The Intersection of Ethics, Law, and Professionalism in Third-Party Reproduction for the Elderly 2 1 2 During our daily practice of providing medical services for infertile couples, we encounter cases where one or both partners are of advanced age, but still they solicit assisted reproduction to achieve a pregnancy using donated gametes or embryo. As a proven scientific fact, male and female fertility is age dependent. Women in their early to mid-20s generally experience the peak of their fertility potential, with an estimated monthly conception rate of 25-30%. The decline in female fertility begins gradually in the early 30s and accelerates significantly in their 40s. As a result, most of women experience menopause after the age of 45. For example, the chance of pregnancy for a 40-year-old woman is less than 5% per month (1). Male fertility also follows the similar progression. Aging has a significant effect on fertility, sexual performance, and sperm parameters, which ultimately leads to a decrease in fertility potential, often evidenced by longer time to achieve pregnancy. Male fertility decline begins around the ages of 40 to 45 with decreasing sperm quality, as well as libido and sexual desire. However, due to life style changes in industrial and urban societies, it has become increasingly common for men over the age of 50 to have children. The above trend will change when it comes to using assisted reproductive technologies (ARTs) and third-party reproduction. These methods have the ability to bypass the limitations imposed by the biological clock, thereby offering opportunities for individuals to achieve pregnancy. As a result, this question arises whether these methods have unlimited power to enable pregnancy at any age or not. Case reports of men and women in their 50s, 60s, 70s, and even 80s, giving birth to children, are the frequent topics in the daily headlines in the congresses, scholarly journals, and articles. Taking a close look at these reports of pregnancy in advanced age, it becomes evident that such pregnancies represent high-risk situations which are not similar to the typical process experienced by young mothers. Despite the provision of full-time rest and special care, most pregnancies at an advanced age threaten the life of the mother and the child. Therefore, after delivery, premature neonates often require long-term care in NICU. Furthermore, the evaluation of various aspects of health of children resulting from pregnancies at an advanced age differs from pregnancies at a younger age (2). Recent data show that as women age, the use of donated eggs for fertility and childbearing also increases. This rate is 10% for women under 40 years and 18% for women aged 41 to 42 years. It increases to 34% at the age of 43-44 and to 71% at the age of 44 and finally 100% for women after the age of 45. It is estimated that more than 75% of all live births among women over 40 are the result of third-party reproduction (3). As a result of fundamental advancements and rapid development in technologies and innovative treatment strategies in reproductive medicine, there is great interest and willingness among couples to delay childbearing, even missing their golden time for fertility, with the hope that ART can help them in the future. Furthermore, the proliferation of excessive and misleading advertising by service providers leads couples to develop misconceptions about the safety and potential outcomes of these technologies. Therefore, the use of ART for elderly couples has provoked wide discussions in the medical, legal, ethical, and social communities, with varying perspectives in both support and opposition to its use. Some individuals advocate for reproductive freedom and rights of people, asserting that there should not be limitation on pregnancy at an advanced age. This viewpoint imparts that individuals who have achieved the necessary financial, educational, familial, and social stability should have the right to make their own decision regarding pregnancy. On the other hand, there are women who are willing to voluntarily donate their oocytes to aged couples for pregnancy and childbearing. They support their viewpoints with scientific evidence, which suggests that with regular prenatal care, the uterus of older women just like that of young women, is capable of completing a full term pregnancy, resulting in a healthy delivery. Also, the oocyte of a young woman in the uterus of an elderly mother can result in the birth of a healthy baby, comparable to the birth of the same baby from a younger mother at her reproductive age. The advocates believe that due to the busy work schedules and limitations faced by parents, the common practice of grandparents in taking care of children has become prevalent in today's society. They also acknowledge that children cared for by their own grandparents do not face any significant problems compared to children raised by their own young parents. Grandparents spend more time and energy for grandchildren compared to young parents because they have fewer commitments and more leisure time. Now, if these grandmothers and grandfathers become the primary parents of these children, they will invest much more energy and time in their upbringing compared to young parents since they have more financial stability and necessary resources to focus on the growth of their children (4). On the contrary, the opponents of pregnancy in elderly individuals believe that with increasing age of women, the success rate of pregnancy following embryo transfer significantly decreases, even when using donated oocytes. Additionally, there is higher rate of pregnancy loss among elderly mothers compared to young mothers. Furthermore, the complications of pregnancy such as high blood pressure, gestational diabetes, pre-eclampsia, and eclampsia are significantly more prevalent in elderly mothers, which in many cases lead to the elective termination of pregnancy to protect the mother's life. Also, the rate of morbidity and mortality in pregnancies among elderly women is higher compared to younger women. From a moral standpoint, the opposition to pregnancy in older women stems from the fact that a child needs healthy parents for balanced growth and development across all physical, mental, social, cultural, and economic dimensions. The early death of one or both parents, which is more likely in aged individuals, cause irreparable damage to the children (4). As a result of the aforementioned viewpoints and in order to reduce the associated risks of pregnancy in elderly women, many countries have implemented restrictions and regulations on third party reproduction for aged couples. The primary factor for imposing such restrictions is the age limitation in undergoing ART. The majority of the executive regulations are established based on the health indicators and life expectancy specific to each country. Out of 43 European countries, 34 have implemented age limitations for infertility treatment. Regarding the age of women, there is a legal limit in 18 countries. For instance, in Denmark and Belgium, the limit is 47 years, while in the Netherlands it is 49 years. Bulgaria specified the limit at 51 years and Greece recently increased the age for pregnancy up to 54 years. The maximum legal age for men to undergo ART treatment is 60 years in Portugal and Finland, while the limit is set at 56 years in Sweden (5). Therefore, in conclusion, it should be stated that the consequences of ART in elderly individuals, both at the individual and the societal levels, can be positive. However, there are important debates about the ethical implications of third-party reproduction after the reproductive period. Such practices have the potential to change patterns of life and community relationships fundamentally. Apart from the medical and scientific advancements enabling childbearing in individuals who have passed their youth and entered grandparenthood, it is crucial to consider this phenomenon from the perspective of ethics, religion, and philosophy. In all our endeavors, we must have a reverent attitude toward God and refrain from exceeding human boundaries to manipulate the fabric of life, as such actions impose unpredictable consequences on the future of society and humanity. Despite the profound ethical questions and challenges surrounding these actions, many philosophers hold the view that helping in childbearing and improving the quality and meaning of life for elderly couples is highly valuable and praiseworthy. Scientists and their supporters argue in response to criticisms by asserting that their work should not be equated to "playing God", as its purpose is not to alter the system of creation. Instead, such innovative actions show the ingenuity and continuous efforts of human to overcome biological limitations and foster the development of life which were not possible in the past. 217 219 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 140208.pdf Steiner AZ, Jukic AMZ. The impact of female age and nulligravidity on fecundity in an older reproductive age cohort. Fertil Steril. 2016;105(6):1584-8.e1.##Bouzaglou A, Aubenas I, Abbou H, Rouanet S, Carbonnel M, Pirtea P, et al. Pregnancy at 40 years old and above: obstetrical, fetal, and neonatal outcomes. is age an independent risk factor for those complications? Front Med (Lausanne). 2020;7:208.##Fritz R, Jindal S. Reproductive aging and elective fertility preservation. J Ovarian Res. 2018;11(1):66.##Harrison BJ, Hilton TN, Rivière RN, Ferraro ZM, Deonandan R, Walker MC. Advanced maternal age: ethical and medical considerations for assisted reproductive technology. Int J Womens Health. 2017;9:561-70.##Calhaz-Jorge C, De Geyter CH, Kupka MS, Wyns C, Mocanu E, Motrenko T, et al. Survey on ART and IUI: legislation, regulation, funding and registries in European countries: the European IVF-monitoring consortium (EIM) for the European society of human reproduction and embryology (ESHRE). Hum Reprod Open. 2020(1):hoz044.##

Maternal and Fetal Factors Affecting Cell-Free Fetal DNA (cffDNA) Fraction: A Systematic Review 2 1 2 Background: Cell-free fetal DNA (cffDNA) is a novel screening method for fetal aneuploidy that facilitated non-invasive prenatal testing (NIPT) through analysis of cffDNA in maternal plasma. However, despite increased sensitivity, it has a number of limitations that may complicate of its results interpretation. Therefore, elucidating factors affecting fetal fraction, as a critical limitation, guides its clinical application. Methods: In this report, systematic search was carried out through PubMed, Web of Science, and Scopus databases until February 11, 2022 by using keywords consist of "noninvasive prenatal screening", "NIPT", "noninvasive prenatal", "cell free DNA" and "fetal fraction". The articles were screened for eligibility criteria before data extraction. Results: A total of 39 eligible studies, most published between 2010 and 2020, were included. Based on the results of studies, a negative correlation between maternal age and BMI/body weight with fetal fraction was found. Furthermore, LDL, cholesterol, triglyceride level, metformin, heparin and enoxaparin therapy, hemoglobin-related hemoglobinopathies, and physical activity showed to have negative associations. Interestingly, it seems the ethnicity of patients from South and East Asia has a correlation with fetal fraction compared to Caucasians. Positive correlation was observed between gestational age, free β-hCG, PAPP-A, living in high altitude, and twin pregnancy. Conclusion: Considering each factor, there was significant inconsistency and controversy regarding their impact on outcomes. Indeed, multiple factors can influence the accuracy of NIPS results, and it is worth noting that the impact of these factors may vary depending on the individual’s ethnic background. Therefore, it is important to recognize that NIPS remains a screening test, and comprehensive pre- and post-NIPS counseling should be conducted as part of standard clinical practice. 219 232 Majid M Zaki-Dizaji Human Genetics Research Center, Baqiyatallah University of Medical Sciences Human Genetics Research Center, Baqiyatallah University of Medical Sciences Iran Arman A Shafiee Student Research Committee, School of Medicine, Alborz University of Medical Sciences Student Research Committee, School of Medicine, Alborz University of Medical Sciences Iran Omid O Kohandel Gargari Student Research Committee, School of Medicine, Alborz University of Medical Sciences Student Research Committee, School of Medicine, Alborz University of Medical Sciences Iran Haniyeh H Fathi Student Research Committee, School of Medicine, Alborz University of Medical Sciences Student Research Committee, School of Medicine, Alborz University of Medical Sciences Iran 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 z.heidary2016@gmail.com Cell-free DNAFetal fractionGestational ageMaternal ageNon-invasive prenatal testing 140205.pdf Warsof SL, Larion S, Abuhamad AZ. 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Size distributions of maternal and fetal DNA in maternal plasma. Clin Chem. 2004;50(1):88-92.##Alberry M, Maddocks D, Jones M, Abdel Hadi M, Abdel-Fattah S, Avent N, et al. Free fetal DNA in maternal plasma in anembryonic pregnancies: confirmation that the origin is the trophoblast. Prenat Diagn. 2007;27(5):415-8.##Scott FP, Menezes M, Palma-Dias R, Nisbet D, Schluter P, da Silva Costa F, et al. Factors affecting cell-free DNA fetal fraction and the consequences for test accuracy. J Matern Fetal Neonatal Med. 2018;31(14):1865-72.##Illanes S, Denbow M, Kailasam C, Finning K, Soothill PW. Early detection of cell-free fetal DNA in maternal plasma. Early Hum Dev. 2007;83(9):563-6.##Lo YM, Zhang J, Leung TN, Lau TK, Chang AM, Hjelm NM. Rapid clearance of fetal DNA from maternal plasma. Am J Hum Genet. 1999;64(1):218-24.##Guy GP, Hargrave J, Dunn R, Price K, Short J, Thilaganathan B. Secondary non-invasive prenatal screening for fetal trisomy: an effectiveness study in a public health setting. BJOG. 2021;128(2):440-6.##Fiorentino F, Bono S, Pizzuti F, Mariano M, Polverari A, Duca S, et al. Response to "the importance of determining the limit of detection of non-invasive prenatal testing methods". Prenatal Diagn. 2016;36(9):898-9.##Scheffer PG, Wirjosoekarto SAM, Becking EC, Weiss MM, Bax CJ, Oepkes D, et al. Association between low fetal fraction in cell-free DNA testing and adverse pregnancy outcome: a systematic review. Prenatal Diagn. 2021;41(10):1287-95.##Deng C, Liu S. Factors affecting the fetal fraction in noninvasive prenatal screening: a review. Front Pediatr. 2022;10:812781.##Mousavi S, Shokri Z, Bastani P, Ghojazadeh M, Riahifar S, Nateghian H. Factors affecting low fetal fraction in fetal screening with cell-free DNA in pregnant women: a systematic review and meta-analysis. BMC Pregnancy Childbirth. 2022;22(1):918.##Deng C, Liu J, Liu S, Liu H, Bai T, Jing X, et al. Maternal and fetal factors influencing fetal fraction: A retrospective analysis of 153,306 pregnant women undergoing noninvasive prenatal screening. Front Pediatr. 2023;11:1066178.##Santoro G, Lapucci C, Giannoccaro M, Caporilli S, Rusin M, Seidenari A, et al. Abnormal circulating maternal miRNA expression is associated with a low (<4%) cell-free DNA fetal fraction. Diagnostics (Basel). 2021;11(11):2108.##Dolatkhah M, Rahnamaye Farzami M, Khavari-Nejad R-A, Noori SJIJoNI. Correlation of maternal age, weight, pregnancy-associated plasma protein a, free beta-human chorionic gonadotropin, fetal crown-rump length, and fetal gender with fetal DNA fraction in non-invasive prenatal testing: an experiment on Iranian pregnant women. Iran J Neonatol. 2021;12(1):26-32.##Talbot AL, Ambye L, Hartwig TS, Werge L, Sørensen S, Stormlund S, et al. Fetal fraction of cell-free DNA in pregnancies after fresh or frozen embryo transfer following assisted reproductive technologies. Hum Reprod. 2020;35(6):1267-75.##Serapinas D, Boreikaitė E, Bartkevičiūtė A, Norvilaitė K, Narbekovas A, Bartkevičienė DJM. The level of free fetal DNA as precise noninvasive marker for chromosomal aneuploidies: first results from BALTIC region. medicina (Kaunas). 2020;56(11):579.##Miltoft CB, Rode L, Bundgaard JR, Johansen P, Tabor A. Cell-free fetal DNA in the early and late first trimester. Fetal Diagn Ther. 2020;47(3):228-36.##Lopes JL, Lopes GS, Enninga EAL, Kearney HM, Hoppman NL, Rowsey RA. Most noninvasive prenatal screens failing due to inadequate fetal cell free DNA are negative for trisomy when repeated. Prenat Diagn. 2020;40(7):831-7.##Zhao Q, HuoJiaBieKe J, Du SJJoGO, Reproduction H. The influence of fetal gender and maternal characteristics on fetal cell-free DNA in maternal plasma. J Gynecol Obstet Hum Reprod. 2019;48(8):653-656.##Qiao L, Yu B, Liang Y, Zhang C, Wu X, Xue Y, et al. Sequencing shorter cfDNA fragments improves the fetal DNA fraction in noninvasive prenatal testing. Am J Obstet Gynecol. 2019;221(4):345.e1-345.e11.##Hou Y, Yang J, Qi Y, Guo F, Peng H, Wang D, et al. Factors affecting cell-free DNA fetal fraction: statistical analysis of 13,661 maternal plasmas for non-invasive prenatal screening. Hum Genomics. 2019;13(1):62.##Guo FF, Yang JX, Huang YL, Qi YM, Hou YP, Peng HS, et al. Association between fetal fraction at the second trimester and subsequent spontaneous preterm birth. Prenat Diagn. 2019;39(13):1191-7.##Rolnik DL, da Silva Costa F, Lee TJ, Schmid M, McLennan ACJUiO, Gynecology. Association between fetal fraction on cell‐free DNA testing and first‐trimester markers for pre‐eclampsia. Ultrasound Obstet Gynecol. 2018;52(6):722-7.##Miltoft CB, Rode L, Ekelund CK, Sundberg K, Kjaergaard S, Zingenberg H, et al. Contingent first‐trimester screening for aneuploidies with cell‐free DNA in a Danish clinical setting. Ultrasound Obstet Gynecol. 2018;51(4):470-9.##Sarno L, Revello R, Hanson E, Akolekar R, Nicolaides KH. Prospective first-trimester screening for trisomies by cell-free DNA testing of maternal blood in twin pregnancy. Ultrasound Obstet Gynecol. 2016;47(6):705-11.##Revello R, Sarno L, Ispas A, Akolekar R, Nicolaides KH. Screening for trisomies by cell‐free DNA testing of maternal blood: consequences of a failed result. Ultrasound Obstet Gynecol. 2016;47(6):698-704.##Krishna I, Badell M, Loucks TL, Lindsay M, Samuel A. Adverse perinatal outcomes are more frequent in pregnancies with a low fetal fraction result on noninvasive prenatal testing. Prenat Diagn. 2016;36(3):210-5.##Zhou Y, Zhu Z, Gao Y, Yuan Y, Guo Y, Zhou L, et al. Effects of maternal and fetal characteristics on cell-free fetal DNA fraction in maternal plasma. Reprod Sci. 2015;22(11):1429-35.##Ashoor G, Syngelaki A, Poon L, Rezende JC, Nicolaides KH. Fetal fraction in maternal plasma cell‐free DNA at 11–13 weeks' gestation: relation to maternal and fetal characteristics. Ultrasound Obstet Gynecol. 2013;41(1):26-32.##Suzumori N, Ebara T, Yamada T, Samura O, Yotsumoto J, Nishiyama M, et al. Fetal cell-free DNA fraction in maternal plasma is affected by fetal trisomy. J Hum Genet. 2016;61(7):647-52.##Rava RP, Srinivasan A, Sehnert AJ, Bianchi DW. Circulating fetal cell-free DNA fractions differ in autosomal aneuploidies and monosomy X. Clin Chem. 2014;60(1):243-50.##Wang E, Batey A, Struble C, Musci T, Song K, Oliphant A. Gestational age and maternal weight effects on fetal cell-free DNA in maternal plasma. Prenat Diagn. 2013;33(7):662-6.##Poon LCY, Musci T, Song K, Syngelaki A, Nicolaides KH. Maternal plasma cell-free fetal and maternal DNA at 11-13 weeks' gestation: relation to fetal and maternal characteristics and pregnancy outcomes. Fetal Diagn Ther. 2013;33(4):215-23.##Lee TJ, Rolnik DL, Menezes MA, McLennan AC, da Silva Costa F. Cell-free fetal DNA testing in singleton IVF conceptions. Hum Reprod. 2018;33(4):572-8.##Kuhlmann‐Capek M, Chiossi G, Singh P, Monsivais L, Lozovyy V, Gallagher L, et al. Effects of medication intake in early pregnancy on the fetal fraction of cell‐free DNA testing. Prenat Diagn. 2019;39(5):361-8.##Kinnings SL, Geis JA, Almasri E, Wang H, Guan X, McCullough RM, et al. Factors affecting levels of circulating cell‐free fetal DNA in maternal plasma and their implications for noninvasive prenatal testing. Prenat Diagn. 2015;35(8):816-22.##Hedriana H, Martin K, Saltzman D, Billings P, Demko Z, Benn P. Cell-free DNA fetal fraction in twin gestations in single-nucleotide polymorphism-based noninvasive prenatal screening. Prenat Diagn. 2020;40(2):179-84.##Cao J, Qiao L, Jin J, Zhang S, Chen P, Tang H, et al. 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The Impact of Endometriosis on Intracellular Calcium Levels, Cyclic Dependent Kinase 1 (Cdk1) Expression, and Cyclin B Expression in Post-Ovulation Oocytes of Mice Model 2 1 2 Background: Since endometriosis causes a decrease in oocyte quality, the success rate of in vitro fertilization cycles decreases. The purpose of the current study was to analyze the effect of endometriosis on intracellular calcium levels, Cdk1 expression, and cyclin B expression in oocytes. Methods: Thirty-two mice (Mus musculus) were divided into control and endo-metriosis groups. The cumulus oocyte complex (COC) were obtained in all groups. Denudated cells were assessed for calcium levels by calorimetric examinations. Complex oocytes were examined for Cdk1 and cyclin B expression by immune-cytochemistry and were read under a microscope. Results: Intercellular calcium levels, Cdk1, and cyclin B expression were signifi-cantly lower in the endometriosis group than in the control group. There was a signi-ficant relationship between calcium levels and Cdk1 expression (p<0.05, r=0.659), a significant relationship between calcium levels and cyclin B expression (p<0.05, r=0.885), and also a significant correlation between Cdk1 and cyclin B expression (p<0.05, r=0.537). Conclusion: The data presented in this study suggested that the intracellular oocyte calcium level, Cdk1 expression, and cyclin B expression were lower in mice with endometriosis. A positive correlation was observed between calcium levels and the expression of Cdk1 and cyclin B. Furthermore, a positive correlation was also found between Cdk1 and cyclin B expression. 232 240 Teguh T Wiyono Department of Obstetrics and Gynecology, Faculty of Medicine, Airlangga University Department of Obstetrics and Gynecology, Faculty of Medicine, Airlangga University Indonesia Sri Ratna SR Dwiningsih Department of Obstetrics and Gynecology, Faculty of Medicine, Airlangga University Department of Obstetrics and Gynecology, Faculty of Medicine, Airlangga University Indonesia Widjiati W Widjiati Department of Veterinary Embryology, Faculty of Veterinary Medicine, Airlangga University Department of Veterinary Embryology, Faculty of Veterinary Medicine, Airlangga University Indonesia widjiati@fkh.unair.ac.id CalciumCdk1Cyclin BEndometriosisOocyte 140207.pdf Gupta S, Goldberg JM, Aziz N, Goldberg E, Krajcir N, Agarwal A. Pathogenic mechanisms in endome-triosis-associated infertility. Fertil Steril. 2008;90 (2):247-57.##Carlberg M, Nejaty J, Froysa B, Guan Y, Soder O, Berqvist A. Elevated expression of tumour necrosis factor alpha in cultured granulosa cells from women with endometriosis. Hum Reprod. 2000;15(6):1250-5.##Hendarto H. Pathomechanism of infertility in endo-metriosis. In: Chaudhury K, Chakravarty B, editors. Endometr: Basic concepts and current research trend; 2012. p. 343-54. ##Harada T, Iwabe T, Terakawa N. Role of cytokines in endometriosis. Fertil Steril. 2001;76(1):1-10.##Mulayim N, Arici A. The relevance of the peritoneal fluid in endometriosis-associated infertility. Hum Reprod. 1999;14 Suppl 2:67-76.##Hogg C, Horne AW, Greaves E. Endometriosis-as-sociated macrophages: origin, phenotype, and func-tion. Front Endocrinol (Lausanne). 2020;11:7.##Aghajanpour S, Ghaedi K, Salamian A, Deemeh MR, Tavalaee M, Moshtaghian J, et al. Quantitative expression of phospholipase C zeta, as an index to assess fertilization potential of a semen sample. Hum Reprod. 2011;26(11):2950-6.##Rubino P, Viganò P, Luddi A, Piomboni P. The ICSI procedure from past to future: a systematic review of the more controversial aspects. Hum Re-prod Update. 2016;22(2):194-227.##Cardona Barberán A, Boel A, Vanden Meerschaut F, Stoop D, Heindryckx B. Diagnosis and treatment of male infertility-related fertilization failure. J Clin Med. 2020;9(12):3899.##Nanda A, Thangapandi K, Banerjee P, Dutta M, Wangdi T, Sharma P, et al. Cytokines, angiogenesis, and extracellular matrix degradation are augmented by oxidative stress in endometriosis. Ann Lab Med. 2020;40(5):390-7.##Winterhager E, Kidder GM. Gap junction connexins in female reproductive organs: implications for women’s reproductive health. Hum Reprod Update. 2015;21(3):340-52.##Chiaratti MR, Garcia BM, Carvalho KF, Macabelli CH, da Silva Ribeiro FK, Zangirolamo AF, et al. Oocyte mitochondria: role on fertility and disease transmission. Anim Reprod. 2018;15(3):231-8.##Annas JY, Hendy Hendarto, Widjiati. Efficacy of various doses of curcumin supplementation on progessive endometriosis in mice. Maj Obstet Gine-kol. 2014;22(3):118-25.##Dwiningsih SR, Darmosoekarto S, Hendarto H, Dachlan EG, Rantam FA, Sunarjo S, et al. Effects of bone marrow mesenchymal stem cell transplantation on tumor necrosis factor-alpha receptor 1 expression, granulosa cell apoptosis, and folliculogenesis repair in endometriosis mouse models. Vet World. 2021;14(07):1788-96.##Salomon J, Piotrowska A, Matusiak L, Dzięgiel P, Szepietowski JC. Chitinase-3-like protein 1 (YKL-40) expression in squamous cell skin cancer. Anticancer Res. 2018;38(8):4753-8.##Tacheau C, Laboureau J, Mauviel A, Verrecchia F. TNF-alpha represses connexin43 expression in HaCat keratinocytes via activation of JNK signaling. J Cell Physiol. 2008;216(2):438-44.##Schwarz DS, Blower MD. The endoplasmic reticulum: structure, function and response to cellular signaling. Cell Mol Life Sci. 2016;73(1):79-94.##Bedaiwy MA, Falcone T, Sharma RK, Goldberg JM, Attaran M, Nelson DR, et al. Prediction of endometriosis with serum and peritoneal fluid markers: a prospective controlled trial. Hum Reprod. 2002;17(2):426-31.##Chen CW, Chavez J, Lin LL, Wang CM, Hsu YT, Hart MJ, et al. Endometrial gap junction expression-early indicators of endometriosis and integral to invasiveness. bioRxiv. 2021. ##Zhang FF, Morioka N, Kitamura T, Hisaoka-Nakashima K, Nakata Y. Proinflammatory cytokines downregulate connexin 43-gap junctions via the ubiquitin-proteasome system in rat spinal astrocytes. Biochem Biophys Res Commun. 2015;464 (4):1202-8.##Arroyo A, Kim B, Yeh J. Luteinizing hormone action in human oocyte maturation and quality: signaling pathways, regulation, and clinical impact. Reprod Sci. 2020;27(6):1223-52.##Hagting A, Jackman M, Simpson K, Pines J. Translocation of cyclin B1 to the nucleus at prophase requires a phosphorylation-dependent nuclear import signal. Curr Biol. 1999;9(13):680-9.##Holesh JE, Bass AN, Lord M. Physiology, Ovulation. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023. ##McIntyre MH. The use of digit ratios as markers for perinatal androgen action. Reprod Biol Endo-crinol. 2006;4:10.##Adeldust H, Zeinoaldini S, Kohram H, Amiri Roudbar M, Daliri Joupari M. In vitro maturation of ovine oocyte in a modified granulosa cells co-culture system and alpha-tocopherol supplementation: effects on nuclear maturation and cleavage. J Anim Sci Technol. 2015;57:27.##Enserink JM, Kolodner RD. An overview of Cdk1-controlled targets and processes. Cell Div. 2010;5:11.##Campbell KH, Loi P, Otaegui PJ, Wilmut I. Cell cycle co-ordination in embryo cloning by nuclear transfer. Rev Reprod. 1996;1(1):40-6.##Hoffmann I, Clarke PR, Marcote MJ, Karsenti E, Draetta G. Phosphorylation and activation of human Cdc25-C by Cdc2–Cyclin B and its involvement in the self-amplification of MPF at mitosis. EMBO J. 1993;12(1):53-63.##Haccard O, Jessus C. Oocyte maturation, nos and cyclins--a matter of synthesis: two functionally redundant ways to induce meiotic maturation. Cell Cycle. 2006;5(11):1152-9.##

The Effect of FSHR (G2039A) Polymorphism on Müllerian Duct Development and Hormonal Profile of Women with Primary Amenorrhea 2 1 2 Background: The function of follicle-stimulating hormone (FSH) is mediated by binding to its G-protein coupled receptor (GPCR) which is expressed on granulosa cells of the ovary. The purpose of the current study was to examine the impact of FSHR G2039A polymorphism (rs6166; Ser680Asn) on clinical and radiology profiles of women with primary amenorrhea (PA) in Gujarat, India. Methods: A total of 90 women (45 controls and 45 cases) were recruited for the study after obtaining informed consent. The DNA extraction was performed on the venous blood samples collected from the participants, followed by polymerase chain reaction (PCR). The presence of polymorphism was then analyzed using restriction fragment polymorphism (RFLP) with the BSeNI enzyme. The statistical analysis was conducted using an independent t-test, chi-square test, and ANOVA. Significance was determined by a p<0.05. Results: Results revealed that homozygous wild type genotype was observed in 46.7% (n=21) of the control group and 11.1% (n=5) of the case group. Heterozygous genotype was observed in 33.3% (n=15) of the control group and 55.6% (n=25) of the case group (p<0.001). Homozygous mutant genotype was observed in 20% (n=9) of the control group and 33.3% (n=15) of the case group (p<0.01). The hormonal profile revealed that serum levels of FSH and luteinizing hormone (LH) were significantly higher (p<0.05) in the AA and AG genotypes compared to the GG genotypes. Conclusion: The FSHR rs6166 G2039A was associated with PA in women, and the A allele could be considered a causative risk factor in developing the condition. 240 248 Priyanka P Sanghavi Department of Zoology, Biomedical Technology and Human Genetics, School of Sciences, Gujarat University, Navrangpura Department of Zoology, Biomedical Technology and Human Genetics, School of Sciences, Gujarat University, Navrangpura India Divya D Chandel Department of Zoology, Biomedical Technology and Human Genetics, School of Sciences, Gujarat University, Navrangpura Department of Zoology, Biomedical Technology and Human Genetics, School of Sciences, Gujarat University, Navrangpura India divya_chandel@yahoo.com, divyachandel@gujaratuniversity.ac.in FSHRGene polymorphismHormonesInfertilityMüllerian ductPCR-RFLPPrimary amenorrhea 140199.pdf Gromoll J, Simoni M, Nordhoff V, Behre HM, De Geyter C, Nieschlag E. Functional and clinical consequences of mutations in the FSH receptor. Mol Cell Endocrinol. 1996;125(1-2):177-82.##Doherty E, Pakarinen P, Tiitinen A, Kiilavuori A, Huhtaniemi I, Forrest S, et al. A novel mutation in the FSH receptor inhibiting signal transduction and causing primary ovarian failure. J Clin Endocrinol Metab. 2002;87(3):1151-5.##Aittomäki K, Dieguez Lucena J, Pakarinen P, Sistonen P, Tapanainen J, Gromoll J, et al. Mutation in the follicle-stimulating hormone receptor gene causes hereditary hypergonadotropic ovarian failure. Cell. 1995;82(6):959-68.##Beau I, Touraine P, Meduri G, Gougeon A, Desroches A, Matuchansky C, et al. A novel phenotype related to partial loss of function mutations of the follicle stimulating hormone receptor. J Clin Invest. 1998;102(7):1352-9.##Touraine P, Beau I, Gougeon A, Meduri G, Desroches A, Pichard C, et al. New natural inactivating mutations of the follicle-stimulating hormone receptor: correlations between receptor function and phenotype. Mol Endocrinol. 1999;13(11):1844-54.##Allen LA, Achermann JC, Pakarinen P, Kotlar TJ, Huhtaniemi IT, Jameson JL, et al. A novel loss of function mutation in exon 10 of the FSH receptor gene causing hypergonadotrophic hypogonadism: clinical and molecular characteristics. Hum Reprod. 2003;18(2):251-6.##Meduri G, Touraine P, Beau I, Lahuna O, Desroches A, Vacher-Lavenu MC, et al. Delayed puberty and primary amenorrhea associated with a novel mutation of the human follicle-stimulating hormone receptor: clinical, histological, and molecular studies. J Clin Endocrinol Metab. 2003;88(8):3491-8.##Nakamura Y, Maekawa R, Yamagata Y, Tamura I, Sugino N. A novel mutation in exon8 of the follicle-stimulating hormone receptor in a woman with primary amenorrhea. Gynecol Endocrinol. 2008;24(12):708-12.##Achrekar SK, Modi DN, Desai SK, Mangoli VS, Mangoli RV, Mahale SD. Poor ovarian response to gonadotrophin stimulation is associated with FSH receptor polymorphism. Reprod Biomed Online. 2009;18(4):509-15.##Conway GS, Conway E, Walker C, Hoppner W, Gromoll J, Simoni M. Mutation screening and isoform prevalence of the follicle stimulating hormone receptor gene in women with premature ovarian failure, resistant ovary syndrome and polycystic ovary syndrome. Clin Endocrinol (Oxf). 1999;51(1):97-9.##Cordts EB, Santos MC, Bianco B, Barbosa CP, Christofolini DM. Are FSHR polymorphisms risk factors to premature ovarian insufficiency? Gynecol Endocrinol. 2015;31(8):663-6.##da Fonte Kohek MB, Batista MC, Russell AJ, Vass K, Giacaglia LR, Mendonca BB, et al. No evidence of the inactivating mutation (C566T) in the follicle-stimulating hormone receptor gene in Brazilian women with premature ovarian failure. Fertil Steril. 1998;70(3):565-7.##Du J, Zhang W, Guo L, Zhang Z, Shi H, Wang J, et al. Two FSHR variants, haplotypes and meta-analysis in Chinese women with premature ovarian failure and polycystic ovary syndrome. Mol Genet Metab. 2010;100(3):292-5.##Sudo S, Kudo M, Wada SI, Sato O, Hsueh AJ, Fujimoto S. Genetic and functional analyses of polymorphisms in the human FSH receptor gene. Mol Hum Reprod. 2002;8(10):893-9.##Sundblad V, Chiauzzi VA, Escobar ME, Dain L, Charreau EH. Screening of FSH receptor gene in Argentine women with premature ovarian failure (POF). Mol Cell Endocrinol. 2004;222(1-2):53-9.##Tong Y, Liao WX, Roy AC, Ng SC. Absence of mutations in the coding regions of follicle-stimulating hormone receptor gene in Singapore Chinese women with premature ovarian failure and polycystic ovary syndrome. Horm Metab Res. 2001;33(4):221-6.##Woad KJ, Prendergast D, Winship IM, Shelling AN. FSH receptor gene variants are rarely associated with premature ovarian failure. Reprod Biomed Online. 2013;26(4):396-9.##Achrekar SK, Modi DN, Meherji PK, Patel ZM, Mahale SD. Follicle stimulating hormone receptor gene variants in women with primary and secondary amenorrhea. J Assist Reprod Genet. 2010;27(6):317-26.##Thomas M, Srivastava S, Devi R. Presence of the ASN680SER Polymorphism in women with Primary Amenorrhea from South India. Indian J Basic Appl Med Res. 2014;3(3):358-67.##John SW, Weitzner G, Rozen R, Scriver CR. A rapid procedure for extracting genomic DNA from leukocytes. Nucleic Acids Res. 1991;19(2):408.##Sujatha T, Jayashankar E, Addepally U, Vijayalakshmi K, Hasan QA. Association of follicle-stimulating hormone receptor gene ser680 asn (rs6166) polymorphism with polycystic ovarian syndrome. Int J Reprod Contracept Obstet Gynecol. 2016;5(9):3127.##Bhagavath B, Layman LC. Genetics of female infertility in humans. In: Rimoin D, Pyeritz R, Korf B, editors. Emery and Rimoin’s Principles and practice of medical genetics. USA: Academic Press; 2013. p. 1-24.##Desai SS, Roy BS, Mahale SD. Mutations and polymorphisms in FSH receptor: functional implications in human reproduction. Reproduction. 2013;146(6):R235-48.##Kuijper EAM, Blankenstein MA, Luttikhof LJ, Roek SJM, Overbeek A, Hompes PG, et al. Frequency distribution of polymorphisms in the FSH receptor gene in infertility patients of different ethnicity. Reprod Biomed Online. 2011;22 Suppl 1:S60-5.##Desai SS, Achrekar SK, Sahasrabuddhe KA, Meharji PK, Desai SK, Mangoli VS, et al. Functional characterization of two naturally occurring mutations (Val514Ala and Ala575Val) in follicle-stimulating hormone receptor. J Clin Endocrinol Metab. 2015;100(4):E638-45.##Bramble MS, Goldstein EH, Lipson A, Ngun T, Eskin A, Gosschalk JE, et al. A novel follicle-stimulating hormone receptor mutation causing primary ovarian failure: a fertility application of whole exome sequencing. Hum Reprod. 2016;31(4):905-14.##Mayorga MP, Gromoll J, Behre HM, Gassner C, Nieschlag E, Simoni M. Ovarian response to follicle-stimulating hormone (FSH) stimulation depends on the FSH receptor genotype. J Clin Endocrinol Metab. 2000;85(9):3365-9.##

Establishment of Cut-off Values for Uterine and Peripheral Blood Natural Killer Cells During the Peri-implantation Period in Fertile Controls and Women with Unexplained Recurrent Implantation Failure 2 1 2 Background: The purpose of the study was to determine the cut-off values for peripheral and uterine natural killer (pNK, uNK) cells in fertile controls and in women with recurrent implantation failure (RIF). Methods: In this study, 50 women with RIF and 50 fertile controls were enrolled. Midluteal endometrial biopsy samples from both cases and controls were obtained for CD 56+ cell immunohistochemistry labeling to identify uNK cells. Peripheral venous blood was also taken during the biopsy to detect pNK cells in peripheral blood mononuclear cells using flow cytometry. Cut-off values were obtained from fertile controls. Using a non-parametric Mann-Whitney U-test, the medians of the data sets were compared. Results: The median values for uNK and pNK cell levels in the control group were 7% and 11.6%, respectively. The median value for uNK cells in RIF patients was 9%, which was higher than the one in controls but not statistically significant (p-value of 0.689). The median pNK levels (11.6% vs. 12.4%) were comparable between the RIF group and the controls. Moreover, it was found that 68% of individuals had uNK cell counts below the reference value, while 32% had excessive levels exceeding 7%. Additionally, only 51.4% of the RIF group had increased pNK cells. Conclusion: The pNK cell cut-off values need to be used with caution because there was no difference between fertile controls and RIF women. If immunotherapy is recommended for RIF women, uNK cell testing should be used as the preferred approach. 248 257 Neeta N Singh All India Institute of Medical Sciences All India Institute of Medical Sciences India Yogita Y Dogra Arriva IVF Superspeciality Centre Shimla Himachal Pradesh Arriva IVF Superspeciality Centre Shimla Himachal Pradesh India dryogitadogra@gmail.com Pawan P Kumar All India Institute of Medical Sciences All India Institute of Medical Sciences India Sandeep S Mathur All India Institute of Medical Sciences All India Institute of Medical Sciences India Ashok A Sharma All India Institute of Medical Sciences All India Institute of Medical Sciences India Garima G Patel All India Institute of Medical Sciences All India Institute of Medical Sciences India CD56 antigen EndometriumImmunohistochemistryNatural killer cellsRecurrent miscarriage 140201.pdf Polanski LT, Baumgarten MN, Quenby S, Brosens J, Campbell BK, Raine-Fenning NJ. What exactly do we mean by “recurrent implantation failure”? a systematic review and opinion. Reprod Biomed Online. 2014;28(4):409-23.##Laufer N, Simon A. Recurrent implantation failure: current update and clinical approach to an ongoing challenge. Fertil Steril. 2012;97(5):1019-20.##Tuckerman E, Laird SM, Prakash A, Li TC. Prognostic value of the measurement of uterine natural killer cells in the endometrium of women with recurrent miscarriage. Hum Reprod. 2007;22(8):2208-13.##Tuckerman E, Mariee N, Prakash A, Li TC, Laird S. Uterine natural killer cells in peri-implantation endometrium from women with repeated implantation failure after IVF. J Reprod Immunol. 2010;87(1-2):60-6.##Tang AW, Alfirevic Z, Quenby S. Natural killer cells and pregnancy outcomes in women with recurrent miscarriage and infertility: a systematic review. Hum Reprod. 2011;26(8):1971-80.##Quenby S, Kalumbi C, Bates M, Farquharson R, Vince G. Prednisolone reduces preconceptual endometrial natural killer cells in women with recurrent miscarriage. Fertil Steril. 2005;84(4):980-4.##Olanski LT, Barbosa MA, Martins WP, Baumgarten MN, Campbell B, Brosens J, et al. Interventions to improve reproductive outcomes in women with elevated natural killer cells undergoing assisted reproduction techniques: a systematic review of literature. Hum Reprod. 2014;29(1):65-75.##Lash GE, Bulmer JN, Li TC, Innes BA, Mariee N, Patel G, et al. Standardisation of uterine natural killer (uNK) cell measurements in the endometrium of women with recurrent reproductive failure. J Reprod Immunol. 2016;116:50-9.##Robertson MJ, Ritz J. Biology and clinical relevance of human natural killer cells. Blood. 1990;76(12):2421-38.##Kwak-Kim J, Gilman-Sachs A. Clinical implication of natural killer cells and reproduction. Am J Reprod Immunol. 2008;59(5):388-400.##Bulmer JN, Lash GE. Human uterine natural killer cells: a reappraisal. Mol Immunol. 2005;42(4):511-21.##Kitaya K, Yamaguchi T, Yasuo T, Okubo T, Honjo H. Post-ovulatory rise of endometrial CD16(-) natural killer cells: in situ proliferation of residual cells or selective recruitment from circulating peripheral blood? J Reprod Immunol. 2007;76(1-2):45-53.##Joseph-Horne R, Mason H, Batty S, White D, Hil-lier S, Urquhart M, et al. Luteal phase progesterone excretion in ovulatory women with polycystic ovaries. Hum Reprod. 2002;17(6):1459-63.##Chen X, Mariee N, Jiang L, Liu Y, Wang CC, Li TC, et al. Measurement of uterine natural killer cell percentage in the periimplantation endometrium from fertile women and women with recurrent reproductive failure: establishment of a reference range. Am J Obstet Gynecol. 2017;217(6):680.e1-680.e6.##Donoghue JF, Paiva P, Teh WT, Cann LM, Nowell C, Rees H, et al. Endometrial uNK cell counts do not predict successful implantation in an IVF population. Hum Reprod. 2019;34(12):2456-66.##Beer AE, Kwak JY, Ruiz JE. Immunophenotypic profiles of peripheral blood lymphocytes in women with recurrent pregnancy losses and in infertile women with multiple failed in vitro fertilization cycles. Am J Reprod Immunol. 1996;35(4):376-82.##Sacks G, Yang Y, Gowen E, Smith S, Fay L, Chapman M. Detailed analysis of peripheral blood natural killer cells in women with repeated IVF failure. Am J Reprod Immunol. 2012;67(5):434-42.##Santillán I, Lozano I, Illán J, Verdú V, Coca S, Bajo-Arenas JM, et al. Where and when should natural killer cells be tested in women with repeated implantation failure? J Reprod Immunol. 2015;108:142-8.##Shivalingegowda DP, Rao V, Rathinam K, Pranesh GT, Dwarakanath M, Mummadi S, et al. Uterine natural killer cell density as a predictor for implantation success or failure in fertile surrogates and in women with implantation failure. Fertil Steril. 2018;110(4):e129.##Von Woon E, Greer O, Shah N, Nikolaou D, Johnson M, Male V. Number and function of uterine natural killer cells in recurrent miscarriage and implantation failure: a systematic review and meta-analysis. Hum Reprod Update. 2022;28(4):548-82.##Sfakianoudis K, Rapani A, Grigoriadis S, Pantou A, Maziotis E, Kokkini G, et al. The role of uterine natural killer cells on recurrent miscarriage and recurrent implantation failure: from pathophysiology to treatment. Biomedicines. 2021;9(10):1425.##

The Amino Acid Profile in Seminal Plasma of Normozoospermic Men: A Correlation Analysis with Spermiogram Parameters and Total Antioxidant Capacity 2 1 2 Background: Male infertility is usually determined by the manual evaluation of the semen, namely the standard semen analysis. It is currently impossible to predict sperm fertilizing ability based on the semen analysis alone. Therefore, a more sensitive and selective diagnosis tool is required. Methods: Twelve fresh semen samples were collected from fertile volunteers attending the Avicenna Fertility Center (Tehran, Iran). The seminal plasma (SP) was prepared and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the total antioxidant capacity (TAC) was analysis. Thirty-four amino acids including essential amino acids (EAA), non-essential amino acids (NEAA), and non-proteinogenic amino acids (NPAA) relative concentration were determined, and the correlation between their concentration with spermiogram parameters and TAC of the SP was analyzed. Results: Significant positive correlations have been found between selected amino acids with the motility (Met and Gln, rs=0.92; Cys, rs=0.72; and Asn, rs=0.82), normal sperm morphology (Met, rs=0.92; Cys, rs=0.72; Glu, rs=0.92; and Asn, rs=0.82), and sperm concentration (Trp, Phe, and Ala). In contrast, several AAs, including Gly, Ser, and Ile showed negative correlations with sperm concentration (rs=-0.93, r=-0.92, and r=-0.89, respectively). Furthermore, TAC showed a positive association only with Tyr (rs=0.79). Conclusion: The strong positive/negative correlations between the seminal metabolic signature and spermiogram demonstrate the significance of determining metabolite levels under normal conditions for normal sperm functions. Combining the metabolome with the clinical characteristics of semen would enable clinicians to look beyond biomarkers toward the clinical interpretation of seminal parameters to explain the biological basis of sperm pathology. 257 269 Naser N Amirjannati ناصر امیرجنتی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Ralf R Henkel LogixX Pharma, Theale LogixX Pharma, Theale United Kingdom Elham E Hosseini Department of Obstetrics and Gynecology, School of Medicine, Zanjan University of Medical Sciences Department of Obstetrics and Gynecology, School of Medicine, Zanjan University of Medical Sciences Iran Peyman P Choopanian Department of Applied Mathematics, Faculty of Mathematical Sciences, Tarbiat Modares University Department of Applied Mathematics, Faculty of Mathematical Sciences, Tarbiat Modares University Iran Hanieh H Moghadasfar Avicenna Fertility Center, Avicenna Research Institute, ACECR Avicenna Fertility Center, Avicenna Research Institute, ACECR Iran Babak B Arjmand Cell Therapy and Regenerative Medicine Research Center, Endocrinology and Metabolism Molecular Cellular Sciences Institute, Tehran University of Medical Sciences Cell Therapy and Regenerative Medicine Research Center, Endocrinology and Metabolism Molecular Cellular Sciences Institute, Tehran University of Medical Sciences Iran Lima L Asgharpour Sarouey Avicenna Fertility Center, Avicenna Research Institute, ACECR Avicenna Fertility Center, Avicenna Research Institute, ACECR Iran Azadeh A Haji Parvaneh Avicenna Fertility Center, Avicenna Research Institute, ACECR Avicenna Fertility Center, Avicenna Research Institute, ACECR Iran Kambiz K Gilany کامبیز گیلانی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran k.gilany@avicenna.ac.ir Amino acidsHuman seminal plasmaLC-MS/MSSpermiogram parametersTotal antioxidant capacity 140202.pdf Thonneau P, Marchand S, Tallec A, Ferial M-L, Ducot B, Lansac J, et al. Incidence and main causes of infertility in a resident population (1 850 000) of three French regions (1988-1989). Hum Reprod. 1991;6(6):811-6.##Akhondi MM, Kamali K, Ranjbar F, Shirzad M, Shafeghati S, Ardakani ZB, et al. Prevalence of primary infertility in Iran in 2010. Iran J Public Health. 2013;42(12):1398-404.##Lue JC, Huang YF, Lu NQ. [WHO laboratory manual for the examination and processing of human semen: its applicablity to andrology laboratories in China]. Zhonghua Nan Ke Xue. 2010;16(10):867-71.##Jafarzadeh N, Mani-Varnosfaderani A, Minai-Tehrani A, Savadi-Shiraz E, Sadeghi MR, Gilany K. Metabolomics fingerprinting of seminal plasma from unexplained infertile men: a need for novel diagnostic biomarkers. Mol Reprod Dev. 2015;82(3):150.##Panner Selvam MK, Finelli R, Agarwal A, Henkel R. Proteomics and metabolomics—current and future perspectives in clinical andrology. Andrologia. 2021;53(2):e13711.##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.##Gilany K, Minai-Tehrani A, Savadi-Shiraz E, Rezadoost H, Lakpour N. Exploring the human seminal plasma proteome: an unexplored gold mine of biomarker for male infertility and male reproduction disorder. J Reprod Infertil. 2015;16(2):61-71.##Mehrparavar B, Minai-Tehrani A, Arjmand B, Gilany K. Metabolomics of male infertility: a new tool for diagnostic tests. J Reprod Infertil. 2019;20(2):64-9.##Minai‐Tehrani A, Jafarzadeh N, Gilany K. Metabolomics: a state‐of‐the‐art technology for better understanding of male infertility. Andrologia. 2016;48(6):609-16.##Gilany K, Lakpour N, Vafakhah M, Sadeghi MR. The profile of human sperm proteome; a mini-review. J Reprod Infertil. 2011;12(3):193-9.##Wishart DS, Feunang YD, Marcu A, Guo AC, Liang K, Vázquez-Fresno R, et al. HMDB 4.0: the human metabolome database for 2018. Nucleic Acids Res. 2018;46(D1):D608-D17.##Bender DA. Amino acid metabolism. 3rf ed. UK: John Wiley & Sons; 2012. 480 p.##Gfeller D, Michielin O, Zoete V. SwissSidechain: a molecular and structural database of non-natural sidechains. Nucleic Acids Res. 2013;41(Database issue):D327-32.##Massey KA, Blakeslee CH, Pitkow HS. A review of physiological and metabolic effects of essential amino acids. Amino Acids. 1998;14(4):271-300.##Hou Y, Wu G. Nutritionally nonessential amino acids: a misnomer in nutritional sciences. Adv Nutr. 2017;8(1):137-9.##Choi BH, Coloff JL. The diverse functions of non-essential amino acids in cancer. Cancers (Basel). 2019;11(5):675.##Hedges JB, Ryan KS. Biosynthetic pathways to nonproteinogenic α-amino acids. Chem Rev. 2019;120(6):3161-209.##Piraud M, Vianey‐Saban C, Petritis K, Elfakir C, Steghens JP, Bouchu D. Ion‐pairing reversed‐phase liquid chromatography/electrospray ionization mass spectrometric analysis of 76 underivatized amino acids of biological interest: a new tool for the diagnosis of inherited disorders of amino acid metabolism. Rapid Commun Mass Spectrom. 2005;19(12):1587-602.##Engel KM, Baumann S, Rolle-Kampczyk U, Schiller J, von Bergen M, Grunewald S. Metabolomic profiling reveals correlations between spermiogram parameters and the metabolites present in human spermatozoa and seminal plasma. PloS One. 2019;14(2):e0211679.##Parkhitko AA, Jouandin P, Mohr SE, Perrimon N. Methionine metabolism and methyltransferases in the regulation of aging and lifespan extension across species. Aging Cell. 2019;18(6):e13034.##Rubinstein S, Breitbart H. Role of spermine in mammalian sperm capacitation and acrosome reaction. Biochem J. 1991;278(Pt 1)(Pt 1):25-8.##Gilany K, Jafarzadeh N, Mani-Varnosfaderani A, Minai-Tehrani A, Sadeghi MR, Darbandi M, et al. Metabolic fingerprinting of seminal plasma from non-obstructive Azoospermia patients: positive versus negative sperm retrieval. J Reprod Infertil. 2018;19(2):109-14.##Gholinezhad M, Aliarab A, Abbaszadeh-Goudarzi G, Yousefnia-Pasha Y, Samadaian N, Rasolpour-Roshan K, et al. Nitric oxide, 8-hydroxydeoxyguanosine, and total antioxidant capacity in human seminal plasma of infertile men and their relationship with sperm parameters. Clin Exp Reprod Med. 2020;47(1):54-60.##van Overveld FW, Haenen GR, Rhemrev J, Vermeiden JP, Bast A. Tyrosine as important contributor to the antioxidant capacity of seminal plasma. Chem Biol Interact. 2000;127(2):151-61.##Mehrparvar B, Chashmniam S, Nobakht F, Amini M, Javidi A, Minai-Tehrani A, et al. Metabolic profiling of seminal plasma from teratozoospermia patients. J Pharm Biomed Anal. 2020;178:112903.##Xu Y, Lu H, Wang Y, Zhang Z, Wu Q. Comprehensive metabolic profiles of seminal plasma with different forms of male infertility and their correlation with sperm parameters. J Pharm Biomed Anal. 2020;177:112888.##Zhang X, Diao R, Zhu X, Li Z, Cai Z. Metabolic characterization of asthenozoospermia using nontargeted seminal plasma metabolomics. Clin Chim Acta. 2015;450:254-61.##Qiao S, Wu W, Chen M, Tang Q, Xia Y, Jia W, et al. Seminal plasma metabolomics approach for the diagnosis ofunexplained male infertility. PLoS One. 2017;12(8):e0181115.##Murgia F, Corda V, Serrenti M, Usai V, Santoru ML, Hurt KJ, et al. Seminal fluid metabolomic markers of oligozoospermic infertility in Human. Metabolites. 2020;10(2):64.##Hosseini E, Amirjannati N, Henkel R, Bazrafkan M, Moghadasfar H, Gilany K. Targeted amino acids profiling of human seminal plasma from teratozoospermia patients using LC–MS/MS. Reprod Sci. 2023:1-11.##

Comparison of the Administration of 150 or 75 IU of Recombinant LH in Agonist ICSI Cycles Stimulated with Recombinant FSH in Women Aged 35-39: A Comparative Study 2 1 2 Background: The purpose of the study was to assess whether the coadministration of 150 IU of recombinant LH instead of 75 IU in women aged 35-39 improves the results in agonist ICSI cycles stimulated with 300 IU of recombinant FSH. Methods: In this study, two ovarian stimulation protocols coexisted which were identical except in the administered dose of recombinant LH, for which some patients received 150 IU (n=231) and some received 75 IU (n=216). Both groups received 300 IU of recombinant FSH. There were no differences in the demographic characteristics of both groups. Gonadotropins were reimbursed by the National Health System. Statistical analysis was performed by Student’s t test, χ2, and ANCOVA. Significance level was established at p=0.05. Results: The number of retrieved oocytes was slightly higher in the 300/150 group (9.06±5.53 vs. 8.61±5.11), but the differences were not significant. Results were similar with the number of metaphase II oocytes (7.18±4.86 vs. 6.72±4.72) and the number of fertilized oocytes (4.64±3.2 vs. 4.23±2.72). The per-transfer clinical pregnancy rates exhibited close similarity between both groups (32.84% vs. 32.46%), as did the per-transfer live birth rates (29.90% vs. 30.37%) and the implantation rate. The rate of hyperstimulation syndrome (OHSS) as well as the rate of cancellation due to OHHS risk was similar in both groups. There was also no difference in the miscarriage rate. When results were expressed by per started cycle or by oocyte pick- up, the results remained very similar in both groups. Conclusion: In women aged 35-39 undergoing ovarian stimulation with recombinant FSH in agonist cycles, the coadministration of 75 or 150 UI of recombinant LH did not influence pregnancy rates. However, a slight increase in the number of retrieved oocytes should not be disregarded. 269 279 Roberto R Matorras Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Fermin F Aspichueta Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Begoña B Prieto Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Rosario R Mendoza Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Iker I Malaina Department of Mathematics, Faculty of Science and Technology, University of the Basque Country Department of Mathematics, Faculty of Science and Technology, University of the Basque Country Spain Blanca B Corral Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Lorena L Crisol Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Alberto A Vendrell Biocruces Bizkaia Health Research Institute Biocruces Bizkaia Health Research Institute Spain alberto.venber@gmail.com Antonia A Exposito Human Reproduction Unit, Cruces University Hospital Human Reproduction Unit, Cruces University Hospital Spain Agonist ICSI cyclesCoadministrationLuteinizing hormoneOocyteOvarian stimulationPregnancy rate 140206.pdf Conforti A, Esteves SC, Humaidan P, et al. Recombinant human luteinizing hormone co-treatment in ovarian stimulation for assisted reproductive technology in women of advanced reproductive age: a systematic review and meta-analysis of randomized controlled trials. Reprod Biol Endocrinol. 2021;19(1):91.##Wang Y, Li L, Deng K, Liu J, Liu Y, Zou K, et al. Comparison of the combination of recombinant follicle-stimulating hormone and recombinant luteinizing hormone protocol versus human menopausal gonadotropin protocol in controlled ovarian stimulation: a systematic review and meta-analysis. J Evid Based Med. 2020;13(3):215-26.##Bosch E, Labarta E, Crespo J, Simón C, Remohí J, Pellicer A. Impact of luteinizing hormone administration on gonadotropin-releasing hormone antagonist cycles: an age-adjusted analysis. Fertil Steril. 2011;95(3):1031-6.##Humaidan P, Bungum M, Bungum L, Andersen CY. Effects of recombinant LH supplementation in women undergoing assisted reproduction with GnRH agonist down-regulation and stimulation with recombinant FSH: an opening study. Reprod Biomed Online. 2004;8(6):635-43.##Matorras R, Prieto B, Exposito A, Mendoza R, Crisol L, Herranz P, et al. Mid-follicular LH supplementation in women aged 35-39 years undergoing ICSI cycles: a randomized controlled study. Reprod Biomed Online. 2011;22 Supp 1;S43-51.##NyboeAndersen A, Humaidan P, Fried G, Hausken J, Antila L, Bangsbøll S, et al. Recombinant LH supplementation to recombinant FSH during the final days of controlled ovarian stimulation for in vitro fertilization. a multicentre, prospective, randomized, controlled trial. Hum Reprod. 2008;23 (2):427-34.##Filicori M. The role of luteinizing hormone in folliculogenesis and ovulation induction. Fertil Steril. 1999;71(3):405-14.##Freis A, Germeyer A, Jauckus J, Capp E, Strowitzki T, Zorn M, et al. Endometrial expression of receptivity markers subject to ovulation induction agents. Arch Gynecol Obstet. 2019;300(6):1741-50.##Marchiani S, Tamburrino L, Benini F, Pallecchi M, Bignozzi C, Conforti A, et al. LH supplementation of ovarian stimulation protocols influences follicular fluid steroid composition contributing to the improvement of ovarian response in poor responder women. Sci Rep. 2020;10(1):12907.##Roman R, Mussarat N, Detti L. Ovarian stimulation in poor responders: have we made Progress? Curr Pharm Biotechnol. 2017;18(8):614-8.##Van Tilborg TC, Torrance HL, Oudshoorn SC, Eijkemans MJC, Koks CAM, Verhoeve HR, et al. Individualized versus standard FSH dosing in women starting IVF/ICSI: an RCT. Part 1: the predicted poor responder. Hum Reprod. 2017;32 (12):2496-505.##Chappel SC, Howles C. Re-evaluation of the roles of luteinizing hormone and follicle-stimulating hormone in the ovulatory process. Hum Reprod. 1991;6(9):1206-12.##Balasch J, Fábregues F. LH in the follicular phase: neither too high nor too low. Reprod BioMed Online. 2006;12(4):406-15.##Behre HM, Howles CM, Longobardi S. Randomized trial comparing luteinizing hormone supplementation timing strategies in older women undergoing ovarian stimulation. Reprod Biomed Online. 2015;31(3);339-46.##Hillier SG. Current concepts of the roles of follicle stimulating hormone and luteinizing hormone in folliculogenesis. Hum Reprod. 1994;9(2):188-91.##Shoham Z. The clinical therapeutic window for luteinizing hormone in controlled ovarian stimulation. Fertil Steril. 2002;77(6):1170-7.##Matorras R, Pérez-Sanz J, Corcóstegui B, Perez-Ruiz I, Malaina I, Quevedo S, et al. Effect of vitamin E administered to men in infertile couples on sperm and assisted reproduction outcomes: a double-blind randomized study. F S Rep. 2020;1(3):219-26.##Matorras R, Exposito A, Ferrando M, Mendoza R, Larreategui Z, Laínz L, et al. Oocytes of women who are obese or overweight have lower levels of n-3 polyunsaturated fatty acids compared with oocytes of women with normal weight. Fertil Steril. 2020;113(1):53-61.##Matorras R, Urquijo E, Mendoza R, Corcóstegui B, Expósito A, Rodríguez-Escudero FJ. Ultrasound-guided embryo transfer improves pregnancy rates and increases the frequency of easy transfers. Hum Reprod. 2002;17(7):1762-6.##Ibañez-Perez J, Díaz-Nuñez M, Clos-García M, Lainz L, Iglesias M, Díez-Zapirain M, et al. microRNA-based signatures obtained from endometrial fluid identify implantative endometrium. Hum Reprod. 2022;37(10):2375-91.##van der Linden M, Buckingham K, Farquhar C, Kremer JA, Metwally M. Luteal phase support for assisted reproduction cycles. Cochrane Database Syst Rev. 2015;2015(7):CD009154.##Watters M, Noble M, Child T, Nelson S. Short versus extended progesterone supplementation for luteal phase support in fresh IVF cycles: a systematic review and meta-analysis. Reprod Biomed Online. 2020;40(1):143-50.##Mochtar MH, Danhof NA, Ayeleke RO, Van der Veen F, van Wely M. Recombinant luteinizing hormone (rLH) and recombinant follicle stimulating hormone (rFSH) for ovarian stimulation in IVF/ICSI cycles. Cochrane Database Syst Rev. 2017;5(5):CD005070.##Xiong Y, Bu Z, Dai W, Zhang M, Bao X, Sun Y. Recombinant luteinizing hormone supplementation in women undergoing in vitro fertilization/ intracytoplasmic sperm injection with gonadotropin releasing hormone antagonist protocol: a systematic review and meta-analysis. Reprod Biol Endocrinol. 2014;12:109.##Zhang Y, Zhang C, Shu J, Guo J, Chang HM, Leung PCK, et al. Adjuvant treatment strategies in ovarian stimulation for poor responders undergoing IVF: a systematic review and network meta-analysis. Hum Reprod Update. 2020;26(2):247-63.##Fábregues F, Iraola A, Casals G, Creus M, Carmona F, Balasch J. Evaluation of two doses of recombinant human luteinizing hormone supplementation in down-regulated women of advanced reproductive age undergoing follicular stimulation for IVF: a randomized clinical study. Eur J Obstet Gynecol Reprod Biol. 2011;158(1):56-61.##Lisi F, Rinaldi L, Fishel S, Caserta D, Lisi R, Campbell A. Evaluation of two doses of recombinant luteinizing hormone supplementation in an unselected group of women undergoing follicular stimulation for in vitro fertilization. Fertil Steril. 2005;83(2):309-15.##Vuong TNL, Phung HT, Ho MT. Recombinant follicle-stimulating hormone and recombinant luteinizing hormone versus recombinant follicle-stimulating hormone alone during GnRH antagonist ovarian stimulation in patients aged ≥35 years: a randomized controlled trial. Hum Reprod. 2015;30(5):1188-95.##Humaidan P, Chin W, Rogoff D, D’Hooghe T, Longobardi S, Hubbard J, et al. Efficacy and safety of follitropin alfa/lutropin alfa in ART: a randomized controlled trial in poor ovarian responders. Hum Reprod. 2017;32(3):544-55.##König TE, Van der Houwen LEE, Overbeek A, Hendriks ML, Beutler-Beemsterboer SN, Kuchenbecker WKH, et al. Recombinant LH supplementation to a standard GnRH antagonist protocol in women of 35 years or older undergoing IVF/ICSI: a randomized controlled multicentre study. Hum Reprod. 2013;28(10):2804-12.##Marrs R, Meldrum D, Muasher S, Schoolcraft W, Werlin L, Kelly E. Randomized trial to compare the effect of recombinant human FSH (follitropin alfa) with or without recombinant human LH in women undergoing assisted reproduction treatment. Reprod Biomed Online. 2004;8(2):175-82.##Barrenetxea G, Agirregoikoa JA, Jiménez MR, de Larruzea AL, Ganzabal T, Carbonero K. Ovarian response and pregnancy outcome in poor-responder women: a randomized controlled trial on the effect of luteinizing hormone supplementation on in vitro fertilization cycles. Fertil Steril. 2008;89(3):546-53.##Younis JS, Izhaki I, Ben-Ami M. The effect of LH supplementation to the GnRH antagonist protocol in advanced reproductive ageing women: a prospective randomized controlled study. Clin Endocrinol (Oxf). 2016;84(1):99-106.##Boothroyd C, Karia S, Andreadis N, Rombauts L, Johnson N, Chapman M, et al. Consensus statement on prevention and detection of ovarian hyperstimulation syndrome. Aust N Z J Obstet Gynaecol. 2015;55(6):523-34.##Koloda Y, Korsak V, Rozenson O, Anshina M, Sagamonova K, Baranov I, et al. Use of a recombinant human follicle-stimulating hormone: recombinant human luteinizing hormone (r-hFSH:r-hLH) 2:1 combination for controlled ovarian stimulation during assisted reproductive technology treatment: A real-world study of routine practice in the Russian Federation. Best Pract Res Clin Obstet Gynaecol. 2022;85(Pt B):134-48.##Canosa S, Carosso AR, Mercaldo N, Ruffa A, Evangelista F, Bongioanni F, et al. Effect of rLH Supplementation during Controlled Ovarian Stimulation for IVF: Evidence from a Retrospective Analysis of 1470 Poor/Suboptimal/ Normal Responders Receiving Either rFSH plus rLH or rFSH Alone. J Clin Med. 2022;11(6):1575.##Chung K, Krey L, Katz J, Noyes N. Evaluating the role of exogenous luteinizing hormone in poor responders undergoing in vitro fertilization with gonadotropin-releasing hormone antagonists. Fertil Steril. 2005;84(2):313-8.##

The Impact of Growth Hormone Co-Treatment Duration on Outcomes in IVF/ICSI Cycles Among Poor Ovarian Responders 2 1 2 Background: The efficiency of in vitro fertilization is improved by growth hormone (GH) during ovarian stimulation. Additionally, patients with diabetes experience impaired insulin resistance and compromised glucose tolerance, which further exacerbate their condition. Due to these side effects, in this study, the duration of GH treatment was compared in IVF/ICSI cycles among poor ovarian responders. Methods: In this study, POSEIDON criteria were used to choose patients. Subcutaneous administration of gonadotropin-releasing hormone (GnRH) antagonist was done beginning on the sixth day of the cycle and continuing through the day of human chorionic gonadotropin (hCG) injection. In one group, GH was administered 4 units/day from the 2 nd day of the cycle until hCG injection, and in another group, the first dose was administered on the 6th day of the cycle. Following the administration of hCG, which lasted from 24 to 36 hr, oocytes were retrieved with the support of B-mode sonography. Results: In our analysis, no significant differences were observed between the two groups in terms of the number of retrieved oocytes, metaphase II oocytes, and quality of grade A and B embryos. The results show that the treatment or conditions did not have a significant impact on the outcomes among the studied groups. Conclusion: Our findings indicate that a shorter duration of GH administration can yield similar outcomes compared to a longer duration in IVF/ICSI cycles involving poor ovarian responders. This result holds the potential for a more cost-effective and patient-friendly approach in managing assisted reproductive technology procedures. It may lead to reduced side effects and improved adherence to medication regimens in patients. 279 287 Zahra Z Mohammadshirazi Department of Obstetrics and Gynecology, Shariati Hospital, Tehran University of Medical Science Department of Obstetrics and Gynecology, Shariati Hospital, Tehran University of Medical Science Iran Ashraf A Alyasin اشرف آل یاسین Department of Obstetrics and Gynecology, Faculty of Medicine, Tehran University of Medical Sciences Department of Obstetrics and Gynecology, Faculty of Medicine, Tehran University of Medical Sciences Iran Marzieh M Agha Hosseini Department of Obstetrics and Gynecology, Faculty of Medicine, Tehran University of Medical Sciences Department of Obstetrics and Gynecology, Faculty of Medicine, Tehran University of Medical Sciences Iran Vajihe V Hazari Department of Obstetrics and Gynecology, Faculty of Medicine, Birjand University of Medical Sciences Department of Obstetrics and Gynecology, Faculty of Medicine, Birjand University of Medical Sciences Iran Hazarivajihe10@gmail.com Assisted reproductive technologyGrade A and B embryosGrowth hormoneMetaphase II oocytesPoor ovarian responderRetrieved oocytes 140204.pdf Ramanaviciene A, Popov A, Baliunaite E, Brasiunas B, Kausaite-Minkstimiene A, Tamer U, et al. Magneto-immunoassay for the detection and quantification of human growth hormone. Biosensors (Basel). 2022;12(2):65.##Sadiq NM, Tadi P. Physiology, Pituitary Hormones. [Updated 2023 May 1]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from: https://www.ncbi.nlm.nih.gov/ books/NBK557556/##Hart RJ, Rombauts L, Norman RJ. Growth hormone in IVF cycles: any hope? Curr Opin Obstet Gynecol. 2017;29(3):119-25.##Xu YM, Hao GM, Gao BL. Application of growth hormone in in vitro fertilization. Front Endocrinol (Lausanne). 2019;10:502.##Shakkarpude J, Mishra A, Lakhani P, Jain A, Kumari M, Caesar DD. Role of growth hormone in reproduction. Int J Chem Stud. 2019;7(3):4683-92.##Zafardoust S, Ansaripor S, Karimi A, Hosseinirad H, Ataei M. Effects of adjuvant growth Hormone therapy on poor ovarian responders in assisted reproductive technology. Maedica (Bucur). 2022;17 (2):336-43.##Yousef MS, Rezk WR, El-Naby A-sA-H, Mahmoud KGM, Takagi M, Miyamoto A, et al. In vitro effect of zearalenone on sperm parameters, oocyte maturation and embryonic development in buffalo. Reprod Biol. 2023;23(1):100732.##Herrick JR. Assisted reproductive technologies for endangered species conservation: developing sophisticated protocols with limited access to animals with unique reproductive mechanisms. Biol Reprod. 2019;100(5):1158-70.##Wang R, Pan W, Jin L, Li Y, Geng Y, Gao C, et al. Artificial intelligence in reproductive medicine. Reproduction. 2019;158(4):R139-54.##Yang JY, Li H, Lu N, Li L, Sun X-X. Influence of growth hormone supplementation in patients with thin endometrium undergoing frozen embryo transfer. Reprod Dev Med. 2019;3(01):49-53.##Chen Y, Tao L, Lin Y, Li X, Ma C. Outcomes of in vitro fertilization–embryo transfer in women with diminished ovarian reserve after growth hormone pretreatment. Gynecol Endocrinol. 2020;36(11):955-8.##Cai MH, Liang XY, Wu YQ, Huang R, Yang X. Six‐week pretreatment with growth hormone improves clinical outcomes of poor ovarian responders undergoing in vitro fertilization treatment: A self‐controlled clinical study. J Obstet Gynaecol Res. 2019;45(2):376-81.##Xia L, Tian L, Zhang S, Huang J, Wu Q. Hormonal replacement treatment for frozen-thawed embryo transfer with or without GnRH agonist pretreatment: a retrospective cohort study stratified by times of embryo implantation failures. Front Endo-crinol (Lausanne). 2022;13:803471.##Liu FT, Hu KL, Li R. Effects of growth hormone supplementation on poor ovarian responders in assisted reproductive technology: a systematic review and meta-analysis. Reprod Sci. 2021;28(4):936-48.##Yang P, Wu R, Zhang H. The effect of growth hormone supplementation in poor ovarian responders undergoing IVF or ICSI: a meta-analysis of randomized controlled trials. Reprod Biol Endocrinol. 2020;18(1):76.##Liu FT, Wu Z, Yan J, Norman RJ, Li R. The potential role of growth hormone on the endometrium in assisted reproductive technology. Front Endocrinol (Lausanne). 2020;11:49.##Cai MH, Gao LZ, Liang XY, Fang C, Wu YQ, Yang X. The effect of growth hormone on the clinical outcomes of poor ovarian reserve patients undergoing in vitro fertilization/intracytoplasmic sperm injection treatment: A retrospective study based on POSEIDON criteria. Front Endocrinol (Lausanne). 2019;10:775.##Taulbee JD, Symons MJ. Sample size and duration for cohort studies of survival time with covariables. Biometrics. 1983;39(2):351-60.##Tesarik J, Mendoza C, Greco E. Paternal effects acting during the first cell cycle of human preimplantation development after ICSI. Hum Reprod. 2002;17(1):184-9.##Gong Y, Zhang K, Xiong D, Wei J, Tan H, Qin S. Growth hormone alleviates oxidative stress and improves the IVF outcomes of poor ovarian responders: a randomized controlled trial. Reprod Biol Endocrinol. 2020;18(1):91##Safdarian L, Aghahosseini M, Alyasin A, Samaei-Nouroozi A, Rashidi S, Shabani-Nashtaei M, et al. Growth hormone (GH) improvement of ovarian responses and pregnancy outcome in poor ovarian responders: a randomized study. Asian Pac J Cancer Prev. 2019;20(7):2033-7.##Zhang Y, Zhang C, Shu J, Guo J, Chang HM, Leung PC, et al. Adjuvant treatment strategies in ovarian stimulation for poor responders undergoing IVF: a systematic review and network meta-analysis. Hum Reprod Update. 2020;26(2):247-63.##Cozzolino M, Cecchino GN, Troiano G, Romanelli C. Growth hormone cotreatment for poor responders undergoing in vitro fertilization cycles: a systematic review and meta-analysis. Fertil Steril. 2020;114(1):97-109.##Jeve YB, Bhandari HM. Effective treatment protocol for poor ovarian response: a systematic review and meta-analysis. J Hum Reprod Sci. 2016;9(2):70-81.##Polyzos NP, Devroey P. A systematic review of randomized trials for the treatment of poor ovarian responders: is there any light at the end of the tunnel? Fertil Steril. 2011;96(5):1058-61.e7.##Li XL, Wang L, Lv F, Huang XM, Wang LP, Pan Y, et al. The influence of different growth hormone addition protocols to poor ovarian responders on clinical outcomes in controlled ovary stimulation cycles: a systematic review and meta-analysis. Medicine (Baltimore). 2017;96(12):e6443.##Yovich JL, Ye Y, Regan SL, Keane KN. The evolving concept of poor-prognosis for women undertaking IVF and the notion of growth hormone as an adjuvant; a single-center viewpoint. Front Endocrinol (Lausanne). 2019;10:808.##Keane KN, Hinchliffe PM, Rowlands PK, Borude G, Srinivasan S, Dhaliwal SS, et al. DHEA supplementation confers no additional benefit to that of growth hormone on pregnancy and live birth rates in IVF patients categorized as poor prognosis. Front Endocrinol (Lausanne). 2018;9:14.##Dakhly DM, Bassiouny YA, Bayoumi YA, Hassan MA, Gouda HM, Hassan AA. The addition of growth hormone adjuvant therapy to the long down regulation protocol in poor responders undergoing in vitro fertilization: randomized control trial. Eur J Obstet Gynecol Reprod Biol. 2018;228:161-5.##Norman RJ, Alvino H, Hull LM, Mol BW, Hart RJ, Kelly TL, et al. Human growth hormone for poor responders: a randomized placebo-controlled trial provides no evidence for improved live birth rate. Reprod Biomed Online. 2019;38(6):908-15.##Kyrou D, Kolibianakis EM, Venetis CA, Papanikolaou EG, Bontis J, Tarlatzis BC. How to improve the probability of pregnancy in poor responders undergoing in vitro fertilization: a systematic review and meta-analysis. Fertil Steril. 2009;91(3):749-66.##Kutluk Oktay, Samir Babayev. Approach to ovarian stimulation and in vitro fertilization in patients with transplanted ovarian tissue. In: Kutluk Oktay, editor. Principles and practice of ovarian tissue cryopreservation and transplantation; 2022. p. 185-92.##Li J, Chen Q, Wang J, Huang G, Ye H. Does growth hormone supplementation improve oocyte competence and IVF outcomes in patients with poor embryonic development? A randomized controlled trial. BMC Pregnancy Childbirth. 2020;20 (1):310.##Mohammad EH, Abou El Serour AG, Mohamed EAH, Abbasy AH, Zaatar M, Rageh KA, et al. Efficacy of growth hormone supplementation with gonadotropins in IVF/ICSI for poor responders: randomised controlled trial. Taiwan J Obstet Gynecol. 2021;60(1):51-5.##

Infertility Burden Across Indian States: Insights from a Nationally Representative Survey Conducted During 2019-21 2 1 2 Background: Infertility is an escalating global concern, impacting approximately one-sixth of the reproductive age population worldwide. Employing data from the National Family Health Survey-5 (NFHS-5, 2019-21), this study assessed the prevalence of primary infertility at both national and state levels in India. Methods: The data of the study was extracted from the National Family Health Survey and Individual file (women file) of the fifth round of NFHS encompassing a sample of 491,484 currently married women in the age group of 15–49 years. Results: The findings showed that the prevalence of infertility is 18.7 per 1,000 women among those married for at least five years and currently in union. This prevalence increases as the duration of marriage decreases. On a state-level analysis, regions such as Goa, Lakshadweep, and Chhattisgarh exhibit the highest burdens. Conclusion: These findings underscore the growing challenge posed by primary infertility in India, calling for targeted interventions and policy measures. The establishment of a national infertility surveillance system is of pivotal importance in addressing this pressing public health issue. 287 294 Varun V Agiwal Lecturer, Indian Institute of Public Health Lecturer, Indian Institute of Public Health India R. Sai RS Madhuri Research Assistant, Indian Institute of Public Health Research Assistant, Indian Institute of Public Health India Sirshendu S Chaudhuri Assistant Professor, Indian Institute of Public Health Assistant Professor, Indian Institute of Public Health India sirshendusisu@gmail.com Female infertilityIndiaInfertilityNFHSReproductive health 140200.pdf World Health Organization: Infertility prevalence estimates, 1990–2021 [Internet]. Geneva: World Health Organization; cited 2023 Nov 13. Available from: https://www.who.int/publications-detail-redirect/97-8920068315. ##Patel M. The socioeconomic impact of infertility on women in developing countries. Facts Views Vis Obgyn. 2016;8(1):59-61.##World Health Organization: Infertility [Internet]. Geneva: World Health Organization; cited 2023 Nov 13. Available from: https://www.who.int/news-room /fact-sheets/detail/infertility.##Wang GT. Population control policies and implementations in India. J Sociol Soci Work. 2019;7(2):135-44.##Purkayastha N, Sharma H. Prevalence and potential determinants of primary infertility in India: Evidence from Indian demographic health survey. Clin Epidemiol Global Health. 2021;9:162-70.##Amodini KN, Chaudhuri S. Infertility management in India: Issues and potential solutions. J Obstet Gynecol India. 2023;73(4):368–9.##Ganguly S, Unisa S. Trends of infertility and childlessness in India: findings from NFHS data. Facts Views Vis Obgyn. 2010;2(2):131-8.##Hazlina NHN, Norhayati MN, Bahari IS, Arif NANM. Worldwide prevalence, risk factors and psychological impact of infertility among women: a systematic review and meta-analysis. BMJ Open. 2022;12(3):e057132.##Katole A, Saoji AV. Prevalence of primary infertility and its associated risk factors in urban population of central India: a community-based cross-sectional study. Indian J Community Med. 2019;44(4):337-4.##

A Rare Case of 45,X/46,X,del(Y)(q12→qter) Mosaicism in An Infertile Male with Y Chromosome Microdeletion 2 1 2 Background: Males with 45,X/46,XY karyotype have two different types of cells. This condition is associated with a wide range of clinical phenotypes. In infertile males, the mosaic 45,X/46,XY karyotype is a frequent sex chromosome defect and they might be able to conceive with the help of assisted reproductive technology; nevertheless, there is a potential risk of transmission of azoospermia factor (AZF) microdeletions in addition to 45,X to all the male progeny. In this case report, the purpose was to present a rare sex chromosomal mosaicism of an infertile man. Case Presentation: Comprehensive molecular and cytogenetic analysis of an infertile male was performed in this case study. A 27-year-old male was presented with history of azoospermia and was unable to conceive after being involved in five years of marriage. Cytogenetic investigation revealed a rare mosaic karyotype pattern of 45,X/46,X,del(Y)(q12→qter). Y chromosome microdeletion (YMD) analysis revealed notable deletions of 06 loci. Comparative genomic hybridization (CGH) microarray was performed to investigate probable functional genetic associations. Conclusion: Deletion of Y-linked genes leads to different testicular pathological conditions contributing to male infertility. Individuals with normal male phenotype harbor YMD, although size and location of the deletion do not always correspond well with quality of sperm. Therefore, in addition to semen analysis, identification of genetic variables is important which will play a crucial role in proper diagnosis and management of infertile couples. The present case study demonstrates the significance of comprehensive molecular testing and cytogenetic screening for individuals with idiopathic infertility. 293 301 Patel P Sunny Kumar Jignesh Kumar inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited India Rahul R Kabir inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited India Ruchismita R Nayak inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited India Indira I Palo Department of Obstetrics and Gynecology, Amit Hospital Department of Obstetrics and Gynecology, Amit Hospital India Birendranath B Banerjee inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited inDNA Center for Research and Innovation in Molecular Diagnostics, inDNA Life Sciences Private Limited India : biren.banerjee@indnalife.com Azoospermia factor (AZF)Chromosomal microarray analysis (CMA)Comparative genomic hybridization (CGH)Fluorescence in situ hybridization (FISH)InfertilityY-chromosome microdeletion (YMD) 140198.pdf Arafa MM, Majzoub A, AlSaid SS, ElAnsari W, Al Ansari A, Elbardisi Y, et al. Chromosomal abnormalities in infertile men with azoospermia and severe oligozoospermia in Qatar and their association with sperm retrieval intracytoplasmic sperm injection outcomes. Arab J Urol. 2017;16(1):132-9.##Elsaid HOA, Gadkareim T, Abobakr T, Mubarak E, Abdelrhem MA, Abu D, et al. Detection of AZF microdeletions and reproductive hormonal profile analysis of infertile sudanese men pursuing assisted reproductive approaches. BMC Urol. 2021;21(1):69.##Pryor JL, Kent-First M, Muallem A, Van Bergen AH, Nolten WE, Meisner L, et al. Microdeletions in the Y chromosome of infertile men. New Engl J Med. 1997;336(8):534-9.##Poongothai J, Gopenath TS, Manonayaki S. Genetics of human male infertility. Singapore Med J. 2009;50(4):336-47.##Chakraborty A, Palo I, Roy S, Koh SW, Hande MP, Banerjee B. A novel balanced chromosomal translocation in an azoospermic male: a case report. J Reprod Infertil. 2021;22(2):133-7.##Li L, Zhang H, Yang Y, Zhang H, Wang R, Jiang Y, et al. High frequency of y chromosome microdeletions in male infertility patients with 45,x/46,xy mosaicism. Brazilian J Med Biol Res. 2020;53(3):e8980.##Rosa RFM, Bartel D’Ecclesiis WF, Dibbi RP, Rosa RCM, Trevisan P, Graziadio C, et al. 45,X/46,XY mosaicism: report on 14 patients from a Brazilian hospital. a retrospective study. Sao Paulo Med J. 2014;132(6):332-8.##Thangaraj K, Gupta NJ, Pavani K, Reddy AG, Subramainan S, Rani DS, et al. Y chromosome deletions in azoospermic men in India. J Androl. 2003;24(4):588-97.##Krausz C, Hoefsloot L, Simoni M, Tüttelmann F, European Academy of Andrology; European Molecular Genetics Quality Network. EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: State-of-the-art 2013. Andrology. 2014;2(1):5-19.##dos Santos AP, Ribeiro Andrade JG, Piveta CSC, de Paulo J, Guerra-Junior G, de Mello MP, et al. Screening of Y chromosome microdeletions in 46,XY partial gonadal dysgenesis and in patients with a 45,X/46,XY karyotype or its variants. BMC Med Genet. 2013;14:115.##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.##Siffroi JP, Le Bourhis C, Krausz C, Barbaux S, Quintana-Murci L, Kanafani S, et al. Sex chromosome mosaicism in males carrying Y chromosome long arm deletions. Hum Reprod. 2000;15(12):2559-62.##Kim SY, Kim HJ, Lee BY, Park SY, Lee HS, Seo JT. Y chromosome microdeletions in infertile men with non-obstructive azoospermia and severe oligozoospermia. J Reprod Infertil. 2017;18(3):307-15.##Stahl PJ, Mielnik AN, Barbieri CE, Schlegel PN, Paduch DA. Deletion or underexpression of the Y-chromosome genes CDY2 and HSFY is associated with maturation arrest in American men with nonobstructive azoospermia. Asian J Androl. 2012;14(5):676-82.##Colaco S, Modi D. Genetics of the human Y chromosome and its association with male infertility. Reprod Biol Endocrinol. 2018;16(1):14.##Lopes AM, Miguel RN, Sargent CA, Ellis PJ, Amorim A, Affara NA. The human RPS4 paralogue on Yq11.223 encodes a structurally conserved ribosomal protein and is preferentially expressed during spermatogenesis. BMC Mol Biol. 2010;11:33.##Elliott DJ, Oghene K, Makarov G, Makarova O, Hargreave TB, Chandley AC, et al. Dynamic changes in the subnuclear organisation of pre-mRNA splicing proteins and RBM during human germ cell development. J Cell Sci. 1998;111(Pt 9):1255-65.##Vogt PH. AZF deletions and Y chromosomal haplogroups: history and update based on sequence. Hum Reprod Update. 2005;11(4):319-36.##Stouffs K, Lissens W, Verheyen G, Van Landuyt L, Goossens A, Tournaye H, et al. Expression pattern of the Y-linked PRY gene suggests a function in apoptosis but not in spermatogenesis. Mol Hum Reprod. 2004;10(1):15-21.##Ferlin A, Raicu F, Gatta V, Zuccarello D, Palka G, Foresta C. Male infertility: role of genetic background. Reprod Biomed Online. 2007;14(6):734-45.##Sen S, Ambulkar P, Hinduja I, Zaveri K, Gokral J, Pal A, et al. Susceptibility of gr/gr rearrangements to azoospermia or oligozoospermia is dependent on DAZ and CDY1 gene copy deletions. J Assist Reprod Genet. 2015;32(9):1333-41.##Saut N, Terriou P, Navarro A, Lévy N, Mitchell MJ. The human Y chromosome genes BPY2, CDY1 and DAZ are not essential for sustained fertility. Mol Hum Reprod. 2000;6(9):789-93.##Lu C, Jiang J, Zhang R, Wang Y, Xu M, Qin Y, et al. Gene copy number alterations in the azoospermia-associated AZFc region and their effect on spermatogenic impairment. Mol Hum Reprod. 2014;20(9):836-43.##

Homozygosity for Robertsonian Translocation (14q;15q) in a Newborn with a Familial History of Recurrent Abortion and Newborns Affected by Hepatosplenomegaly: A Case Report 2 1 2 Background: Robertsonian translocations (RobTs) are one of the major chromosomal abnormalities which lead to spontaneous abortion. They occur in the human population at the rate of 1 in 1000 live infants. In this paper, a family carrying one of the rare RobTs was presented and some features of all kinds of RobTs were reviewed. Case Presentation: A couple with a history of three miscarriages was referred to Omid Health Clinic of Hamadan, Iran. The karyotype of the woman was 45,XX, rob(14;15)(q10;q10) and she exhibited phenotypically good health. Karyotype analysis of proband’s uncle and his wife with a consanguineous marriage revealed that they were both carriers of rob(14;15). This couple had six offspring, three of which were dead, and the other three were alive with a normal phenotype. Besides, this couple had an unborn child, with a karyotype of 44,XX,rob(14;15)(q10;q10). Conclusion: These observation showed that genetic counseling, pedigree, and chromosomal analysis are needed to discover the cause of spontaneous abortion, stillbirth, congenital anomalies, sudden infant death syndrome (SIDS), etc. Moreover, families carrying RobTs would be offered prenatal diagnosis screening tests and, if necessary, assisted reproductive technology methods to assist with preimplantation genetic test for structural rearrangement (PGT-SR) reproduction. 301 306 Sahra S Sahraeean Dr. F. Nabipour Pathobiology laboratory Dr. F. Nabipour Pathobiology laboratory Iran Asiyeh A Jebelli Department of Biological Sciences, Faculty of Basic Sciences, Higher Education Institute of Rab-Rashid Department of Biological Sciences, Faculty of Basic Sciences, Higher Education Institute of Rab-Rashid Iran Zahra Z Shahbazi Molecular Medicine Department, Pasteur Institute of Iran Molecular Medicine Department, Pasteur Institute of Iran Iran Fahimeh F Piryaei Research Center for Molecular Medicine, School of Medicine, Hamadan University of Medical Sciences Research Center for Molecular Medicine, School of Medicine, Hamadan University of Medical Sciences Iran f.piryaei@umsha.ac.ir AneuploidyChromosomal translocationGenetic disordersInfertilityPrenatal diagnosisSpontaneous abortion 140203.pdf Song J, Sun L, Xu S, Liu N, Yao Y, Liu Z, et al. A family with Robertsonian translocation: a potential mechanism of speciation in humans. Mol Cytogenet. 2016;9:48.##Tunç E, Ilgaz S. Robertsonian translocation (13; 14) and its clinical manifestations: a literature review. Reprod Biomed Online. 2022;45(3):563-73.##Wiland E, Olszewska M, Woźniak T, Kurpisz M. How much, if anything, do we know about sperm chromosomes of Robertsonian translocation carriers? Cell Mol Life Sci. 2020;77(23):4765-85.##Sasi R, Senft J, Spruill M, Rej S, Perrotta PL. Double robertsonian translocations in an infertile patient with macrocytic anemia: a case report. Mol Cytogenet. 2020;13:14.##Vozdova M, Oracova E, Kasikova K, Prinosilova P, Rybar R, Horinova V, et al. Balanced chromosomal translocations in men: relationships among semen parameters, chromatin integrity, sperm meiotic segregation and aneuploidy. J Assist Reprod Genet. 2013;30(3):391-405.##Huang S, Juneau K, Bogard PE, Davies KA, Wang ET, Kingsley CB, et al. Identifying Robertsonian translocation carriers by microarray-based DNA analysis. Fetal Diagn Ther. 2016;40(1):59-62.##Soltani N, Mirzaei F, Ayatollahi H. Cytogenetic studies of 608 couples with recurrent spontaneous abortions in northeastern Iran. Iran J Pathol. 2021;16(4):418-25.##Moradkhani K, Puechberty J, Bhatt S, Lespinasse J, Vago P, Lefort G, et al. Rare robertsonian translocations and meiotic behaviour: sperm FISH analysis of t (13; 15) and t (14; 15) translocations: a case report. Hum Reprod. 2006;21(12):3193-8.##Kocaaga A, Kilic H, Gulec S. The pattern of chromosomal abnormalities in recurrent miscarriages: a single center retrospective study. Ann Saudi Med. 2022;42(6):385-90.##Shaffer LG, Agan N, Goldberg JD, Ledbetter DH, Longshore JW, Cassidy SB. American college of medical genetics statement on diagnostic testing for uniparental disomy. Genet Med. 2001;3(3):206-11.##Kim SR, Shaffer LG. Robertsonian translocations: mechanisms of formation, aneuploidy, and uniparental disomy and diagnostic considerations. Genet Test. 2002;6(3):163-8.##Ko DS, Cho JW, Lee HS, Kim JY, Kang IS, Yang KM, et al. Preimplantation genetic diagnosis outcomes and meiotic segregation analysis of robertsonian translocation carriers. Fertil Steril. 2013;99(5):1369-76.##Rogenhofer N, Dürl S, Ochsenkühn R, Neusser M, Aichinger E, Thaler C, et al. Case report: elevated sperm aneuploidy levels in an infertile Robertsonian translocation t (21; 21) carrier with possible interchromosomal effect. J Assist Reprod Genet. 2012;29(4):343-6.##Kovaleva NV, Shaffer LG. Under‐ascertainment of mosaic carriers of balanced homologous acrocentric translocations and isochromosomes. Am J Med Genet Part A. 2003;121(2):180-7.##Acar H, Yildirim MS, Çora T, Ceylaner S. Evaluation of segregation patterns of 21; 21 Robertsonian translocation along with sex chromosomes and interchromosomal effects in sperm nuclei of carrier by FISH technique. Mol Reprod Dev. 2002;63(2):232-6.##Venkateshwari A, Srilekha A, Sunitha T, Pratibha N, Jyothy A. A Robertsonian Translocation rob (14; 15)(q10: q10) in a patient with recurrent abortions: a case report. J Reprod Infertil. 2010;11(3):197-200.##Katagiri Y, Tamaki Y. Genetic counseling prior to assisted reproductive technology. Reprod Med Biol. 2021;20(2):133-43.##Poornima S, Daram S, Devaki RK, Qurratulain H. Chromosomal abnormalities in couples with primary and secondary infertility: Genetic counseling for assisted reproductive techniques (ART). J Reprod Infertil. 2020;21(4):269-74.##