3-β gene expression was assessed quantitatively by real time-PCR. Results: The WNT3a protein concentration had no significant difference between the two test groups and controls. Expression of GSK3-β was down-regulated in non-obstructive azoospermia (3.10±0.19) compared with normal (7.12±0.39) and obstructive azoospermia (6.32±0.42) groups (p=0.001). Conclusion: Down-regulation of GSK-3β may cause to non-obstructive azoospermia. Regulation and modification of GSK-3β gene expression by drugs could be used as a therapeutic solution. 313 0 2017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/4 1396/11/15 حمید نظریان Hamid Nazarian Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران معرفت غفاری نوین Marefat Ghaffari Novin Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران mghaffarin@yahoo.com محمدرضا جلیلی Mohammad Reza Jalili Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران رضا میرفخرایی Reza Mirfakhraie Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران محمد حسن حیدری Mohammad Hassan Heidari Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران سید جلیل حسینی Seyed Jalil Hosseini Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران محسن نوروزیان Mohsen Norouzian Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران ناهید احسانی Nahid Ehsani Infertility and Reproductive Health Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran Azoospermia Wnt signaling pathway glycogen synthase kinase 3 beta Spermatogenesis. آزوسپرمی مسیر پیام رسانی Wnt گلیکوژن سنتاز کیناز 3بتا اسپرماتوژنز. 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Assessemnt of nasal bone in first trimester screening for chromosomal abnormalities in Khuzestan بررسی میزان وجود استخوان بینی در سه ماه اول بارداری در غربال‌گری ناهنجاری‌های کروموزومی در جمعیت بومی منطقه خوزستان  2 1 مقدمه: یکی از فاکتورهایی که همراه با دیگر فاکتورهای سونوگرافی جهت تشخیص ناهنجاری­های کروموزومی مطرح است، وجود یا عدم وجود استخوان بینی جنین در سونوگرافی سه ماهه اول بارداری می­باشد. هدف: هدف از انجام این مطالعه بررسی میزان وجود این فاکتور در سه ماهه اول بارداری در جمعیت بومی منطقه خوزستان و بررسی ارزش آن در تشخیص ناهنجاری کروموزومی می­باشد. مواد و روش­ها: این مطالعه بر روی 2314 خانم باردار در سن بارداری 13-11 هفته که جهت غربال­گری سه ماهه اول بارداری مراجعه نموده بودند انجام شد. کلیه افراد از نظر وجود یا عدم وجود استخوان بینی جنین بررسی شدند. همچنین کلیه افراد تحت غربال­گری از نظر ناهنجاری کروموزومی با توجه به فاکتورهای سونوگرافی روشنی پشت گردن (NT)، تعداد ضربان قلب جنین (FHR)، طول سری-دمی جنین (CRL) و سن مادر و فاکتورهای سرمی خون مادر             Free βHCG] و پلاسما پروتیین A وابسته به حاملگی (PAPP_A) ]قرار گرفته و افراد در سه گروه کم خطر، پرخطر و با خطر متوسط قرار داده شدند و سرانجام بارداری در همه افراد تحت مطالعه، از نظر ابتلا یا عدم ابتلای جنین به ناهنجاری کروموزومی مورد بررسی قرار گرفتند. سپس بر اساس تست فیشر ارتباط بین ناهنجاری کروموزومی و وجود یا عدم وجود استخوان بینی بررسی شد. نتایج: از تعداد 2180 مورد خانم تحت مطالعه که در آن­ها فاکتور استخوان بینی ارزیابی شده بود، 2173 مورد دارای جنین با فنوتیپ نرمال و 7 مورد دارای جنین با ناهنجاری کروموزومی (3مورد تریزومی 21، 2مورد تریزومی 18، 1مورد سندرم ترنر استخوان بینی منفی و تریزومی 13، مثبت بود. از 2173 مورد با فنوتیپ نرمال نیز در 6 مورد فاکتور استخوان بینی منفی بود. براساس تست دقیق فیشر با0/0001=p ارتباط معنی­داری بین عدم وجود استخوان بینی و ابتلا به ناهنجاری کروموزومی حاصل شد. نتیجه­گیری: غربال­گری سه ماهه اول بارداری در تشخیص ناهنجاری­های جنینی بسیار مؤثر می­باشد و فاکتور استخوان بینی در منطقه تحت مطالعه دارای ارزش قابل توجه در تشخیص ناهنجاری­های کروموزومی می­باشد. 2 Background: Fetal nasal bone assessment is a non-invasive procedure that helps provide even greater assurance to patients undergoing their first trimester risk assessment for aneuploidies. Absence or presence of this factor is different in some races. Objective: The study was aimed to evaluate nasal bone in the first trimester of pregnancy in the indigenous population of Khuzestan Province, and to monitor its value in the diagnosis of chromosomal abnormalities. Materials and Methods: This study was conducted on 2314 pregnant women between 17-43 years old who referred for first trimester screening for chromosomal abnormalities. Gestational age was between 11-13w + 6 days. Nuchal translucency (NT), fetal heart rate (FHR), crown rump length (CRL), and maternal age and maternal blood serum factors (Free βHCG) and pregnancy-associated plasma protein-A (PAPP-A) and nasal bone were assessed. Finally the risk of trisomies was calculated. The statistical tests are based on the relationship between chromosomal abnormality and the presence or absence of the nasal bone. Results: In 114 cases we could not examine the nasal bone. Also, in 20 cases missed abortion happened without knowing the karyotype. 2173 cases were delivered normal baby, and in seven cases chromosomal abnormalities were diagnosed. Nasal bone was absent in all three cases with trisomy 21 and six of 2173 cases with normal phenotype (0.3%). With use of the Fisher exact test (p=0.0001), a significant correlation was found between the absence of the nasal bone and the risk of chromosomal abnormality. Conclusion: Inclusion of the nasal bone in first-trimester combined screening for aneuploidies achieves greater detection rate especially in Down syndrome 321 0 2017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/4 1396/11/15 سارا مسیحی Sara Masihi Depatment of Obstetric and Gynecology, Fertility Infertility and Perinatology Research Center, Ahvaz Jundishapour University of Medical Sciences, Ahvaz, Iran ایران sara.masihi@yahoo.com مژگان براتی Mojgan Barati Depatment of Obstetric and Gynecology, Fertility Infertility and Perinatology Research Center, Ahvaz Jundishapour University of Medical Sciences, Ahvaz, Iran ایران راضیه محمدجعفری Razieh Mohamadjafari Depatment of Obstetric and Gynecology, Fertility Infertility and Perinatology Research Center, Ahvaz Jundishapour University of Medical Sciences, Ahvaz, Iran ایران مرضیه هاشمی Marzieh Hashemi Department of obstetrics and Gynecology, Ahvaz Jundishapour University of Medical Sciences, Ahvaz, Iran ایران Prenatal diagnosis Nuchal translucency measurement Maternal serum screening test Nasal bone غربال گری فاکتور استخوان بینی ناهنجاری کروموزومی فاکتور روشنی پشت گردن. Adzick NS, Harrison MR, Cunningham F, Levavano J. Williams's obstetrics. 23PthP Ed. New York, MC Graw Hill; 2010: 287-311.##Bindra R, Heath V, Liao A, Spencer K, Nicolaides KH. One stop clinic for assessment of risk for trisomy 21 at 11-14 weeks: A prospective study of 15030 pregnancies. Ultrasound Obstet Gynecol 2002; 20: 219-225.##Nicolaides KH. Screening for fetal aneuploidies at 11-13 weeks. Prenat Diagn 2011; 31: 7-15.##Mogra R, Schluter PJ, Ogle RF, O'Connell J, Fortus L, Hyett JA. A prospective cross-sectional study to define racial variation in fetal Nasal bone length through ultrasound assessment at 11-13 weeks' gestation. Aust N Z J Obstet Gynaecol 2010; 50: 528-533.##Ozer A, Ozaksit G, Kanat Pektas M, Ozers: First trimester examination of fetal Nasal bone in the Turkish population. J Obstet Gynaecol Res 2010; 36: 739-744.##58TKagan KO58T, 58TCicero S58T, 58TStaboulidou I58T, 58TWright D58T, 58TNicolaides KH58T. Fetal Nasal bone in screening for trisomies 21, 18 and 13 and turner syndrome at 11-13 weeks of gestation. Ultrasound Obstet Gynecol 2009; 33: 259-264.##58TBunduki V58T, 58TRuano R58T, 58TMiguelez J58T, 58TYoshizaki CT58T, 58TKahhale S58T, 58TZugaib M58T. Fetal nasal bone length: reference range and clinical application in ultrasound screening for trisomy 21. Ultrasound Obstet Gynecol 2003; 21: 156-160.##58TYeo GS58T, 58TLai FM58T, 58TWei X58T, 58TLata P58T, 58TTan DT58T, 58TYong MH58T, et al. Validation of first trimester screening for trisomy21 in Singapore with reference to performance of nasal bone. Fetal Diagn Ther 2012; 32: 166-170.##58TGhaffari SR58T, 58TTahmasebpour AR58T, 58TJamal A58T, 58THantoushzadeh S58T, 58TEslamian L58T, 58TMarsoosi V58T, et al. First-trimester screening for chromosomal abnormalities by integrated application of nuchal translucency, nasal bone, tricuspid regurgitation and ductus venosus flow combined with maternal serum free βhCG and PAPP-A: a 5-year prospective study. Ultrasound Obstet Gynecol 2012; 39: 528-534.##Nicolaides KH. Nuchal translucency and other first-trimester sonographic markers of chromosomal abnormalities. Am J Obstet Gynecol 2004; 191: 45-67.##Nicolaides KH. Screening for chromosomal defects. Ultrasound Obstet Gynecol 2003; 21: 313-321.## ##

Molecular and serologic diagnostic approaches; the prevalence of herpes simplex in idiopathic men infertile تشخیص مبتنی بر روش های مولکولی و سرولوژیک: میزان شیوع هرپس سیمپلکس در موارد ناباروری ایدیوپاتیک مردان  2 1 مقدمه: امروزه ناباروری یکی از مهمترین مشکلات بهداشتی در جوامع مختلف در دنیا است. به نظر می­رسد که برخی پاتوژن­های انسانی می­توانند بر کیفیت، مورفولوژی و تعداد اسپرم­ها تأثیر داشته باشند. هدف: به منظور بررسی شیوع ویروس هرپس سیمپلکس انسانی در مایع منی مراجعه­کنندگان به کلینیک ناباروری ابن سینا و مقایسه آن با نتایج سرولوژی این تحقیق صورت گرفته است. مواد و روش­ها: در این مطالعه مقطعی مردانی که دچار ناباروری ایدیوپاتیک بودند و در طی سال­های 2008 تا 2009 به مرکز ناباروری ابن سینا مراجعه کرده بودند انتخاب شدند. این بیماران هیچ گونه علائم بالینی ناشی از آلودگی با این ویروس را نداشتند و هیچ­گونه علت خاص پزشکی برای ناباروری آن­ها متصور نبود. بعد از اخذ رضایت اگاهانه این افراد وارد مطالعه شدند. نمونه­های خون و مایع منی از تعداد 217 نفر به روش­های سرولوژیک و مولکولی مورد بررسی قرار گرفتند. سپس با استفاده از آزمون آماری اسپیرمن و پیرسون نتایج مورد مقایسه و بررسی قرار گرفتند. نتایج: حدود 4% افراد واجد IgM مثبت بودند. با انجام تست­های PCR مشخص شد که شیوع این ویروس در خون و مایع منی به ترتیب برابر با 3/2% و 12%  می­باشد. 10% از افراد IgM منفی و 3% از افراد IgM مثبت دارای تست PCR مثبت بودند. نتیجه­گیری: در صورتی که شک بالینی و تشخیصی برای وجود ویروس هرپس به عنوان عاملی برای ناباروری مطرح باشد بهتر است که از PCR روی مایع منی به جای سرولوژی الایزا استفاده نمود.  2 Background: Human pathogens that can cause infertility may also affect sperm count and quality. Viral infections can be considered as direct and/or indirect cause of male factor infertility. Objective: Our goal was to investigate the prevalence of herpes simplex virus in the semen of infertile men attending the Avicenna Infertility Clinic, and to compare it with the herpes virus serology results. Materials and Methods: This cross sectional study was conducted during 2009-2010. Infertile men participating without any clinical signs of infection with herpes simplex virus, and no obvious cause for their infertility were included. Semen and blood samples were used for Polymerase Chain Reaction (PCR) and serologic testing for these people. Two samples were collected: one ml semen sample to verify the existence of genital herpes simplex virus in infertile men, and blood samples of 217 individuals tested for antibodies to herpes simplex virus. Data were analyzed by SPSS 16. Results: According to the PCR results of semen samples the prevalence of herpes simplex in semen was 12% and serologic test showed 3.2% prevalence within blood. Nine to 10% of IgM negative were PCR positive and only 2-3% of IgM positive were PCR positive. Between herpes serologic studies with positive controls and negative controls by using both tests, there was a significant positive relationship (r=0.718 and p<0.001). The relationship between semen PCR test results and serological survey of herpes patients with a negative control in both Pearson and Spearman tests was positive and significant (r=0.229 and p=0.001). Correlation between the PCR results of semen samples with two positive control subjects and a positive IgM test was statistically confirmed (r=0.235 and p<0.001). Conclusion: We recommend that if there is suspicion to herpes simplex as a microorganism that theoretically could impact semen parameters and cause infertility it is prudent to use PCR technique on semen sample rather than ELISA on serum. 327 0 2017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/4 1396/11/15 ناصر امیرجنتی Nasser Amirjannati Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ایران فرهاد یغمایی Farhad Yaghmaei Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ایران yaghmaie@avicenna.ac.ir محمدمهدی آخوندی Mohammad Mehdi Akhondi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ایران محبوبه نصیری Mahboubeh Nasiri Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ایران حامد حیدری والا Hamed Heidari-Vala Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ایران زهرا صحت Zahra Sehhat Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ایران Herpes virus Infertility IgM Polymerase Chain Reaction Semen Serologic Sexually transmitted diseases (STD). هرپس ویروس ناباروری IgM واکنش زنجیره ای پلیمراز منی سرولوژیک STD. 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Semen washing rocedures do not eliminate human papilloma virus sperm infection in infertile patients. Fertil Steril 2011; 96: 1077-1082.##Dulioust E, Du AL, Costagliola D, Guibert J, Kunstmann JM, Heard I, et al. Semen alterations in HIV-1 infected men. Hum Reprod 2002; 17: 2112-2118.##Dias CF, Nunes CC, Freitas IO, Lamego IS, Oliveira IM, Gilli S, et al. High prevalence and association of HIV-1 non-B subtype with specific sexual transmission risk among antiretroviral naïve patients in Porto Alegre, RS, Brazil. Rev Inst Med Trop Sao Paulo 2009; 51: 191-196.##Bezold G, Schuster-Grusser A, Lange M, Gall H, Wolff H, Peter RU. Prevalence of human herpesvirus types 1-8 in the semen of infertility patients and correlation with semen parameters. Fertil Steril 2001; 76: 416-418.##Tremellen K, Oxidative stress and male infertility-a clinical perspective. Hum Reprod Update 2008; 14: 243-258.##Asgari S, Chamani-Tabriz L, Asadi S, Fatemi F, Zeraati H, Akhondi MM, et al. HSV-2 Seroepidemiology and Risk Factors among Iranian Women: A Time to New Thinking. Iran Red Crescent Med J 2011; 13: 818-823.##Salehi-vaziri M, Monavari SH, Khalili M, Shamsi-Shahrabadi M, Keyvani H, Mollaei H, et al. Detection of HSV-1 DNA in the Semen of Infertile Men and Evaluation of its Correlation with Semen Parameters in Iran. Iran J Virol 2010; 4: 1-6.##Kailash U, Hedau S, Gopalkrishna V, Katiyar S, Das BC. A simple 'paper smear' method for dry collection, transport and storage of cervical cytological specimens for rapid screening of HPV infection by PCR. J Med Microbiol 2002; 51: 606-683.##Wysocka J, Herr W. The herpes simplex virus VP16-induced complex: the makings of a regulatory switch. Trends Biochem Sci 2003; 28: 294-304.##Smith JS, Robinson NJ. Age-specific prevalence of infection with herpes simplex virus types 2 and 1: a global review. J Infect Dis 2002; 186 (Suppl.): 3-28.##Brooks GF. Medical Microbiology. 25th Ed. London, Appleton & Lange; 2010.##Collier LH, Oxford JS. Human Virology. 2nd Ed. Oxford, Oxford University Press; 1999.##Zaravinos A, Bizakis J, Spandidos DA. Prevalence of human papilloma virus and human herpes virus types 1-7 in human nasal polyposis. J Med Virol 2009; 81: 1613-1619.##Alberts CJ, Schim van der Loeff MF, Papenfuss MR, da Silva RJ, Villa LL, Lazcano-Ponce E, et al. Association of Chlamydia trachomatis Infection and Herpes Simplex Virus Type 2 Serostatus With Genital Human Papillomavirus Infection in Men: The HPV in Men Study. Sex Transm Dis 2013; 40: 508-515.##Aynaud O, Poveda JD, Huynh B, Guillemotonia A, Barrasso R. Frequency of herpes simplex virus, cytomegalovirus and human papillomavirus DNA in semen. Int J STD AIDS 2002; 13: 547-550.##Wald A, Matson P, Ryncarz A, Corey L. Detection of herpes simplex virus DNA in semen of men with genital HSV-2 infection. Sex Transm Dis 1999; 26: 1-3.##el Borai N, Inoue M, Lefèvre C, Naumova EN, Sato B, Yamamura M. Detection of herpes simplex DNA in semen and menstrual blood of individuals attending an infertility clinic. J Obstet Gynaecol Res 1997; 23: 17-24.##Foresta C, Patassini C, Bertoldo A, Menegazzo M, Francavilla F, Barzon L, et al. Mechanism of human papillomavirus binding to human spermatozoa and fertilizing ability of infected spermatozoa. PLoS One 2011; 6: e15036.##Todd J, Riedner G, Maboko L, Hoelscher M, Weiss HA, Lyamuya E, et al. Effect of genital herpes on cervicovaginal HIV shedding in women co-infected with HIV AND HSV-2 in Tanzania. PLoS One 2013; 8: e59037.##Bezold G, Politch JA, Kiviat NB, Kuypers JM, Wolff H, Anderson DJ. Prevalence of sexually transmissible pathogens in semen from asymptomatic male infertility patients with and without leukocytospermia. Fertil Steril 2007; 87: 1087-1097.##Smith JS, Rosinska M, Trzcinska A, Pimenta JM, Litwinska B, Siennicka J. Type specific seroprevalence of HSV-1 and HSV-2 in four geographical regions of Poland. 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Effects of acrylamide on sperm parameters, chromatin quality, and the level of blood testosterone in mice تأثیرآکریلامید بر پارامترهای اسپرم،کیفیت کروماتین و هورمون تستوسترون درموش سوری 2 1 مقدمه: آکریلامید (AA) ،یک ماده شیمیایی مهم صنعتی است که در بعضی غذاهای انسان از جمله غذاهای فرآوری نشاسته­دار مثل چیپس سیب زمینی و نان که در دمای بالا تولید می­شوند، وجود دارد. آکریلامید باعث القاء آسیب اسپرم در حیوانات می­گردد. هدف: هدف از این مطالعه، مشاهده اثرات AA بر پارامترهای اسپرم و ارزیابی کیفیت کروماتین اسپرم و هورمون تستوسترون در موش بود. مواد و روش­ها: 16 موش نر بالغ به دو گروه تقسیم شدند. 1- گروه کنترل: این گروه غذای معمول جوندگان را دریافت کردند. 2- گروه اکریلامید: این گروه علاوه بر غذای معمول جوندگان، اکریلامید را با دوز mg/kg 10 به مدت 35 روز به صورت محلول در آب دریافت کردند. سرانجام پس از بیهوشی و خونگیری، دم اپیدیدیم سمت راست جدا و در محلول Ham’s F10 قرار داده شد. سپس نمونه­های اسپرم از نظر حرکت، تعداد (با لنز 40×)، حیات (رنگ­آمیزی ائوزین) و مورفولوژی (رنگ­آمیزی پاپانیکولا) مورد بررسی میکروسکوپی قرار گرفتند. همچنین برای بررسی کیفیت و تراکم کروماتین از تست­های اکریدین ارنج، انیلین بلو، تولوئیدین بلو وکرومومایسین استفاده شد. نتایج: درگروه اکریلامید، کاهش قابل ملاحظه­ای در تعداد، تحرک، حیات و اسپرم­های با مورفولوژی نرمال نسبت به گروه کنترل مشاهده شد. با توجه به ارزیابی کروماتین اسپرم ،به جز تست تولوئیدین بلو در همه تست­های دیگر تفاوت قابل ملاحظه­ای در بین دو گروه یافت شد. همچنین کاهش معنی­داری را در غلظت تستوسترون خون در حیوانات تحت درمان با آکریلامید مشاهده شد. نتیجه­گیری: بر طبق نتایج مطالعه ما اکریلامید می­تواند بر روی پارامترهای اسپرم و کیفیت DNA اسپرم تأثیر بگذارد که این ناهنجاری­ها ممکن است به علت کاهش سطح تستوسترون درموش­های تحت درمان با اکریلامید باشد. 2 Background: Acrylamide (AA) is an important industrial chemical primarily. AA is also found in carbohydrate-rich foods that are prepared at high temperatures, such as French fries and potato chips. It is demonstrated that AA is a carcinogen and reproductive toxin and has ability to induce sperm damage. Objective: The aim of this study was to observe the effects of AA on sperm parameters and evaluation of sperm chromatin quality and testosterone hormone in mice. Materials and Methods: Totally, 16 adult male mice were divided into two groups. Mice of group A fed on basal diet; group B received basal diet and AA (10 mg/kg, water solution) for 35 days. The right cauda epididymis was incised and then placed in Ham’s F10 culture media at 37oC for 15 min. Released spermatozoa were used to analyze count, motility, morphology and viability. To determine the sperm DNA integrity and chromatin condensation, the cytochemical techniques including Aniline blue, Acridine orange and Chromomycin A3 staining were used. Results: AA-treated mice had poor parameters in comparison with control animals. In sperm chromatin assessments, except TB (p=0.16), significant differences were found in all of the tests between two groups. It was also seen a significant decrease in concentration of blood testosterone in AA-treated animals when compared to controls (p<0.001). Conclusion: According to our results, AA can affect sperm parameters as well as sperm chromatin condensation and DNA integrity in mice. These abnormalities may be related to the reduction in blood testosterone. 335 0 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/42018/02/4 1396/11/15 مجید پورانتظاری Majid Pourentezari Department of Biology and Anatomy, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران علیرضا طالبی Alireza Talebi Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران ابوالقاسم عباسی Abulghasem Abbasi Department of Biology and Anatomy, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران محمدعلی خلیلی Mohammad Ali Khalili Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران عصمت منگلی Esmat Mangoli Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران مرتضی انوری Morteza Anvari Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran ایران moanvari@yahoo.com Mice Acrylamide Sperm Chromatin Testosterone موش اکریلامید کروماتین اسپرم پارامترهای اسپرم تستوسترون Friedman M. Chemistry, biochemistry, and safety of acrylamide. A review. J Agric Food Chem 2003; 51: 4504-4526.##Tareke E, Rydberg P, Karlsson P, Eriksson S, Törnqvist M. Analysis of acrylamide, a carcinogen formed in heated foodstuffs. J Agric Food Chem 2002; 50: 4998-5006.##World Health Organization. FAO/WHO Consultations on the health implications of acrylamide in food. Summary report of a meeting held in Geneva. Geneva; 2002.##LoPachin RM, Balaban CD, Ross JF. Acrylamide axonopathy revisited. Toxicol Appl Pharmacol 2003; 188: 135-153.##Ghanayem BI, McDaniel LP, Churchwell MI, Twaddle NC, Snyder R, Fennell TR, et al. Role of CYP2E1 in the epoxidation of acrylamide to glycidamide and formation of DNA and hemoglobin adducts. Toxicol Sci 2005; 88: 311-318.##Ghanayem BI, Witt KL, El-Hadri L, Hoffler U, Kissling GE, Shelby MD, et al. Comparison of germ cell mutagenicity in male CYP2E1-null and wild-type mice treated with acrylamide: evidence supporting a glycidamide-mediated effect. Biol Reprod 2005; 72: 157-163.##Ghanayem BI, Witt KL, Kissling GE, Tice RR, Recio L. Absence of acrylamideinducedgenotoxicity in CYP2E1-null mice: evidence consistent with a glycidamide-mediated effect. Mutat Res 2005; 578: 284-297.##IARC. Acrylamide. IARC Monogr Eval Carcinog Risks Hum. Institute of Laboratory Animal Resources (1996) Guide for the Care and Use of Laboratory Animals 7th Ed. Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council, Washington DC; 1994: 389-4334.##Sumner SC, MacNeela JP, Fennell TR. Characterization and quantitation of urinary metabolites of [1, 2, 3-13C] acrylamide in rats and mice using 13C nuclear magnetic resonance spectroscopy. Chem Res Toxicol 1992; 5: 81-89.##Manjanatha MG, Aidoo A, Shelton SD, Bishop ME, McDaniel LP, Lyn-Cook LE, et al. Genotoxicity of acrylamide and its metabolite glycidamide administered in drinking water to male and female Big Blue mice. Environ Mol Mutagen 2006; 47: 6-17.##Shelby MD, Cain KT, Cornett CV, Generoso WM. Acrylamide: induction of heritable translocation in male mice. Environ Mutagen 1987; 9: 363-368.##Dobrzyn´ska MM, Gajewski AK. Induction of micronuclei in bone marrow and sperm head abnormalities after combined exposure of mice to low doses of X-rays and acrylamide. Teratogen Carcinogen Mutagen 2000; 20: 133-140.##https://doi.org/10.1002/(SICI)1520-6866(2000)20:3<133::AID-TCM4>3.0.CO;2-B##Collins, BW, Howard, DR, Allen JW. Kinetochore-staining of spermatid micronuclei: studies of mice treated with X-radiation or acrylamide. Mutat Res 1992; 281: 287-294.##Tyl RW, Friedman MA. Effects of acrylamide on rodent reproductive performance. Reprod Toxicol 2003; 17: 1-13.##Tyl RW, Marr MC, Myers CB, Ross WP, Friedman M. A. Relationship between acrylamide reproductive and neurotoxicity in male rats. Reprod Toxicol 2000; 14: 147-157.##Adler ID, Baumgartner A, Gonda H, Friedman MA, Skerhut M. 1-Aminobenzotriazole inhibits acrylamide-induced dominant lethal effects in spermatids of male mice. Mutagenesis 2000; 15: 133-136.##Talebi AR. Sperm nuclear maturation. 1thEd. New York, Nova Science Publishers Inc.; 2011: 45-55.##Talebi AR, Vahidi S, Aflatoonian A, Ghasemi N, Ghasemzadeh J, Firoozabadi RD, et al. Cytochemical evaluation of sperm chromatin and DNA integrity in couples with unexplained recurrent spontaneous abortions. J Androl 2012; 44: 462-470.##Talebi AR, Khalili MA, Nahangi H, Abbasi AG, Anvari M. Evaluation of epididymal necrospermia following experimental chronic spinal cord injury in rat. Iran J Reprod Med 2007; 5: 171-176.##Carrell DT, Emery BR, Hammoud S. Altered protamine expression and diminished spermatogenesis: What is link? Hum Reprod 2007; 13: 313-327.##Kermani-Alghoraishi M, Anvari M, Talebi AR, Amini-Rad O, Ghahramani R, Miresmaili SM. The effects of acrylamide on sperm parameters and membrane integrity of epididymal spermatozoa in mice. Eur J Obstet Gynecol Reprod Biol 2010; 153: 52-55.##Amaral S, Oliveira PJ, Ramalho-Santos J. Diabetes and the impairment of reproductive function: possible role of mitochondria and reactive species. Curr Diabetes Rev 2008; 4: 46-54.##Talebi AR, Moein MR, Tabibnejad N, Ghasemzadeh J. Effect of varicocele on chromatin condensation and DNA integrity of ejaculated spermatozoa using cytochemical tests. Andrologia 2008; 40: 245-251.##Yang HJ, Lee SH, Jin Y, Choi JH, Han CH, Lee MH. Genotoxicity and toxicological effects of acrylamide on reproductive system in male rats. J Vet Sci 2005; 6: 103-109.##Wang H, Huang P, Lie T, Li J, Hutz RJ, Li K, et al. Reproductive toxicity of acrylamide-treated male rats. Reprod Toxicol 2010; 29: 225-230.##Song HX, Wang R, Geng ZM, Cao SX, Liu TZ. [Subchronic exposure to acrylamide affects reproduction and testis endocrine function of rats]. 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Effect of saffron on rat sperm chromatin integrity تأثیر زعفران بر تمامیت ساختار کروماتین اسپرم رت 2 1 مقدمه: ارتباط بین میزان ROS و کیفیت سمن مشخص گردیده است. زعفران نه تنها به عنوان چاشنی غذایی، بلکه به عنوان یک گیاه دارویی از زمان­های گذشته مورد توجه بوده است. هدف: هدف از این مطالعه، بررسی تأثیر زعفران و ویتامین E در حفظ تمامیت کروماتین اسپرم رت بود. مواد و روش­ها: تعداد 30 رت نر بالغ نژاد  Wistarبه طور تصادفی به 3 گروه زعفران، ویتامین E و کنترل تقسیم شدند (در هر گروه 10 رت). گروه زعفران روزانه  mg/kg100زعفران، گروه ویتامین E روزانه mg/kg100 ویتامین E و گروه کنترل روزانه ml 0/5 آب مقطر بصورت گاواژ دریافت نمودند. پس از 60 روز رت­ها کشته شده و بخش دیستال دم اپی­دیدیم آن­ها جدا شد و از اسپرم بدست آمده جهت بررسی تراکم کروماتین رنگ آمیزی کرومومایسین A3 (CMA3)، همچنین بررسی حساسیت DNA به دناتوراسیون  توسط اسید توسط رنگ آمیزی آکریدین اورانژ (AO) استفاده شد. نتایج: بررسی­ها نشان داد که میانگین درصد اسپرم­های CMA3+ در گروه کنترل 0/36 به صورت معناداری به­ترتیب نسبت به گروه­های زعفران و ویتامین E کاهش داشت. بررسی­ها نشان داد که میانگین درصد اسپرم­های CMA3 مثبت به صورت معناداری در گروه­های زعفران و ویتامین E نسبت به گروه کنترل کاهش معناداری دارد (0/001>p ). نتیجه­گیری: این نتایج نشان داد که زعفران و ویتامین E می­توانند اسپرم را در برابر آسیب­های DNA و اختلالات کروماتین محافظت نمایند. 2 Background: Currently, relation between reactive oxygen species (ROS) ROS concentration and semen quality was indicated. Saffron has traditionally been not only considered as a food additive but also as a medicinal herb, which has a good antioxidant properties. Objective: The aim of this study was to evaluate the protection potency of saffron and vitamin E on sperm chromatin integrity. Materials and Methods: Thirty adult male Wistar rats divided equally into saffron (100 mg/kg), vitamin E (100 mg/kg) and control (0.5cc distilled water /day) groups. After 60 days, cauda epididymis dissected and sperm cells were used for analysis of sperm chromatin packaging by chromomycin A3 (CMA3) staining, and sperm chromatin susceptibility to acid denaturation by acridine orange (AO) staining. Results: The mean percentage of CMA3 positive sperm was significantly decreased in saffron and vitamin E groups relative to control group (p<0.001). Moreover, the AO staining results showed that the mean percentage of sperm with DNA damage was significantly decreased in saffron and vitamin E groups as compared with control group (p<0.001). Conclusion: Our results purposed that saffron can protect sperm against DNA damage and chromatin anomalies. 343 0 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/4 1396/11/15 محمد مردانی Mohammad Mardani Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran ایران احمد واعظ Ahmad Vaez Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran ایران شهناز رضوی Shahnaz Razavi Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran ایران razavi@med.mui.ac.ir Antioxidants Saffron Vitamin E DNA damage Sperm chromatin. آنتی اکسیدان زعفران ویتامین E آسیب DNA کروماتین اسپرم Dohle GR, Colpi GM, Hargreave TB, Papp GK, Jungwirth A, Weidner W, et al. EAU guidelines on male infertility. 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Behav Brain Res 2011; 219: 197-204.##Mansoori P, Akhondzadeh S, Raisi F, Ghaeli P, Jamshidi AH, Nasehi AA, et al. A randomized, double-blind, placebo-controlled study of safety of the adjunctive saffron on sexual dysfunction induced by a selective serotonin reuptake inhibitor. J Med Plants 2011; 10: 121-130.##Assimopoulou AN, Sinakos Z, Papageorgiou VP. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytother Res 2005; 19: 997-1000.##Fernández JA, Pandalai S. Biology, biotechnology and biomedicine of saffron. Recent Res Dev Plant Sci 2004; 2: 127-159.##Premkumar K, Thirunavukkarasu C, Abraham S, Santhiya S, Ramesh A. Protective effect of saffron (Crocus sativus L.) aqueous extract against genetic damage induced by anti-tumor agents in mice. Hum Experiment Toxicol 2006; 25: 79-84.##Heidary M, Vahhabi S, Reza Nejadi J, Delfan B, Birjandi M, Kaviani H, et al. Effect of saffron on semen parameters of infertile men. Urology J 2008; 5: 255-259.##Khayatnouri M, Safavi SE, Safarmashaei S, Babazadeh D, Mikailpourardabili B. The effect of Saffron Orally Administration on Spermatogenesis Index in Rat. Adv Environ Biol 2011; 5: 1514-1521.##El-Demerdash FM, Yousef MI, Kedwany FS, Baghdadi HH. Cadmium-induced changes in lipid peroxidation, blood hematology, biochemical parameters and semen quality of male rats: protective role of vitamin E and b-carotene. Food Chem Toxicol 2004; 42: 1563-1571.##World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. Cambridge university press, Cambridge; 1992.##Razavi S, Nasr Esfahani MH, Mardani M, Mafi A, Moghdam A. Effect of human sperm chromatin anomalies on fertilization outcome post ICSI. Andrologia 2003; 35: 238-243.##Lolis D, Georgiou I, Syrrou M, Zikopoulos K, Konstantelli M, Messinis I. Chromomycin A3-staining as an indicator of protamine deficiency and fertilization. Int J Androl 1996; 19: 23-27.##Nasr-Esfahani MH, Razavi S, Mardani M. Relation between different human sperm nuclear maturity tests and in vitro fertilization. J Assist Reprod Genet 2001; 18: 219-225.##Manicardi GC, Bianchi PG, Pantano S, Azzoni P, Bizzaro D, Bianchi U, et al. Presence of endogenous nicks in DNA of ejaculated human spermatozoa and its relationship to chromomycin A3 accessibility. Biol Reprod 1995; 52: 864-867.##Hoshi K, Katayose H, Yanagida K, Kimura Y, Sato A. The relationship between acridine orange fluorescence of sperm nuclei and the fertilizing ability of human sperm. Fertil Steril 1996; 66: 634-639.##Razavi SH, Nasr-Esfahani MH, Deemeh MR, Shayesteh M, Tavalaee M. Evaluation of zeta and HA‐binding methods for selection of spermatozoa with normal morphology, protamine content and DNA integrity. Andrologia 2010; 42: 13-19.##Kemal Duru N, Morshedi M, Oehninger S. Effects of hydrogen peroxide on DNA and plasma membrane integrity of human spermatozoa. Fertil Steril 2000; 74: 1200-1207.##Aitken RJ, Krausz C. Oxidative stress, DNA damage and the Y chromosome. Reproduction 2001; 122: 497-506.##Suleiman SA, Ali ME, Zaki ZM, el-Malik EM, Nasr MA. Lipid peroxidation and human sperm motility: protective role of vitamin E. J Androl 1996; 17: 530.##Kumar V, Bhat ZA, Kumar D, Khan NA, Chashoo IA, Shah MY. Pharmacological profile of crocus sativus-a comprehe sive review. Pharmacol Online 2011; 3: 799-811.##Hosseinzadeh H, Ziaee T, Sadeghi A. The effect of saffron, Crocus sativus stigma, extract and its constituents, safranal and crocin on sexual behaviors in normal male rats. Phytomedicine 2008; 15: 491-495.##Dominguez-Rebolledo AE, Fernandez-Santos MR, Bisbal A, Ros-Santaella JL, Ramon M, Carmona M, et al. Improving the effect of incubation and oxidative stress on thawed spermatozoa from red deer by using different antioxidant treatments. Reprod Fertil Dev 2010; 22: 856-870.##Safarinejad MR, Shafiei N, Safarinejad S. A prospective double-blind randomized placebo- controlled study of the effect of saffron (Crocus sativus Linn.) on semen parameters and seminal plasma antioxidant capacity in infertile men with idiopathic oligoasthenoteratozoospermia. Phytother Res 2011; 25: 508-516.##Hosseinzadeh H, Abootorabi A, Sadeghnia HR. Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate-Induced DNA Damage in Mice Organs. DNA Cell Biol 2008; 27: 657-664.##Uzunhisarcikli M, Kalender Y, Dirican K, Kalender S, Ogutcu A, Buyukkomurcu F. Acute, subacute and subchronic administration of methyl parathion-induced testicular damage in male rats and protective role of vitamins C and E. Pesticide Biochem Physiol 2007; 87: 115-122.##Keskes-Ammar L, Feki-Chakroun N, Rebai T, Sahnoun Z, Ghozzi H, Hammami S, et al. Sperm oxidative stress and the effect of an oral vitamin E and selenium supplement on semen quality in infertile men. 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Improvement in sperm DNA quality using an oral antioxidant therapy. Reprod Biomed Online 2009; 18: 761-768.##Menezo YJ, Hazout A, Panteix G, Robert F, Rollet J, Cohen-Bacrie P, et al. Antioxidants to reduce sperm DNA fragmentation: an unexpected adverse effect. Reprod Biomed Online 2007; 14: 418-421.##Piomboni P, Gambera L, Serafini F, Campanella G, Morgante G, De Leo V. Sperm quality improvement after natural anti-oxidant treatment of asthenoteratospermic men with leukocytospermia. Asian J Androl 2008; 10: 201-206.## ##

A newborn with ambiguous genitalia and a complex X;Y rearrangement نوزادی با ژنیتالیای مبهم و جابجایی کروموزومی X;Y پیچیده 2 1 مقدمه: در اغلب پستانداران تعیین جنسیت در ابتدای بارداری با توجه به وضعیت کروموزوم­های جنسی، XY در جنس مذکر و XX در جنس مؤنث صورت   می­گیرد. مورد: در اینجا نمونه جالبی با ژنیتالیای مبهم و بافت ترکیبی تخمدان و بیضه با کاریو تایپ مونث گزارش می­گردد. ریز آرایه ژنومی مقایسه­ای یک باز­آرایی ناپایدار در نتیجه حذف در ناحیه انتهایی بازوی کوتاه کروموزوم X و مضاعف شدگی قسمت پروگزیمال ناحیه بهم پیوسته و وجود کروموزم Y ازY11-Ypte. دورگه سازی فلوروسانت درجا نشان داد که این قسمت از کروموزوم Y باقسمت انتهایی بازوی کوتاه کروموزوم X غیر طبیعی جابجا شده و قسمت مضاعف شده کروموزوم X واژگون شده است. با همه این موارد کروموزوم X غیر طبیعی دارای دو سانترومر بوده که سانترومر مربوط به کروموزوم Y غیر فعال گردیده است. نتیجه­گیری: وجود کروموزوم Y همراه با ژن SRY در این جابجایی می­تواند توجیه کننده تکامل بیضه باشد هرچند دلیل ابهام در دستگاه تناسلی چندان روشن نمی­باشد. 2 Background: In most mammals, sex is determined at the beginning of gestation by the constitution of the sex chromosomes, XY in males and XX in females. Case: Here we report an interesting case characterized by ambiguous genitalia and ovotestis in a newborn carrying an apparently female karyotype (46 XX). Array Comparative Genomic Hybridization (Array-CGH) revealed an unbalanced rearrangement resulting in the deletion of the distal Xp and the duplication of the proximal Xp contiguous region with presence of the Y chromosome from Ypter to Yq11. Fluorescent in situ hybridization (FISH) showed that this portion of the Y was translocated to the tip of the abnormal X and that the duplicated portion of chromosome X was inverted. Altogether, the abnormal chromosome was a dicentric one with the centromere of the Y chromosome apparently inactivated. Conclusion: The presence within the translocated Y chromosome of the SRY gene explains the devolopment of testes although it is not clear the reason for the genitalia ambiguity. 351 0 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/5 1396/11/16 Mohammadreza Dehghani Mohammadreza Dehghani Department of Molecular Medicine, University of Pavia, Pavia, Italy Elena Rossi Elena Rossi Department of Molecular Medicine, University of Pavia, Pavia, Italy Annalisa Vetro Annalisa Vetro Biotechnology Research Laboratories, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy Gianni Russo Gianni Russo Department of Pediatrics, Endocrine Unit, University Vita-Salute, San Raffaele Hospital, Italy Zahra Hashemian Zahra Hashemian Shahid Sadoughi Hospital, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Orsetta Zuffardi Orsetta Zuffardi Department of Molecular Medicine, University of Pavia, Pavia, Italy zuffardi@unipv.it Ambiguous genitalia 46 XX testicular DSD Inverted duplication and Xp terminal Deletion(Invdup del) Rearrangement Array Comparative Genomic Hybridization FISH. ژنیتالیای مبهم Invdup del بازآرایی ناپایدار ریزآرایه ژنومیک مقایسه ای هیبریدیزاسیون فلوروسنت درجا. Eggers S, Sinclair A. Mammalian sex determination- insights from humans and mice. Chromosome Res 2012; 20: 215-238.##Blakless M CA. How Sexually Dimorphic Are We? Review and Synthesis. Am J Hum Biol 2000; 12: 151-166.##https://doi.org/10.1002/(SICI)1520-6300(200003/04)12:2<151::AID-AJHB1>3.0.CO;2-F##Kim Y CB. Balancing the bipotential gonad between alternative organ fates: a new perspective on an old problem. Dev Dyn 2006; 235: 2292-2300.##Achermann JCH IA. Williams Textbook of Endocrinology. Saunders, Philadelphia; 2011: 869-934.##Rey RAGRP. Normal male sexual differentiation and aetiology of disorders of sex development. Best Pract Res Clin Endocrinol Metab 2011; 25: 221-238.##Achermann JC, Hughes IA. Disorders of Sex Development. Williams Textbook of Endocrinology. 12th Ed. Saunders, Philadelphia; 2011.##Temel SG ea. Extended pedigree with multiple cases of XX sex reversal in the absence of SRY and of a mutation at the SOX9 locus. Sex Dev 2007; 1: 24-34.##Sutton EHJ. Identification of SOX3 as an XX male sex reversal gene in mice and humans. J Clin Invest 2011; 121: 328-341.##Hughes IA HC, Ahmed SF, Lee PA Consensus statement on management of intersex disorders. J Pediatr Urol 2006; 2: 148-162.##Abusheikha N, Lass A, Brinsden P. XX males without SRY gene and with infertility. Hum Reprod 2001; 16: 717-718.##Harley MOaVR. Disorders of sex development: new genes, new concepts. Nat Gene 2013; 9: 79- 91.##Zenteno JC, Lopez M, Vera C, Mendez JP, Kofman-Alfaro S. Two SRY-negative XX male brothers without genital ambiguity. Hum Genet 1997; 100: 606-610.##de la Chapelle A. The Y-chromosomal and autosomal testis-determining genes. Development 1987; 101 (Suppl.): 33-38.##Faith Y, Dorsey MHH, David R. 46,xx SRY negative as a true hermapherodism. Urology 2009; 73: 529-531.##Parma P, Radi O, Vidal V, Chaboissier MC, Dellambra E, Valentini S, et al. R-spondin1 is essential in sex determination, skin differentiation and malignancy. Nat Genet 2006; 38: 1304-1309.##Queipo G, Zenteno JC, Pena R, Nieto K, Radillo A, Dorantes LM, et al. Molecular analysis in true hermaphroditism: demonstration of low-level hidden mosaicism for Y-derived sequences in 46,XX cases. Hum Genet 2002; 111: 278-283.##Zenteno-Ruiz JC, Kofman-Alfaro S, Mendez JP. 46,XX sex reversal. Arch Med Res 2001; 32: 559-566.##de la Chapelle A. Analytic review: nature and origin of males with XX sex chromosomes. Am J Hum Genet 1972; 24: 71-105.##Ferguson-Smith MA, Cooke A, Affara NA, Boyd E, Tolmie JL. Genotype-phenotype correlations in XX males and their bearing on current theories of sex determination. Hum Genet 1990; 84: 198-202.##Boucekkine C, Toublanc JE, Abbas N, Chaabouni S, Ouahid S, Semrouni M, et al. Clinical and anatomical spectrum in XX sex reversed patients. Relationship to the presence of Y specific DNA-sequences. Clin Endocrinol 1994; 40: 733-742.##Ergun-Longmire B, Vinci G, Alonso L, Matthew S, Tansil S, Lin-Su K, et al. Clinical, hormonal and cytogenetic evaluation of 46,XX males and review of the literature. J Pediatric Endocrinol Metab 2005; 18: 739-748.##State-specific rates of mental retardation--United States, 1993. MMWR Morbidity and mortality weekly report 1996; 45: 61-65.##Kusz K, Kotecki M, Wojda A, Szarras-Czapnik M, Latos-Bielenska A, Warenik-Szymankiewicz A, et al. Incomplete masculinisation of XX subjects carrying the SRY gene on an inactive X chromosome. J Med Genet 1999; 36: 452-456.##Jobling MA. A selective difference between human Y-chromosomal DNA haplotypes. Current Biol 1998; 8: 4.##Rosser ZH, Balaresque P, Jobling MA. Gene conversion between the X chromosome and the male-specific region of the Y chromosome at a translocation hotspot. Am J Hum Genet 2009; 85: 130-134.##Bonaglia MC, Giorda R, Massagli A, Galluzzi R, Ciccone R, Zuffardi O. A familial inverted duplication/deletion of 2p25.1-25.3 provides new clues on the genesis of inverted duplications. Eur J Hum Genet 2009; 17: 179-186.##Zuffardi O, Bonaglia M, Ciccone R, Giorda R. Inverted duplications deletions: underdiagnosed rearrangements? Clin Genet 2009; 75: 505-513.## ##

Association of heteromorphism of chromosome 9 and recurrent abortion (ultrasound diagnosed blighted ovum): A case report ارتباط هترومورفیسم کروموزوم شماره 9 و سقط های مکرر (ساک حاملگی بدون جنین) 2 1 مقدمه: ناهنجاری­های کروموزومی شایع­ترین علت سقط­ها در سه ماهه اول بارداری می­باشند. در میان کروموزوم­های انسان، کروموزوم شماره 9 دارای بیشترین تغییرات ساختاری است که به آن هترومورفیسم نیز اطلاق می­شود و به نظر می­رسد فاقد اهمیت بالینی بوده و حتی به عنوان واریانت نرمال  نیز در میان        پلی­مورفیسم­های ساختاری در نظر گرفته می­شوند، با این وجود در بعضی مطالعات، ارتباط معنی­داری بین هترومورفیسم کروموزوم شماره 9 و سقط­های تکراری گزارش شده است. مورد: به منظور بررسی ارتباط بین هترومورفیسم کروموزوم شماره 9 و سقط­های مکرر، آنالیز کروموزومی با استفاده از تکنیک G- بندینگ از زوجی با سابقه 4 مورد تشخیص ساک حاملگی بدون جنین (blighted ovum) انجام شد تا مشخص شود که آیا هترومورفیسم ساختاری کروموزومی می­تواند مسئول چنین پیامدهای حاملگی باشد. نتایج بدست آمده از آنالیز کروموزومی، در 30 سلول متافازی هترومورفیسم کروموزوم شماره 9 در هر والد را نشان داد. نتیجه­گیری: اگرچه هترومورفیسم کروموزوم شماره 9 پدیده­ای شایع است که اغلب آن را به عنوان یک واریانت نرمال فاقد علائم بالینی درنظر می­گیرند، اما با توجه به این نکته که محل این توالی پلی­مورف در کروموزوم شماره 9 می­تواند با مداخله در فرایند کراسینگ اور یا جدایی میوزی در سلول تخم، اثرگذار بوده و منجر به ایجاد باروری تواٌم با ناهنجاری کروموزومی شود، در نهایت احتمال بارداری بدون جنین دور از انتظار نخواهد بود. بنابراین در صورت گزارش در زوج­هایی با سابقه سقط­های تکراری (blighted ovum)، بررسی کروموزومی به منظور جستجوی هترومورفیسم کروموزوم شماره 9 قویاً پیشنهاد می­گردد.   2 Background: Chromosomal disorders are the most common cause of first trimester spontaneous abortion. Among the human chromosomes, chromosome no.9 was the most common structural chromosomal variant and it is not thought to be of any functional importance, which often considers as a normal variation in structural polymorphisms, nevertheless there are some studies which claim that there is an association between heteromorphism of chromosome no.9 and some pregnancy complication. Case: To postulate any correlation between chromosome no. 9 heteromorphism and recurrent abortion, chromosomal analysis was performed on the basis of G-banding technique at high resolution for a couple with the history of 4 ultrasound diagnosed blighted ovum and Chromosome constitution appeared with chromosome no.9 heteromorphism in all 30 metaphases screened for both partners (9p11-q13). Conclusion: Observation of reproductive failure in couples with heteromorohic pattern of chromosome no.9 suggests that, although the heteromorphism of chromosome no.9 is not a rare condition which often consider as a normal variation with no evidence of any phenotypic effect of patient, nevertheless it seems as if the location of heteromorphic region maybe interfere with meiotic events like the phenomenon of crossing over or miotic segregation of fertilized egg that eventually lead to the development of fertilized eggs with chromosomal abnormalities leading to the possibility of anemberyonic pregnancy, therefore chromosomal analysis for detecting of chromosome no.9 heteromorphism for couples with the history of ultrasound diagnosed blighted ovum will be strongly suggested. 357 0 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/52018/02/5 1396/11/16 فاطمه باغبانی Fatemeh Baghbani Department of Medical Genetics, Faculty of Medicine, Pardis Campus, Mashhad, Iran ایران سلمه میرزایی Salmeh Mirzaee Department of Medical Genetics, Faculty of Medicine, Pardis Campus, Mashhad, Iran ایران محمد حسن زاده نظرآبادی Mohammad Hassanzadeh-Nazarabadi Department of Medical Genetics, Faculty of Medicine, Pardis Campus, Mashhad, Iran ایران nazarabadim@mums.ac.ir Recurrent abortion Heteromorphism Chromosome no. 9 Pregnancy complication Blighted ovum هترومورفیسم کروموزوم شماره 9 سقط مکرر ساک حاملگی بدون جنین. Sotiriadis A, Makrydimas G, Papatheodorou S, Ioannidis JP. Expectant, medical, or surgical management of first-trimester miscarriage: a meta-analysis. Obstet Gynecol 2005; 105: 1104-1113.##Albayram F, Hamper UM. "First‐Trimester obstetric emergencies: Spectrum of sonographic findings." J Clin Ultrasound 2002; 30: 161-177.##Carrell DT, Liu L, Peterson CM, Jones KP, Hatasaka HH, Erickson L, et al. Sperm DNA fragmentation is increased in couples with unexplained recurrent pregnancy loss. Arch Androl 2003; 49: 49-55.##Azmanov DN, Milachich TV, Zaharieva BM, Michailova GI, Dimitrova VG, Karagiozova ZH, et al. Profile of chromosomal aberrations in different gestational age spontaneous abortions detected by comparative genomic hybridization. Eur J Obstet Gynecol Reprod Biol 2007; 131: 127-131.##Brajenović-Milić B, Petrović O, Krasević M, Ristić S, Kapović M. Chromosomal anomalies in abnormal human pregnancies. Fetal Diagn Ther 1998; 13: 187-191.##Tibiletti MG, Simoni G, Terzoli GL, Romitti L, Fedele L, Candiani GB. Pericentric inversion of chromosome 9 in couples with repeated spontaneous abortion. Acta Eur Fertil 1981; 12: 245-248.##Pandya PP, Snijders RJ, Psara N, Hilbert L, Nicolaides KH. The prevalence of non-viable pregnancy at 10-13 weeks of gestation. Ultrasound Obstet Gynecol 1996; 7: 170-173.##Goddijn M, Leschot NJ. Genetic aspects of miscarriage. Baillieres Best Pract Res Clin Obstet Gynaecol 2000; 14: 855-865.##Franssen MT, Korevaar JC, Leschot NJ, Bossuyt PM, Knegt AC, Gerssen-Schoorl KB, et al. Selective chromosome analysis in couples with two or more miscarriages: case-control study. BMJ 2005; 331: 137-141.##Humphray SJ, Oliver K, Hunt AR, Plumb RW, Loveland JE, Howe KL, et al. DNA sequence and analysis of human chromosome 9. Nature 2004; 429: 369-374.##Babu V Rao, Lily Kerketta, Seema Korgaonkar, Kanjaksha Ghosh. Pericentric inversion of chromosome 9[inv(9)(p12q13)]: Its association with genetic diseases. Indian J Hum Genet 2006; 12: 129-132.##Kosyakova N, Grigorian A, Liehr T, Manvelyan M, Simonyan I, Mkrtchyan H, et al. Heteromorphic variants of chromosome 9. Mol Cytogenet 2013; 6: 14.##Elghezal H, Hidar S, Mougou S, Khairi H, Saâd A. Prevalence of chromosomal abnormalities in couples with recurrent miscarriage. Fertil Steril 2007; 88: 721-723.##Dana M, Stoian V. Association of pericentric inversion of chromosome 9 and infertility in romanian population. Maedica (Buchar) 2012; 7: 25-29.##Madon PF, Athalye AS, Parikh FR. Polymorphic variants on chromosomes probably play a significant role in infertility. Reprod Biomed Online 2005; 11: 726-732.##Gianaroli L, Magli MC, Ferraretti AP, Munné S, Balicchia B, Escudero T, et al. Possible interchromosomal effect in embryos generated by gametes from translocation carriers. Hum Reprod 2002;##17: 3201-3207. 17.Dewald GW, Michels VV. Recurrent miscarriages: cytogenetic causes and genetic counseling of affected families. Clin Obstet Gynecol 1986; 29: 865-885.##Craig-Holmes AP, Moore FB, Shaw MW. Polymporphism of human C-band heterochromatin. II. Family studies with suggestive evidence for somatic crossing over. Am J Hum Genet 1975; 27: 178-189.##Franssen MT, Korevaar JC, Tjoa WM, Leschot NJ, Bossuyt PM, Knegt AC, et al. Inherited unbalanced structural chromosome abnormalities at prenatal chromosome analysis are rarely ascertained through recurrent miscarriage. Prenat Diagn 2008; 28: 408-411.##Farcaş S, Crişan CD, Andreescu N, Stoian M, Motoc AG. Structural chromosomal anomalies detected by prenatal genetic diagnosis: our experience. Rom J Morphol Embryol 2013; 54: 377-383.## ##

Tuboplasty as a reversal macrosurgery for tubal ligation, is pregnancy possible? A case series توبوپلاستی به روش ماکروسرجری بعد از بستن لوله های رحمی، آیا حاملگی امکان پذیر است؟  2 1 مقدمه: بازکردن لوله­های رحمی بعد از بستن آن یکی از درخواست­های زنان به دلایل مختلف می­باشد. هدف این مطالعه تعیین میزان حاملگی بعد از باز کردن لوله­های رحمی است. موارد: این مطالعه به صورت آینده نگر بر روی ١٥ بیمار که خواهان باز کردن لوله­های رحمی بعد از بستن آن­ها بودند، از فروردین ١٣٧٥لغایت فروردین ١٣٩١ انجام گرفت و آناستوموز لوله­های رحمی انجام شد. چهار نفر بعد از آناستوموز لوله­های رحمی باردار شدند (26/6%) که ٢ نفر حاملگی ترم داشتند و ٢ نفر سقط کردند. نتیجه­گیری: میزان موفقیت بارداری بعد از باز کردن و آناستوموز لوله­های رحمی به روش ماکروسرجری خوب است و قبل از انجام لقاح مصنوعی (IVF)     می­توان این روش را در نظر داشت. نوع بستن لوله­های رحمی در موفقیت عمل آناستوموز لوله­ها اثر عمده و مهمی دارد. 2 Background: Reversal of tubal ligation is requested by some women for various reasons. The present study aims to determine the rate of pregnancy after tubal ligation reversal. Case: In these case series, we reported fifteen women who requested tuboplasty after tubal ligation. In these cases reversal of tubal ligation was done in 15 women. Pregnancy occurred in 4 women (26.6%). Two term pregnancy and 2 abortions were determined. Conclusion: The success rate of pregnancy after macro surgical reversal of tubal ligation is good and can be considered before in vitro fertilization. The type of tubal ligation and the procedure used will determine the best procedure for reversal and have a major impact on chance of success for reversal surgery. 361 0 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/52018/02/52018/02/5 1396/11/16 فخرالملوک یاسایی Fakhrolmolouk Yassaee Department of Obstetrics and Gynecology, Genomic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran ایران dr_fyass@yahoo.com Tubal ligation Reversal macro surgery Rate of pregnancy Fallopian tube reanastomoses. بستن لوله ها برگشت پذیری لوله ها میزان بارداری آناستوموز لوله های فالوپ. Boeckxtaens A, Devroey P, Collins J, Tourneys H. Getting pregnant after tubal sterilization: surgical reversal or IVF? Hum Reprod 2007; 22: 2660-2664.##Promila J, Bhupinder Kaur G, Shweta G. Reversal of tubal ligation under 4x magnification. J Obstet Gynecol India 2005; 55: 448-450.##Narvekar AN. Reversal of sterilization using microsurgical techniques. J Obstet Gynecol India 1988; 38: 211-213.##Marshall S, O'Brien BD, Jones K: Women Health Tubal ligation reversal. Health Wise Staff. Updated 2012. Available at: http://www.healthlinkbc.ca.##John A Rock, Howard W, Jones III. Reconstructive tubal surgery. Victor Gomel and Elizabeth Taylor. Telind`s Operative Gynecology. 10th Ed. 2008.##Sanger K, Rana Ak, Sharma S. Tuboplasty or recanalization: nursing care case study. Nursing J India 1991; 82: 327-328.##Timonen S, Tuominen J, Irjala K, Mäenpää J. Ovarian function and regulation of the hypothalamic-pituitary-ovarian axis after tubal sterilization. J Reprod Med 2002; 47: 131-136.##Trussell J, Guibert E, Hedley A. Sterilization failures, sterilization reversal and pregnancy after sterilization reversal of Quebec. Obstet Gynecol 2003; 101: 677-684.##Brar MK, Kaur J, Kaur S. A study of microsurgical reanastomosis of the fallopian tubes for reversal of sterilization. J Obstet Gynecol India 2000; 50: 75-78.##Singhal V, Li TC, Cooke ID. An analysis of factors in fluencing the outcome of 232 consecutive tubal microsurgery cases. Br J Obstet Gynecol 1991; 98: 628-636.##Silber SJ, Cohen R. Micro surgical reversal of tubal sterilization: factors affecting pregnancy rate, with long-term-follow up. Obstet Gynecol 1984; 64: 679-682.##Spivak MM, Librach CL, Rosenthal DM. Microsurgical reversal of sterilization: a six year study. Am J Obstet Gynecol 1986; 154: 355-361.##Putman JM, Holden AEC, Olive DL. Pregnancy rates following tubal anastomosis: Pomeroy partial salpingectomy versus electrocautery. J Gynecol Surg 1990; 6: 173-178.##Paterson PJ. Factors influencing the success of microsurgical tuboplasty for sterilization. Clin Reprod Fertil 1985; 3: 57-64.##Mc Comb P. Microsurgical tubocornual anastomosis for occlusive corneal disease: reproducible results without the need for tubouterine implantation. Fertil Steril 1986; 46: 571.## ##

Protective effects of celery (Apium Graveolens) on testis and cauda epididymal spermatozoa in rat 2 2 Infertility is one of the health problems that will have a negative impact on the individual, social and economic and is seen in 10-15% of couples (1). About 40 % of the issues involved with infertility are due to the man. Male sperm cells count nowadays has decreased dramatically in comparison with those who lived 50 years ago (2). Causes of infertility in men are included: oligozoospermia, immaturity of sperm, sperm deformity, and sperm non-motility. Spermatogenesis takes place within the testes under control of testosterone secreted by the testes and secretory activity of the testes controlled by the hypothalamic-pituitary-testicle axis. Due to adverse effects and side effects of chemical drugs today, the use of traditional medicine, especially herbal therapy is taken into consideration. In traditional medicine, it has been pointed to therapeutic properties of celery. Celery has anti-fungal, anti-bacterial, and anti-cancer properties (3). Also this plant is an appetite stimulant and sexual booster (4). Previous studies have shown that sperm cells are largely vulnerable to oxidative stress but celery is rich in antioxidant compounds such as flavonoids (apiin and apigenin), vitamins E and C that can reduce oxidative stress (5, 6). So, in the present study the protective effect of celery was investigated on the cauda epididymal spermatozoa and testis in rat. A total number of 32 male Wistar rats (weighting 170-220 g) were prepared from animal house central of Ahvaz Jundishapur University of Medical Sciences. Animals were maintained in plastic cages with 12/12 h light/dark cycle at 21±2oC. All experimental animals were carried out in accordance with Ahvaz University Ethical Committee. Hydro-alcoholic extract of celery was prepared by maceration method. The rats were divided into four groups of 8 animals each: control, did not receive anything; vehicle, received propylene glycol; experimental groups, and received hydro-alcoholic extract of celery with doses of 100 and 200 (mg/kg) with solvent of propylene glycol by gavage once every 48 hours for twenty days. At the end of 20th day, rats were scarified under ketamine and xylazine anesthesia then the epididymis and testes were carefully separated. The epididymis was used for sperm count and testes were prepared for morphometric and histologic evaluation. Statistical significance of differences were assessed with one-way ANOVA by SPSS for windows (version 15) followed by LSD test. P<0.05 was assumed as statistically significant. Results of morphometric studies indicated a decrease in number of primary spermatocytes, Sertoli cells and sperm as well as an increase of lumen diameter of seminiferous tubules in vehicle group when compared to the control (p<0.05), but there was not different between the experimental groups and control (p>0.05). Evaluation of tissue sections showed that germinal epithelium in the control group was normal and tissue damage was not observed in epithelial tissue. However, in the vehicle group, epithelium was destroyed and arrangement of epithelial cells was disordered, and fluid aggregation is seen into the epithelial cell and also, reduction of epithelium thickness was observed. In the experimental group (100 mg/kg), there was arrangement in germinal epithelium cells but fluid aggregation was observed into the epithelial cells. A reduced epithelial thickness was seen only in some tubules. However, all these histological changes were less than the vehicle group. In the experimental group (200 mg/kg), tissue destruction was largely improved, and there was an arrangement of the epithelial cells, there was not fluid aggregation into the epithelium, and the thickness of the epithelium was returned almost too normal state. The results of the present study showed that hydro-alcoholic extract of celery improved the destructive effects of propylene glycol on the testes and sexual cells. These findings are similar to previous studied (7, 8). Previous studies have demonstrated that excessive alcohol consumption in men can cause a deficiency in testosterone production and testicular atrophy. Testicular atrophy results primarily from the loss of spermatogenic cells of the seminiferous tubules that this can be caused by oxidative stress generated by alcohol (9). Researches also indicated alcohol with involvement of phase system and activation of caspases induced apoptosis in testicular cells (10). Spermatogenesis and maturation of sexual cells depends on protection of cytotoxic and pathologic lesions that threatens these events. Free radicals due to a strong desire to get electrons induce damage to molecules such as fatty acid of biological membranes and its oxidation. Celery is a strong antioxidant due to flavonoids such as apiein and apigenin (5, 6). Antioxidant compounds are able to protect cell membranes against damage. Antioxidants directly or indirectly impact on hypothalamic-pituitary-testicular axis thus increase sperm count and fertility (5, 6). So celery can be considered as a medicinal herb for infertility. However, further clinical studies are recommended. The study in the form of a research plan was approved with no 91s8 of Research Deputy of Ahvaz Jundishapur University of Medical Sciences. Finally, we acknowledge deputy vice-chancellor for research affairs of AJUMS for financial support, and particularly Research Consultation Center (RCC) for technical support. 365 0 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/42018/02/52018/02/52018/02/52018/02/5 1396/11/16 Wesam Kooti Wesam Kooti Department of Laboratory Sciences, School of Paramedicine, Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Esrafil Mansori Esrafil Mansori Department of Anatomical Sciences, Cellular and Molecular Research Center, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran esrafilmansori@yahoo.com Maryam Ghasemiboroon Maryam Ghasemiboroon Department of Public Health, School of Health, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Mahmoud Harizi Mahmoud Harizi Department of Laboratory Sciences, School of Paramedicine, Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Ashraf Amirzarga Ashraf Amirzarga Department of Physiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Ikechebula JI, Adinma JI, Orie EF, Ikegwuonu SO. High prevalence of male infertility in southeastern. J Obstet Gynecol 2003; 23: 657-659.##Fisch H. Declining worldwide sperm count: disproviding a myth. Urol Clin North Am 2008; 35: 137-146.##Zidorn C, Johrer K, Ganzera M, Schubert B, Sigmund EM, Mader J, et al. Polyacetylenes from the apiaceae vegetables carrot, celery, fennel, parsley, and parsnip and their cytotoxic activities. J Agric Food chem 2005; 53: 2518-2523.##Khosravi M. Medicinal Herb. 3rd Ed. Tehran, Mohammed Publicathion; 2006.##Fazal SS, Ansari MM, Singla RK, Khan S. Isolation of 3-n-Butyl Phthalide and Sedanenolide from Apium graveolens Linn. IGJPS 2012; 2: 258-261.##Fazal SS, Singla RK. Review on the Pharmacognostical and Pharmacological Characterization of Apium Graveolens Linn. IGJPS 2012; 2: 36-42.##Hamza AA, Amin A. Apium graveolens modulates sodium valproate-induced reproductive toxicity in rats. J Exp Zool Ecol Genet Physiol 2007; 307: 199-206.##Madkour NK. The beneficial role of celery oil in lowering of di (2-ethylhexyl) phthalate-induced Testicular damage. Toxicol Ind Health 2012; 19: 1-12.##Albadri CT, Al Ani IM, Hiba HMA. Alcohol consumption and its effect on testicular structure and on sperm count and motility in parent mice and their offspring. IMJM 2013; 12: 43-48.##Eid NA, Shibata MA, Ito Y, Kusakabe K, Hammad H, Otsuki Y. Involvement of Fas system and active caspases in apoptotic signalling in testicular germ cells of ethanol-treated rats. Int J Androl 2002; 25: 159-167.##Ikechebula JI, Adinma JI, Orie EF, Ikegwuonu SO. High prevalence of male infertility in southeastern. J Obstet Gynecol 2003; 23: 657-659.##Fisch H. Declining worldwide sperm count: disproviding a myth. Urol Clin North Am 2008; 35: 137-146.##Zidorn C, Johrer K, Ganzera M, Schubert B, Sigmund EM, Mader J, et al. Polyacetylenes from the apiaceae vegetables carrot, celery, fennel, parsley, and parsnip and their cytotoxic activities. J Agric Food chem 2005; 53: 2518-2523.##Khosravi M. Medicinal Herb. 3rd Ed. Tehran, Mohammed Publicathion; 2006.##Fazal SS, Ansari MM, Singla RK, Khan S. Isolation of 3-n-Butyl Phthalide and Sedanenolide from Apium graveolens Linn. IGJPS 2012; 2: 258-261.##Fazal SS, Singla RK. Review on the Pharmacognostical and Pharmacological Characterization of Apium Graveolens Linn. IGJPS 2012; 2: 36-42.##Hamza AA, Amin A. Apium graveolens modulates sodium valproate-induced reproductive toxicity in rats. J Exp Zool Ecol Genet Physiol 2007; 307: 199-206.##Madkour NK. The beneficial role of celery oil in lowering of di (2-ethylhexyl) phthalate-induced Testicular damage. Toxicol Ind Health 2012; 19: 1-12.##Albadri CT, Al Ani IM, Hiba HMA. Alcohol consumption and its effect on testicular structure and on sperm count and motility in parent mice and their offspring. IMJM 2013; 12: 43-48.##Eid NA, Shibata MA, Ito Y, Kusakabe K, Hammad H, Otsuki Y. Involvement of Fas system and active caspases in apoptotic signalling in testicular germ cells of ethanol-treated rats. Int J Androl 2002; 25: 159-167.## ##