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The State of Semen Analysis over Time 2 1 564 2 Contrary to public opinion on the causes of infertility, male factor accounts for over 50% of infertility during reproductive age of couples and even there are several reports on its increase during the recent years. Semen analysis as one of the main laboratory tests provides valuable information about male fertility status. In spite of huge development in diagnostic tests and laboratory innovation, this test has never lost its place and importance during the last decades and many researchers focused on its standardization and accuracy. Furthermore, World Health Organization (WHO) has emphasized its importance and published the book entitled "WHO Laboratory Manual for the Examination of Human Semen and Semen-Cervical Mucus Interaction" for standard methods in semen analysis. This book is upgraded periodically and its fifth edition was published in 2010. Although there is consensus on the importance and position of semen analysis in diagnosis of male infertility, numerous reports have challenged the practice due to its lack of compliance with spermogram results. In fact, in less than 10% of men with normal semen parameters, there is no ability to conceive naturally. On the other hand, equally the same number of men with abnormal semen parameters are quite fertile and have no problem to have a child through natural pregnancy. The low sensitivity and specificity of semen analysis results led to a new direction for studies and  researches on cellular and molecular aspects of human and mammalian spermatozoa to find  a more sensitive and specific biomarker  of sperm quality during the last decades. The outcome of these attempts was the introduction of several sperm function tests including  zona-free hamster oocyte penetration assay, sperm zona pellucida binding assay, acrosome reaction, sperm migration assay, computer aided sperm analysis (CASA), hyaluronon binding assay (HBA), sperm chromatin integrity and maturity (1).<br>In spite of the benefits and efficient performance of above assays in comparison with semen analysis, the low sensitivity and specificity of the first four assays and the need for salt-stored zona pellucida, hamester oocytes or human donated eggs led to their decreased use in sperm evaluation over time. Therefore, the older tests were gradually replaced by more accurate tests such as sperm nucleus and chromatin evaluation. Sperm DNA and chromatin evaluation has attracted more attention during the recent decade due to its high correlation with sperm quality, fertilizing ability, embryo quality and pregnancy outcome. Therefore, along with increased clinical research on the role of sperm chromatin, a broad range of researches were focused on innovating new methods and equipments for assessment of sperm chromatin quality. Currently, different types of cyotchemical tests such as aniline blue, toluiden blue, chromomycine A3 and acridine orang stainings, sperm chromatin dispersion (SCD),  sperm chromatin structure assay (SCSA), comet assay and TUNEL assay are available for sperm DNA and chromatin evaluation and each of them has its own advantages and disadvantages (2). <br>Although more sensitive and accurate tests are available for sperm nucleus evaluation, the main problem of these methods is their damage to the sperm during evaluation, so the evaluated normal sperm cannot be used in IVF procedures.  Moreover, it is probable that the obtained results of diagnostic assays would not be consistent with the outcomes of treatment plan due to the extremely heterogeneous sperm population of semen. Consequently, this defect has led to a new generation of assays that select sperm based on surface charge (electrophoresis and zeta potential), apoptosis (magnetic cell sorting and glass wool), membrane maturity (hyaluronic acid binding) and ultramorphology (high magnification) without damaging the selected spermatozoa. In these cases, selection of sperm via hyaluronane binding or annexine V binding or electrical charge provides functional sperm with good quality which can be used for oocyte fertilization (3).<br>Although the initial results of these assays are satisfactory, the findings are preliminary and further clinical trials are needed to validate their safety and efficacy before their implementation in ART practice. In addition, these assays do not provide direct assessment of sperm chromatin. Consequently, future research should be focused on promotion and optimization of sperm chromatin assays without any damage to sperm quality for its subsequent use during in vitro fertilization. The review article of professor Evgeni and his colleagues entitled "Human Sperm DNA Fragmentation and its Correlation with Conventional Semen Parameters" discussed this subject in detail in current issue. We hope it will be welcomed and used by clinicians, scientists and other audiences of journal of reproduction and infertility. 01 2 Mohammad Reza MR Sadeghi محمدرضا صادقی Editor-in-chief Editor-in-chief Iran No Keyword 564.pdf Franken DR, Oehninger S. Semen analysis and sperm function testing. Asian J Androl. 2012;14(1):6-13.##Shamsi MB, Imam SN, Dada R. Sperm DNA integrity assays: diagnostic and prognostic challenges and implications in management of infertility. J Assist Reprod Genet. 2011;28(11):1073-85.##Said TM, Land JA. Effects of advanced selection methods on sperm quality and ART outcome: a systematic review. Hum Reprod Update. 2011;17(6):719-33.##

Human Sperm DNA Fragmentation and Its Correlation with Conventional Semen Parameters 2 1 548 2 Background: The initial step in the diagnostic investigation of male infertility has been traditionally based on the conventional seminal profile. However, there are significant limitations regarding its ability to determine the underlying mechanisms that cause the disorder. Sperm DNA fragmentation has emerged as a potential causative factor of reproductive failure and its assessment has been suggested as a useful adjunct to the laboratory methodology of male infertility evaluation, especially before the application of assisted reproduction technology (ART). Methods: A review of recent bibliography was carried out in PubMed by the use of relevant keywords, in order to evaluate the possible correlation between the conventional seminal parameters and sperm DNA fragmentation assessment as diagnostic tools in male infertility evaluation. Results: A comprehensive diagnostic approach of male infertility should be based on a combination of diagnostic attributes, derived from the conventional semen analysis, as well as the investigation of genomic integrity testing. Conclusion: Due to its strong correlation with several aspects of ART procedures and further consequences for the offspring, sperm DNA fragmentation is a parameter worth integrating in routine clinical practice. However, additional large scale studies focusing on specific subgroups of infertile men who may benefit from an efficient therapeutic management based on the optimization of sperm DNA integrity are needed. 02 15 Evangelini E Evgeni Evangelini Evgeni Seminology Laboratory Seminology Laboratory Greece lina_evgeni@yahoo.gr Konstantinos K Charalabopoulos Konstantinos Charalabopoulos Laboratory of Physiology, School of Medicine, Democritus University of Thrace Laboratory of Physiology, School of Medicine, Democritus University of Thrace Greece Byron B Asimakopoulos Byron Asimakopoulos Laboratory of Physiology, School of Medicine, Democritus University of Thrace Laboratory of Physiology, School of Medicine, Democritus University of Thrace Greece Assisted reproductionConventional semen parametersDNA fragmentation 548.pdf Saleh RA, Agarwal A, Nelson DR, Nada EA, El-Tonsy MH, Alvarez JG, et al. Increased sperm nuclear DNA damage in normozoospermic infertile men: a prospective study. 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Sperm DNA damage assessment: a test whose time has come. Fertil Steril. 2005;84(4):850-3.##Spano M, Seli E, Bizzaro D, Manicardi GC, Sakkas D. The significance of sperm nuclear DNA strand breaks on reproductive outcome. Curr Opin Obstet Gynecol. 2005;17(3):255-60.##Lin MH, Kuo-Kuang Lee R, Li SH, Lu CH, Sun FJ, Hwu YM. Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertil Steril. 2008;90(2):352-9.##Lewis SE, Agbaje I, Alvarez J. Sperm DNA tests as useful adjuncts to semen analysis. Syst Biol Reprod Med. 2008;54(3):111-25.##Benchaib M, Lornage J, Mazoyer C, Lejeune H, Salle B, Francois Guerin J. Sperm deoxyribonucleic acid fragmentation as a prognostic indicator of assisted reproductive technologyoutcome. Fertil Steril. 2007;87(1):93-100.##Collins JA, Barnhart KT, Schlegel PN. Do sperm DNA integrity tests predict pregnancy with in vitro fertilization?. Fertil Steril. 2008;89(4):823-31.##Muriel L, Garrido N, Fernández JL, Remohi J, Pellicer A, de los Santos MJ, et al. Value of the sperm deoxyribonucleic acid fragmentation level, as measured by the sperm chromatin dispersion test, in the outcome of in vitro fertilization and intracytoplasmic sperm injection. Fertil Steril. 2006;85(2):371-83.##Schirren C. Practical Andrology. 1st ed. Berlin: Verlag Bruder Hartman; 1972.##Menkveld R, editor. The basic semen analysis. Oxon: Informa UK Ltd; c2007. 141 p. (Oehninger SC, Kruger TF, editors. Male Infertility. Diagnosis and Treatment; vol. 9).##McDonough R. Editorial comment: has traditional sperm analysis lost its clinical relevance?. Fertil Steril. 1997;67(3):585.##Haidl G, Allam JP, Schuppe HC. Chronic epididymitis: impact on semen parameters and therapeutic options. Andrologia. 2008;40(2):92-6.##World Health Organization. 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Asian J Androl. 2011;13(1):31-5.##Leduc F, Nkoma GB, Boissonneault G. Spermiogenesis and DNA repair: a possible etiology of human infertility and genetic disorders. Syst Biol Reprod Med. 2008;54(1):3-10.##Alvarez JG. DNA fragmentation in human spermatozoa: significance in the diagnosis and treatment of infertility. Minerva Ginecol. 2003;55(3):233-9.##De Palma A, Vicari E, Palermo I, D'Agata R, Calogero AE. Effects of cancer and anti-neoplastic treatment on the human testicular function. J Endocrinol Invest. 2000;23(10):690-6.##French DB, Desai NR, Agarwal A. Varicocele repair: does it still have a role in infertility treatment?. Curr Opin Obstet Gynecol. 2008;20(3):269-74.##Sergerie M, Mieusset R, Croute F, Daudin M, Bujan L. High risk of temporary alteration of semen parameters after recent acute febrile illness. Fertil Steril. 2007;88(4):970.e1-7.##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(6):1215-24.##Young KE, Robbins WA, Xun L, Elashoff D, Rothmann SA, Perreault SD. Evaluation of chromosome breakage and DNA integrity in sperm: an investigation of remote semen collection conditions. J Androl. 2003;24(6):853-61.##Xia Y, Cheng S, Bian Q, Xu L, Collins MD, Chang HC, et al. Genotoxic effects on spermatozoa of carbaryl-exposed workers. Toxicol Sci. 2005;85(1):615-23.##Rubes J, Selevan SG, Evenson DP, Zudova D, Vozdova M, Zudova Z, et al. Episodic air pollution is associated with increased DNA fragmentation in human sperm without other changes in semen quality. Hum Reprod. 2005;20(10):2776-83.##Rubes J, Selevan SG, Sram RJ, Evenson DP, Perreault SD. GSTM1 genotype influences the susceptibility of men to sperm DNA damage associated with exposure to air pollution. Mutat Res. 2007;625(1-2):20-8.##Wyrobek AJ, Eskenazi B, Young S, Arnheim N, Tiemann-Boege I, Jabs EW, et al. Advancing age has differential effects on DNA damage, chromatin integrity, gene mutations, and aneuploidies in sperm. Proc Natl Acad Sci USA. 2006;103(25):9601-6.##Cocuzza M, Sikka SC, Athayde KS, Agarwal A. Clinical relevance of oxidative stress and sperm chromatin damage in male infertility: an evidence based analysis. Int Braz J Urol. 2007;33(5):603-21.##Ji G, Long Y, Zhou Y, Huang C, Gu A, Wang X. Common variants in mismatch repair genes associated with increased risk of sperm DNA damage and male infertility. BMC Med. 2012;10:49.##Jaiswal D, Sah R, Agrawal NK, Dwivedi US, Trivedi S, Singh K. Combined effect of GSTT1 and GSTM1 polymorphisms on human male infertility in north Indian population. Reprod Sci. 2012;19(3):312-6.##Ji G, Gu A, Wang Y, Huang C, Hu F, Zhou Y, et al. Genetic variants in antioxidant genes are associated with sperm DNA damage and risk of male infertility in a Chinese population. Free Radic Biol Med. 2012;52(4):775-80.##Perrin A, Nguyen MH, Bujan L, Vialard F, Amice V, Gueganic N, et al. DNA fragmentation is higher in spermatozoa with chromosomally unbalanced content in men with a structural chromosomal rearrangement. Andrology. 2013;1(4):632-8.##Rouen A, Pyram K, Pollet-Villard X, Hyon C, Dorna M, Marques S, et al. Simultaneous cell by cell study of both DNA fragmentation and chromosomal segregation in spermatozoa from chromosomal rearrangement carriers. J Assist Reprod Genet. 2013;30(3):383-90.##Shamsi MB, Kumar R, Malhotra N, Singh N, Mittal S, Upadhyay AD, et al. Chromosomal aberrations, Yq microdeletion, and sperm DNA fragmentation in infertile men opting for assisted reproduction. Mol Reprod Dev. 2012; 79(9):637-50.##Aitken RJ, Koppers AJ. Apoptosis and DNA damage in human spermatozoa. Asian J Androl. 2011;13(1):36-42.##Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl. 2002;23(1):25-43.##Schlegel PN, Paduch DA. Yet another test of sperm chromatin structure. Fertil Steril. 2005;84(4):854-9.##Erenpreiss J, Spano M, Erenpreisa J, Bungum M, Giwercman A. Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl. 2006;8(1):11-29.##Sakkas D, Alvarez JG. Sperm DNA fragmentation: mechanisms of origin, impact on reproductive outcome, and analysis. Fertil Steril. 2010;93(4):1027-36.##Younglai EV, Holt D, Brown P, Jurisicova A, Casper RF. Sperm swim-up techniques and DNA fragmentation. Hum Reprod. 2001;16(9):1950-3.##Liu CH, Tsao HM, Cheng TC, Wu HM, Huang CC, Chen CI, et al. DNA fragmentation, mitochondrial dysfunction and chromosomal aneuploidy in the spermatozoa of oligoasthenoteratozoospermic males. 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Study of two markers of apoptosis and meiotic segregation in ejaculated sperm of chromosomal translocation carrier patients. Hum Reprod. 2006;21(3):685-93.##Muriel L, Goyanes V, Segrelles E, Gosalvez J, Alvarez JG, Fernandez JL. Increased aneuploidy rate in sperm with fragmented DNA as determined by the sperm chromatin dispersion (SCD) test and FISH analysis. J Androl. 2007;28(1):38-49.##Caglar GS, Koster F, Schopper B, Asimakopoulos B, Nehls B, Nikolettos N, et al. Semen DNA fragmentation index, evaluated with both TUNEL and Comet assay, and the ICSI outcome. In Vivo. 2007;21(6):1075-80.##Garolla A, Fortini D, Menegazzo M, De Toni L, Nicoletti V, Moretti A, et al. High-power microscopy for selecting spermatozoa for ICSI by physiological status. Reprod Biomed Online. 2008;17(5):610-6.##Nikolettos N, Asimakopoulos B, Papastefanou IS. Intracytoplasmic sperm injection—an assisted reproduction technique that should make us cautious about imprinting deregulation. 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Relation of Cumulus Cell Status with Single Oocyte Maturity, Fertilization Capability and Patient Age 2 1 549 2 Background: The production of competent oocytes depends on a bi-directional communication between the oocyte and cumulus cells. The goal of this study was to determine whether simple parameters monitored in cumulus cells from individual human oocytes have any predictive value, and thus correlate with clinically relevant parameters. Methods: 97 cumulus-oocyte complexes were recovered from 31 patients undergoing ICSI treatment. After the oocytes were denuded, cumulus cell density from individual oocytes was determined. Cells were probed for viability using propidium iodide and for apoptosis by Annexin V staining or by monitoring caspase activity. These parameters were correlated with oocyte status, fertilization ability and patient age (≤29 years old and ≥30 years old). All variables were checked for normal distribution and then compared by Kruskal-Wallis, Mann-Whitney or one-way ANOVA tests. Results: Mature oocytes were surrounded by more cumulus cells (16073±2595, p=0.026), which were also more viable and less apoptotic than atretic or degenerated oocytes. Mature oocytes that fertilized had higher caspase activity in the surrounding cumulus cells than those that did not fertilize. Younger patients presented lower cumulus cells density (8882±2380 vs. 15036±2143 cells; p=0.034); and cumulus cells had higher apoptosis levels in younger patients than older ones (6775.5±1831.6 RLU vs. 2591±46.5 RLU, p=0.002 for caspase activity). Conclusion: The data suggests that high density and apoptosis of cumulus cells are promising parameters to indirectly predict individual oocyte status. Although more studies and a larger data set are needed, cumulus cells presented the potential to be used as simple predictors of female fertility and/or ovarian ageing. 15 22 Bárbara B Lourenço Bárbara Lourenço Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra Portugal Ana AP Sousa Ana Sousa Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra Portugal Teresa T Almeida-Santos Teresa Almeida-Santos Human Reproduction Service, University Hospitals of Coimbra Human Reproduction Service, University Hospitals of Coimbra Portugal João J Ramalho-Santos João Ramalho-Santos Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra Biology of Reproduction and Stem Cell Group, Center for Neuroscience and Cell Biology (CNC), University of Coimbra Portugal jramalho@ci.uc.pt Annexin VApoptosisCaspase activityCumulus cellCumulus-oocyte complexFemale fertilityOocyte biomarkersWoman age 549.pdf Zhuo L, Kimata K. 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J Assist Reprod Genet. 2001;18(9):490-8.##Mikkelsen AL, Host E, Lindenberg S. Incidence of apoptosis in granulosa cells from immature human follicles. Reproduction. 2001;122(3):481-6.##Host E, Gabrielsen A, Lindenberg S, Smidt-Jensen S. Apoptosis in human cumulus cells in relation to zona pellucida thickness variation, maturation stage, and cleavage of the corresponding oocyte after intracytoplasmic sperm injecti on. Fertil Steril. 2002;77(3):511-5.##Corn CM, Hauser-Kronberger C, Moser M, Tews G, Ebner T. Predictive value of cumulus cell apoptosis with regard to blastocyst development of corresponding gametes. Fertil Steril. 2005;84(3): 627-33.##Yuan YQ, Van Soom A, Leroy JL, Dewulf J, Van Zeveren A, de Kruif A, et al. Apoptosis in cumulus cells, but not in oocytes, may influence bovine embryonic developmental competence. Theriogenology. 2005;63(8):2147-63.##Abu-Hassan D, Koester F, Shoepper B, Schultze-Mosgau A, Asimakopoulos B, Diedrich K, et al. Comet assay of cumulus cells and spermatozoa DNA status, and the relationship to oocyte fertilization andembryo quality following ICSI. Reprod Biomed Online. 2006;12(4):447-52.##Han ZB, Lan GC, Wu YG, Han D, Feng WG, Wang JZ, et al. Interactive effects of granulosa cell apoptosis, follicle size, cumulus-oocyte complex morphology, and cumulusexpansion on the developmental competence of goat oocytes: a study using the well-in-drop culture system. Reproduction. 2006;132(5):749-58.##Huang Z, Wells D. The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol Hum Reprod. 2010;16(10):715-25.##Hashimoto S, Saeki K, Nagao Y, Minami N, Yamada M, Utsumi K. Effects of cumulus cell density during in vitro maturation of the developmental competence of bovine oocytes. Theriogenology. 1998;49(8):1451-63.##Ameisen JC. On the origin, evolution, and nature of programmed cell death: a timeline of four billion years. Cell Death Differ. 2002;9(4):367-93.##Varum S, Bento C, Sousa AP, Gomes-Santos CS, Henriques P, Almeida-Santos T, et al. Characterization of human sperm populations using conventional parameters, surface ubiquitination, andapoptotic markers. Fertil Steril. 2007;87(3):572-83.##Budihardjo I, Oliver H, Lutter M, Luo X, Wang X. Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol. 1999;15:269-90.##Yuan YQ, Peelman LJ, Williams JL, Van Zeveren A, de Kruif A, Law A, et al. Mapping and transcription profiling of CASP1, 3, 6, 7 nd 8 in relation to caspase activity in the bovine cumulus-oocyte complex. Anim Genet. 2004;35(3):234-7.##O’Brien M, Moravec R, Riss T. Caspase-GloTM 3/7 assay: use fewer cells and spend less time with this homogeneous assay. Cell Notes. 2003;6:13-5.##Zeuner A, Muller K, Reguszynski K, Jewgenow K. Apoptosis within bovine follicular cells and its effect on oocyte development during in vitro maturation. Theriogenology. 2003;59(5-6):1421-33.##Hussein TS, Froiland DA, Amato F, Thompson JG, Gilchrist RB. Oocytes prevent cumulus cell apoptosis by maintaining a morphogenic paracrine gradient of bonemorphogenetic proteins. J Cell Sci. 2005;118(Pt 22):5257-68.##Baird DT, Collins J, Egozcue J, Evers LH, Gianaroli L, Leridon H, et al. Fertility and ageing. Hum Reprod Update. 2005;11(3):261-76.##Alviggi C, Humaidan P, Howles CM, Tredway D, Hillier SG. Biological versus chronological ovarian age: implications for assisted reproductive technology. Reprod Biol Endocrinol. 2009;7:101.##Balasch J. Ageing and infertility: an overview. Gynecol Endocrinol. 2010;26(12):855-60.##

Protective Effects of Antioxidants on Sperm Parameters and Seminiferous Tubules Epithelium in High Fat-fed Rats 2 1 550 2 Background: Prescription of antioxidants might increase the quality of sperm parameters and improve the rate of pregnancy in obese people who suffer from infertility. Therefore, the present study investigated protective effects of vitamin A, E and astaxanthin on sperm parameters and seminiferous tubules epithelium in high-fat diet model. Methods: Thirty-six numbers of 3 months old albino Wistar rats were divided to control, high-fat diet and high-fat diet with antioxidants groups. After 12 weeks, levels of LDL-C and HDL-C were detected in the groups. Sperm was obtained from the tail of epididymis and its parameters (count, vitality, motility and morphology) were analyzed. Testes were fixed in 10% formalin and after tissue processing, stained with Hematoxylin and Eosine (H&E) for histological evaluation. Data were analyzed by a one-way ANOVA and p<0.05 was considered significant. Results: Our results indicated that viability, motility and normal morphology of sperm in high-fat diet (HFD) decreased significantly compared to high-fat diet with antioxidant (HFD+A) and the control groups (p<0.05). Also spermatogonium and the number of Sertoli cells increased significantly in HFD+A compared to the control (p<0.05). Conclusion: As it is shown in our study, application of antioxidants decreased serum triglyceride, cholesterol and HDL-C/LDL-C in high-fat diet model and improved the semen parameters. Therefore, it is suggested that the low quality of sperm can be improved in obese men through antioxidant prescription. Finally, it seems that the antioxidants in obese patients with subfertility or infertility is a new and efficient strategy with few side effects. 22 29 Motahareh M Mortazavi Motahareh Mortazavi Anatomical Sciences Department, Medicine faculty, Hamadan University of Medical Sciences Anatomical Sciences Department, Medicine faculty, Hamadan University of Medical Sciences Iran Iraj I Salehi Iraj Salehi Neurophysiology Research Center, Hamadan University of Medical Sciences Neurophysiology Research Center, Hamadan University of Medical Sciences Iran Zohreh Z Alizadeh Zohreh Alizadeh Anatomical Sciences Department, Medicine faculty, Hamadan University of Medical Sciences Anatomical Sciences Department, Medicine faculty, Hamadan University of Medical Sciences Iran Mehrangiz M Vahabian Mehrangiz Vahabian Department of Persian Literature and English Language, Medicine faculty, Hamadan University of Medical Sciences Department of Persian Literature and English Language, Medicine faculty, Hamadan University of Medical Sciences Iran Amaneh A Mohammadi Roushandeh Amaneh Mohammadi Roushandeh Research Center for Molecular Medicine, Medicine Faculty, Hamadan University of Medical Sciences Research Center for Molecular Medicine, Medicine Faculty, Hamadan University of Medical Sciences Iran a.mohammadiroshandeh@umsha.ac.ir AntioxidantAstaxanthinHigh-fat DietSpermatogenesisTestisVitamin AVitamin C 550.pdf de Groot PC, Dekkers OM, Romijn JA, Dieben SW, Helmerhorst FM. PCOS, coronary heart disease, stroke and the influence of obesity: a systematic review and meta-analysis. Hum Reprod Update. 2011;17(4):495-500.##Katz DL, O'Connell M, Yeh MC, Nawaz H, Njike V, Anderson LM, et al. Public health strategies for preventing and controlling overweight and obesity in school and worksite settings: a report on recommendations of the Task Force on Community Preventive Services. MMWR Recomm Rep. 2005; 54(RR-10):1-12.##Goldberg RD, Cardamone S, Murthy A. Obesity and contraception: controversy? Open Access J Contracept. 2012;3:1-8.##Fernandez CD, Bellentani FF, Fernandes GS, Perobelli JE, Favareto AP, Nascimento AF, et al. Diet-induced obesity in rats leads to a decrease in sperm motility. Reprod Biol Endocrinol. 2011;9:32.##Robker RL. Evidence that obesity alters the quality of oocytes and embryos. Pathophysiology. 2008;5(2):115-21.##Jensen TK, Heitmann BL, Jensen MB, Halldorsson TI, Andersson AM, Skakkebæk NE, et al. High dietary intake of saturated fat is associated with reduced semen quality among 701 young Danish men from the general population. Am J Clin Nutr. 2013;97(2):411-8.##Ghanayem BI, Bai R, Kissling GE, Travlos G, Hoffler U. Diet-induced obesity in male mice is associated with reduced fertility and potentiation of acrylamide-inducedreproductive toxicity. Biol Reprod. 2010;82(1):96-104.##Nguyen RH, Wilcox AJ, Skjaerven R, Baird DD. Men's body mass index and infertility. Hum Reprod. 2007;22(9):2488-93.##Hammoud AO, Gibson M, Peterson CM, Meikle AW, Carrell DT. Impact of male obesity on infertility: a critical review of the current literature. Fertil Steril. 2008;90(4):897-904.##Aitken RJ, Jones KT, Robertson SA. Reactive oxygen species and sperm function--in sickness and in health. J Androl. 2012;33(6):1096-106.##Vigueras-Villasenor RM, Rojas-Castaneda JC, Chavez-Saldana M, Gutierrez-Perez O, Garcia-Cruz ME, Cuevas-Alpuche O, et al. Alterations in the spermatic function generated by obesity in rats. Acta Histochem. 2011;113(2):214-20.##Agarwal A, Saleh RA, Bedaiwy MA. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril. 2003;79(4):829-43.##Chen XL, Gong LZ, Xu JX. Antioxidative activity and protective effect of probiotics against high-fat diet-induced sperm damage in rats. Animal. 2013;7(2):287-92.##Agarwal A, Allamaneni SS. Free radicals and male reproduction. J Indian Med Assoc. 2011;109(3):184-7.##Chen H, Zhao HX, Huang XF, Chen GW, Yang ZX, Sun WJ, et al. Does high load of oxidants in human semen contribute to male factor infertility?. Antioxid Redox Signal. 2012;16(8):754-9.##Guerin M, Huntley ME, Olaizola M. Haematococcus astaxanthin: applications for human health and nutrition. Trends Biotechnol. 2003;21(5):210-6.##Lennikov A, Kitaichi N, Fukase R, Murata M, Noda K, Ando R, et al. Amelioration of ultraviolet-induced photokeratitis in mice treated with astaxanthin eye drops. Mol Vis. 2012;18:455-64.##Ikeuchi M, Koyama T, Takahashi J, Yazawa K. Effects of astaxanthin in obese mice fed a high-fat diet. Biosci Biotechnol Biochem. 2007;71(4):893-9.##Vinayagamoorthi R, Bobby Z, Sridhar MG. Antioxidants preserve redox balance and inhibit c-Jun-N-terminal kinase pathway while improving insulin signaling in fat-fed rats: evidence for the role of oxidative stress on IRS-1 serine phosphorylation and insulin resistance. J Endocrinol. 2008;197(2):287-96.##Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem. 1996;239(1):70-6.##Bakos HW, Mitchell M, Setchell BP, Lane M. The effect of paternal diet-induced obesity on sperm function and fertilization in a mouse model. Int J Androl. 2011;34(5 Pt 1):402-10.##Agarwal A, Sekhon LH. The role of antioxidant therapy in the treatment of male infertility. Hum Fertil (Camb). 2010;13(4):217-25.##Ross C, Morriss A, Khairy M, Khalaf Y, Braude P, Coomarasamy A, et al. A systematic review of the effect of oral antioxidants on male infertility. Reprod Biomed Online. 2010;20(6):711-23.##Hull MG, North K, Taylor H, Farrow A, Ford WC. Delayed conception and active and passive smoking. The Avon longitudinal study of pregnancy and childhood study team. Fertil Steril. 2000;74(4):725-33.##De Lamirande E, Gagnon C. Reactive oxygen species and human spermatozoa. II. Depletion of adenosine triphosphate plays an important role in the inhibition of sperm motility. J Androl. 1992;13(5):379-86.##Zhang K, Lv Z, Jia X, Huang D. Melatonin prevents testicular damage in hyperlipidaemic mice. Andrologia. 2012;44(4):230-6.##Erdemir F, Atilgan D, Markoc F, Boztepe O, Suha-Parlaktas B, Sahin S. [The effect of diet induced obesity on testicular tissue and serum oxidative stress parameters]. Actas Urol Esp. 2012;36(3):153-9. Spanish.##Fassett RG, Coombes JS. Astaxanthin, oxidative stress, inflammation and cardiovascular disease. Future Cardiol. 2009;5(4):333-42.##

Sexual and Reproductive Outcomes in Early Stage Cervical Cancer Patients after Excisional Cone as a Fertility-sparing Surgery: An Italian Experience 2 1 551 2 Background: The purpose of this study was to analyze the quality of life in terms of sexual and reproductive outcome in patients suffering from early stage cervical cancer, submitted to an excisional cone as fertility-sparing treatment. Methods: A multicenter retrospective analysis about specific dimensions of physical, psychological, reproductive and sexual functions after a cold-knife conization plus pelvic laparoscopic lymphadenectomy was conducted at Division of Gynecologic Oncology, Catholic University of the Sacred Heart, Rome-Italy and at Division of Gynecology, European Institute of Oncology, Milan-Italy. The aim of this study was twofold. It aimed to analyze the quality of life in patients submitted to minimally invasive surgery and to compare these data with radical trachelectomy. Results: Twenty-three patients with an average age of 30 years decided to participate in this study. After the treatment, all women (100%) had regular menstruation, 7 (30.4%) had increased not invalidating dysmenorrhea; 1 (4.4%) experienced a cervical stenosis; 6 among 10 patients that tried to conceive (60%) obtained one spontaneous pregnancy; 4 more (40%) underwent in vitro fertilization and embryo transfer and only 1 of them (25%) was successful. About sexual assessment, 1 patient (4.4%) had trouble in lubricating, 3 (13%) had anxiety about performance, 6 (26.1%) complained of dyspareunia which was resolved within 3 subsequent months. All patients (100%) obtained a complete psychological and physical recovery. Conclusion: This study demonstrated preliminary encouraging data about sexual and reproductive outcome after excisional conization. A comparison with trachelectomy surely needs longer follow-ups, more cases and prospective analyses. 29 35 Francesco F Fanfani Francesco Fanfani Institute for Maternal and Child Health, IRCCS &quot;Burlo Garofolo&quot; Institute for Maternal and Child Health, IRCCS "Burlo Garofolo" Italy francesco.fanfani@rm.unicatt.it Fabio F Landoni Fabio Landoni Division of Gynecology, European Institute of Oncology Division of Gynecology, European Institute of Oncology Italy Maria ML Gagliardi Maria Gagliardi Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Italy Anna A Fagotti Anna Fagotti Division of Minimally Invasive Gynecological Surgery, Maria Hospital-University of Perugia Division of Minimally Invasive Gynecological Surgery, Maria Hospital-University of Perugia Italy Eleonora E Preti Eleonora Preti Division of Gynecology, European Institute of Oncology Division of Gynecology, European Institute of Oncology Italy Maria M Moruzzi Maria Moruzzi Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Italy Giorgia G Monterossi Giorgia Monterossi Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Italy Giovanni G Scambia Giovanni Scambia Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Catholic University of the Sacred Heart Italy Cervical cancerConservative approachExcisional coneFertility-sparing surgeryQuality of life 551.pdf Dargent D, Brun JL, Roy M, Mathevet P, Remy I. La trachelectomie elargie (te), une alternative a l’hysterectomie radicale dans le traitement des cancers infiltrants developp_es sur la face externe du col uterin. J Obstet Gynecol. 1994;2:285-92.##Dursun P, LeBlanc E, Nogueira MC. Radical vaginal trachelectomy (Dargent's operation): a critical review of the literature. Eur J Surg Oncol. 2007;33(8):933-41.##Gien LT, Covens A. Fertility-sparing options for early stage cervical cancer. Gynecol Oncol. 2010;117(2):350-7.##Burnett AF. Radical trachelectomy with laparoscopic lymphadenectomy: review of oncologic and obstetrical outcomes. Curr Opin Obstet Gynecol. 2006;18(1):8-13.##Beiner ME, Covens A. Surgery insight: radical vaginal trachelectomy as a method of fertility preservation for cervical cancer. Nat Clin Pract Oncol. 2007;4(6):353-61.##Maneo A, Chiari S, Bonazzi C, Mangioni C. Neoadjuvant chemotherapy and conservative surgery for stage IB1 cervical cancer. Gynecol Oncol. 2008;111(3):438-43.##Landoni F, Parma G, Peiretti M, Zanagnolo V, Sideri M, Colombo N, et al. Chemo-conization in early cervical cancer. Gynecol Oncol. 2007;107(1 Suppl 1):S125-6.##Fagotti A, Gagliardi ML, Moruzzi C, Carone V, Scambia G, Fanfani F. Excisional cone as fertility-sparing treatment in early-stage cervical cancer. Fertil Steril. 2011;95(3):1109-12.##Wenzel L, DeAlba I, Habbal R, Kluhsman BC, Fairclough D, Krebs LU, et al. Quality of life in long-term cervical cancer survivors. Gynecol Oncol. 2005;97(2):310-7.##Moinpour CM, Lyons B, Grevstad PK, Lovato LC, Crowley J, Czaplicki K, et al. Quality of life in advanced non-small-cell lung cancer: results of a Southwest Oncology Group randomized trial. Qual Life Res. 2002;11(2):115-26.##Beitz J, Gnecco C, Justice R. Quality-of-life end points in cancer clinical trials: the U.S. Food and Drug Administration perspective. J Natl Cancer Inst Monogr. 1996;(20):7-9.##Kiebert GM, Kaasa S. Quality of life in clinical cancer trials: experience and perspective of the European Organization for Research and Treatment of Cancer. J Natl Cancer Inst Monogr. 1996;(20):91-5.##Distefano M, Riccardi S, Capelli G, Costantini B, Petrillo M, Ricci C, et al. Quality of life and psychological distress in locally advanced cervical cancer patients administered pre-operative chemoradiotherapy. Gynecol Oncol. 2008;111(1):144-50.##Carter J, Sonoda Y, Abu-Rustum NR. Reproductive concerns of women treated with radical trachelectomy for cervical cancer. Gynecol Oncol. 2007;105(1):13-6.##Carter J, Sonoda Y, Chi DS, Raviv L, Abu-Rustum NR. Radical trachelectomy for cervical cancer: postoperative physical and emotional adjustment concerns. Gynecol Oncol. 2008;111(1):151-7.##Carter J, Sonoda Y, Baser RE, Raviv L, Chi DS, Barakat RR, et al. A 2-year prospective study assessing the emotional, sexual, and quality of life concerns of women undergoing radical trachelectomy versus radical hysterectomy for treatment of early-stage cervical cancer. Gynecol Oncol. 2010;119(2):358-65.##Meeting report. The new FIGO staging system for cancers of the vulva, cervix, endometrium and sarcomas. Gynecol Oncol. 2009;115:325-8.##Kobayashi Y, Akiyama F, Hasumi K. A case of successful pregnancy after treatment of invasive cervical cancer with systemic chemotherapy and conization. Gynecol Oncol. 2006;100(1):213-5.##Rob L, Pluta M, Strnad P, Hrehorcak M, Chmel R, Skapa P, et al. A less radical treatment option to the fertility-sparing radical trachelectomy in patients with stage I cervical cancer. Gynecol Oncol. 2008;111(2 Suppl):S116-20.##Ferrandina G, Salutari V, Petrillo M, Carbone A, Scambia G. Conservatively treated glassy cell carcinoma of the cervix. World J Surg Oncol. 2008;6:92.##Plante M, Renaud MC, Francois H, Roy M. Vaginal radical trachelectomy: an oncologically safe fertility-preserving surgery. An updated series of 72 cases and review of the literature. Gynecol Oncol. 2004;94(3):614-23.##Maneo A, Sideri M, Scambia G, Boveri S, Dell'anna T, Villa M, et al. Simple conization and lymphadenectomy for the conservative treatment of stage IB1 cervical cancer. An Italian experience. Gynecol Oncol. 2011;123(3):557-60.##Finnstrom O, Kallen B, Lindam A, Nilsson E, Nygren KG, Olausson PO. Maternal and child outcome after in vitro fertilization--a review of 25 years of population-based data from Sweden. Acta Obstet Gynecol Scand. 2011;90(5):494-500.##

Comparing the Roles of Sperm Chromatin Integrity and Apoptosis in Intrauterine Insemination Outcomes of Couples with Mild Male and Female Factor Infertility 2 1 552 2 Background: Intrauterine insemination (IUI) is one of the therapeutic approaches for infertility. The objective of this study was to evaluate DNA integrity and apoptosis role in success of IUI in both mild male and female factor infertility. Methods: Patients were divided into two groups: M (mild male factor; n=29) and F (female factor; n=31) undergoing single IUI. Ejaculates were analyzed and chromatin quality was assessed using chromomycin A3 (CMA3) staining. In addition, spermatozoal apoptosis was recognized using TUNEL assay. Statistical analyses were done using t-test and Mann Whitney test for sperm apoptosis and sperm chromatin by SPSS. Data were expressed in mean ± SD for variables. P<0.05 was considered statistically significant. Results: Sperm concentration and progressive motility were higher in F than M group. Sperm with normal morphology were statistically similar in M and F infertile patients (32.7±15.6% vs. 35.5±9.07%, p=0.39). Sperm chromatin immaturity was higher in patients with mild male infertility, when compared with the other group (p<0.01). Also, 32.0±5.6% and 30.8±6.1% of the spermatozoa showed signs of apoptosis in groups M and F, respectively (p=0.49). Very low (3.4%) clinical pregnancy rates were noticed in patients with mild male factor infertility Conclusion: Defect in sperm motility as well as high rates of DNA damage and apoptosis may be involved with very low rate of pregnancy outcomes in patients with mild male factor infertility. Therefore, it seems the application of IUI may have better outcomes in patients with female infertility compared to mild male factor infertility. 35 41 Mohammad Ali MA Khalili محمد علی خلیلی Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Iran Saeedeh S Nazari Saeedeh Nazari Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Iran nazari.saeedeh65@gmail.com Razieh R Dehghani-Firouzabadi Razieh Dehghani-Firouzabadi Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Iran Alireza A Talebi علیرضا طالبی Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Iran Shekofeh Sh Baghazadeh-Naeini Shekofeh Baghazadeh-Naeini Infertility Center, Isfahan University of Medical Sciences Infertility Center, Isfahan University of Medical Sciences Iran Fatemeh F Sadeghian-Nodoshan Fatemeh Sadeghian-Nodoshan Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Iran Azam A Agha-Rahimi اعظم آقارحیمی Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Infertility Research and Clinical Center, Shahid Sadoughi Medical Sciences University Iran ApoptosisInfertilityIntrauterine transfusionMaleMorphologySperm motilitySpermatozoa 552.pdf Lachaud C, Tesarik J, Canadas ML, Mendoza C. Apoptosis and necrosis in human ejaculated spermatozoa. Hum Reprod. 2004;19(3):607-10.##Custers IM, Steures P, Hompes P, Flierman P, van Kasteren Y, van Dop PA, et al. Intrauterine insemination: how many cycles should we perform?. Hum Reprod. 2008;23(4):885-8.##Kamath MS, Bhave P, Aleyamma T, Nair R, Chandy A, Mangalaraj AM, et al. Predictive factors for pregnancy after intrauterine insemination: A prospective study of factors affecting outcome. J Hum Reprod Sci. 2010;3(3):129-34.##Lucchini C, Volpe E, Tocci A. Comparison of intrafollicular sperm injection and intrauterine insemination in the treatment of subfertility. J Assist Reprod Genet. 2012;29(10):1103-9.##Jayaraman V, Upadhya D, Narayan PK, Adiga SK. Sperm processing by swim-up and density gradient is effective in elimination of sperm with DNA damage. J Assist Reprod Genet. 2012;29(6):557-63.##El-Melegy NT, Ali ME. Apoptotic markers in semen of infertile men: Association with cigarette smoking. Int Braz J Urol. 2011;37(4):495-506.##Taylor SL, Weng SL, Fox P, Duran EH, Morshedi MS, Oehninger S, et al. Somatic cell apoptosis markers and pathways in human ejaculated sperm: potential utility as indicators of sperm quality. Mol Hum Reprod. 2004;10(11):825-34.##Jurisicova A, Lopes S, Meriano J, Oppedisano L, Casper RF, Varmuza S. DNA damage in round spermatids of mice with a targeted disruption of the Pp1cgamma gene and in testicular biopsies of patients with non-obstructive azoospermia. Mol Hum Reprod. 1999;5(4):323-30.##Aziz N, Said T, Paasch U, Agarwal A. The relationship between human sperm apoptosis, morphology and the sperm deformity index. Hum Reprod. 2007;22(5):1413-9.##Angelopoulou R, Plastira K, Msaouel P. Spermatozoal sensitive biomarkers to defective protaminosis and fragmented DNA. Reprod Biol Endocrinol. 2007;5:36.##Nasr-Esfahani MH, Aboutorabi R, Razavi S. Credibility of chromomycin A3 staining in prediction of fertility. Int J Fertil Steril. 2009;9:5-10.##Tarozzi N, Nadalini M, Stronati A, Bizzaro D, Dal Prato L, Coticchio G, et al. Anomalies in sperm chromatin packaging: implications for assisted reproduction techniques. Reprod Biomed Online. 2009;18(4):486-95.##Evenson DP, Darzynkiewicz Z, Melamed MR. Relation of mammalian sperm chromatin heterogeneity to fertility. Science. 1980;210(4474):1131-3.##World Health Organization. WHO laboratory manual for the examination and processing of human semen. 2010.##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(4):245-51.##Kupker W, Schulze W, Diedrich K. Ultrastructure of gametes and intracytoplasmic sperm injection: the significance of sperm morphology. Hum Reprod. 1998;13 Suppl 1:99-106.##Dorjpurev U, Kuwahara A, Yano Y, Taniguchi T, Yamamoto Y, Suto A, et al. Effect of semen characteristics on pregnancy rate following intrauterine insemination. J Med Invest. 2011;58(1-2):127-33.##Abdelkader AM, Yeh J. The potential use of intrauterine insemination as a basic option for infertility: a review for technology-limited medical settings. Obstet Gynecol Int. 2009;2009:584837.##Van Voorhis BJ, Barnett M, Sparks AE, Syrop CH, Rosenthal G, Dawson J. Effect of the total motile sperm count on the efficacy and cost-effectiveness of intrauterine insemination and in vitro fertilization. Fertil Steril. 2001;75(4):661-8.##Lee RK, Hou JW, Ho HY, Hwu YM, Lin MH, Tsai YC, et al. Sperm morphology analysis using strict criteria as a prognostic factor in intrauterine insemination. Int J Androl. 2002;25(5):277-80.##Agarwal A, Said TM. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update. 2003;9(4):331-45.##Saleh RA, Agarwal A, Sharma RK, Said TM, Sikka SC, Thomas AJ Jr. Evaluation of nuclear DNA damage in spermatozoa from infertile men with varicocele. Fertil Steril. 2003;80(6):1431-6.##Bungum M, Bungum L, Humaidan P, Yding Andersen C. Day 3 versus day 5 embryo transfer: a prospective randomized study. Reprod Biomed Online. 2003;7(1):98-104.##Bungum M, Humaidan P, Spano M, Jepson K, Bungum L, Giwercman A. The predictive value of sperm chromatin structure assay (SCSA) parameters for the outcome of intrauterine insemination, IVF and ICSI. Hum Reprod. 2004;19(6):1401-8.##Giwercman A, Richthoff J, Hjollund H, Bonde JP, Jepson K, Frohm B, et al. Correlation between sperm motility and sperm chromatin structure assay parameters. Fertil Steril. 2003;80(6):1404-12.##Ahinko-Hakamaa K, Huhtala H, Tinkanen H. Success in intrauterine insemination: the role of etiology. Acta Obstet Gynecol Scand. 2007;86(7): 855-60.##Zadehmodarres S, Oladi B, Saeedi S, Jahed F, Ashraf H. Intrauterine insemination with husband semen: an evaluation of pregnancy rate and factors affecting outcome. J Assist Reprod Genet. 2009;26(1):7-11.##Nuojua-Huttunen S, Tomas C, Bloigu R, Tuomivaara L, Martikainen H. Intrauterine insemination treatment in subfertility: an analysis of factors affecting outcome. Hum Reprod. 1999;14(3):698-703.##

Effects of Fenugreek Seed on the Severity and Systemic Symptoms of Dysmenorrhea 2 1 553 2 Background: Primary dysmenorrhea is a prevalent disorder and its unfavorable effects deteriorates the quality of life in many people across the world. Based on some evidence on the characteristics of fenugreek as a medical plant with anti-inflammatory and analgesic properties, this double-blind, randomized, placebo controlled trial was conducted. The main purpose of the study was to evaluate the effects of fenugreek seeds on the severity of primary dysmenorrhea among students. Methods: Unmarried Students were randomly assigned to two groups who received fenugreek (n=51) or placebo (n=50). For the first 3 days of menstruation, 23 capsules containing fenugreek seed powder (900 mg) were given to the subjects three times daily for two consecutive menstrual cycles. Pain severity was evaluated using a visual analog scale and systemic symptoms were assessed using a multidimensional verbal scale. Results: Pain severity at baseline did not differ significantly between the two groups. Pain severity was significantly reduced in both groups after the intervention; however, the fenugreek group experienced significantly larger pain reduction (p<0.001). With respect to the duration of pain, there was no meaningful difference between the two cycles in the placebo group (p=0.07) but in the fenugreek group, the duration of pain decreased between the two cycles (p<0.001). Systemic symptoms of dysmenorrhea (fatigue, headache, nausea, vomiting, lack of energy, syncope) decreased in the fenugreek seed group (p<0.05). No side effects were reported in the fenugreek group. Conclusion: These data suggest that prescription of fenugreek seed powder during menstruation can reduce the severity of dysmenorrhea. 41 49 Sima S Younesy Sima Younesy Department of Midwifery, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences Department of Midwifery, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences Iran Sedigheh S Amiraliakbari Sedigheh Amiraliakbari Department of Midwifery, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences Department of Midwifery, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences Iran asa_akbari@yahoo.com Somayeh S Esmaeili Somayeh Esmaeili Department of Traditional Pharmacy, School of Traditional Medicine, Shahid Beheshti University of Medical Sciences Department of Traditional Pharmacy, School of Traditional Medicine, Shahid Beheshti University of Medical Sciences Iran Hamid H Alavimajd حمید علوی‌مجد Department of Biostatistics, Faculty of Paramedical, Shahid Beheshti University of Medical Sciences Department of Biostatistics, Faculty of Paramedical, Shahid Beheshti University of Medical Sciences Iran Soheila S Nouraei Soheila Nouraei Department of Midwifery, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences Department of Midwifery, School of Nursing and Midwifery, Shahid Beheshti University of Medical Sciences Iran DysmenorrheaFenugreekHerbal medicine 553.pdf Shah Hosseini Z, Amin GR, Salehi Sormaghi MH, Danesh MM, Abedian K. 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Mutation Analysis of Exons 10 and 17a of CFTR Gene in Patients with Cystic Fibrosis in Kermanshah Province, Western Iran 2 1 554 2 Background: Cystic fibrosis (CF) is the most common genetic disorder with autosomal recessive inheritance among Caucasian populations. So far, more than 1950 different mutations were identified in cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR gene has 27 exons. The type and distribution of mutations vary widely among different countries and/or ethnic groups. Therefore, a comprehensive analysis was performed on exon10 and exon17a of CFTR gene in CF patients in the Kermanshah province, western Iran. Methods: We tested 27 patients admitted to the medical genetics laboratory of Kermanshah University of Medical Sciences. The patients were from different cities of Kermanshah province. All the patients had the clinical signals and two positive sweat tests. After filling agreement forms and questionnaire, the peripheral blood sampling and DNA extraction were done. DNA samples were extracted. PCR and sequencing special PCR were done. Finally analysis of the results with DNA sequencing analysis version 5.2 software was performed. Results: CFTR mutations analysis identified 4 different mutations in our CF patients. The disease-causing mutations were p.F508del (∆F508) (14.81%), p.S466X (1.85%), and p.T1036I (1.85%). M470V polymorphism with frequency of 74.1% was found in 23 patients (17 homozygous and 6 heterozygous). Conclusion: Three disease-causing mutations in CF patients in the present study account for approximately 18.51% of mutations. The frequency of p.F508del, the most common mutation was 1618.1% in Iranian population. The results of the present study can be applied for genetic counseling, population screening and prenatal diagnosis. 49 57 Abbas A Sahami Abbas Sahami Department of Biology, Faculty of Islamic Azad University, Jahrom Branch Department of Biology, Faculty of Islamic Azad University, Jahrom Branch Iran Reza R Alibakhshi Reza Alibakhshi Nano Drug Delivery Research Centre, Kermanshah University of Medical Sciences Nano Drug Delivery Research Centre, Kermanshah University of Medical Sciences Iran ralibakhshi@yahoo.com Keyghobad K Ghadiri Keyghobad Ghadiri Nosocomial Infectious Disease Research Center, Kermanshah University of Medical Sciences Nosocomial Infectious Disease Research Center, Kermanshah University of Medical Sciences Iran Hamid H Sadeghi Hamid Sadeghi Department of Biology, Faculty of Islamic Azad University, Jahrom Branch Department of Biology, Faculty of Islamic Azad University, Jahrom Branch Iran F508∆Cystic fibrosisDirect sequencingIranKermanshahM470VS466XT1036I 554.pdf Allen J, Panitch H, Rubenstein R, editors. Cystic fibrosis. New York: Informa Healthcare USA; c2010. 1-3 p. (Kreindler JL, Foskett JK, editors. Lung biology in health and disease series; vol. 242).##Geborek A, Hjelte L. Association between genotype and pulmonary phenotype in cystic fibrosis patients with severe mutations. J Cyst Fibros. 2011;10(3): 187-92.##Dorwart M, Thibodeau P, Thomas P. Cystic fibrosis: recent structural insights. J Cyst Fibros. 2004;3 Suppl 2:91-4.##Rommens JM. Cystic Fibrosis Mutation Database [Internet]. Toronto (CA): The Hospital for Sick Children; 1989 [updated 2011 Apr 25; cited 2013 Dec 20). Available from: http://www.genet.sickkids. on.ca/Home.html.##Estivill X, Bancells C, Ramos C. Geographic distribution and regional origin of 272 cystic fibrosis mutations in European populations. The Biomed CF Mutation Analysis Consortium. Hum Mutat. 1997;10 (2):135-54.##Dawson KP, Frossard PM. The geographic distribution of cystic fibrosis mutations gives clues about population origins. Eur J Pediatr. 2000;159(7):496-9.##Shah U, Frossard P, Moatter T. Cystic fibrosis: defining a disease under-diagnosed in Pakistan. Trop Med Int Health. 2009;14(5):542-5.##Alibakhshi R, Kianishirazi R, Cassiman JJ, Zamani M, Cuppens H. Analysis of the CFTR gene in Iranian cystic fibrosis patients: identification of eight novel mutations. J Cyst Fibros. 2008;7(2):102-9.##Elahi E, Khodadad A, Kupershmidt I, Ghasemi F, Alinasab B, Naghizadeh R, et al. A haplotype framework for cystic fibrosis mutations in Iran. J Mol Diagn. 2006;8(1):119-27.##Alibakhshi R, Zamani M. Mutation analysis of CFTR gene in 70 Iranian cystic fibrosis patients. Iran J Allergy Asthma Immunol. 2006;5(1):3-8.##Wikipedia, the free encyclopedia. Kurdish people [Internet]. Wikimedia Foundation, Inc; 2001 Jan 10 [updated 2013 Aug 14; cited 2013 Aug 20]. Available from: http://en.wikipedia.org/wiki/Kurdish_people.##Miller SA, Dykes DD, Polesky HF. 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J Cyst Fibros. 2008;7(2):110-5.##Kilinc MO, Ninis VN, Dagli E, Demirkol M, Ozkinay F, Arikan Z, et al. Highest heterogeneity for cystic fibrosis: 36 mutations account for 75% of all CF chromosomes in Turkish patients. Am J Med Genet. 2002;113(3):250-7.##Rawashdeh M, Manal H. Cystic fibrosis in Arabs: a prototype from Jordan. Ann Trop Paediatr. 2000;20(4):283-6.##Wahab AA, Janahi IA, Hebi S, al-Hamed M, Kambouris M. Cystic fibrosis in a child from Syria. Ann Trop Paediatr. 2002;22(1):53-5.##Kambouris M, Banjar H, Moggari I, Nazer H, Al-Hamed M, Meyer BF. Identification of novel mutations in Arabs with cystic fibrosis and their impact on the cystic fibrosis transmembrane regulator mutation detection rate in Arab populations. Eur J Pediatr. 2000;159(5):303-9.##el-Harith EA, Dork T, Stuhrmann M, Abu-Srair H, al-Shahri A, Keller KM, et al. Novel and characteristic CFTR mutations in Saudi Arab children with severe cystic fibrosis. 1997;34(12):996-9.##Banjar H. Geographic distribution of cystic fibrosis transmembrane regulator gene mutations in Saudi Arabia. East Mediterr Health J. 1999;5(6):1230-5.##Eskandarani HA. Cystic fibrosis transmembrane regulator gene mutations in Bahrain. J Trop Pediatr. 2002;48(6):348-50.##Lucotte G, Barre E, Berriche S. Frequency of the cystic fibrosis mutation delta F508 in Algeria. Hum Genet. 1991;87(6):759.##Messaoud T, Verlingue C, Denamur E, Pascaud O, Quere I, Fattoum S, et al. Distribution of CFTR mutations in cystic fibrosis patients of Tunisian origin: identification of two novel mutations. Eur J Hum Genet. 1996;4(1):20-4.##Saleheen D, Frossard PM. The cradle of the deltaF508 mutation. J Ayub Med Coll Abbottabad. 2008;20(4):157-60.##Schwartz M, Johansen HK, Koch C, Brandt NJ. Frequency of the delta F508 mutation on cystic fibrosis chromosomes in Denmark. Hum Genet. 1990;85(4):427-8.##McIntosh I, Lorenzo ML, Brock DJ. Frequency of delta F508 mutation on cystic fibrosis chromosomes in UK. Lancet. 1989;2(8676):1404-5.##Gradient of distribution in Europe of the major CF mutation and of its associated haplotype. European Working Group on CF Genetics (EWGCFG). Hum Genet. 1990;85 (4):436-45.##Casals T, Ramos MD, Gimenez J, Larriba S, Nunes V, Estivill X. High heterogeneity for cystic fibrosis in Spanish families: 75 mutations account for 90% of chromosomes. Hum Genet. 1997;101(3):365-70.##Tzetis M, Kanavakis E, Antoniadi T, Doudounakis S, Adam G, Kattamis C. Characterization of more than 85% of cystic fibrosis alleles in the Greek population, including five novelmutations. Hum Genet. 1997;99(1):121-5.##Onay T, Zielenski J, Topaloglu O, Gokgoz N, Kayserili H, Apak MY, et al. Cystic fibrosis mutations and associated haplotypes in Turkish cystic fibrosis patients. Hum Biol. 2001;73(2):191-203.##Castellani C, Cuppens H, Macek M Jr, Cassiman JJ, Kerem E, Durie P, et al. Consensus on the use and interpretation of cystic fibrosis mutation analysis in clinical practice. 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Laparoscopic Approach to a Large Ovarian Fibroma: A Case Report 2 1 555 2 Background: Ovarian fibroma is the most common benign solid tumor of the ovary, which is often difficult to diagnose preoperatively. The choice treatment for ovarian fibroma is surgical removal, but discussions for the operative approach, laparoscopic or open, in the literature seem to be scant. Case Presentation: We presented a unique clinical experience of laparoscopic approach to a case of 15 cm unilateral ovarian fibroma in a 24 year old patient, with a successful and complete resection of the tumor beside ovarian tissue preservation in December 2012 in Rasoul-e-Akram hospital, Tehran, Iran. Conclusion: In conclusion, we should not forget the role of laparoscopy as a diagnostic procedure even in suspicious cases of ovarian fibroma with solid tumor, ascites, and pleural effusion. 57 61 Zahra Z Najmi Zahra Najmi Minimally Invasive Surgery Research Center, Iran University of Medical Sciences Minimally Invasive Surgery Research Center, Iran University of Medical Sciences Iran Abolfazl A Mehdizadehkashi Abolfazl Mehdizadehkashi Endometriosis Research Center, Iran University of Medical Sciences Endometriosis Research Center, Iran University of Medical Sciences Iran amehdizadehkashi@yahoo.com Maryam M Kadivar Maryam Kadivar Department of Pathology, Rasoul-e-Akram Hospital, Iran University of Medical Sciences Department of Pathology, Rasoul-e-Akram Hospital, Iran University of Medical Sciences Iran Zeinab Z Tamannaie Zeinab Tamannaie Minimally Invasive Surgery Research Center, Iran University of Medical Sciences Minimally Invasive Surgery Research Center, Iran University of Medical Sciences Iran Shahla Sh Chaichian Shahla Chaichian Endometriosis Research Center, Iran University of Medical Sciences Endometriosis Research Center, Iran University of Medical Sciences Iran Benign ovarian neoplasmLaparoscopyOvarian fibromaThecoma 555.pdf Chechia A, Attia L, Temime RB, Makhlouf T, Koubaa A. Incidence, clinical analysis, and management of ovarian fibromas and fibrothecomas. Am J Obstet Gynecol. 2008;199(5):473.e1-4.##Nocito AL, Sarancone S, Bacchi C, Tellez T. Ovarian thecoma: clinicopathological analysis of 50 cases. Ann Diagn Pathol. 2008;12(1):12-6.##Gargano G, De Lena M, Zito F, Fanizza G, Mattioli V, Schittulli F. Ovarian fibroma: our experience of 34 cases. Eur J Gynaecol Oncol. 2003;24(5):429-32.##Leung SW, Yuen PM. Ovarian fibroma: a review on the clinical characteristics, diagnostic difficulties, and management options of 23 cases. Gynecol Obstet Invest. 2006;62(1):1-6.##Sivanesaratnam V, Dutta R, Jayalakshmi P. Ovarian fibroma--clinical and histopathological characteristics. Int J Gynaecol Obstet. 1990;33(3):243-7.##Patsner B. Meigs syndrome and "false positive" preoperative serum CA-125 levels: analysis of ten cases. Eur J Gynaecol Oncol. 2000;21(4):362-3.##Son CE, Choi JS, Lee JH, Jeon SW, Hong JH, Bae JW. Laparoscopic surgical management and clinical characteristics of ovarian fibromas. JSLS. 2011;15(1):16-20.##Ball A, Wenning J, Van Eyk N. Ovarian fibromas in pediatric patients with basal cell nevus (Gorlin) syndrome. J Pediatr Adolesc Gynecol. 2011;24(1):e5-7.##Aram S, Moghaddam NA. Bilateral ovarian fibroma associated with Gorlin syndrome. J Res Med Sci. 2009;14(1):57-61.##Seracchioli R, Bagnoli A, Colombo FM, Missiroli S, Venturoli S. Conservative treatment of recurrent ovarian fibromas in a young patient affected by Gorlin syndrome. Hum Reprod. 2001;16(6):1261-3.##Leung SW, Yuen PM. Ovarian fibroma: a review on the clinical characteristics, diagnostic difficulties, and management options of 23 cases. Gynecol Obstet Invest. 2006;62(1):1-6.##Moran-Mendoza A, Alvarado-Luna G, Calderillo-Ruiz G, Serrano-Olvera A, Lopez-Graniel CM, Gallardo-Rincon D. Elevated CA125 level associated with Meigs' syndrome: case report and review of the literature. Int J Gynecol Cancer. 2006;16 Suppl 1:315-8.##Sevinc A, Camci C, Turk HM, Buyukberber S. How to interpret serum CA 125 levels in patients with serosal involvement? A clinical dilemma. Oncology. 2003;65(1):1-6.##Tarcoveanu E, Dimofte G, Niculescu D, Vasilescu A, Moldovanu R, Ferariu D, et al. Ovarian fibroma in the era of laparoscopic surgery: a general surgeon's experience. Acta Chir Belg. 2007;107(6): 664-9.##