Ovarian Rejuvenation: Turning Dreams into Reality 2 1 2 Throughout the history, the desire to appear youthful and delay the signs of aging has always been a common aspiration of mankind. A literature review on historical texts shows that different methods and materials have been used by individuals in past centuries to achieve youthfulness. However, most of these approaches were unscientific, immoral, and even illegal. One organ that undergoes a drastic change by aging is the ovary, subsequentially affecting fertility potential and the ability to conceive. In women, the impact of age on fertility is much more evident compared to men, as testicular activity and sperm production in males persist throughout the adulthood following puberty. Women are born with a limited and constant population of oocytes in their ovaries, which gradually diminishes over time until around the age of 50, marking the onset of a new phase called menopause. Premature ovarian insuffi-ciency (POI) affects approximately 1% of women under 40 and 0.4% of women under 35. The ovary undergoes accelerated aging compared to other tissues and organs, likely due to evolutionary factors. The termination of ovarian function serves as a mechanism to prevent fertility and pregnancy at older age, avoiding further risks for both the mother and the fetus. The onset of menopause is characterized by irregular menstruation cycles, attributed to a decrease in the production of steroid hormones by the ovaries. This decline gives rise to the classic symptoms of menopause, including hot flashes, night sweats, heart palpitations, insomnia, headaches, dysphoria, and various mood changes. Menopause also significantly reduces vaginal secretions and tissue elasticity and increases vaginal acidity, contrib-uting to dyspareunia. Additionally, hormonal and physiological changes of menopause also lead to a gradual decrease in bone mass and quality, resulting in development of osteoporosis. Therefore, the proper function of ovaries guarantees the long-term health and fertility of women (1). Aside from declined fertility, the complications of menopause are very similar to other age-related health issues, resulting from accumulation of cellular and molecular damage in tissues and organs over time. In recent times, there have been contemporary efforts to preserve women's fertility and delay or eliminate menopause-related symptoms through new biomedical technologies. These approaches in-volve repairing damaged cells or replacing them with new ones, particularly utilizing stem cells as well as employing ovarian tissue grafts. These advancements have the potential to significantly prevent or reverse menopause-related complications. A new term that has gained popularity in public and profes-sional discussions, as well as scientific literature, is "ovarian rejuvenation". Ovarian rejuvenation is an example of regenerative medicine, and recent studies have explored using bone marrow derived stem cells, menstrual blood stem cells, as well as platelet-rich plasma (PRP) to restore function and rejuvenate the pre-menopausal or menopausal ovary. These treatments are based on successful identification and isolation of germline stem cells (GSCs) from the ovaries of women in their reproductive age. Additionally, about 5 to 10 percent of POI cases may experience spontaneous follicular growth, menstruation, and pregnancy within a year of diagnosis. This occurrence is attributed to the presence of a number of quiescent primordial follicles or even GSCs in the ovary that are una-ble to resume their development to mature oocytes due to endocrine and paracrine changes during the menstrual cycle. Instead, they require a stronger stimulus to trigger a change in the ovarian niche. Alt-hough the occurrence of de novo oogenesis in adult women has not been proven based on current re-search, the administration of stem cells or growth factors and the resumption of ovarian activity after cessation of ovarian function can put an end to the theory of the ovary as a non-renewable pool of female germ cells (1, 2). With the progress in ovarian rejuvenation, different types of stem cells have been used for quiescent follicles recovery and ovarian niche restoration. Among them, mesenchymal stem cells (MSCs) have emerged as the most widely used, demonstrating promising outcomes. The successful and encouraging results from animal studies involving MSC injection into the ovary have provided a strong foundation for their application in human clinical trials. A diverse range of MSCs from different tissue sources, including MSCs of amniotic fluid, umbilical cord MSC (UCMSCs), menstrual blood-derived stromal cells (MenSCs), adipose tissue derived MSC (ADMSCs), bone marrow-derived stem cells (BMDSCs), and even human embryonic stem cells (ESCs) has been employed for ovarian rejuvenation (2). In addition to the aforementioned approaches, other types of stem cell sources are being investigated in several randomized controlled trials (RCTs) for ovarian rejuvenation in different countries. Howev-er, most of them are still ongoing and their results have not yet been reported. In the latest findings re-ported by our team, intraovarian injection of autologous MenSC led to increased rates of spontaneous pregnancy, elevated levels of anti-Müllerian hormone (AMH), and a higher antral follicle count in the experimental group compared to the controls. Furthermore, in cases treated through ICSI/IVF cycles, MenSC injection resulted in higher number of mature oocytes and embryos (3). MSC therapy is a labor-intensive, time-consuming, and expensive process. There are potential risks associated with in vitro MSC expansion, such as the risk of infection, as well as the risk of graft-versus-host disease (GVHD) and tissue specialization of MSCs. Hence, researchers have been explor-ing more effective methods with fewer side effects and complications. One such method is intraovari-an injection of platelet rich plasma (PRP). PRP has a history of at least 70 years and was initially rec-ognized as the standard platelet concentrate for transfusion in the 1960s. Over the past decades, the use of PRP has gradually expanded and it is now employed in various medical fields including derma-tology, orthopedics, sports medicine, hair and beauty, dentistry, urology, and ophthalmology. The use of PRP in reproductive medicine is relatively new and dates back to recent years. In addition to ovari-an rejuvenation, PRP has been investigated in several studies for the treatment of recurrent implanta-tion failure (RIF), thin endometrium, and endometriosis. Intraovarian administration of PRP, similar to stem cells, has shown the potential to improve AMH levels, decrease serum FSH levels, improve oo-cyte retrieval, increase antral follicle count (AFC), and promote spontaneous pregnancy (2,4). Therefore, intraovarian PRP injection serves as a convenient, cost-effective, and applicable alternative to MSC treatments. PRP promises a bright future for improvement of menopause-related symptoms and ovarian rejuvenation. However, our understanding of its long-term side effects, optimal treatment duration, mechanism of action, and other outcomes remains limited. Therefore, it is crucial to conduct several double-blind randomized clinical trials with long-term follow-up to thoroughly assess the out-comes of ovarian rejuvenation before recommending it as a routine treatment for women with POI and diminished ovarian reserve (DOR). 01 3 Mohammad Reza MR Sadeghi محمدرضا صادقی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran sadeghi@avicenna.ac.ir No Keyword 140219.pdf Sills ES. The scientific and cultural journey to ovarian rejuvenation: background, barriers, and beyond the biological clock. Medicines (Basel). 2021;8(6):29.##Polonio AM, García-Velasco JA, Herraiz S. Stem cell paracrine signaling for treatment of premature ovarian insufficiency. Front Endocrinol (Lausanne). 2021;11:626322.##Zafardoust S, Kazemnejad S, Fathi-Kazerooni M, Darzi M, Sadeghi MR, Sadeghi Tabar A, et al. The effects of intraovarian injection of autologous menstrual blood-derived mesenchymal stromal cells on pregnancy outcomes in women with poor ovarian response. Stem Cell Res Ther. 2023;14(1):332.##Serdarogullari M, Raad G, Makieva S, Liperis G, Fraire-Zamora JJ, Celik-Ozenci C. Revitalising female fertility: platelet-rich plasma (PRP) - hype or hope? Reprod Biomed Online. 2024. In press. ##

Metabolic Fingerprinting of Serum and Seminal Plasma of Testicular Cancer Patients Using Raman Spectroscopy: A Pilot Study 2 1 2 Background: Testicular cancer (TC) is a relatively rare type of cancer in men. Early diagnosis of TC remains challenging. Metabolomics holds promise in offering valuable insights in this regard. In this study, a metabolic fingerprinting approach was employed to identify potential biomarkers in both serum and seminal plasma of TC patients. Methods: A total of 9 patients with testicular cancer and 10 controls were included in the study. The metabolic fingerprinting approach was utilized as a rapid diagnostic tool to analyze the metabolome in serum and seminal plasma of TC patients in comparison to fertile men. Raman spectroscopy was applied for the analysis of metabolites in these biological samples. Results: Principal component analysis (PCA) and functional group analysis showed that the differentiation between serum samples from healthy men and TC patients was not possible. However, when analyzing seminal plasma, a significant difference was found between the two groups (p<0.05). Functional group analysis of serum only showed an increase in tryptophan concentration ratio in TC patients as compared to healthy men (p=0.03). In contrast, in seminal plasma of TC patients, this increase was observed in all analyzed compounds, including phenylalanine, tyrosine, lipids, proteins, phenols (p<0.001). Conclusion: Our study highlights the potential of metabolic fingerprinting as a fast diagnostic tool for screening TC patients, with seminal plasma serving as a valuable biological sample. Furthermore, several potential biomarkers, particularly phenylalanine, were identified in seminal plasma. This research contributes to our understanding of TC pathogenesis and has the potential to pave the way for early detection and personalized treatment approaches. 03 12 Niknam N Lakpour نیکنام لک‌پور Oncopathology Research Center, Iran University of Medical Sciences Oncopathology Research Center, Iran University of Medical Sciences Iran Mohammad Reza MR Sadeghi محمدرضا صادقی Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Naser N Jafarzadeh Department of Medical Physics, Tarbiat Modares University Department of Medical Physics, Tarbiat Modares University Iran Ralf R Henkel Department of Metabolism, Digestion and Reproduction, Imperial College London Department of Metabolism, Digestion and Reproduction, Imperial College London UK Azadeh A Hajiparvaneh Avicenna Fertility Center, Avicenna Research Institute, ACECR, Tehran, Iran Avicenna Fertility Center, Avicenna Research Institute, ACECR, Tehran, Iran Iran Zohreh Z Fathi Avicenna Fertility Center, Avicenna Research Institute, ACECR, Tehran, Iran Avicenna Fertility Center, Avicenna Research Institute, ACECR, Tehran, Iran Iran Roya R Ghods رویا قدس Oncopathology Research Center, Iran University of Medical Sciences Oncopathology Research Center, Iran University of Medical Sciences Iran ghods.ro@iums.ac.ir, rghods77@yahoo.com Kambiz K Gilany کامبیز گیلانی Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Iran k.gilany@avicenna.ac.ir Zahra Z Madjd Oncopathology Research Center, Iran University of Medical Sciences Oncopathology Research Center, Iran University of Medical Sciences Iran majdjabari.z@iums.ac.ir, Zahra.madjd@yahoo.com Metabolic fingerprintingRaman spectroscopySeminal plasmaSerumTesticular cancer 140218.pdf Cheng L, Albers P, Berney DM, Feldman DR, Daugaard G, Gilligan T, et al. Testicular cancer. Nat Rev Dis Primers. 2018;4(1):29.##Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69(1):7-34.##Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;136(5):E359-86.##Ferlay J, Soerjomataram I, Ervik M, Dikshit R, Eser S, Mathers C, et al. GLOBOCAN 2012 v1. 0, cancer incidence and mortality worldwide. IARC. Cancer Base. 2013. 11.##Milardi D, Grande G, Vincenzoni F, Pierconti F, Pontecorvi A. Proteomics for the identification of biomarkers in testicular cancer-review. Front Endocrinol (Lausanne). 2019;10:462.##Leman ES, Gonzalgo ML. Prognostic features and markers for testicular cancer management. Indian J Urol. 2010;26(1):76-81.##McGlynn KA, Cook MB. Etiologic factors in testicular germ-cell tumors. Future Oncol. 2009;5(9):1389-402.##Adra N, Einhorn LH. Testicular cancer update. Clin Adv Hematol Oncol. 2017;15(5):386-96.##Koši Kunac A, Gnjidić M, Antunac Golubić Z, Gamulin M. Treatment of germ cell testicular cancer. Acta Clin Croat. 2020;59(3):496-504.##Daugaard G, Gundgaard MG, Mortensen MS, Agerbæk M, Holm NV, Rørth M, et al. Surveillance for stage I nonseminoma testicular cancer: outcomes and long-term follow-up in a population-based cohort. J Clin Oncol. 2014;32(34):3817-23.##Mortensen MS, Lauritsen J, Gundgaard MG, Agerbæk M, Holm NV, Christensen IJ, et al. A nationwide cohort study of stage I seminoma patients followed on a surveillance program. Eur Urol. 2014;66(6):1172-8.##Kier MG, Lauritsen J, Mortensen MS, Bandak M, Andersen KK, Hansen MK, et al. Prognostic factors and treatment results after bleomycin, etoposide, and cisplatin in germ cell cancer: a population-based study. Eur Urol. 2017;71(2):290-8.##Nicholson BD, Jones NR, Protheroe A, Joseph J, Roberts NW, Van den Bruel A, et al. The diagnostic performance of current tumour markers in surveillance for recurrent testicular cancer: a diagnostic test accuracy systematic review. Cancer Epidemiol. 2019;59:15-21.##Lakpour N, Saliminejad K, Ghods R, Reza Sadeghi M, Pilatz A, Khosravi F, et al. Potential biomarkers for testicular germ cell tumour: risk assessment, diagnostic, prognostic and monitoring of recurrence. Andrologia. 2021;53(4):e13998.##Lakpour N, Ghods R, Sadeghi MR, Ranjbar MM, Abolhasani M, Kiani J, et al. Production and characterization of a new specific monoclonal antibody against A‐isoform of SALL4: a novel emerging testicular cancer marker. Andrologia. 2022;54(1):e14608.##Fiehn O. Metabolomics--the link between genotypes and phenotypes. Plant Mol Biol. 2002;48(1-2):155-71.##Weckwerth W. Metabolomics in systems biology. Ann Rev Plant Biol. 2003;54:669-89.##Goodacre R. Metabolomics of a superorganism. J Nutr. 2007;137(1 Suppl):259S-66S.##Shulaev V. Metabolomics technology and bioinformatics. Brief Bioinform. 2006;7(2):128-39.##Liu X, Locasale JW. Metabolomics: a primer. Trends Biochem Sci. 2017;42(4):274-84.##Kumamoto Y, Harada Y, Takamatsu T, Tanaka H. Label-free molecularimaging and analysis by raman spectroscopy. Acta Histochem Cytochem. 2018;51(3):101-10.##Zhang S, Qi Y, Tan SP, Bi R, Olivo M. Molecular fingerprint detection using raman and infrared spectroscopy technologies for cancer detection: a progress review. Biosensors (Basel). 2023;13(5):557.##Kong K, Kendall C, Stone N, Notingher I. Raman spectroscopy for medical diagnostics--from in-vitro biofluid assays to in-vivo cancer detection. Adv Drug Deliv Rev. 2015;89:121-34.##Jones RR, Hooper DC, Zhang L, Wolverson D, Valev VK. Raman techniques: fundamentals and frontiers. Nanoscale Res Lett. 2019;14(1):231.##Ferlay J, Colombet M, Soerjomataram I, Parkin DM, Piñeros M, Znaor A, et al. Cancer statistics for the year 2020: an overview. Int J Cancer. 2021.##Gilany K, Mani‐Varnosfaderani A, Minai‐Tehrani A, Mirzajani F, Ghassempour A, Sadeghi MR, et al. Untargeted metabolomic profiling of seminal plasma in nonobstructive azoospermia men: A noninvasive detection of spermatogenesis. Biomed Chromatogr. 2017;31(8).##Gilany K, Moazeni‐Pourasil RS, Jafarzadeh N, Savadi‐Shiraz E. Metabolomics fingerprinting of the human seminal plasma of asthenozoospermic patients. Mol Reprod Dev. 2014;81(1):84-6.##Yu B, Ge M, Li P, Xie Q, Yang L. Development of surface-enhanced Raman spectroscopy application for determination of illicit drugs: towards a practical sensor. Talanta. 2019;191:1-10.##Ugboma HA, Aburoma HL. Public awareness of testicular cancer and testicular self-examination in academic environments: a lost opportunity. Clinics (Sao Paulo). 2011;66(7):1125-8.##Garolla A, De Giorgi U, Milardi D. Editorial: testicular cancer: new insights on the origin, genetics, treatment, fertility, general health, quality of life and sexual function. Front Endocrinol (Lausanne). 2020;11:41.##Cao D, Li J, Guo CC, Allan RW, Humphrey PA. SALL4 is a novel diagnostic marker for testicular germ cell tumors. Am J Surg Pathol. 2009;33(7):1065-77.##Lakpour N, Ghods R, Sadeghi MR, Ranjbar MM, Abolhasani M, Kiani J, et al. Production and characterization of a new specific monoclonal antibody against A-isoform of SALL4: a novel emerging testicular cancer marker. Andrologia. 2022;54(11):e14608.##Jiang S, Liu Y, Xu Y, Sang X, Lu X. Research on liquid biopsy for cancer: a bibliometric analysis. Heliyon. 2023;9(3):e14145.##Fernández-Lázaro D, García Hernández JL, García AC, Córdova Martínez A, Mielgo-Ayuso J, Cruz-Hernández JJ. Liquid biopsy as novel tool in precision medicine: origins, properties, identification and clinical perspective of cancer's biomarkers. Diagnostics (Basel). 2020;10(4):215.##Siravegna G, Marsoni S, Siena S, Bardelli A. Integrating liquid biopsies into the management of cancer. Nat Rev Clin Oncol. 2017;14(9):531-48.##Beger RD, Dunn W, Schmidt MA, Gross SS, Kirwan JA, Cascante M, et al. Metabolomics enables precision medicine: "a white paper, community perspective". Metabolomics. 2016;12(10):149.##Worley B, Powers R. Multivariate analysis in metabolomics. Curr Metabolomics. 2013;1(1):92-107.##Vasseur S, Guillaumond F. Lipids in cancer: a global view of the contribution of lipid pathways to metastatic formation and treatment resistance. Oncogenesis. 2022;11(1):46.##Platten M, Nollen EA, Röhrig UF, Fallarino F, Opitz CA. Tryptophan metabolism as a common therapeutic target in cancer, neurodegeneration and beyond. Nat Rev Drug Discov. 2019;18(5):379-401.##Drabovich AP, Saraon P, Jarvi K, Diamandis EP. Seminal plasma as a diagnostic fluid for male reproductive system disorders. Nat Rev Urol. 2014;11(5):278-88.##Agarwal A, Saleh RA, Bedaiwy MA. Bedaiwy, Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril. 2003;79(4):829-43.##Aykin-Burns N, Ahmad IM, Zhu Y, Oberley LW, Spitz DR. Increased levels of superoxide and H2O2 mediate the differential susceptibility of cancer cells versus normal cells to glucose deprivation. Biochem J. 2009;418(1):29-37.##Gupta-Elera G, Garrett AR, Robison RA, O’Neill KL. The role of oxidative stress in prostate cancer. Eur J Cancer Prev. 2012;21(2):155-62.##Trachootham D, Lu W, Ogasawara MA, Valle NR, Huang P. Redox regulation of cell survival. Antioxid Redox Signal. 2008;10(8):1343-74.##Akbari Z, Dijojin RT, Zamani Z, Hosseini RH, Arjmand M. Aromatic amino acids play a harmonizing role in prostate cancer: a metabolomics-based cross-sectional study. Int J Reprod Biomed. 2021;19(8):741-50.##Pietkiewicz D, Klupczynska-Gabryszak A, Plewa S, Misiura M, Horala A, Miltyk W, et al. Free amino acid alterations in patients with gynecological and breast cancer: a review. Pharmaceuticals (Basel). 2021;14(8):731.##Neurauter G, Grahmann AV, Klieber M, Zeimet A, Ledochowski M, Sperner-Unterweger B, et al. Serum phenylalanine concentrations in patients with ovarian carcinoma correlate with concentrations of immune activation markers and of isoprostane-8. Cancer Lett. 2008;272(1):141-7.##Sharifi-Rad J, Seidel V, Izabela M, Monserrat-Mequida M, Sureda A, Ormazabal V, et al. Phenolic compounds as Nrf2 inhibitors: potential applications in cancer therapy. Cell Commun Signal. 2023;21(1):89.##

Evaluation of Sperm DNA Fragmentation Using Halosperm Technique after the Freezing–Thawing Process in Men: A Study on the Validation of the SCD Protocol 2 1 2 Background: DNA fragmentation index (DFI) enhances routine semen analysis by providing valuable insights into male reproductive potential. Utilizing Halosperm test, a sperm chromatin dispersion (SCD) assay based on induced condensation. The purpose of this study was to assess sperm DNA damage both before and after freezing. By following the specified kit instructions, an attempt was made to validate the SCD test protocol, with a particular emphasis on the implications of sperm freezing on its DNA integrity. Methods: In total, 380 fresh human semen samples from normozoospermic patients were frozen at -20°C for 10 days, using SCD cryopreservation reagent. Routine semen analysis and DNA fragmentation index (DFI) were determined for each sample before freezing and after thawing. Semen morphology and sperm DFI were compared before and after freezing/thawing process. Results: There was a significant decrease in sperm normal morphology after thawing (9.31±2.42% vs. 7.1±1.53%, p<0.05, respectively). The sperm head, midpiece, and tail defect rate increased after freezing at -20°C. Moreover, DFI was significantly higher after thawing compared to before freezing (20.71±1.61% before freezing vs. 29.1±0.21% after thawing with p<0.001). Conclusion: Cryoconservation of semen samples at -20°C for 10 days using SCD cryopreservation reagent seems to damage sperm morphology, resulting in a reduction in sperm DNA integrity. The measurement of DFI on a fresh sample remains the most reliable technique for obtaining accurate results. 12 20 Chaymae Ch Rochdi Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Morocco c.rochdi@ump.ac.ma Larbi L Allai Laboratory of Sustainable Agriculture Management, Higher School of Technology Sidi Bennour, Chouaib Doukkali University Laboratory of Sustainable Agriculture Management, Higher School of Technology Sidi Bennour, Chouaib Doukkali University Morocco Ibtissam I Bellajdel Medically Assisted Procreation Unit, Central Laboratory, Mohammed VI University Hospital Center Medically Assisted Procreation Unit, Central Laboratory, Mohammed VI University Hospital Center Morocco Hafsa H Taheri Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Morocco Hanane H Saadi Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Morocco Ahmed A Mimouni Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Morocco Mohammed M Choukri Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Maternal-Child and Mental Health Research Laboratory, Faculty of Medicine and Pharmacy, Mohammed First University Morocco CryopreservationDNA fragmentationFreezingHalosperm testSpermatozoaThawing 140215.pdf Björndahl L, Brown JK. The sixth edition of the WHO laboratory manual for the examination and processing of human semen: ensuring quality and standardization in the basic examination of human ejaculates. Fertil Steril. 2022;117(2):246-51.##Sharma A, Minhas S, Dhillo WS, Jayasena CN. Male infertility due to testicular disorders. J Clinl Endocrinol Metab. 2021;106(2):e442-e59.##Bosch E, De Vos M, Humaidan P. The future of cryopreservation in assisted reproductive technologies. Front Endocrinol (Lausanne). 2020;11:67.##Gilbert K, Nangia AK, Dupree JM, Smith JF, Mehta A. Fertility preservation for men with testicular cancer: is sperm cryopreservation cost effective in the era of assisted reproductive technology? Urol Oncol. 2018;36(3):92.e1-92.e9.##Taylor MJ, Weegman BP, Baicu SC, Giwa SE. New approaches to cryopreservation of cells, tissues, and organs. Transfus Med Hemother. 2019;46(3):197-215.##Alahmar AT, Singh R, Palani A. Sperm DNA fragmentation in reproductive medicine: a review. J Hum Reprod Sci. 2022;15(3):206-18.##Le MT, Nguyen TTT, Nguyen TT, Nguyen VT, Nguyen TTA, Nguyen VQH, et al. Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects. Eur J Obstet Gynecol Reprod Biol. 2019;234:14-20.##Upadhyay VR, Ramesh V, Dewry RK, Kumar G, Raval K, Patoliya P. Implications of cryopreservation on structural and functional attributes of bovine spermatozoa: an overview. Andrologia. 2021;53(8):e14154.##Mohammadi F, Varanloo N, Heydari Nasrabadi M, Vatannejad A, Amjadi FS, Javedani Masroor M, et al. Supplementation of sperm freezing medium with myoinositol improve human sperm parameters and protects it against DNA fragmentation and apoptosis. Cell Tissue Bank. 2019;20(10):77-86.##Lu JC, Jing J, Chen L, Ge YF, Feng RX, Liang YJ, et al. Analysis of human sperm DNA fragmentation index (DFI) related factors: a report of 1010 subfertile men in China. Reprod Biol Endocrinol. 2018;16(1):23.##Panner Selvam MK, Sengupta P, Agarwal A. Sperm DNA fragmentation and male infertility. In: Arafa M, Elbardisi H, Majzoub, A, Agarwal A, editors. Genetics of male infertility. Switzerland: Springer, Cham; 2020. p. 155-72.##Chua SC, Yovich SJ, Hinchliffe PM, Yovich JL. How well do semen analysis parameters correlate with sperm DNA fragmentation? a retrospective study from 2567 semen samples analyzed by the Halosperm test. J Pers Med. 2023;13(3):518.##Le MT, Nguyen TT, Nguyen TT, Van Nguyen T, Nguyen TA, Nguyen QH, et al. Does conventional freezing affect sperm DNA fragmentation? Clin Exp Reprod Med. 2019;46(2):67-75.##Rochdi Ch, Bellzjdel I, El Moudane A, El Assri S, Mamri S, Taheri H, et al. The effects of varicocelectomy on sperm DNA fragmentation and conventional semen parameters in men with severe oligoasthenoteratozoospermia. Int J Fertil Steril. 2023. In press.##Zhang F, Li J, Liang Z, Wu J, Li L, Chen C, et al. Sperm DNA fragmentation and male fertility: a retrospective study of 5114 men attending a reproductive center. J Assist Reprod Genet. 2021;38(5):1133-41.##Kim SM, Kim SK, Jee BC, Kim SH. Effect of sperm DNA fragmentation on embryo quality in normal responder women in in vitro fertilization and intracytoplasmic sperm injection. Yonsei Med J. 2019;60(5):461-6.##Lusignan MF, Li X, Herrero B, Delbes G, Chan PTK. Effects of different cryopreservation methods on DNA integrity and sperm chromatin quality in men. Andrology. 2018;6(6):829-35.##Ozimic S, Ban-Frangez H, Stimpfel M. Sperm cryopreservation today: approaches, efficiency, and pitfalls. Curr Issues Mol Biol. 2023;45(6):4716-34.##Raju R, Bryant SJ, Wilkinson BL, Bryant G. The need for novel cryoprotectants and cryopreservation protocols: Insights into the importance of biophysical investigation and cell permeability. Biochim Biophys Acta Gen Subj. 2021;1865(1):129749.##Thomson LK, Fleming SD, Barone K, Zieschang JA, Clark AM. The effect of repeated freezing and thawing on human sperm DNA fragmentation. Fertil Steril. 2010;93(4):1147-56.##Lehti MS, Sironen A. Formation and function of sperm tail structures in association with sperm motility defects. Biol Reprod. 2017;97(4):522-36.##Zandiyeh S, Shahverdi A, Ebrahimi B, Sabbaghian M. A novel approach for human sperm cryopreservation with AFPIII. Reprod Biol. 2020;20(2):169-74.##Spano M, Cordelli E, Leter G, Lombardo F, Lenzi A, Gandini L. Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay. Mol Hum Reprod. 1999;5(1):29-37.##Isachenko E, Isachenko V, Katkov II, Rahimi G, Schondorf T, Mallmann P, et al. DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectant‐free vitrification. Hum Reprod. 2004;19(4):932-9.##Duru NK, Morshedi M, Schuffner A, Oehninger S. Cryopreservation-thawing of fractionated human spermatozoa and plasma membrane translocation of phosphatidylserine. Fertil Steril. 2001;75(2):263-8.##Moghadam MT, Fard YA, Saki G, Nikbakht R. Effect of vitamin D on apoptotic marker, reactive oxygen species and human sperm parameters during the process of cryopreservation. Iran J Basic Med Sci. 2019;22(9):1036-43.##Thu NTM, Trang NT. Comparison of the sperm testing kit with the Halosperm testing kit in an analysis of sperm DNA fragmentation. Vietnam J Sci Technol Eng. 2020;62(2):71-6.##Cankut S, Dinc T, Cincik M, Ozturk G, Selam B. Evaluation of sperm DNA fragmentation via halosperm technique and TUNEL assay before and after cryopreservation. Reprod Sci. 2019;26(12):1575-81.##

DEFB126 2-nt Deletion (rs11467417) as a Potential Risk Factor for Chlamydia Trachomatis Infection and Subsequent Infertility in Iranian Men 2 1 2 Background: Chlamydia trachomatis (CT) is one of the most prevalent sexually transmitted infections, causing genital tract infections and infertility. Defensins have an immunomodulatory function and play an important role in sperm maturation, motility, and fertilization. DEFB126 is present on ejaculated spermatozoa and is essential for them to pass through the female reproductive tract. The purpose of the current study was to determine the frequency of the 2-nt deletion of the DEFB126 (rs11467417) in Iranian infertile males with a recurrent history of CT. Methods: Semen samples of 1080 subfertile males were investigated. Among patients who had CT-positive results, sperm DNA from 50 symptomatic and 50 asymptomatic patients were collected for the DEFB126 genotype analysis. Additionally, a control group comprising 100 DNA samples from individuals with normal spermogram and testing negative for CT was included in the study. The PCR-sequencing technique for detecting the 2-nt deletion of the second exon of the DEFB126 was performed. Results: The Chi-squared test comparing all three groups revealed no significant difference across the different genotypes. Moreover, no significant difference between the symptomatic and asymptomatic groups was seen. However, analysis within CT-positive patients and controls demonstrated significant difference between the frequencies of homozygous del/del. Conclusion: The higher frequency of the 2-nt deletion of the DEFB126 in CT-positive patients suggests that the occurrence of mutations in the DEFB-126 may cause the impairment of the antimicrobial activity of the DEFB126 protein and consequently makes individuals more susceptible to infections such as CT. 20 28 Kaveh K Haratian Department of Pathology and Laboratory Medicine, Western University Department of Pathology and Laboratory Medicine, Western University Canada Parnaz P Borjian Boroujeni Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran Marjan M Sabbaghian Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran Elham E Maghareh Abed Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran Maedeh M Moazenchi Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran Anahita A Mohseni Meybodi Department of Pathology and Laboratory Medicine, Western University Department of Pathology and Laboratory Medicine, Western University Canada Anahita.MohseniMeybodi@lhsc.on.ca Chlamydia infectionDefensin geneInfertility 140212.pdf Aitken RJ. The changing tide of human fertility. Hum Reprod. 2022;37(4):629-38.##World Health Organization. Infections, pregnancies, and infertility: perspectives on prevention. World Health Organization. Fertil Steril. 1987;47(6):964-8.##Brunham RC, Rekart ML. Considerations on Chlamydia trachomatis disease expression. FEMS Immunol Med Microbiol. 2009;55(2):162-6.##Zhai YJ, Feng Y, Ma X, Ma F. Defensins: defenders of human reproductive health. Hum Reprod Update. 2023;29(1):126-54.##Tollner TL, Bevins CL, Cherr GN. Multifunctional glycoprotein DEFB126—a curious story of defensin-clad spermatozoa. Nat Rev Urol. 2012;9 (7):365-75.##Tollner TL, Venners SA, Hollox EJ, Yudin AI, Liu X, Tang G, et al. A common mutation in the defensin DEFB126 causes impaired sperm function and subfertility. Sci Transl Med. 2011;3(92):92ra65.##Patrão MTCC, Silva EJR, Avellar MCWA. Androgens and the male reproductive tract: an overview of classical roles and current perspectives. Arq Bras Endocrinol Metabol. 2009;53(8):934-45.##Yenugu S, Hamil KG, Radhakrishnan Y, French FS, Hall SHJE. The androgen-regulated epididymal sperm-binding protein, human β-defensin 118 (DEFB118)(formerly ESC42), is an antimicrobial β-defensin. Endocrinology. 2004;145(7):3165-73.##Zhao Y, Diao H, Ni Z, Hu S, Yu H, Zhang Y, et al. The epididymis-specific antimicrobial peptide β-defensin 15 is required for sperm motility and male fertility in the rat (Rattus norvegicus). Cell Mol Life Sci. 2011;68(4):697-708.##Fleming DC, King AE, Williams AR, Critchley HOD, Kelly RW. Hormonal contraception can suppress natural antimicrobial gene transcription in human endometrium. Fertil Steril. 2003;79(4):856-63.##Cao D, Li Y, Yang R, Wang Y, Zhou Y, Diao H, et al. Lipopolysaccharide-induced epididymitis disrupts epididymal beta-defensin expression and inhibits sperm motility in rats. Biol Reprod. 2010;83(6):1064-70.##Quayle AJ, Porter EM, Nussbaum AA, Wang YM, Brabec C, Yip KP, et al. Gene expression, immunolocalization, and secretion of human defensin-5 in human female reproductive tract. Am J Pathol. 1998;152(5):1247-58.##Wilson SS, Wiens ME, Smith JG. Antiviral mechanisms of human defensins. J Mol Biol. 2013;425(24):4965-80.##Pivarcsi A, Nagy I, Koreck A, Kis K, Kenderessy-Szabo A, Szell M, et al. Microbial compounds induce the expression of pro-inflammatory cytokines, chemokines and human β-defensin-2 in vaginal epithelial cells. Microbes Infect. 2005;7(9-10):1117-27.##Diao R, Fok KL, Chen H, Yu MK, Duan Y, Chung CM, et al. Deficient human β-defensin 1 underlies male infertility associated with poor sperm motility and genital tract infection. Sci Transl Med. 2014;6(249):249ra108.##Xu D, Lu W. Defensins: a double-edged sword in host immunity. Front Immunol. 2020;11:764.##Yamaguchi Y, Nagase T, Makita R, Fukuhara S, Tomita T, Tominaga T, et al. Identification of multiple novel epididymis-specific β-defensin isoforms in humans and mice. J Immunol. 2002;169(5):2516-23.##Gilks CB, Reid PE, Clement PB, Owen DA. Histochemical changes in cervical mucus-secreting epithelium during the normal menstrual cycle. Fertil Steril. 1989;51(2):286-91.##Tollner TL, Yudin AI, Tarantal AF, Treece CA, Overstreet JW, Cherr GN. Beta-defensin 126 on the surface of macaque sperm mediates attachment of sperm to oviductal epithelia. Biol Reprod. 2008;78(3):400-12.##Toshimori K, Araki S, Öra C, Eddy EJ. Loss of sperm surface sialic acid induces phagocytosis: an assay with a monoclonal antibody T21, which recognizes a 54K sialoglycoprotein. Arch Androl. 1991;27(2):79-86.##Yudin AI, Treece CA, Tollner TL, Overstreet JW, Cherr GN. The carbohydrate structure of DEFB126, the major component of the cynomolgus Macaque sperm plasma membrane glycocalyx. J Membr Biol. 2005;207(3):119-29.##Hollox EJ, Barber JC, Brookes AJ, Armour JAJ. Defensins and the dynamic genome: what we can learn from structural variation at human chromosome band 8p23. Genome Res. 2008;18(11):1686-97.##Frischmeyer PA, Van Hoof A, O'Donnell K, Guerrerio AL, Parker R, Dietz HCJS. An mRNA surveillance mechanism that eliminates transcripts lacking termination codons. Science. 2002;295(5563):2258-61.##Van Hoof A, Frischmeyer PA, Dietz HC, Parker R. Exosome-mediated recognition and degradation of mRNAs lacking a termination codon. Science. 2002;295(5563):2262-4.##Moazenchi M, Totonchi M, Salman Yazdi R, Hratian K, Mohseni Meybodi M, Ahmadi Panah M, et al. The impact of Chlamydia trachomatis infection on sperm parameters and male fertility: A comprehensive study. Int J STD AIDS. 2018;29(5):466-73.##Boroujeni PB, Ebrahimian S, Abedini M, Chayjan MR, Hassani M, Gourabi H, et al. The role of DEFB126 variation in male infertility and medically assisted reproduction technique outcome. Reprod Biomed Online. 2019;39(4):649-57.##Wollenhaupt J, Kolbus F, Weissbrodt H, Schneider C, Krech T, Zeidler HJC, et al. Manifestations of Chlamydia induced arthritis in patients with silent versus symptomatic urogenital chlamydial infection. Clin Exp Rheumatol. 1995;13(4):453-8.##Cunningham KA, Beagley KW. Male genital tract chlamydial infection: implications for pathology and infertility. Biol Reprod. 2008;79(2):180-9.##Matzuk MM, Lamb DJ. Genetic dissection of mammalian fertility pathways. Nat Med. 2002;4 Suppl:s41-9.##Idahl A, Abramsson L, Kumlin U, Liljeqvist J, Olofsson JI. Male serum Chlamydia trachomatis IgA and IgG, but not heat shock protein 60 IgG, correlates with negatively affected semen characteristics and lower pregnancy rates in the infertile couple. Int J Androl. 2007;30(2):99-107.##Veznik Z, Pospisil L, Svecova D, Zajicova A, Unzeitig V. Chlamydiae in the ejaculate: their influence on the quality and morphology of sperm. Acta Obstet Gynecol Scand. 2004;83(7):656-60.##Vigil P, Morales P, Tapia A, Riquelme R, Salgado AM. Chlamydia trachomatis infection in male partners of infertile couples: incidence and sperm function. Andrologia. 2002;34(3):155-61.##Lehrer RI, Lu W. α‐Defensins in human innate immunity. Immunol Rev. 2012;245(1):84-112.##Wiechuła B, Cholewa K, Ekiel A, Romanik M, Doleżych H, Martirosian G. [HBD-1 and hBD-2 are expressed in cervico-vaginal lavage in female genital tract due to microbial infections]. Ginekol Pol. 2010;81(4):268-71. Polish.##Wiechuła BE, Friedek DA, Ekiel AM, Romanik MK, Martirosian G. Human neutrophil peptides in vaginitis/cervicitis of different etiology. Pol J Microbiol. 2007;56(3):185-9.##Wiesenfeld HC, Heine RP, Krohn MA, Hillier SL, Amortegui AA, Nicolazzo M, et al. Association between elevated neutrophil defensin levels and endometritis. J Infect Dis. 2002;186(6):792-7.##Mukura LR, Hickey DK, Rodriguez‐Garcia M, Fahey JV, Wira CR. Chlamydia trachomatis regulates innate immune barrier integrity and mediates cytokine and antimicrobial responses in human uterine ECC‐1 epithelial cells. Am J Reprod Immunol. 2017;78(6).##Noda-Nicolau NM, Bastos LB, Bolpetti AN, Pinto GVS, Marcolino LD, Marconi C, et al. Cervicovaginal levels of human β-defensin 1, 2, 3, and 4 of reproductive-aged women with Chlamydia trachomatis infection. J Low Genit Tract Dis. 2017;21(3):189-92.##Fichorova RN, Morrison CS, Chen PL, Yamamoto HS, Govender Y, Junaid D, et al. Aberrant cervical innate immunity predicts onset of dysbiosis and sexually transmitted infections in women of reproductive age. PLoS One. 2020;15(1):e0224359.##Porter E, Yang H, Yavagal S, Preza GC, Murillo O, Lima H, et al. Distinct defensin profiles in Neisseria gonorrhoeae and Chlamydia trachomatis urethritis reveal novel epithelial cell-neutrophil interactions. Infect Immun. 2005;73(8):4823-33.##Xin A, Cheng L, Diao H, Wu Y, Zhou S, Shi C, et al. Lectin binding of human sperm associates with DEFB126 mutation and serves as a potential biomarker for subfertility. Sci Rep. 2016;6:20249.##Liu H, Yu H, Gu Y, Xin A, Zhang Y, Diao H, et al. Human beta-defensin DEFB126 is capable of inhibiting LPS-mediated inflammation. Appl Microbiol Biotechnol. 2013;97(8):3395-408.##Diao H, Yu HG, Sun F, Zhang YL, Tanphaichitr N. Rat recombinant β-defensin 22 is a heparin-binding protein with antimicrobial activity. Asian J Androl. 2011;13(2):305-11.##Silva EJR, Ribeiro CM, Mirim AF, Silva AA, Romano RM, Hallak J, et al. Lipopolysaccharide and lipotheicoic acid differentially modulate epididymal cytokine and chemokine profiles and sperm parameters in experimental acute epididymitis. Sci Rep. 2018;8(1):103.##He JY, Peng JY, Li QF, Lin XL, Cui YR, Ma SY, et al. DEFB126 polymorphisms and association with idiopathic asthenozoospermia in China. Asian J Androl. 2022;24(6):607-14.##Aram R, Chan PT, Cyr DG. Beta-defensin126 is correlated with sperm motility in fertile and infertile men. Biol Reprod. 2020;102(1):92-101.##

Serum Testosterone-Estradiol Ratio in Toxoplasma-Seropositive Infertile Men: A Prospective, Single-Center Study 2 1 2 Background: The purpose of the current study was to compare the testosterone-estradiol (T:E2) ratio in Toxoplasma gondii (T. gondii) seropositive infertile men with seropositive and seronegative normozoospermic controls. Methods: A total of 200 men with normal virilization, including 100 with idiopathic infertility and 100 normozoospermic men, were included in the study. Participants underwent a comprehensive evaluation including a detailed medical history assessment, physical examination, semen analysis, testing for T. gondii IgM/IgG, and estimation of serum T:E2 ratios. Statistical comparisons between groups were performed using t-test and Chi square (x2) with a significance level set at p<0.05. Results: Infertile cases were diagnosed with oligozoospermia (63%), oligoasthenozoospermia (34%), and oligoasthenoteratozoospermia (3%). Regarding anti-Toxoplasma IgG and IgM antibodies, among infertile men, 34 tested positive for IgG and 8 tested positive for IgM. Among the 34 infertile men who tested positive for IgG antibodies, 13 individuals (38.2%) had disturbed T:E2 ratios. Also, among the 12 IgG-positive controls, 5 individuals (41.7%) had disturbed T:E2 ratios (p=0.834). However, only 2 out of the 83 seronegative controls (2.5%) had disturbed T:E2 ratios, which was statistically significant (p<0.001). Furthermore, 6 out of 8 IgM-positive infertile men had altered T:E2 ratios, compared to 3 out of 5 IgM-positive controls (p=0.568) and 2 out of 83 seronegative controls (p<0.001). The T:E2 ratio was significantly lower (8.68±1.95) among IgM-positive infertile men and higher (13.04±3.78) among IgG-positive infertile men when compared to seronegative controls (10.45±0.54) (p<0.001). There were no significant differences in the testosterone-estradiol (T:E2) ratios between infertile men with positive IgM or IgG serology and the control group with the same serology. Conclusion: A significant proportion of infertile men with toxoplasmosis exhibit altered T:E2 ratios. These findings emphasize the importance of conducting anti-T. gondii-IgG testing in individuals with abnormal T:E2 ratios. 28 38 Ahmed A Ragab Department of Andrology, Sexology and STIs, Faculty of Medicine, Beni-Suef University Department of Andrology, Sexology and STIs, Faculty of Medicine, Beni-Suef University Egypt drahmedragab1981@ gmail.com, ahmed.abdeltawab@ med.bsu.edu.eg Doaa DA Hamdy Department of Medical Parasitology, Faculty of Medicine, Beni-Suef University Department of Medical Parasitology, Faculty of Medicine, Beni-Suef University Egypt Shimaa ShS Ibrahim Department of Medical Parasitology, Faculty of Medicine, Beni-Suef University Department of Medical Parasitology, Faculty of Medicine, Beni-Suef University Egypt AsthenozoospermiaEstradiolImmunoglobulin GImmunoglobulin MMale infertilityOligospermiaTestosteroneToxoplasma 140217.pdf Devroey P, Fauser BC, Diedrich K; Evian annual reproduction (EVAR) workshop group 2008. Approaches to improve the diagnosis and management of infertility. Hum Reprod Update. 2009;15(4):391-408.##Peterson BD, Gold L, Feingold T. The experience and influence of infertility: considerations for couple Counselors. Fam J. 2007;15(3):251-7.##Krausz C. Male infertility: pathogenesis and clinical diagnosis. Best Pract Res Clin Endocrinol Metab. 2011;25(2):271-85.##Aquila S, Sisci D, Gentile M, Carpino A, Middea E, Catalano S, et al. Towards a physiological role for cytochrome P450 aromatase in ejaculated human sperm. Hum Reprod. 2003;18(8):1650-9.##Luboshitzky R, Kaplan-Zverling M, Shen-Orr Z, Nave R, Herer P. Seminal plasma androgen/oestrogen balance in infertile men. Int J Androl. 2002;25(6):345-51.##Yamamoto M, Hibi H, Katsuno S, Miyake K. Serum estradiol levels in normal men and men with idiopathic infertility. Int J Urol. 1995;2(1):44-6.##Schlegel PN. Aromatase inhibitors for male infertility. Fertil Steril. 2012;98(6):1359-62.##Castelló-Porcar AM, Martínez-Jabaloyas JM. Testosterone/estradiol ratio, is it useful in the diagnosis of erectile dysfunction and low sexual desire? Aging Male. 2016;19(4):254-8.##Gregoriou O, Bakas P, Grigoriadis C, Creatsa M, Hassiakos D, Creatsas G. Changes in hormonal profile and seminal parameters with use of aromatase inhibitors in management of infertile men with low testosterone to estradiol ratios. Fertil Steril. 2012;98(1):48-51.##Hlaváčová J, Flegr J, Řežábek K, Calda P, Kaňková Š. Association between latent toxoplasmosis and fertility parameters of men. Andrology. 2021;9(3):854-62.##Singh S. Congenital toxoplasmosis: clinical features, outcomes, treatment, and prevention. Trop Parasitol. 2016;6(2):113-22.##Flegr J, Klapilová K, Kaňková S. Toxoplasmosis can be a sexually transmitted infection with serious clinical consequences. Not all routes of infection are created equal. Med Hypotheses. 2014;83(3):286-9.##Kaňková Š, Hlaváčová J, Flegr J. Oral sex: a new, and possibly the most dangerous, route of toxoplasmosis transmission. Med Hypotheses. 2020;141:109725.##Robert-Gangneux F, Dardé ML. Epidemiology of and diagnostic strategies for toxoplasmosis. Clin Microbiol Rev. 2012;25(2):264-96.##Garg M, Stern D, Groß U, Seeberger PH, Seeber F, Varón Silva D. Detection of anti-toxoplasma gondii antibodies in human sera using synthetic glycosylphosphatidylinositol glycans on a bead-based multiplex assay. Anal Chem. 2019;91(17):11215-22.##Dalimi A, Abdoli A. Toxoplasma gondii and male reproduction impairment: A new Aspect of Toxoplasmosis Research. Jundishapur J Microbiol. 2013;6(8):e7184.##Dongmei X, Zhou Y, Diao W. Preliminary investigation on relationship between spermatogenic cells apoptosis and infection of toxoplasma gondii in male infertility. Chinese J Schistomiasis. 2005;17(6):451-53.##Mohammed NS, Al-Muhsin A, Al-khayat FA, Hussien SK. The impact of Toxoplasma gondii on mitochondrial DNA of sub-fertile men sperms. Biomed Pharmacol J. 2017;10(2).##Dardona Z, Amane M, Boussaa S. Toxoplasmosis-related psychological, behavioral, neurological, and hormonal changes: a literature review. Eur J Med Health Sci. 2023;5(5):128-44.##Shirbazou S, Abasian L, Meymand FT. Effects of Toxoplasma gondii infection on plasma testosterone and cortisol level and stress index on patients referred to Sina hospital, Tehran. Jundishapur J Microbiol. 2011;4(3):167-73.##Hodková H, Kolbeková P, Skallová A, Lindová J, Flegr J. Higher perceived dominance in Toxoplasma infected men--a new evidence for role of increased level of testosterone in toxoplasmosis-associated changes in human behavior. Neuro Endocrinol Lett. 2007;28(2):110-4.##Flegr J, Lindová J, Pivoňková V, Havlíček J. Brief communication: latent toxoplasmosis and salivary testosterone concentration-important confounding factors in second to fourth digit ratio studies. Am J Phys Anthropol. 2008;137(4):479-84.##Oktenli C, Doganci L, Ozgurtas T, Araz RE, Tanyuksel M, Musabak U, et al. Transient hypogonadotrophic hypogonadism in males with acute toxoplasmosis: suppressive effect of interleukin-1 on the secretion of GnRH. Hum Reprod. 2004;19(4):859-66.##Zhang X, Liu J, Li M, Fu Y, Zhang T, Han Q, et al. Role of an estradiol regulatory factor-hydroxysteroid dehydrogenase (HSD) in Toxoplasma gondii infection and pathogenicity. J Steroid Biochem Mol Biol. 2017;174:176-82.##World medical association. World medical association declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA. 2013;310(20):2191-4.##World Health Organization (WHO). WHO laboratory manual for the examination and processing of human semen. 5th ed. Geneva: WHO; 2010. 271 p.##Farhan BA. Seroprevelance of toxoplasma gondii (IgM, IgG) among aborted women in some Karbala hospital by using ELISA technique and microscopic examination. Indian J Foren Med Toxicol. 2020;14.1.##van Koeverden ID, de Bakker M, Haitjema S, van der Laan SW, de Vries JPM, Hoefer IE, et al. Testosterone to oestradiol ratio reflects systemic and plaque inflammation and predicts future cardiovascular events in men with severe atherosclerosis. Cardiovasc Res. 2019;115(2):453-62.##Pavlovich CP, King P, Goldstein M, Schlegel PN. Evidence of a treatable endocrinopathy in infertile men. J Urol. 2001;165(3):837-41.##Abdoli A. Toxoplasma, testosterone, and behavior manipulation: The role of parasite strain, host variations, and intensity of infection. Front Biol. 2014;9(2):151-60.##Zghair KH, Al-Qadhi BN, Mahmood SH. The effect of toxoplasmosis on the level of some sex hormones in males blood donors in Baghdad. J Parasit Dis. 2015;39(3):393-400.##Al-Kurdy MJ, A’aiz NN, Jawad TI. Study the Effect of Chronic Toxoplasmosis on Serum Testosterone Level in Men. Indian J Forensic Med Toxicol. 2020;14(4):3132-7.‏##Qi R, Su XP, Gao XL, Liang XL. [Toxoplasma infection in males with sterility in Shenyang, China]. Zhonghua Nan Ke Xue. 2005;11(7):503-4. Chinese.##Zhou YH, Lu YJ, Wang RB, Song LM, Shi F, Gao QF, et al. [Survey of infection of Toxoplasma gondii in infertile couples in Suzhou countryside]. Zhonghua Nan Ke Xue. 2002;8(5):350-2. Chinese.##Muhsin SS, Mamdooh AM, Baseer AN. Seroprevalance study of toxoplasmosis among males in Al-Ruasfa institute of management in Baghdad province-Iraq. J Pharm Biol Sci. 2018;13(2):22-6.##Rostami A, Seyyedtabaei SJ, Aghamolaie S, Behniafar H, Lasjerdi Z, Abdolrasouli A, et al. Seroprevalence and risk factors associated with Toxoplasma gondii infection among rural communities in northern Iran. Rev Inst Med Trop Sao Paulo. 2016;58:70.##Kawashima T, Khin-Sane-Win, Kawabata M, Barzaga N, Matsuda H, Konishi E. Prevalence of antibodies to Toxoplasma gondii among urban and rural residents in the Philippines. Southeast Asian J Trop Med Public Health. 2000;31(4):742-6.##Abdulla HE, Al-bashier NM, Al-kawaz U, Al-Shuwaikh AM, Abood AS, Cross-sectional study of infertile males with toxoplasmosis in Baghdad province. Int J Sci Eng Res. 2015;6(1):254.##Eslamirad Z, Hajihossein R, Ghorbanzadeh B, Alimohammadi M, Mosayebi M, Didehdar M. Effects of toxoplasma gondii infection in level of serum testosterone in males with chronic toxoplasmosis. Iran J Parasitol. 2013;8(4):622-6.##Obaid HM, Ahmed FF, Ismaeol AN. Some hormonal assay in toxoplasma infected university students. Kirkuk Univ J Sci Stud. 2016;11(3):66-80.##Abbasian L. Role of toxoplasma gondii infection in serum level of testosterone. Kowsar Med J. 2011;16(2):123-7.##Kanková S, Kodym P, Flegr J. Direct evidence of toxoplasma induced changes in serum testosterone in mice. Exp Parasitol. 2011;128(3):181-3.##James WH: Potential solutions to problems posed by the offspring sex ratio of people with parasitic and viral infection. Folia Parasitol (Praha). 2010;57(2):114-20.##Alardi MH. The role of toxoplasma gondii in concentration of some sex hormones in infertile individual. J Adv Med Pharm Sci. 2011;5(1):72-82.##Flegr J, Lindová J, Kodym P. Sex-dependent toxoplasmosis-associated differences in testosterone concentration in humans. Parasitology. 2008;135(4):427-31.##Khan AA, Araujo FG, Brighty KE, Gootz TD, Remington JS. Anti-toxoplasma gondii activities and structure-activity relationships of novel fluoroquinolones related to trovafloxacin. Antimicrob Agents Chemother. 1999;43(7):1783-7.##Babu SR, Sadhnani MD, Swarna M, Padmavathi P, Reddy PP. Evaluation of FSH, LH and testosterone levels in different subgroups of infertile males. Indian J Clin Biochem. 2004;19(1):45-9.##Roberts CW, Walker W, Alexander J. Sex-associated hormones and immunity to protozoan parasites. Clin Microbiol Rev. 2001;14(3):476-88.##Harper JM, Huynh MH, Coppens I, Parussini F, Moreno S, Carruthers VB. A cleavable propeptide influences toxoplasma infection by facilitating the trafficking and secretion of the TgMIC2-M2AP invasion complex. Mol Biol Cell. 2006;17(10):4551-63.##Montoya JG, Liesenfeld O. Toxoplasmosis. Lancet. 2004;363(9425):1965-76.##

The Association of AMH Level with the Number and Quality of Oocytes in Women Undergoing IVF/ICSI: A Single-Center Study 2 1 2 Background: The recognized role of Anti-Müllerian hormone (AMH) as a marker for women's biological age and ovarian reserve prompts debate on its efficacy in predicting oocyte quality during IVF/ICSI. Recent findings challenging this view compelled us to conduct this study to examine the correlation between AMH levels and quantity/quality of oocytes in IVF/ICSI procedures. Methods: The data were collected retrospectively from the medical records of 320 women between 25-42 years old. The included patients were divided into two groups: the high AMH group (>1.1 ng/ml) and the low AMH (=<1.1 ng/ml) group. The high AMH group comprised 213 patients, while the low AMH group consisted of 107 patients. Spearman's correlation coefficient and Multinomial logistic regression were computed to assess the relationships between different variables. Results: Significant positive correlations were detected between AMH level and the number of aspirated follicles (rho=0.741, p<0.001), retrieved oocytes (rho=0.659, p<0.001), M2 oocytes (rho=0.624, p<0.001), grade A embryos (rho=0.419, p<0.001), and grade AB embryos (rho=0.446, p<0.001. In contrast, AMH levels had negative associations with the number and duration of cycles (p<0.05). AMH emerged as a statistically significant independent predictor of the number of M2 oocytes. Conclusions: Serum AMH level could represent the quantity and quality of oocytes following IVF/ICSI treatments. Future studies should aim to delve deeper into the correlations between AMH levels and both the quality and quantity of embryos. Additionally, it would be beneficial to consider the influence of sperm factors, as well as assess pregnancy rates. 38 46 Zohreh Z Heidary Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Iran Masoumeh M Masoumi Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Iran Mohadese M Dashtkoohi Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Iran Niusha N Sharifinejad Non-communicable Diseases Research Center, Alborz University of Medical Sciences Non-communicable Diseases Research Center, Alborz University of Medical Sciences Iran Masoumeh M Dehghan Tarzjani Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Iran Marjan M Ghaemi Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Iran Batool B Hossein Rashidi بتول حسین رشیدی Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences Iran bhrashidi@gmail.com Anti- mullerian hormoneIn vitro fertilizationIntracytoplasmic sperm injectionOocytes 140214.pdf Patrizio P, Sakkas D. From oocyte to baby: a clinical evaluation of the biological efficiency of in vitro fertilization. Fertil Steril. 2009;91(4):1061-6.##Farhi J, Cohen K, Mizrachi Y, Weissman A, Raziel A, Orvieto R. Should ICSI be implemented during IVF to all advanced-age patients with non-male factor subfertility? Reprod Biol Endocrinol. 2019;17(1):30.##Cai QF, Wan F, Huang R, Zhang HW. Factors predicting the cumulative outcome of IVF/ICSI treatment: a multivariable analysis of 2450 patients. Hum Reprod. 2011;26(9):2532-40.##Ingilizova G, Ivanov D, Kovachev E, Evrev M, Kostov I, Necheva V. [Oocyte quality as a predictive marker for assessment of IVF/ICSI procedure outcome]. Akush Ginekol (Sofiia). 2014;53(6):41-6.##Li A, Zhang J, Kuang Y, Yu C. Analysis of IVF/ICSI-FET outcomes in women with advanced endometriosis: influence on ovarian response and oocyte competence. Front Endocrinol (Lausanne). 2020;11:427.##Lasiene K, Lasys V, Glinskyte S, Valanciute A, Vitkus A. Relevance and methodology for the morphological analysis of oocyte quality in IVF and ICSI. J Reprod Stem Cell Biotechnol. 2011;2(1):1-13.##Weenen C, Laven JS, Von Bergh AR, Cranfield M, Groome NP, Visser JA, et al. Anti-Müllerian hormone expression pattern in the human ovary: potential implications for initial and cyclic follicle recruitment. Mol Hum Reprod. 2004;10(2):77-83.##Wiweko B, Prawesti DMP, Hestiantoro A, Sumapraja K, Natadisastra M, Baziad A. Chronological age vs biological age: an age-related normogram for antral follicle count, FSH and anti-Mullerian hormone. J Assisted Reprod Genet. 2013;30(12):1563-7.##La Marca A, Stabile G, Artenisio AC, Volpe A. Serum anti-Mullerian hormone throughout the human menstrual cycle. Hum Reprod. 2006;21(12):3103-7.##Broekmans FJ, Faddy MJ, Scheffer G, te Velde ER. Antral follicle counts are related to age at natural fertility loss and age at menopause. Menopause. 2004;11(6 Pt 1):607-14.##Smeenk JM, Sweep FC, Zielhuis GA, Kremer JA, Thomas CM, Braat DD. Antimüllerian hormone predicts ovarian responsiveness, but not embryo quality or pregnancy, after in vitro fertilization or intracyoplasmic sperm injection. Fertil Steril. 2007;87(1):223-6.##Yılmaz T, Tavukçuoğlu Ş, Taşdemir S, Esenkaya S, İrez T. Can follicular fluid anti-mullerian hormone level be a determinant of pregnancy in women under 35 years of age? Int J Womens Health Reprod Sci. 2018;6(1):6-10.##Papas M, Govaere J, Peere S, Gerits I, Van de Velde M, Angel-Velez D, et al. Anti-Müllerian hormone and OPU-ICSI outcome in the mare. Animals (Basel). 2021;11(7):2004.##Morin SJ, Patounakis G, Juneau CR, Neal SA, Scott RT, Seli E. Diminished ovarian reserve and poor response to stimulation in patients <38 years old: a quantitative but not qualitative reduction in performance. Hum Reprod. 2018;33(8):1489-98.##Chang CC, Shapiro D, Toledo AA, Sadruddin S, Wright G, Nagy ZP. Does AMH correlate with oocyte quality obtained from donors? Fertil Steril. 2018;110(4):e228.##Park HJ, Lyu SW, Seok HH, Yoon TK, Lee WS. Anti-Müllerian hormone levels as a predictor of clinical pregnancy in in vitro fertilization/intracytoplasmic sperm injection-embryo transfer cycles in patients over 40 years of age. Clin Exp Reprod Med. 2015;42(4):143-8.##Harris ID, Roth L, Brinker K, McShane P, Wang S, Alvero R. Determining the optimal cut-off value for antimullerian hormone in ovarian reserve testing. Fertil Steril. 2011;96(3):S193.##Zhang JJ, Yang M, Merhi Z. Efficiency of metaphase II oocytes following minimal/mild ovarian stimulation in vitro fertilization. Fertil Res Pract. 2016;2:2.##Tesarik J, Junca AM, Hazout A, Aubriot FX, Nathan C, Cohen-Bacrie P, et al. Embryos with high implantation potential after intracytoplasmic sperm injection can be recognized by a simple, non-invasive examination of pronuclear morphology. Hum Reprod. 2000;15(6):1396-9.##Sun XY, Lan YZ, Liu S, Long XP, Mao XG, Liu L. Relationship between anti-müllerian hormone and in vitro fertilization-embryo transfer in clinical pregnancy. Front Endocrinol (Lausanne). 2020;11:595448.##Zhao D, Fan J, Wang P, Jiang X, Yao J, Li X. Age-specific definition of low anti-Mullerian hormone and associated pregnancy outcome in women undergoing IVF treatment. BMC Pregnancy Childbirth. 2021;21(1):186.##Irez T, Ocal P, Guralp O, Cetin M, Aydogan B, Sahmay S. Different serum anti-Müllerian hormone concentrations are associated with oocyte quality, embryo development parameters and IVF-ICSI outcomes. Arch Gynecol Obstet. 2011;284(5):1295-301.##Park HJ, Lyu SW, Seok HH, Yoon TK, Lee WS. Anti-müllerian hormone levels as a predictor of clinical pregnancy in in vitro fertilization/intracytoplasmic sperm injection-embryo transfer cycles in patients over 40 years of age. Clin Exp Reprod Med. 2015;42(4):143-8.##Lawal OI, Yusuff J-dO. Demographic, lifestyle, and reproductive determinants of serum anti-Müllerian hormone levels in adult women of reproductive age in Ilorin, North-Central Nigeria. Middle East Fertil Soci J. 2021;26(1):23.##Ghomian N, Lotfalizadeh M, Malihe A, Keshtan F. Association between Anti-Mullerian Hormone and Ovarian Response and Pregnancy in Polycystic Ovary Syndrome Patients. Int J Women's Health Reprod Sci. 2018;7:49-53.##Bakhtiari M, Babaei T, Safarpour M, Esmaeili M, Esmaeili M, Asgari R, et al. Investigating the factors affecting the ICSI (microinjection) success in infertile people referred to an infertility treatment center in western Iran from 2011 to 2017. J Obstet Gynecol Cancer Res. 2020;5(3):115-25.##Parsanezhad ME, Jahromi BN, Rezaee S, Kooshesh L, Alaee S. The effect of four different gonadotropin protocols on oocyte and embryo quality and pregnancy outcomes in IVF/ICSI cycles; a randomized controlled trial. Iran J Med Sci. 2017;42(1):57-65.##Yang YC, Li YP, Pan SP, Chao KH, Chang CH, Yang JH, et al. The different impact of stimulation duration on oocyte maturation and pregnancy outcome in fresh cycles with GnRH antagonist protocol in poor responders and normal responders. Taiwan J Obstet Gynecol. 2019;58(4):471-6.##Dai X, Wang Y, Yang H, Gao T, Yu C, Cao F, et al. AMH has no role in predicting oocyte quality in women with advanced age undergoing IVF/ICSI cycles. Sci Rep. 2020;10(1):19750.##Dahan MH, Tannus S. Believing that transferring more embryos will result in increased pregnancy rates: a flawed concept: a SWOT analysis. Middle East Fertil Soci J. 2020;25(1):32.##McLernon DJ, Harrild K, Bergh C, Davies MJ, de Neubourg D, Dumoulin JCM, et al. Clinical effectiveness of elective single versus double embryo transfer: meta-analysis of individual patient data from randomised trials. BMJ. 2010;341:c6945.##Sun TC, Zhou SJ, Song LL, Li JH, Chen X, Tian L. High anti-müllerian hormone levels might not reflect the likelihood of clinical pregnancy rate in IVF/ICSI treatment. JBRA Assist Reprod. 2021;25(2):266-71.##Sahmay S, Oncul M, Tuten A, Tok A, Acıkgoz AS, Cepni I. Anti-müllerian hormone levels as a predictor of the pregnancy rate in women of advanced reproductive age. J Assist Reprod Genet. 2014;31(11):1469-74.##

Dose Sildenafil Citrate Reduce the Incidence of Emergency Cesarean Section and Fetal Distress During Labor? A Randomized Double-Blinded Clinical Trial 2 1 2 Background: Fetal distress (FD) is one of the most frequent causes of emergency cesarean section (CS) due to the insufficient uteroplacental blood supply during labor. There is a theory that Sildenafil citrate (SC) may improve the uteroplacental blood supply and decrease fetal hypoxia and FD. In this study, the effect of SC on the incidence of FD and emergency C/S was analyzed. Methods: In a randomized double-blinded clinical trial, a total of 208 low-risk subjects who met our stringent inclusion criteria were randomly assigned into two groups: the Sildenafil citrate group (n=104) and the placebo group (n=104). These participants were referred to our referral gynecology and obstetrics department for delivery between July 2022 to September 2022. The SC group received oral SC at a dose of 50 mg every 6 hr, up to a maximum of three times. The final maternal-fetal-neonatal results were recorded and all data were analyzed using SPSS version 23. Results: The mean age of mothers was 28.98±5.6 years and 120 cases were primigravid (57.7%). Out of a total of 208 pregnant subjects, 168 subjects delivered through normal vaginal delivery (80.8%) and 40 cases underwent emergency CS (19.2%). The number of NVD in Sildenafil group was significantly more than placebo group (87.5% versus 74%) and SC decreased the rate of emergency CS to 87.5% (RR=2.46%, 95%CI 1.19-5.08). Also, SC decreased the rate of FD to 53.8% (RR=2.83%, 95%CI of 1-8.24). The hemoglobin level was significantly higher in SC group 6 hr after delivery (11.06±1.26 versus 10.75±1.14, respectively; p=0.049). Also, higher Apgar scores at both the 1st and 5th min after delivery were observed in SC group (p=0.001). Conclusion: The results showed that SC can effectively decrease the rate of emergency CS and FD during labor. 46 56 Seyedeh Hajar SH Sharami Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Iran Forozan F Milani Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Iran forozan.pnc@gmail.com Roya R Kabodmehri Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Iran Misa M Naghdipour Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Iran Azade A Mahmoudi Isaabadi Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Iran Zahra Z Haghparast Ghadim-Limudahi Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Reproductive Health Research Center, Department of Obstetrics and Gynecology, School of Medicine, Al-Zahra Hospital, Guilan University of Medical Sciences Iran Cesarean sectionFetal distressLaborSildenafil citrate 140213.pdf Kale I. Does continuous cardiotocography during labor cause excessive fetal distress diagnosis and unnecessary cesarean sections? J Matern Fetal Neonatal Med. 2022;35(6):1017-22.##Lakhno IV, Uzel K. Diagnosing antenatal fetal distress. Ginekol Pol. 2021.##Maybin JA. Hypoxia and reproductive health. Reproduction. 2021;161(1):E1-2.##Turner JM, Mitchell MD, Kumar SS. The physiology of intrapartum fetal compromise at term. Am J Obstet Gynecol. 2020;222(1):17-26.##Evans MI, Britt DW, Eden RD, Evans SM, Schifrin BS. Earlier and improved screening for impending fetal compromise. J Matern Fetal Neonatal Med. 2022;35(15):2895-903.##Dunn L, Flenady V, Kumar S. Reducing the risk of fetal distress with sildenafil study (RIDSTRESS): a double-blind randomised control trial. J Transl Med. 2016;14:15.##Umar BU, Haque M. Growing concern over rising caesarean section rates: Is it a problem for low-and middle-income countries only? Adv Hum Biol. 2022;12(2):93.##Betran AP, Ye J, Moller AB, Souza JP, Zhang J. Trends and projections of caesarean section rates: global and regional estimates. BMJ Glob Health. 2021;6(6):e005671.##Eyi EGY, Mollamahmutoglu L. An analysis of the high cesarean section rates in Turkey by Robson classification. J Matern Fetal Neonatal Med. 2021;34(16):2682-92.##Buhimschi CS, Garfield RE, Weiner CP, Buhimschi IA. The presence and function of phosphodiesterase type 5 in the rat myometrium. Am J Obstet Gynecol. 2004;190(1):268-74.##Hafeez MA, Kortam AM, Youssef AM, Reda A, Abdelrahman RM. Effect of nitric oxide donors on uterine and sub-endometrial blood flow in patients with unexplained infertility: a randomized controlled trial. Int J Reprod Contraception Obstet Gynecol. 2021;10(6):2178-85.##Bahaa HA. Efficacy of sildenafil citrate in women with unexplained recurrent miscarriage preconceptional and during 1st trimester of pregnancy. Evid Based Women's Health J. 2018;8(1):138-44.##Russo FM, Benachi A, Van Mieghem T, De Hoon J, Van Calsteren K, Annaert P, et al. Antenatal sildenafil administration to prevent pulmonary hypertension in congenital diaphragmatic hernia (Stop-PH): study protocol for a phase I/IIb placenta transfer and safety study. Trials. 2018;19(1):524.##Ghaleb MM, Labib YS, Wahba KA. Sildenafil plus low dose aspirin for prevention of preeclampsia: a randomized controlled trial. Open J Obstet Gynecol. 2021;11(02):189.##Mohammadi E, Teymoordash SN, Norouzi AR, Norouzi F, Norouzi HR. Comparison of the effect of nifedipine alone and the combination of nifedipine and sildenafil in delaying preterm labor: a randomized clinical trial. J Family Report Health. 2021;15(2):112-7.##Maher MA, Sayyed TM, El-Khadry SW. Nifedipine alone or combined with sildenafil citrate for management of threatened preterm labor: a randomized trial. BJOG. 2018;126(6):729-35.##Maged M, Wageh A, Shams M, Elmetwally A. Use of sildenafil citrate in cases of intrauterine growth restriction (IUGR); a prospective trial. Taiwan J Obstet Gynecol. 2018;57(4):483-6.##Hessami K, Cozzolino M, Shamshirsaz AA. The effect of phosphodiesterase-5 inhibitors on uteroplacental and fetal cerebral perfusion in pregnancies with fetal growth restriction: a systematic review and meta-analysis. Eur J Obstet Gynecol Reprod Biol. 2021;267:129-36.##Nath J, Chaube S, Rani N, Verma P. A study on sildenafil citrate in the treatment of IUGR & Oligohydramnios. Invest Gynecol Res Women’s Health. 2021;4(2):431-5.##Russo FM, De Bie F, Hodges R, Flake A, Deprest J. Sildenafil for antenatal treatment of congenital diaphragmatic hernia: from bench to bedside. Curr Pharm Des. 2019;25(5):601-8.##Turner J, Kumar S. Neurodevelopmental outcomes in infants following intrapartum maternal oral sildenafil citrate treatment. Am J Obstet Gynecol. 2021;224(3):316-7.##Turner J, Dunn L, Tarnow-Mordi W, Flatley C, Flenady V, Kumar S. Safety and efficacy of sildenafil citrate to reduce operative birth for intrapartum fetal compromise at term: a phase 2 randomized controlled trial. Am J Obstet Gynecol. 2020;222(5):401-14.##Turner JM, Russo F, Deprest J, Mol BW, Kumar S. Phosphodiesterase-5 inhibitors in pregnancy: Systematic review and meta-analysis of maternal and perinatal safety and clinical outcomes. BJOG. 2022;129(11):1817-31.##Ahmed WS, Geethakumari AM, Biswas KH. Phosphodiesterase 5 (PDE5): Structure-function regulation and therapeutic applications of inhibitors. Biomed Pharmacother. 2021;134:111128.##G Shilkrut A, C Hsu R, M Fuks A. Fetal heart rate tracing category II: a broad category in need of stratification. Neoreviews. 2021;22(2):e88-94.##Krause BJ. Novel insights for the role of nitric oxide in placental vascular function during and beyond pregnancy. J Cell Physiol. 2021;236(12):7984-99.##Wareing M, Myers JE, O’Hara M, Baker PN. Sildenafil citrate (Viagra) enhances vasodilatation in fetal growth restriction. J Clin Endocrinol Metab. 2005;90(5):2550-5.##Dastjerdi MV, Hosseini S, Bayani L. Sildenafil citrate and uteroplacental perfusion in fetal growth restriction. J Res Med Sci. 2012;17(7):632-6.##Dunn L, Greer R, Flenady V, Kumar S. Sildenafil in pregnancy: a systematic review of maternal tolerance and obstetric and perinatal outcomes. Fetal Diagn Ther. 2017;41(2):81-8.##

Evaluating the Plausibility of Euploid Embryos Transfer on Day-5 by Reanalysis of Day-3 Single Aneuploid Embryos: A Case Series 2 1 2 Background: During preimplantation development, single aneuploidies are more commonly tolerated than complex aneuploidies. Some studies have reported that blastocysts with aneuploid karyotypes on Day-3 embryo biopsy can exhibit a normal karyotype on Day-5 rebiopsy, suggesting that single aneuploidies may have a higher likelihood of presenting a normal karyotype on Day-5. The purpose of the current study was to assess the benefit of reanalyzing the karyotypes of Day-3 single aneuploid embryos on Day-5. Methods: Day-3 and Day-5 biopsies of preimplantation embryos were subjected to array comparative genomic hybridization (aCGH). A proof of concept case series study was conducted involving 13 Day-5 embryos from 4 couples across 3 ART centers, collected between October 2019 and June 2020. Each center provided one normal embryo and 3-4 embryos with single aneuploidy based on Day-3 aCGH results. The karyotype of each Day-5 embryo was compared with its corresponding Day-3 karyotype. Results: Among the 10 embryos with single aneuploidy on Day-3, 3 (30%) exhibited discordant karyotypes on Day-5, while the remaining 7 single aneuploid embryos and 3 normal embryos maintained the same karyotype from Day-3 to Day-5. None of the Day-3 single aneuploid embryos displayed a normal karyotype on Day-5. Conclusion: Contrary to previous reports suggesting the potential correction of single aneuploidies in some embryos, the findings of this study did not support such a possibility in the analyzed embryos. Genomic reanalysis of Day-3 single aneuploid embryos on Day-5 does not appear to be a reliable method for identifying euploid embryos suitable for transfer. 56 60 Masood M Bazrgar Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran mbazrgar@royaninstitute.org Roxana R Kariminejad Kariminejad-Najmabadi Pathology and Genetics Center Kariminejad-Najmabadi Pathology and Genetics Center Iran Poopak P Eftekhari-Yazdi Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran Hamid H Gourabi Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Iran AneuploidyBiopsyBlastocystCleavage stagePreimplantation 140209.pdf Van Montfoort A, Carvalho F, Coonen E, Kokkali G, Moutou C, Rubio C, et al. ESHRE PGT consortium data collection XIX–XX: PGT analyses from 2016 to 2017. Hum Reprod Open. 2021;2021(3):hoab024.##Bolton H, Graham SJ, Van der Aa N, Kumar P, Theunis K, Fernandez Gallardo E, et al. Mouse model of chromosome mosaicism reveals lineage-specific depletion of aneuploid cells and normal developmental potential. Nat Commun. 2016;7:11165.##Bazrgar M, Gourabi H, Valojerdi MR, Yazdi PE, Baharvand H. Self-correction of chromosomal abnormalities in human preimplantation embryos and embryonic stem cells. Stem Cells Dev. 2013;22(17):2449-56.##McCoy RC, Demko ZP, Ryan A, Banjevic M, Hill M, Sigurjonsson S, et al. Evidence of selection against complex mitotic-origin aneuploidy during preimplantation development. PLoS Genet. 2015;11(10):e1005601.##Vega M, Breborowicz A, Moshier EL, McGovern PG, Keltz MD. Blastulation rates decline in a linear fashion from euploid to aneuploid embryos with single versus multiple chromosomal errors. Fertil Steril. 2014;102(2):394-8.##Capalbo A, Bono S, Spizzichino L, Biricik A, Baldi M, Colamaria S, et al. Sequential comprehensive chromosome analysis on polar bodies, blastomeres and trophoblast: insights into female meiotic errors and chromosomal segregation in the preimplantation window of embryo development. Hum Reprod. 2013;28(2):509-18.##Liñán A, Lawrenz B, El Khatib I, Bayram A, Arnanz A, Rubio C, et al. Clinical reassessment of human embryo ploidy status between cleavage and blastocyst stage by Next Generation Sequencing. PLoS One. 2018;13(8):e0201652.##Bazrgar M, Gourabi H, Eftekhari-Yazdi P, Vazirinasab H, Fakhri M, Hassani F, et al. The effect of prolonged culture of chromosomally abnormal human embryos on the rate of diploid cells. Int J Fertil Steril. 2016;9(4):563-73.##Huang J, Zhao N, Wang X, Qiao J, Liu P. Chromosomal characteristics at cleavage and blastocyst stages from the same embryos. J Assist Reprod Genet. 2015;32(5):781-7.##Munné S, Alikani M, Ribustello L, Colls P, Martínez-Ortiz PA, McCulloh DH, et al. Euploidy rates in donor egg cycles significantly differ between fertility centers. Hum Reprod. 2017;32(4): 743-9.##

Successful Treatment of Sperm DNA Fragmentation Through Ayurveda Rasayana Therapy: A Case Study 2 1 2 Background: Sperm DNA fragmentation (SDF) can affect fertilization rate and embryo development, making it a useful measure for assessing male fertility. Available evidence supports the association between high sperm DNA fragmentation and poor outcomes, with regard to natural conception. Several treatment options are being adopted with varying degrees of success. Some of the commonly used treatment options are the intake of oral antioxidants, varicocele repair, and techniques like micromanipulation-based sperm selection and use of testicular sperm for intracytoplasmic sperm injection. Case Presentation: Studies have shown that around 29% of couples depend on complementary and alternative medicine (CAM) modality for the treatment of infertility. However, there is a lack of substantial evidence regarding its efficacy in treating various aspects of infertility in couples. The current case report is about a 44 year-old male patient with infertility, who has a known diagnosis of sex chromosome abnormalities. Meanwhile, the SDF study reports indicated the presence of chromosomal abnormalities. This patient was treated exclusively with Ayurveda therapy aimed towards qualitative improvement in reproductive tissues (Shukra Dhatu as per Ayurveda). Patient was assessed periodically for changes in chromosomal abnormality. After four months of treatment, the evaluations demonstrated the presence of completely normal chromosomes. Conclusion: This case study indicates the potential of Ayurveda therapy in treating cases of male infertility caused by DNA fragmentation. Furthermore, observations and systematically designed clinical trials are warranted to establish a stronger level of evidence before making further clinical recommendations. 60 66 Vineeta V Bendale Rasayu Ayurved Clinic Rasayu Ayurved Clinic India rasayupublication2020@gmail.com Sreelakshmi S Chaganti Ojas Multispeciality Ayurveda and Panchakarma Centre Ojas Multispeciality Ayurveda and Panchakarma Centre India Rutuja R Pandav Rasayu's Dyumna Women's Clinic Rasayu's Dyumna Women's Clinic India Deepali D Pawar Rasayu's Dyumna Women's Clinic Rasayu's Dyumna Women's Clinic India Complementary therapiesDNA fragmentationIntegrative medicineMale infertility 140210.pdf Jiji V, Priyanka R, Asha ST, Asha Sreedhar. Ayurvedic management of male infertility due to oligospermia and varicocele: a case study. Int J Ayurveda Res. 2020;8(12):11-4.##Agarwal A, Samanta L, Bertolla RP, Durairajanayagam D, Intasqui P. Proteomics in human reproduction- biomarkers for millennials. 1st ed. New York: Springer Cham; 2016. Springer briefs in reproductive biology; p. 21-43.##Alvarez JG. DNA fragmentation in human spermatozoa: significance in the diagnosis and treatment of infertility. Minerva Ginecol. 2003;55(3):233-9.##Agarwal A, Majzoub A, Baskaran S, Selvam MK, Cho CL, Henkel R, et al. Sperm DNA fragmentation: a new guideline for clinicians. World J Mens Health. 2020;38(4):412-71##Tahmasbpour E, Balasubramanian D, Agarwal A. A multi-faceted approach to understanding male infertility: gene mutations, molecular defects and assisted reproductive techniques (ART). J Assist Reprod Genet. 2014;31(9):1115-37.##Choy JT, Eisenberg ML. Male infertility as a window to health. Fertil Steril. 2018;110(5):810-4.##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.##Zini A, Dohle G. Are varicoceles associated with increased deoxyribonucleic acid fragmentation? Fertil Steril. 2011;96(6):1283-7.##Ghazeeri GS, Nassar AH, Younes Z, Awwad JT. Pregnancy outcomes and the effect of metformin treatment in women with polycystic ovary syndrome: an overview. Acta Obstet Gynecol Scand. 2012;91(6):658-78.##Smith JF, Eisenberg ML, Millstein SG, Nachtigall RD, Shindel AW, Wing H, et al. The use of complementary and alternative fertility treatment in couples seeking fertility care: data from a prospective cohort in the United States. Fertil Steril. 2010;93(7):2169-74.##Sharma M, Arya D, Bhagour K, Gupta RS. Natural aphrodisiac and fertility enhancement measures in males: A review. Curr Med Res Pract. 2017;7(2):51-8.##Ruidas B, Sur TK, Pal K, Som Chaudhury S, Prasad P, Sinha K, et al. Herbometallic nano-drug inducing metastatic growth inhibition in breast cancer through intracellular energy depletion. Mol Biol Rep. 2020;47(5):3745-63.##Paladhi A, Rej A, Sarkar D, Singh R, Bhattacharyya S, Sarkar PK, et al. Nanoscale diamond-based formulation as an immunomodulator and potential therapeutic for lymphoma. Front Pharmacol. 2022;13:852065##Guruprasad K, Mascarenhas R, Satyamoorthy K. Studies on brahma rasayana in male swiss albino mice: chromosomal aberrations and sperm abnormalities. J Ayurveda Integr Med. 2010;1:40-4.##Fainberg J, Kashanian JA. Recent advances in understanding and managing male infertility. F1000Res. 2019;8:F1000 Faculty Rev-670.##Sharma R, Agarwal A, Rohra VK, Assidi M, Abu Elmagd M, Turki RF. Effects of increased paternal age on sperm quality, reproductive outcome and associated epigenetic risks to offspring. Reprod Biol Endocrinol. 2015;13:35.##Sarkar PK, Das S, Prajapati PK. Ancient concept of metal pharmacology based on Ayurvedic literature. Anc Sci Life. 2010;29(4):1-6.##Godatwar P, Prasad B, Skandhan K, Mehra B, Singh G, Mounika B. Effect of Suvarna Bhasma (Gold Calx) on Reproductive System of Male Albino Rats. Int J Life Sci Pharma Res. 2021;11(6): 61-5.##Gopinath H, Shivashankar M. A study on toxicity and anti-hyperglycemic effects of Abhrak Bhasma in rats. J Ayurveda Integr Med. 2021;12(3):443-51.##Khoobchandani M, Katti KK, Karikachery AR, Thipe VC, Srisrimal D, Dhurvas Mohandoss DK, et al. New approaches in breast cancer therapy through green nanotechnology and nano-ayurvedic medicine–pre-clinical and pilot human clinical investigations. Int J Nanomedicine. 2020;15:181-97.##Bendale NY, Kadam A, Patil A, Birari‐Gawande P. Complete tumor regression with exclusive Ayurvedic rasayana regimen in high‐grade diffuse large B‐cell lymphoma: a case report. Clin Case Rep. 2022;10(4):e0596.##Arnold JT. Integrative ayurvedic medicine into cancer research programmes part 2: ayurvedic herbs and research opportunities. J Ayurveda Integr Med. 2023;14(2):100677.##Gupta S, Patil V, Tiwari S. Principle and practice of Yapana basti-A critical review. Ayushdhara. 2016;3(6):929-35.##Shukla KK, Mahdi AA, Ahmad MK, Jaiswar SP, Shankwar SN, Tiwari SC. Mucuna pruriens reduces stress and improves the quality of semen in infertile men. Evid Based Complement Alternat Med. 2010;7(1):137-44.##Moichela FT, Adefolaju GA, Henkel RR, Opuwari CS. Aqueous leaf extract of Moringa oleifera reduced intracellular ROS production, DNA fragmentation and acrosome reaction in Human spermatozoa in vitro. Andrologia. 2021;53(1): e13903.##

Detection of Germline Mosaicism for Robertsonian Translocation 14;14: A Case Report 2 1 2 Background: Chromosomal structural rearrangements can lead to fertility prob-lems and recurrent miscarriages. The intricate interplay of genetics during hu-man development can lead to subtle anomalies that may affect reproduction. Case Presentation: A 33-year-old woman sought fertility treatment after expe-riencing six miscarriages. Products of conception from the final pregnancy loss had been karyotyped, revealing a Robertsonian translocation (RT), involving chromosome 14. Fertility investigations showed low anti-Mullerian hormone (AMH) levels but otherwise normal female characteristics with normal sperm parameters of her husband were observed and both partners having a normal karyotype. Two embryos were transferred in an IVF cycle but neither resulted in a successful pregnancy. Subsequently, preimplantation genetic testing for an-euploidy (PGT-A) was applied to trophectoderm biopsy specimens from 4 em-bryos, which revealed abnormalities involving chromosome 14. Sperm aneu-ploidy testing failed to detect any increase in the incidence of aneuploidy af-fecting chromosome 14. Further embryos genetic testing indicated that all iden-tified chromosome 14 abnormalities in the embryos had a maternal (oocyte) origin. Conclusion: This case underscores challenges in diagnosing and managing germline mosaicism in fertility. A maternal 14;14 Robertsonian translocation, undetected in the patient's blood but impacting oocytes, likely explains recur-rent miscarriage and observed embryo aneuploidies. Genetic mosaicism in re-productive medicine highlights the necessity for advanced testing and personal-ized treatments. Data integration from various genetic analyses could enhance managing treatment expectations and improving fertility experiences. 66 72 Xavier XV Gonzalez Embryology Department, Aria Fertility Embryology Department, Aria Fertility UK xavier@ariafertility.co.uk Francisca F Mora Embryology Department, Aria Fertility Embryology Department, Aria Fertility UK Falak F Arshad Reproductive Genetics Department, Juno Genetics Reproductive Genetics Department, Juno Genetics UK Yiping Y Zhang Reproductive Genetics Department, Juno Genetics Reproductive Genetics Department, Juno Genetics UK Dhruti D Babariya Reproductive Genetics Department, Juno Genetics Reproductive Genetics Department, Juno Genetics UK Dagan D Wells Reproductive Genetics Department, Juno Genetics Reproductive Genetics Department, Juno Genetics UK Amanda A Tozer Clinical Department, Aria Fertility Clinical Department, Aria Fertility UK In vitro fertilizationMiscarriageMosaicism 140211.pdf Delhanty JD, SenGupta SB, Ghevaria H. How common is germinal mosaicism that leads to premeiotic aneuploidy in the female? J Assist Reprod Genet. 2019;36(12):2403-18.##Hardy PJ, Hardy K. Chromosomal instability in first trimester miscarriage: a common cause of pregnancy loss? Transl Pediatr. 2018;7(3):211-8.##Sudha T, Gopinath PM. Homologous robertsonian translocation (21q21q) and abortions. Hum Genet. 1990;85(2):253-5.##Scriven PN, Flinter FA, Braude PR, Ogilvie CM. Robertsonian translocations--reproductive risks and indications for preimplantation genetic diagnosis. Hum Reprod. 2001;16(11):2267-73.##Tiegs AW, Tao X, Zhan Y, Whitehead C, Kim J, Hanson B, et al. A multicenter, prospective, blinded, nonselection study evaluating the predictive value of an aneuploid diagnosis using a targeted next-generation sequencing-based preimplantation genetic testing for aneuploidy assay and impact of biopsy. Fertil Steril. 2021;115(3):627-37.##Kim J, Tao X, Cheng M, Steward A, Guo V, Zhan Y, Scott RT Jr, et al. The concordance rates of an initial trophecto-derm biopsy with the rest of the embryo using PGTseq, a targeted next-generation sequencing platform for preim-plantation genetic testing-aneuploidy. Fertil Steril. 2022;117(2):315-23.##Anton E, Blanco J, Egozcue J, Vidal F. Sperm FISH studies in seven male carriers of Robertsonian translocation t(13;14)(q10;q10). Hum Reprod. 2004;19(6):1345-51.##Caspersson T, Hultén M, Lindsten J, Therkelsen AJ, Zech L. Identification of different Robertsonian translocations in man by quinacrine mustard fluorescence analysis. Hereditas. 1971;67(2):213-20.##Gracias-Espinal R, Roberts SH, Duckett DP, Laurence KM. Recurrent spontaneous abortions due to a homologous Robertsonian translocation (14q14q). J Med Genet. 1982;19(6):465-7.##Pentao L, Lewis RA, Ledbetter DH, Patel PI, Lupski JR. Maternal uniparental isodisomy of chromosome 14: asso-ciation with autosomal recessive rod monochromacy. Am J Hum Genet. 1992;50(4):690-9.##Papenhausen PR, Mueller OT, Sutcliffe M, Diamond TM, Kousseff BG, Johnson VP. Uniparental isodisomy of chromosome 14 in two cases: an abnormal child and a normal adult. Am J Med Genet. 1996;66(1):90.##Cinar C, Beyazyurek C, Ekmekci CG, Aslan C, Kahraman S. Sperm fluorescence in situ hybridization analysis re-veals normal sperm cells for 14;14 homologous male Robertsonian translocation carrier. Fertil Steril. 2011;95(1):289.e5-9.##Kotzot D, Utermann G. Uniparental disomy (UPD) other than 15: phenotypes and bibliography updated. Am J Med Genet A. 2005;136(3):287-305.##

Mixed Gonadal Dysgenesis with 45,X/46,X,idic(Y)/46,XY Karyotype: A Case Report 2 1 2 Background: The purpose of the current study was to report a case with 45,X/ 46,XY/46,X,idic(Yp) mosaicism showing the male phenotype with mixed gonadal dysgenesis. Case Presentation: A 27 year-old individual, phenotypically male, presented with azoospermia and a micropenis. Both testes were not visualized in the scrotal sac. Due to the presence of a small-sized uterus, the individual was referred to the KSHEMA Center for Genetic Services for chromosomal analysis. Karyotyping revealed a mosaic karyotype of 45,X[44]/46,XY[5]/46,X,idic(Yp)[1]. This finding was further confirmed through fluorescent in situ hybridization (FISH) analysis. The individual's mosaic karyotype consisted of three cell lines, with a higher proportion of the 45,X cell line and lower proportions of the idic(Yp) and 46,XY cell lines. It is worth noting that this mosaic condition in postnatal peripheral blood has not been reported in the literature thus far. Conclusion: The case report demonstrated the importance of performing karyotype and FISH analysis in understanding genetic defects including mosaicism and other chromosomal aberrations, which can influence not only growth and puberty but also sexual development and maturation. Hence, performing cytogenetic and molecular cytogenetic analysis will help clinicians to take a further step in understanding and managing the condition. 72 77 Reshma RA Shetty KSHEMA Center for Genetic Services, K.S. Hegde Medical Academy, Nitte University KSHEMA Center for Genetic Services, K.S. Hegde Medical Academy, Nitte University India Deyyanthody DP Shetty KSHEMA Center for Genetic Services, K.S. Hegde Medical Academy, Nitte University KSHEMA Center for Genetic Services, K.S. Hegde Medical Academy, Nitte University India dprashanthshetty@gmail. com, drprashanth@nitte.edu.in Pooja PS Kulshreshtha Center for Human Genetics Center for Human Genetics India Jayarama JS Kadandale Center for Human Genetics Center for Human Genetics India AzoospermiaFluorescence in situ hybridization (FISH)Isodicentric YKaryotypingMosaicism 140216.pdf Telvi L, Lebbar A, Del Pino O, Barbet JP, Chaussain JL. 45,X/46,XY mosaicism: report of 27 cases. Pediatrics. 1999;104(2 Pt):304-8.##Ketheeswaran S, Alsbjerg B, Christensen P, Gravholt CH, Humaidan P. 45,X/46,XY Mosaicism and normozoospermia in a patient with male phenotype. Case Rep Med. 2019;2019:2529080.##Layman LC, Tho SP, Clark AD, Kulharya A, McDonough PG. Phenotypic spectrum of 45,X/46,XY males with a ring Y chromosome and bilaterally descended testes. Fertil Steril. 2009;91(3):791-7.##Mohammadpour Lashkari F, Sadighi Gilani MA, Ghaheri A, Zamanian MR, Borjian Boroujeni P, Mohseni Meybodi A, et al. Clinical aspects of 49 infertile males with 45,X/46,XY mosaicism karyotype: a case series. Andrologia. 2018;50(5):e13009.##Akinsal EC, Baydilli N, Bayramov R, Ekmekcioglu O. A rare cause of male infertility: 45,X/46,XY mosaicism. Urol Int. 2018;101(4):481-5.##Jacobs PA, Ross A. Structural abnormalities of the Y chromosome in man. Nature. 1966;210(5034):352-4.##Hsu LY. Phenotype/karyotype correlations of Y chromosome aneuploidy with emphasis on structural aberrations in postnatally diagnosed cases. Am J Med Genet. 1994;53(2):108-40.##Pasantes JJ, Wimmer R, Knebel S, Munch C, Kelbova C, Junge A, et al. 47,X,idic(Y),inv dup(Y): a non-mosaic case of a phenotypically normal boy with two different Y isochromosomes and neocentromere formation. Cytogenet Genome Res. 2012;136(2):157-62.##Robinson DO, Dalton P, Jacobs PA, Mosse K, Power MM, Skuse DH, et al. A molecular and FISH analysis of structurally abnormal Y chromosomes in patients with turner syndrome. J Med Genet. 1999;36(4):279-84.##Bergeron MB, Brochu P, Lemyre E, Lemieux N. Correlation of intercentromeric distance, mosaicism, and sexual phenotype: molecular localization of breakpoints in isodicentric Y chromosomes. Am J Med Genet A. 2011;155A(11):2705-12.##Sarrate Z, Vidal F, Blanco J. Role of sperm fluorescent in situ hybridization studies in infertile patients: indications, study approach, and clinical relevance. Fertil Steril. 2010;93(6):1892-902.##Yang Y, Hao W. Clinical, cytogenetic, and molecular findings of isodicentric Y chromosomes. Mol Cytogenet. 2019;12:55.##Forabosco A, Carratu A, Assuma M, Pol AD, Dutrillaux B, Cheli E. Male with 45,X karyotype. Clin Genet. 1977;12(2):97-100.##Cools M, Pleskacova J, Stoop H, Hoebeke P, Van Laecke E, Drop SL, et al. Gonadal pathology and tumor risk in relation to clinical characteristics in patients with 45, X/46, XY mosaicism. J Clin Endocrinol Metab. 2011;96(7):E1171-80.##Bernstein R, Steinhaus KA, Cain MJ. Prenatal application of fluorescent in situ hybridization (FISH) for identification of a mosaic Y‐chromosome marker, IDIC (Yp). Prenatal Diagn. 1992;12(9):709-16.##Hernando C, Carrera M, Ribas I, Parear N, Baraibar R, Egocue J, et al. Prenatal and postnatal characterization of Y chromosome structural anomalies by molecular cytogenetic analysis. Prenat Diagn. 2002;22(9):802-5.##Kotzot D, Dufke A, Tzschach A, Baeckert‐Sifeddine IT, Geppert M, Holland H, et al. Molecular breakpoint analysis and relevance of variable mosaicism in a woman with short stature, primary amenorrhea, unilateral gonadoblastoma, and a 46,X, del (Y)(q11)/45,X karyotype. Am J Med Genet. 2002;112(1):51-5.##DesGroseilliers M, Beaulieu Bergeron M, Brochu P, Lemyre E, Lemieux N. Phenotypic variability in isodicentric Y patients: study of nine cases. Clin Genet. 2006;70(2):145-50.##Jiang Y, Wang R, Li L, Xue L, Deng S, Liu R. Molecular cytogenetic study of de novo mosaic karyotype 45,X/46,X,i(Yq)/46,X,idic(Yq) in an azoospermic male: case report and literature review. Mol Med Rep. 2017;16(3):3433-8.##Guevarra FM, Nimkarn S, New MI, Lin-Su K. Long-term growth hormone therapy in an adolescent boy with 45,X/46,XidicY(p11). J Pediatr. 2009;155(5):752-5.e1-3.##Bruyere H, Speevak MD, Winsor EJ, de Freminville B, Farrell SA, McGowanJordan J, et al. IsodicentricYp: prenatal diagnosis and outcome in 12 cases. Prenat Diagn. 2006;26(4):324-9.##