Technical Problems and Ethical Concerns Regarding Gene Editing in Human Germlines and Embryos 2 1 2 In vitro fertilization was initially introduced by Patrick Steptoe and Robert Edwards in 1978 in England with the aim of treating infertility of Lesley Brown due to damage or blockage of her fallopian tubes. Later, a wide range of techniques were developed for treatment of infertility such as IVF, GIFT, ZIFT, SUZI, ICSI, TESE, and PESA based on its many potential causes. According to the global ART monitoring report, presented by David Adamson on behalf of International Committee for Monitoring Assisted Reproductive Technologies (ICMART) at the annual meeting of ESHRE in 2023, more than 12 million babies have been born around the world since the first IVF birth about 45 years ago. One such example of the ever-escalating trend in application and expansion of ART is the 3.5 million IVF cycles in 2019, that led to the birth of 750,000 babies. It is estimated that by the year 2100, the number of babies born from IVF will reach 400 million or 3% of the world's population (1). In addition, in vitro fertilization of human and other mammalian oocytes and embryo development to blastocyst stage result in IVF advancement and introduction of treatment possibilities in other fields of medicine and biology such as embryonic stem cells, cloning, genetic manipulations of animal embryos (knockout and knock-in animals), and preimplantation genetic testing of human embryo for monogenic disorders (PGT-M) or aneuploidy (PGT-A) or structural rearrangements (PGT-SR). Therefore, the use of PGT techniques added to the applications of IVF for fertile couples who can produce numerous embryos despite carrying defective genes or suffering from genetic diseases; in fact, PGT, evolving as an essential procedure in ART, paves the way for successful IVF procedures by selecting unaffected embryos and transferring them to the uterus. Also, these methods are frequently used to select euploid embryos for fertile women with recurrent pregnancy loss (RPL) and infertile women with repeated implantation failures (RIF). Moreover, advances in gene technology and application of cutting-edge approaches such as NGS, CGH array, and SNP array improved accuracy and efficiency of PGT. However, the main problem of PGT is that all affected embryos should be discarded with no ability for editing their genetic errors; this problem is more evident when all tested embryos are affected and there is no possibility to take any embryos in other cycles (2). The first genome editing technology was introduced in 1984 with successful insertion of the new genetic material into pluripotent hematopoietic stem cells in mice, which showed that the virus could transfer desired genes into the target cells. This was the starting point for the possibility of gene editing and recent gene therapy technology for a wide range of genetic and non-genetic diseases. The first effective gene therapy was performed in a 4-year-old girl with severe combined immunodeficiency (SCID) due to adenosine deaminase (ADA) deficiency in 1990. The Necker–Enfants Malades Hospital reported the first clinical trial of gene therapy for SCID-X1 in France in 2000, but five of 20 treated children died of cancer. The viral vector for gene insertion into T cells activated proto-oncogene and led to leukemia. At the same time, an 18-year-old boy died in the United States following gene therapy for a rare metabolic disorder of ornithine transcarbamylase (OTC) deficiency. In this tragic story that shocked the world, the viral vector induced a lethal immune response with multiple organ failure and brain death. Therefore, the flames of fear, hope, and desire for opening the horizons in treatment of many rare genetic diseases has been extinguished and the incident halted all gene therapy trials (3). Therefore, research switched its focus from old methods of using viral vectors to finding new technologies. This field has fundamentally changed in less than ten years following abandonment of gene editing research. Emergence of several new tools including ZFNs, TALENs and CRISPR/Cas9 has translated the idea of gene editing into clinical practice. CRISPR/Cas9, the so-called "genetic scissors", has many advantages over ZFNs and TALENs due to its great accuracy, excellent efficiency, and high specificity. Consequently, only eight years later, Emmanuelle Charpentier and Jennifer A. Doudna were jointly awarded the Nobel Prize in Chemistry (2020) for the development of CRISPR-Cas9 which allows the researchers to precisely cut and edit human genome (4). After confirming the effectiveness of CRISPR/Cas9 through experimental studies, the first ex vivo clinical trial using CRISPR/Cas9 to inactivate the PD-1 gene in blood cells was conducted in China in 2016. The engineered cells returned to circulation to attack non-small cell lung cancers (NSCLCs). CRISPR/Cas9 was subsequently used to treat a patient with sickle cell anemia in 2019 in the United State. Despite solid evidence on the efficiency of this system, following publication of the preliminary results of clinical trials, there were widespread discussions and concerns about the safety, and ethical and technical challenges in CRISPR/Cas9 manipulations of human genome. Numerous conferences and articles published in this field basically recommended prohibition of using genetic editing in human embryonic cells. In fact, the research on CRISPR/Cas9 technology is still in its infancy and its lack of efficiency and specificity raises uncertainties regarding the application of the technology in human embryo gene editing (3, 5). Despite the concern of the scientific community and societies on prohibition of gene editing in human embryos, a Chinese scientist, He Jiankui, used CRISPR/Cas9 system through in vitro fertilization (IVF) process to delete the gene copies responsible for the HIV receptor in human immune cells. He publicized the birth of two girls in 2018 while global protests and public pressure caused him to be imprisoned for 3 years. A few months later, a second scientific scandal was revealed. Denis Rebrikov, a Russian scientist who initially planned to edit the same CCR5 gene responsible for HIV receptor on human cells, changed the target gene to GJB2 as the single most frequent cause of genetic hearing loss due to the pressure of scientific community and protests (5). At present, the legal frameworks for gene editing of human germline and embryo in different countries are not the same. Therefore, the gene editing of the embryo for non-reproduction purposes is allowed in at least 11 countries, including China, United States, and UK. Nineteen countries, including Belarus, Canada, Sweden, and Switzerland have banned gene editing trials on human embryo. In other countries (including Russia), a neutral position is adopted (5). Recently, during congress of the European Society of Human Reproduction and Embryology (ESHRE) in 2023 in Copenhagen, Nada Kubikova presented the findings of her research team at Oxford University. The primary objective of their research was to determine the feasibility of using CRISPR/Cas9 to correct genetic errors in human preimplantation embryos, as well as to investigate the potential short-term and long-term effects of utilizing this tool for genome editing of early embryos. In their study, 84 embryos were produced using donated sperm and oocytes through ICSI. Following embryo development for 60 hr, blastomeres were subjected to whole genome amplification (WGA) and next-generation sequencing (NGS) to identify segmental aneuploidy and evaluate DNA repair after DNA breakage through CRISPR/Cas9 system. In total, 53 double-strand breaks were created by CRISPR/Cas9, of which 32 (60%) were repaired and 21 (40%) remained unrepaired leading to segmental aneuploidy in the embryo. Therefore, they concluded that CRISPR/Cas9 has the ability to induce DNA breakage with high efficiency but most of embryos are unable to repair DNA damage (6). Despite the potential of CRISPR/Cas9 to revolutionize human gene editing, there are many drawbacks and ethical concerns surrounding its application in human embryos and germlines. Its recent potential shortcomings including off-target effects, increased mosaicism, the unknown nature of all genes and genetic diseases, and further ethical concerns call for caution in using CRISPR/Cas9 technology in embryos and germline gene editing. Therefore, it is necessary to conduct further research on the risks and potential benefits of gene editing in human embryos and germlines before proceeding with clinical applications. 145 147 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 140197.pdf European society of human reproduction and embryology [Internet]. Belgium: European society of human reproduc-tion and embryology: Focus on reproduction At least 12 million babies’ since the first IVF birth in 1978; 2023 Jun 28 [Ccited 2023 Jul 29]; [about 2 screens]. Availble from: https://www.focusonreproduction.eu/article/ESHRE-News-COP23_adamson##Greco E, Litwicka K, Minasi MG, Cursio E, Greco PF, Barillari P. Preimplantation genetic testing: where we are today. Inte J Mol Sci. 2020;21(12):4381.##Secord E, Hartog NL. Review of treatment for adenosine deaminase deficiency (ADA) severe combined immuno-deficiency (SCID). Ther Clin Risk Manag. 2022;18:939-44.##Farhud DD, Zarif-Yeganeh M. CRISPR pioneers win 2020 nobel prize for chemistry. Iran J Public Health. 2020;49(12):2235-9.##Gostimskaya I. CRISPR–Cas9: A History of Its Discovery and Ethical Considerations of Its Use in Genome Editing. Biochemistry (Moscow). 2022;87(8):777-88.##Kubikova N, Esbert M, Titus S, Coudereau C, Savash M, Fagan J, et al. O-075 Deficiency of DNA double-strand break repair in human preimplantation embryos revealed by CRISPR-Cas9. Hum Reprod. 2023;38(Supplement_1):dead093-089.##

The Role of MicroRNAs in Development of Endometrial Cancer: A Literature Review 2 1 2 Endometrial cancer (EC) ranks as the second most common gynaecological cancer worldwide. EC patients are diagnosed at an early clinical stage and generally have a good prognosis. Therefore, there is a dire need for development of a specific marker for early detection of endometrial adenocarcinoma. The development of EC is conditioned by a multistep process of oncogenic upregulation and tumor suppressor downregulation as shown by molecular genetic evidence. In this setting, microRNAs appear as significant regulators of gene expression and several variations in the expression of microRNAs have been implicated in normal endometrium, endometrial tissue, metrorrhagia, and endometrial cancer. Furthermore, microRNAs act as highly precise, sensitive, and robust molecules, making them potential markers for diagnosing specific cancers and their progression. With the rising incidence of EC, its management remains a vexing challenge and diagnostic methods for the disease are limited to invasive, expensive, and inaccurate tools. Therefore, the prospect of exploiting the utility of microRNAs as potential candidates for diagnosis and therapeutic use in EC seems promising. 147 166 Somasundaram S Indumati Department of Stem Cell and Regenerative Medicine, D.Y. Patil Education Society Department of Stem Cell and Regenerative Medicine, D.Y. Patil Education Society India drindumathisomasundaram@gmail.com Birajdar B Apurva Department of Stem Cell and Regenerative Medicine, D.Y. Patil Education Society Department of Stem Cell and Regenerative Medicine, D.Y. Patil Education Society India Gaur G Gaurav Logical Life science Logical Life science India Singh S Nehakumari Logical Life science Logical Life science India Vyas V Nishant Logical Life science Logical Life science India BiomarkersEndometrial neoplasmsGene therapyMicroRNAsSignal transduction 140193.pdf de Haydu C, Black JD, Schwab CL, English DP, Santin AD. An update on the current pharmacotherapy for endometrial cancer. Expert Opin Pharmacother. 2016;17(4):489-99.##Ferlay J, Laversanne M, Ervik M, Lam F, Colombet M, Mery L, et al. The global cancer observatory-all cancer. Int Agency Res Cancer. 2020;419:199-200.##Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin. 2019;68(1):7-30. ##Jick H, Walker AM, Rothman KJ. The epidemic of endometrial cancer: a commentary. Am J Public Health. 1980;70(3):264-7. ##Spurdle AB, Bowman MA, Shamsani J, Kirk J. Endometrial cancer gene panels: clinical diagnostic vs research germline DNA testing. Mod Pathol. 2017;30(8):1048-68.##Evans J, Salamonsen LA, Winship A, Menkhorst E, Nie G, Gargett CE, et al. Fertile ground: human endometrial programming and lessons in health and disease. Nat Rev Endocrinol. 2016;12(11):654-7.##Gargett CE, Masuda H. Adult stem cells in the endometrium. Mol Hum Reprod. 2010;16(11):818-34.##Emons G, Fleckenstein G, Hinney B, Huschmand A, Heyl W. Hormonal interactions in endometrial cancer. Endocr Relat Cancer. 2000;7(4):227-42.##Jurcevic S, Klinga-Levan K, Olsson B, Ejeskär K. Verification of microRNA expression in human endometrial adenocarcinoma. BMC Cancer. 2016;16:261.##Cancer Genome Atlas Research Network, Kandoth C, Schultz N, Cherniack AD, Akbani R, Liu Y, et al. Integrated genomic characterization of endometrial carcinoma. Nature. 2013;497(7447):67-73.##Ritterhouse LL, Howitt BE. Molecular pathology: predictive, prognostic, and diagnostic markers in uterine tumors. Surg Pathol Clin. 2016;9(3):405-26. ##Yu S, Shao H, Ban X, Zhang H, You Y, Zhou N, et al. Detection of POLE Subtypes in High-Grade Endometrioid Carcinoma by BaseScope-ISH Assay. Front Oncol. 2019;9:831.##A Berchuck A, Boyd J. Molecular basis of endometrial cancer. Cancer. 1995;76(10 Suppl):2034-40.##Okuda T, Sekizawa A, Purwosunu Y, Nagatsuka M, Morioka M, Hayashi M, et al. Genetics of endometrial cancers. Obstet Gynecol Int. 2010;2010:984013.##Bansal N, Yendluri V, Wenham RM. The molecular biology of endometrial cancers and the implications for pathogenesis, classification, and targeted therapies. Cancer Control. 2019;16(1):8-13.##Mahajan N. Endometrial receptivity array: clinical application. J Hum Reprod Sci. 2015;8(3):121-9.##Ruiz Alonso M, Blesa D, Simón C. The genomics of the human endometrium. Biochim Biophys Acta. 2012;1822(12):1931-42.##Yanokura M, Banno K, Kobayashi Y, Kisu I, Ueki A, Ono A, et al. MicroRNA and endometrial cancer: roles of small RNAs in human tumors and clinical applications (Review). Oncol Lett. 2010;1(6):935-40.##Peng Y; Croce CM. The role of MicroRNAs in human cancer. Signal Transduct Target Ther. 2016;1:15004.##Pełka K, Klicka K, Grzywa TM, Gondek A, Marczewska JM, Garbicz F, et al. miR-96-5p, miR-134-5p, miR-181b-5p and miR-200b-3p heterogenous expression in sites of prostate cancer versus benign prostate hyperplasia-archival samples study. Histochem Cell Biol. 2021;155(3):423-33.##Uzuner E, Ulu GT, Gürler SB, Baran Y. The role of MiRNA in Cancer: pathogenesis, diagnosis, and treatment. Methods Mol Biol. 2022;2257:375-422. ##Xing Y, Ruan G, Ni H, Qin H, Chen S, Gu X, et al. Tumor immune microenvironment and its related miRNAs in tumor progression. Front Immunol. 2021;12:624725. ##Tang Y, Zong S, Zeng H, Ruan X, Yao L, Han S, et al. MicroRNAs and angiogenesis: a new era for the management of colorectal cancer. Cancer Cell Int. 2021;21(1):221. ##Kunej T, Godnic I, Horvat S, Zorc M, Calin GA. Cross talk between microRNA and coding cancer genes. Cancer J. 2012;18(3):223-31.##Zhou XC, Dowdy SC, Podratz KC, Jiang SW. Epigenetic considerations for endometrial cancer prevention, diagnosis and treatment. Gynecol Oncol. 2007;107(1):143-53.##Wang H, Peng R, Wang J, Qin Z, Xue L. Circulating microRNAs as potential cancer biomarkers: the advantage and disadvantage. Clin Epigenetics. 2018;10:59.##Cui M, Wang H, Yao X, Zhang D, Xie Y, Cui R, et al. Circulating microRNAs in cancer: potential and challenge. Front Genet. 2019;10:626.##Condrat CE, Thompson DC, Barbu MG, Bugnar OL, Boboc A, Cretoiu D, et al. miRNAs as biomarkers in disease: latest findings regarding their role in diagnosis and prognosis. Cells. 2020;9(2):276. ##Pal MK, Jaiswar SP, Dwivedi VN, Tripathi AK, Dwivedi A, Sankhwar P. MicroRNA: a new and promising potential biomarker for diagnosis and prognosis of ovarian cancer. Cancer Biol Med. 2015;12(4):328-41. ##Iorio MV, Croce CM. MicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. a comprehensive review. EMBO Mol Med. 2012;4(3):143-59. ##Hammond SA. An overview of microRNAs. Adv Drug Deliv Rev. 2015;87:3-14.##Kim VN, Han J, Siomi MC. Biogenesis of small RNAs in animals. Nat Rev Mol Biol. 2019;10(2):126-39.##Tauro BJ, Greening DW, Mathias RA, Ji H, Mathivanan S, Scott AM, et al. Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes. Methods. 2012;56(2):293-304.##Tian C, Gao L, Zimmerman MC, Zucker IH. Myocardial infarction-induced microRNA-enriched exosomes contribute to cardiac Nrf2 dysregulation in chronic heart failure. Am J Physiol Heart Circ Physiol. 2018;314(5):H928-39. ##Wang X, Chen Y, Zhao Z, Meng Q, Yu Y, Sun J, et al. Engineered exosomes with ischemic myocardium-targeting peptide for targeted therapy in myocardial infarction. J Am Heart Assoc. 2018;7(15):e008737.##Mott JL, Kobayashi S, Bronk SF, Gores GJ. mir-29 regulates Mcl-1 protein expression and apoptosis. Oncogene. 2007;26(42):6133-40.##Rajewsky N, Socci ND. Computational identification of microRNA targets. Genome Biol. 2004;267(2):529-35. ##Schmidt MF. Drug Target miRNA: methods and protocols. Methods Mol Biol. 2017;1517:3-22.##Pavet V, Portal MM, Moulin JC, Herbrecht R, Gronemeyer H. Towards novel paradigms for cancer therapy. Oncogene. 2011;30(1):1-20.##Zhang P, Wu W, Chen Q, Chen M. Non-coding RNAs and their integrated networks. J Integr Bioinform. 2019;16(3):20190027. ##Reddy KB. MicroRNA (miRNA) in cancer. Cancer Cell Int. 2015;15:38.##Calin GA, Dumitru CD, Shimizu M, Bichi R, Zupo S, Noch E, et al. Frequent deletions and down-regulation of micro-RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc Natl Acad Sci USA. 2002;99(24):15524-9.##Tüfekci KU, Oner MG, Meuwissen RL, Genç S. The role of microRNAs in human diseases. Methods Mol Biol. 2014;1107:33-50. ##Grzywa TM, Klicka K, Włodarski PK. Regulators at every step-how microRNAs drive tumor cell invasiveness and metastasis. Cancers (Basel). 2020;12(2):3709.##Supernat A, Lapinska-Szumczyk S, Majewska H, Gulczynski J, Biernat W, Wydra D. et al. Epithelial-mesenchymal transition and cancer stem cells in endometrial cancer. Anticancer Res. 2013;33(12):5461-9##Liu P, Wang, C, Ma C, Wu Q, Zhang W, Lao G. MicroRNA-23a regulates epithelial-to-mesenchymal transition in endometrial endometrioid adenocarcinoma by targeting SMAD3. Cancer Cell Int. 2016;16(1):67.##Hrzenjak A, Dieber-Rotheneder M, Moinfar F, Petru E, Zatloukal K. Molecular mechanisms of endometrial stromal sarcoma and undifferentiated endometrial sarcoma as premises for new therapeutic strategies. Cancer Lett. 2014;354(1):21-7.##Wu X, Han Y, Liu F, Ruan L. Downregulations of miR-449a and miR-145-5p Act as prognostic biomarkers for endometrial cancer. J Comput Biol. 2020;27(5):834-44.##Liu Y, Chang Y, Cai Y. Hsa_circ_0061140 promotes endometrial carcinoma progression via regulating miR-149-5p/STAT3. Gene. 2020;745:144625. ##Su Y, Wang J, Ma Z, Gong W, Yu L. miR-142 suppresses endometrial cancer proliferation in vitro and in vivo by targeting cyclin D1. DNA Cell Biol. 2019;38(2):144-50.##Favier A, Rocher G, Larsen AK, Delangle R, Uzan C, Sabbah M, et al. MicroRNA as epigenetic modifiers in endometrial cancer: a systematic review. Cancers (Basel). 2021;13(5):1137. ##Montagnana M, Benati M, Danese E, Giudici S, Perfranceschi M, Ruzzenenete O, et al. Aberrant microRNA expression in patients with endometrial cancer. Int J Gynecol Cancer. 2017;27(3):459-66. ##Wilczynski M, Senderowska D, Krawczyk T, Szymanska B, Malinowski A. MiRNAs in endometrioid endometrial cancer metastatic loci derived from positive lymph nodes. Acta Obstet Gynecol Scand. 2020;99(8):1085-1091.##Ali Syeda Z, Langden SSS, Munkhzul C, Lee M, Song SJ. Regulatory mechanism of microRNA expression in cancer. Int J Mol Sci. 2020;21(5):1723. ##Chen P, Xing T, Wang Q, Liu A, Liu H, Hu Y, et al. MicroRNA-202 inhibits cell migration and invasion through targeting FGF2 and inactivating Wnt/β-catenin signaling in endometrial carcinoma. Biosci Rep. 2019;39(10):BSR20190680. ##Li W, Meng Z, Zou T, Wang G, Su Y, Yao S, et al. MiR-374a activates Wnt/beta-catenin signaling to promote osteosarcoma cell migration by targeting WIF-1. Pathol Oncol Res. 2020;26(1):533-9.##van der Zee M, Jia Y, Wang Y, Heijmans‐Antonissen C, Ewing PC, Franken P, et al. Alterations in Wnt-beta-catenin and Pten signalling play distinct roles in endometrial cancer initiation and progression. J Pathol. 2013;230(1):48-58.##Sun X, Hou L, Qiu C, Kong B. MiR-501 promotes tumor proliferation and metastasis by targeting HOXD10 in endometrial cancer. Cell Mol Biol Lett. 2021;26(1):20.##Yuan YH, Wang HY, Lai Y, Zhong W, Liang WL, Yan FD, et al. Epigenetic inactivation of HOXD10 is associated with human colon cancer via inhibiting the RHOC/AKT/MAPK signaling pathway. Cell Commun Signal. 2019;17(1):9.##Lin Z, Hu Y, Lin R, Ye H. The efect of overexpression of the HOXD10 gene on the malignant proliferation, invasion, and tumor formation of pancreatic cancer cell PANC-1. Gland Surg. 2020;9(2):385-91.##Nakayama I, Shibazaki M, Yashima-Abo A, Miura F, Sugiyama T, Masuda T, et al. Loss of HOXD10 expression induced by upregulation of miR-10b accelerates the migration and invasion activities of ovarian cancer cells. Int J Oncol. 2013;43(1):63-71.##Yachi K, Tsuda M, Kohsaka S, Wang L, Oda Y, Tanikawa S, et al. miR-23a promotes invasion of glioblastoma via HOXD10-regulated glial-mesenchymal transition. Signal Transduct Target Ther. 2018;3:33.##Yang F, Liu D, Deng Y, Wang J, Mei S, Ge S, et al. Frequent promoter methylation of HOXD10 in endometrial carcinoma and its pathological signifcance. Oncol Lett. 2020;19(5):3602-8.##Zhang J, Liu S, Zhang D, Ma Z, Sun L. Homeobox D10, a tumor suppressor, inhibits the proliferation and migration of esophageal squamous cell carcinoma. J Cell Biochem. 2019;120(8):13717-25.##Xiao H, Li H, Yu G, Xiao W, Hu J, Tang K, et al. MicroRNA-10b promotes migration and invasion through KLF4 and HOXD10 in human bladder cancer. Oncol Rep. 2014;31(4):1832-8.##Ma J, Li D, Kong FF, Yang D, Yang H, Ma XX, miR-302a-5p/367-3p-HMGA2 axis regulates malignant processes during endometrial cancer development. J Exp Clin Cancer Res. 2018;37(1):19.##Wei L, Liu X, Zhang W, Wei Y, Li Y, Zhang Q, et al. Overexpression and oncogenic function of HMGA2 in endometrial serous carcinogenesis. Am J Cancer Res. 2016;6(2):249-59.##Kong J, He X, Wang Y, Li J. Effect of microRNA-29b on proliferation, migration, and invasion of endometrial cancer cells. J Int Med Res. 2019;47(8):3803-17.##Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol. 2014;740:364-78.##Moxley KM, McMeekin DS. Endometrial carcinoma: a review of chemotherapy, drug resistance, and the search for new agents. Oncologist. 2010;15(10):1026-33.##Chen HX, Xu XX, Tan BZ, Zhang Z, Zhou XD. MicroRNA-29b inhibits angiogenesis by targeting VEGFA through the MAPK/ERK and PI3K/Akt signaling pathways in endometrial carcinoma. Cell Physiol Biochem. 2017;41(3):933-46.##Xiong H, Wang N, Chen H, Zhang M, Lin Q. MicroRNA 199a/b 5p inhibits endometrial cancer cell metastasis and invasion by targeting FAM83B in the epithelial to mesenchymal transition signaling pathway. Mol Med Rep. 2021;23(5):304.##Shen CQ, Yan TT, Liu W, Zhu XQ, Tian XL, Fu XL, et al. High expression of FAM83B predicts poor prognosis in patients with pancreatic ductal adeno carcinoma and correlates with cell cycle and cell proliferation. J Cancer. 2017;8(16):3154-65.##Zou Z, Ma T, He X, Zhou J, Ma H, Xie M, et al. Long intergenic non coding RNA 00324 promotes gastric cancer cell proliferation via binding with HuR and stabilizing FAM83B expression. Cell Death Dis. 2018;9(7):717.##Wang J, Zhang L, Jiang W, Zhang R, Zhang B, Silayiding A, et al. MicroRNA-135a promotes proliferation, migration, invasion and induces chemoresistance of endometrial cancer cells. Eur J Obstet Gynecol Reprod Biol. 2019;5:100103.##Kong D, Suzuki A, Zou TT, Sakurada A, Kemp LW, Wakatsuki S, et al. PTEN1 is frequently mutated in primary endometrial carcinomas. Nat Genet. 1997;17(2):143-4.##Zhang W, Chen JH, Shan T, Aguilera-Barrantes I, Wang LS, Huang TH, et al. miR-137 is a tumor suppressor in endometrial cancer and is repressed by DNA hypermethylation. Lab Invest. 2018;98(11):1397-407.##Wang J, Yu L, Cai J, Jia J, Gao Y, Liang M, et al. The role of EZH2 and DNA methylation in hMLH1 silencing in epithelial ovarian cancer. Biochem Biophys Res Commun. 2013;433(4):470-6.##Lu Y, Wajapeyee N, Turker MS, Glazer PM. Silencing of the DNA mismatch repair gene MLH1 induced by hypoxic stress in a pathway dependent on the histone demethylase LSD1. Cell Rep. 2014;8(2):501-13.##Yang Q, Laknaur A, Elam L, Ismail N, Gavrilova-Jordan L, Lue J, et al. Identification of polycomb group protein EZH2-mediated DNA mismatch repair gene MSH2 in human uterine fibroids. Reprod Sci. 2016;23(10):1314-25.##Snowdon J, Zhang X, Childs T, Tron VA, Feilotter H. The microRNA-200 family is upregulated in endometrial carcinoma. PloS One. 2011;6(8):e22828.##Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, et al. The miR200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 2008;10(5):593-601.##Park SM, Gaur AB, Lengyel E, Peter ME. The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev. 2008;22(7):894-907.##Korpal M, Lee ES, Hu G, Kang Y. The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. J Biol Chem. 2008;283(22):14910-4. ##Nam EJ, Yoon H, Kim SW, Kim H, Kim YT, et al. MicroRNA expression profiles in serous ovarian carcinoma. Clin Cancer Res. 2008;14(9):2690-5.##Hu X, Macdonald DM, Huettner PC, Feng Z, El Naqa IM, Schwarz JK, et al. A miR200 microRNA cluster as prognostic marker in advanced ovarian cancer. Gynecol Oncol. 2009;114(3):457-64. ##Cochrane DR, Cittelly DM, Richer JK. Steroid receptors and microRNAs: Relationships revealed. Steroids. 2010;76(1-2):2-10.##Krasner C. Aromatase inhibitors in gynecologic cancers. J Steroid Biochem Mol Biol. 2007;106(1-5):76-80.##Wilczynski M, Danielska J, Dzieniecka M, Szymanska B, Wojciechowski M, Malinowski A. Prognostic and Clinical Significance of miRNA-205 in Endometrioid Endometrial Cancer. PLoS One. 2016;11(10):e0164687.##Matias-Guiu X, Prat J. Molecular pathology of endometrial carcinoma. Histopathology. 2013;62(1):111-23.##Zhang S, Yu D. PI(3)king apart PTEN’s role in cancer. Clin Cancer Res. 2010;16(17):4325-30.##Karaayvaz M, Zhang C, Liang S, Shroyer KR, Ju J. Prognostic significance of miR-205 in endometrial cancer. PLoS One. 2012;7(4):e35158.##Wu W, Lin Z, Zhuang Z, Liang X. Expression profile of mammalian microRNAs in endometrioid adenocarcinoma. Eur J Cancer Prev. 2009;18(1):50-5.##He S, Zeng S, Zhou ZW, He ZX, Zhou SF. Hsa-microRNA-181a is a regulator of a number of cancer genes and a biomarker for endometrial carcinoma in patients: a bioinformatic and clinical study and the therapeutic implication. Drug Des Devel Ther. 2015;9:1103-75. ##Chen Y, Ke G, Han D, Liang S, Yang G, Wu X. MicroRNA-181a enhances the chemoresistance of human cervical squamous cell carcinoma to cisplatin by targeting PRKCD. Exp Cell Res. 2014;320(1):12-20.##Panda H, Chuang TD, Luo X, Chegini N. Endometrial miR-181a and miR-98 expression is altered during transition from normal into cancerous state and target PGR, PGRMC1, CYP19A1, DDX3X, and TIMP3. J Clin Endocrinol Metab. 2012;97(7):E1316-26.##Parikh A, Lee C, Peronne J, Marchini S, Baccarini A, Kolev V, et al. microRNA-181a has a critical role in ovarian cancer progression through the regulation of the epithelial-mesenchymal transition. Nat Commun. 2014;5:2977.##Meng F, Henson R, Lang M, Wehbe H, Maheshwari S, Mendel JT, et al. Involvement of human micro-RNA in growth and response to chemotherapy in human cholangiocarcinoma cell lines. Gastroenterology. 2006;130(7):2113-29.##Velasco A, Bussaglia E, Pallares J, Dolcet X, Llobet D, Encinas M, et al. PIK3CA gene mutations in endometrial carcinoma: correlation with PTEN and K-RAS alterations. Hum Pathol. 2006;37(11):1465-72.##Strickland AL, Rivera G, Lucas E, John G, Cuevas I, Castrillon DH. PI3K Pathway effectors pAKT and FOXO1 as novel markers of endometrioid intraepithelial neoplasia. Int J Gynecol Pathol. 2019;38(6):503-13.##Catasus L, Gallardo A, Cuatrecasas M, Prat J. PIK3CA mutations in the kinase domain (exon 20) of uterine endometrial adenocarcinomas are associated with adverse prognostic parameters. Mod Pathol. 2008;21(2):131-9.##Salvesen HB, Carter SL, Mannelqvist M, Dutt A, Getz G, Stefansson IM, et al. Integrated genomic profiling of endometrial carcinoma associates aggressive tumors with indicators of PI3 kinase activation. Proc Natl Acad Sci USA. 2009;106(12):4834-9.##Gao Q, Ye F, Xia X, Xing H, Lu Y, Zhou J, et al. Correlation between PTEN expression and PI3K/Akt signal pathway in endometrial carcinoma. J Huazhong Univ Sci Technol Med Sci. 2009;29(1):59-63. ##Maxwell GL, Risinger JI, Gumbs C, Shaw H, Bentley RC, Barrett JC, et al. Mutation of the PTEN tumor suppressor gene in endometrial hyperplasias. Cancer Res. 1998;58(12):2500-3.##Levine RL, Cargile CB, Blazes MS, van Rees B, Kurman RJ, Ellenson LH. PTEN mutations and microsatellite instability in complex atypical hyperplasia, a precursor lesion to uterine endometrioid carcinoma. Cancer Res. 1998;58(15):3254-8.##Mutter GL, Lin MC, Fitzgerald JT, Kum JB, Baak JP, Lees JA, et al. Altered PTEN expression as a diagnostic marker for the earliest endometrial precancers. J Natl Cancer Inst. 2000;92(11):924-30.##Zhou XP, Kuismanen S, Nystrom-Lahti M, Peltomaki P, Eng C. Distinct PTEN mutational spectra in hereditary non-polyposis colon cancer syndrome-related endometrial carcinomas compared to sporadic microsatellite unstable tumors. Hum Mol Genet. 2002;11(4):445-50.##Cheung LW, Hennessy BT, Li J, Yu S, Myers AP, Djordjevic B, et al. High frequency of PIK3R1 and PIK3R2 mutations in endometrial cancer elucidates a novel mechanism for regulation of PTEN protein stability. Cancer Discov. 2011;1(2):170-85.##Urick ME, Rudd ML, Godwin AK, Sgroi D, Merino M, Bell DW. PIK3R1 (p85α) is somatically mutated at high frequency in primary endometrial cancer. Cancer Res. 2011;71(12):4061-7.##Matsushima K, Isomoto H, Yamaguchi N, Inoue N, Machida H, Nakayama T, et al. MiRNA-205 modulates cellular invasion and migration via regulating zinc finger E-box binding homeobox 2 expression in esophageal squamous cell carcinoma cells. J Transl Med. 2011;9:30.##Luo X, Dong Z, Chen Y, Yang L, Lai D. Enrichment of ovarian cancer stem-like cells is associated with epithelial to mesenchymal transition through an miRNA-activated AKT pathway. Cell Prolif. 2013;46(4):436-46.##Qin X, Yan L, Zhao X, Li C, Fu Y. microRNA-21 overexpression contributes to cell proliferation by targeting PTEN in endometrioid endometrial cancer. Oncol Lett. 2012;4(6):1290-6.##Sayed D, Abdellatif M. AKT-ing via microRNA. Cell Cycle. 2010;9(16):3213-7.##Fang Y, Xue JL, Shen Q, Chen J, Tian L. MicroRNA-7 inhibits tumor growth and metastasis by targeting the phosphoinositide 3-kinase/Akt pathway in hepatocellular carcinoma. Hepatology. 2012;55(6):1852-62.##Dedes KJ, Wetterskog D, Mendes-Pereira AM, Natrajan R, Lambros MB, Geyer FC, et al. PTEN deficiency in endometrioid endometrial adenocarcinomas predicts sensitivity to PARP inhibitors. Sci Transl Med. 2010;2(53):53ra75.##Cancer genome atlas research network, Kandoth C, Schultz N, Cherniack AD, Akbani R, Liu Y, et al. Integrated genomic characterization of endometrial carcinoma. Nature. 2013;497(7447):67-73.##Dravid G, Ye Z, Hammond H, Chen G, Pyle A, Donovan P, et al. Defining the role of Wnt/beta-catenin signaling in the survival, proliferation, and selfrenewal of human embryonic stem cells. Stem Cells. 2005;23(10):1489-501.##Nusse R, Clevers H. Wnt/beta-catenin signaling, disease, and emerging therapeutic modalities. Cell. 2017;169(6):985-99.##Gumbiner BM. Signal transduction of beta-catenin. Curr Opin Cell Biol. 1995;7(5):634-40.##Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B, et al. Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. Science. 1997;275(5307):1787-90.##Kim G, Kurnit KC, Djordjevic B, Singh C, Munsell MF, Wang WL, et al. Nuclear β-catenin localization and mutation of the CTNNB1 gene: a context-dependent association. Mod Pathol. 2018;31(10):1553-9.##Wang Y, Yu M, Yang JX, Cao DY, Zhang Y, Zhou HM, et al. Genomic comparison of endometrioid endometrial carcinoma and its precancerous lesions in Chinese patients by high-depth next generation sequencing. Front Oncol. 2019;9:123. ##Lei Y, Chen L, Zhang G, Shan A, Ye C, Liang B, et al. MicroRNAs target the Wnt/β catenin signaling pathway to regulate epithelial mesenchymal transition in cancer (Review). Oncol Rep. 2020;44(4):1299-1313. ##Li Y, Sun D, Gao J, Shi Z, Chi P, Meng Y, et al. MicroRNA-373 promotes the development of endometrial cancer by targeting LATS2 and activating the Wnt/β-Catenin pathway. J Cell Biochem. 2019;120(5):8611-8.##Axelson H. Notch signaling and cancer: emerging complexity. Semin Cancer Biol. 2004;14(5):317-9.##Bigas A, Robert-Moreno A, Espinosa L. The Notch pathway in the developing hematopoietic system. Int J Dev Biol. 2010;54(6-7):1175-88.##Bolos V, Grego-Bessa J, de la Pompa JL. Notch signaling in development and cancer. Endocr Rev. 2007;28(3):339-63. ##Artavanis-Tsakonas S, Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development. Science. 1999;284(5415):770-6. ##Wei Y, Zhang Z, Liao H, Wu L, Wu X, Zhou D, et al. Nuclear estrogen receptor-mediated Notch signaling and GPR30-mediated PI3K/AKT signaling in the regulation of endometrial cancer cell proliferation. Oncol Rep. 2012;27(2):504-10. ##Rose SL. Notch signaling pathway in ovarian cancer. Int J Gynecol Cancer. 2009;19(4):564-6.##Gao Y, Liu T, Huang Y. MicroRNA-134 suppresses endometrial cancer stem cells by targeting POGLUT1 and Notch pathway proteins. FEBS lett. 2015;589(2):207-14. ##Schubbert S, Shannon K, Bollag G. Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer. 2007;7(4):295-308. ##Berg A, Hoivik EA, Mjos S, Holst F, Werner HM, Tangen IL, et al. Molecular profiling of endometrial carcinoma precursor, primary and metastatic lesions suggests different targets for treatment in obese compared to non-obese patients. Oncotarget. 2015;6(2):1327-39. ##Rudd ML, Price JC, Fogoros S, Godwin AK, Sgroi DC, Merino MJ, et al. A unique spectrum of somatic PIK3CA (p110alpha) mutations within primary endometrial carcinomas. Clin Cancer Res. 2011;17(6):1331-40. ##Oda K, Okada J, Timmerman L, Rodriguez-Viciana P, Stokoe D, Shoji K, et al. PIK3CA cooperates with other phosphatidylinositol 3′- kinase pathway mutations to effect oncogenic transformation. Cancer Res. 2008;68(19):8127-36.##Chen HX, Xu XX, Tan BZ, Zhang Z, Zhou XD. MicroRNA-29b inhibits angiogenesis by targeting VEGFA through the MAPK/ERK and PI3K/Akt signaling pathways in endometrial carcinoma. Cell Physiol Biochem. 2017;41(3):933-46.##Mutlu M, Saatci Ö, Ansari SA, Yurdusev E, Shehwana H, Konu Ö, et al. miR-564 acts as a dual inhibitor of PI3K and MAPK signaling networks and inhibits proliferation and invasion in breast cancer. Sci Rep. 2016;6:32541.##Owen KL, Brockwell NK, Parker BS. JAK-STAT Signaling: a double-edged sword of immune regulation and cancer progression. Cancers (Basel). 2019;11(12):2002.##Subramaniam KS, Omar IS, Kwong SC, Mohamed Z, Woo YL, Adenan NA, et al. Cancer-associated fibroblasts promote endometrial cancer growth via activation of interleukin-6/STAT-3/c-Myc pathway. Am J Cancer Res. 2016;6(2):200-13.##Huang Y, Yang N. MicroRNA-20a-5p inhibits epithelial to mesenchymal transition and invasion of endometrial cancer cells by targeting STAT3. Int J Clin Exp Pathol. 2018;11(12):5715-24.##Li Y, Zhang Z, Liu X, Huang T, He W, Shen Y, et al. miR-124 functions as a tumor suppressor in the endometrial carcinoma cell line HEC-1B partly by suppressing STAT3. Mol Cell Biochem. 2014;388(1-2):219-31.##Dong P, Xiong Y, Yue J, Xu D, Ihira K, Konno Y, et al. Long noncoding RNA NEAT1 drives aggressive endometrial cancer progression via miR-361-regulated networks involving STAT3 and tumor micro-environment related genes. J Exp Clin Cancer Res. 2019;38(1):295.##Hutt S, Tailor A, Ellis P, Michael A, Butler-Manuel S, Chatterjee J. The role of biomarkers in endometrial cancer and hyperplasia: a literature review. Acta Oncol. 2019;58(3):342-52.##Moreno-Moya JM, Vilella F, Simón C. MicroRNA: key gene expression regulators. Fertil Steril. 2014;101(6):1516-23.##Boren T, Xiong Y, Hakam A, Wenham R, Apte S, Wei Z, et al. MicroRNAs and their target messenger RNAs associated with endometrial carcinogenesis. Gynecol Oncol. 2008;110(2):206-15.##Torres A, Torres K, Pesci A, Ceccaroni M, Paszkowski T, Cassandrini P, et al. Diagnostic and prognostic significance of miRNA signatures in tissues and plasma of endometrioid endometrial carcinoma patients. Int J Cancer. 2013;132(7):1633-45.##Bao W, Zhang Y, Li S, Fan Q, Qiu M, Wang Y, et al. miR-107-5p promotes tumor proliferation and invasion by targeting estrogen receptor-α in endometrial carcinoma. Oncol Rep. 2019;41(3):1575-85. ##Tsukamoto O, Miura K, Mishima H, Abe S, Kaneuchi M, Higashijima A, et al. Identification of endometrioid endometrial carcinoma-associated microRNAs in tissue and plasma. Gynecol Oncol. 2014;132(3):715-21. ##Zhang X, Dong Y, Ti H, Zhao J, Wang Y, Li T, et al. Down-regulation of miR-145 and miR-143 might be associated with DNA methyltransferase 3B overexpression and worse prognosis in endometrioid carcinomas. Hum Pathol. 2013;44(11):2571-80.##De Foucher T, Sbeih M, Uzan J, Bendifallah S, Lefevre M, Chabbert-Buffet N, et al. Identification of micro-RNA expression profile related to recurrence in women with ESMO low-risk endometrial cancer. J Transl Med. 2018;16(1):131. ##Yu S, Liu Y, Wang J, Guo Z, Zhang Q, Yu F, et al. Circulating microRNA profiles as potential biomarkers for diagnosis of papillary thyroid carcinoma. J Clin Endocrinol Metab. 2012;97(6):2084-92.##Srivastava A, Moxley K, Ruskin R, Dhanasekaran DN, Zhao YD, Ramesh RA. Non-invasive Liquid Biopsy Screening of Urine-Derived Exosomes for miRNAs as Biomarkers in Endometrial Cancer Patients. AAPS J. 2018;20(5):82. ##Mall C, Rocke D, Durbin-Johnson B, Weiss R. Stability of miRNA in human urine supports its biomarker potential. Biomark Med. 2013;7(4):623-31. ##Tan ZQ, Liu FX, Tang HL, Su Q. [Expression and its clinical significance of hsa-miR-155 in serum of endometrial cancer]. Zhonghua Fu Chan Ke Za Zhi. 2010;45(10):772-4. Chinese.##Torres A, Torres K, Pesci A, Ceccarino M, Paszkowski T, Cassandrini P, et al. Deregulation of miR-100, miR-99a and miR-199b in tissues and plasma coexists with increased expression of mTor kinase in endometrioid endometrial carcinoma. BMC Cancer. 2012;12:369.##Jurcevic S, Olsson B, Klinga-Levan K. MicroRNA expression in human endometrial adenocarcinoma. Cancer Cell Int. 2014;14(1):88.##Wang F, Zhong S, Zhang H, Zhang W, Zhang H, Wu X, et al. Prognostic value of MicroRNA-182 in cancers: a meta-analysis. Dis Markers. 2015;2015:482146.##Chung TK, Lau TS, Cheung TH, Yim SF, Lo KW, Siu NS, et al. Dysregulation of microRNA-204 mediates migration and invasion of endometrial cancer by regulating FOXC1. Int J Cancer. 2012;130(5):1036-45.##Zhai H, Karaayvaz M, Dong P, Sakuragi N, Ju J. Prognostic significance of miR-194 in endometrial cancer. Biomark Res. 2013;1:12.##Dong P, Kaneuchi M, Watari H, Hamada J, Sudo S, Ju J, et al. MicroRNA-194 inhibits epithelial to mesenchymal transition of endometrial cancer cells by targeting oncogene BMI-1. Mol Cancer. 2011;10:99.##Garzon R, Marcucci G, Croce CM. Targeting microRNAs in cancer: rationale, strategies and challenges. Nat Rev Drug Discov. 2010;9(10):775-89, ##Xiao F, Bai Y, Chen Z, Li Y, Luo L, Huang J, et al. Downregulation of HOXA1 gene affects small cell lung cancer cell survival and chemoresistance under the regulation of miR-100. Eur J Cancer. 2014;50(8):1541-54.##Liu MX, Siu MK, Liu SS, Yam JW, Ngan HY, Chan DW. Epigenetic silencing of microRNA-199b-5p is associated with acquired chemoresistance via activation of JAG1-Notch1 signaling in ovarian cancer. Oncotarget. 2014;5(4):944-58.##Lee YS, Dutta A. The tumor suppressor microRNA let-7 represses the HMGA2 oncogene. Genes Dev. 2007;21(9):1025-30.##Wu Q, Lu RL, Li JX, Rong LJ. MiR-200a and miR-200b target PTEN to regulate the endometrial cancer cell growth in vitro. Asian Pac J Trop Med. 2017;10(5):498-502.##Zhuo Z, Yu H. miR-205 inhibits cell growth by targeting AKT-mTOR signaling in progesterone-resistant endometrial cancer Ishikawa cells. Oncotarget. 2017;8(17):28042-51. ##Weigelt B, Warne PH, Lambros MB, Reis-Filho JS, Downward J. PI3K pathway dependencies in endometrioid endometrial cancer cell lines. Clin Cancer Res. 2013;19(13):3533-44. ##Shen Y, Lu L, Xu J, Meng W, Qing Y, Liu Y, et al. Bortezomib induces apoptosis of endometrial cancer cells through microRNA-17-5p by targeting p21. Cell Biol Int. 2013;37(10):1114-21.##Jiang L, Meng W, Zeng J, Hu H, Lu L. MiR-34c oligonucleotide enhances chemosensitivity of Ishikawa cell to cisplatin by inducing apoptosis. Cell Biol Int. 2013;37(6):577-83.##Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA. 2008;105(30):10513-8.##Chen Y, Zhu X, Zhang X, Liu B, Huang L. Nanoparticles modified with tumor-targeting scFv deliver siRNA and miRNA for cancer therapy. Mol Ther. 2010;18(9):1650-6.##Sharma S, Kelly TK, Jones PA. Epigenetics in cancer. Carcinogenesis. 2009;31(1):27-36.##Chung TK, Cheung TH, Huen NY, Wong KW, Lo KW, Yim SF, et al. Dysregulated microRNAs and their predicted targets associated with endometrioid endometrial adenocarcinoma in Hong Kong women. Int J Cancer. 2009;124(6):1358-65.##Chan E, Prado DE, Weidhaas JB. Cancer microRNAs: from subtype profiling to predictors of response to therapy. Trends Mol Med. 2011;17(5):235-43.##Chung TK, Lau TS, Cheung TH, Yim SF, Lo KW, Siu NS, et al. Dysregulation of microRNA‐204 mediates migration and invasion of endometrial cancer by regulating FOXC1. Int J Cancer. 2012;130(5):1036-45. ##Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA. 2008;105(30):10513-8. ##Tsuruta T, Kozaki KI, Uesugi A, Furuta M, Hirasawa A, Imoto I, et al. miR-152 is a tumor suppressor microRNA that is silenced by DNA hypermethylation in endometrial cancer. Cancer Res. 2011;71(20):6450-62.##

Comparison of Diff-Quick and Spermac Staining Methods for Sperm Morphology Evaluation 2 1 2 Background: The objective of the current study was comparing the impact of two staining techniques on semen morphological parameters and their influence on patient diagnosis. The ideal staining method should preserve cell integrity while providing detailed information. Methods: Semen samples from fifty men were stained using Diff-Quick or Spermac methods. Morphological parameters were classified based on the Tygerberg criteria, and final diagnosis was according to WHO manual guidelines. Statistical analysis was performed through conducting paired t-tests or Wilcoxon rank-sum tests, with GLIMMIX and Fisher's exact test for determining the significance (p≤0.05). Results: Both staining methods highlighted head and tail regions, with Spermac offering better visualization of the midpiece. Spermac demonstrated fewer normal spermatozoa (2.8±0.3%) compared to Diff-Quick (3.98±0.4%; p=0.0385). Midpiece abnormalities were more evident with Spermac (55.7±2.1%) than Diff-Quick (24.8±2.0%; p<0.0001). No significant difference was found in head and tail abnormalities (p>0.05). Conclusion: Diff-Quick staining resulted in a higher proportion of normal spermatozoa, primarily due to its midpiece evaluation. The choice of staining method significantly impacts the diagnosis of infertile males. These findings have important implications for clinical practice and future research, suggesting the need for further investigations to assess different staining methods and determine optimal diagnostic thresholds. 166 171 Lincoln LBF Junior Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro) Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro) Brazil André AR da Cunha Barreto-Vianna Federal University of Paraná-Palotina Federal University of Paraná-Palotina Brazil Mariana MD de Mello Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro) Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro) Brazil Alexandre AL dos Santos Federal University of Paraná-Palotina Federal University of Paraná-Palotina Brazil Cristiane C da Fonte Ramos State University of Rio de Janeiro State University of Rio de Janeiro Brazil Paula P Fontoura Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro) Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro) Brazil contato@bsrj.com.br HumanMorphologySpermatozoaStaining 140192.pdf World Health organization. WHO laboratory manual for the examination and processing of human semen. 6th ed. Geneva: World Health Organization; 2021. 292 p.##Gatimel N, Moreau J, Parinaud J, Léandri RD. Sperm morphology: assessment, pathophysiology, clinical relevance, and state of the art in 2017. Andrology. 2017;5(5):845-62.##Maree L, Plessis SS Du, Menkveld R, Horst G Van Der. Morphometric dimensions of the human sperm head depend on the staining method used. Hum Reprod. 2010;25(6):1369-82.##Horst G Van Der, Maree L. SpermBlue: a new universal stain for human and animal sperm which is also amenable to automated sperm morphology analysis. Biotech Histochem. 2009;84(6):299-308.##Mortimer D, Björndahl L, Barratt CL, Castilla JA, Menkveld R, Kvist U, et al. A practical guide to basic laboratory andrology. 2nd ed. UK: Cambridge university press; 2022. 352 p.##Tuset VM, Dietrich GJ, Wojtczak M, Słowińska M, Monserrat J de, Ciereszko A. Comparison of three staining techniques for the morphometric study of rainbow trout (Oncorhynchus mykiss) spermatozoa. Theriogenology. 2008;69(8):1033-8.##Runcan EE, Pozor MA, Zambrano GL, Benson S, Macpherson ML. Use of two conventional staining methods to assess the acrosomal status of stallion spermatozoa. Equine Vet J. 2014;46(4):503-6.##World Health organization. WHO laboratory manual for the examination and processing of human semen. 5nd ed. Geneva: World Health Organization; 2010. 271 p.##Menkveld R, Stander FS, Kotze TJ, Kruger TF, Zyl JA van. The evaluation of morphological characteristics of human spermatozoa according to stricter criteria. Hum Reprod. 1990;5(5):586-92.##Gonçalves J, Pizzichini E, Pizzichini MMM, Steidle LJM, Rocha CC, Ferreira SC, et al. Reliability of a rapid hematology stain for sputum cytology. J Bras Pneumol. 2014;40(3):250-8.##Henkel R, Schreiber G, Sturmhoefel A, Hipler UC, Zermann DH, Menkveld R. Comparison of three staining methods for the morphological evaluation of human spermatozoa. Fertil Steril. 2008;89(2):449-55.##Natali I, Muratori M, Sarli V, Vannuccini M, Cipriani S, Niccoli L, et al. Scoring human sperm morphology using Testsimplets and Diff-Quik slides. Fertil Steril. 2013;99(5):1227-32.e2.##Chan PJ, Corselli JU, Jacobson JD, Patton WC, King A. Spermac stain analysis of human sperm acrosomes. Fertil Steril. 1999;72(1):124-8.##Schaäfer S, Holzmann A. The use of transmigration and Spermac stain to evaluate epididymal cat spermatozoa. Anim Reprod Sci. 2000;59(3-4):201-11.##Agarwal A, Sharma R, Gupta S, Finelli R, Parekh N, Selvam MKP, et al. Sperm morphology assessment in the era of intracytoplasmic sperm injection: Reliable results require focus on standardization, quality control, and training. World J Men's Health. 2022;40(3):347-60.##Chemes HE, Alvarez Sedo C. Tales of the tail and sperm head aches: changing concepts on the prognostic significance of sperm pathologies affecting the head, neck and tail. Asian J Androl. 2012;14(1):14-23.##Nakamura S, Terada Y, Rawe VY, Uehara S, Morito Y, Yoshimoto T, et al. A trial to restore defective human sperm centrosomal function. Hum Reprod. 2005;20(7):1933-7.##Ugajin T, Terada Y, Hasegawa H, Nabeshima H, Suzuki K, Yaegashi N. The shape of the sperm midpiece in intracytoplasmic morphologically selected sperm injection relates sperm centrosomal function. J Assist Reprod Genet. 2010;27(2-3):75-81.##

Sox2 Localization During Spermatogenesis and Its Association with other Spermatogenesis Markers Using Protein-Protein Network Analysis 2 1 2 Background: Sox2 (SRY box2) is an essential transcription factor that plays a vital role in spermatogenesis and regulates the genes in this process. Sox2 is important for pluripotency, self-renewal, and even spermatogonial stem cell differentiation. This gene is found in pluripotent and specialized cells, and it is involved in their biological activities. Methods: Protein-protein interaction (PPI) network analysis was performed during spermatogenesis using NCBI, STRING, and Cytoscape databases. Then, after isolating spermatogonial stem cells from 6 C57BL/6 mice, mouse embryonic stem cells and ES-like cells were prepared. In the following, Sox2 expression was examined in differentiated and undifferentiated spermatogonia by immunohistochemistry (IMH), immunocytochemistry (ICC), and Fluidigm PCR (polymerase chain reaction). Finally, the results were compared using the Kruskal-Wallis and Dunn tests at the significance level of p<0.05. Results: The results of this experiment showed that contrary to expectations, Sox2 has cytoplasmic expression in undifferentiated cells and nuclear expression in differentiated cells in in vitro conditions. In addition, the expression of Sox2 increased during differentiation. Fluidigm PCR showed a significantly higher expression of Sox2 (p<0.05) in differentiated compared to undifferentiated spermatogonia. Sox2 has an interaction with other genes during spermatogenesis such as Oct4, Nanog, Klf4, Stra8, Smad1, Tcf3, and Osm. Conclusion: Sox2, which is known as a pluripotency marker, has a vital role in spermatogenesis and could be a differential marker. Sox2 has strong connections with other genes such as Oct4, Nanog, Klf4, Tcf3, Osm, Stra8, Lim2, Smad1, Gdnf, and Kit. 171 181 Emad E Reza Department of Nanobiotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies Department of Nanobiotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies Iran Hossein H Azizi Department of Nanobiotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies Department of Nanobiotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies Iran h.azizi@ausmt.ac.ir Thomas T Skutella Institute for Anatomy and Cell Biology, Medical Faculty, University of Heidelberg Institute for Anatomy and Cell Biology, Medical Faculty, University of Heidelberg Germany Adult germline stem cellsCell differentiationSeminiferous tubulesSox2 proteinTranscription factors 140190.pdf Kubota H, Brinster RL. Spermatogonial stem cells. Biol Reprod. 2018;99(1):52-74.##Potter SJ, DeFalco T. Role of the testis interstitial compartment in spermatogonial stem cell function. Reproduction. 2017;153(4):R151-62.##Oatley JM. Recent advances for spermatogonial stem cell transplantation in livestock. Reprod Fertil Dev. 2018;30(1):44-9.##Jung D, Xiong J, Ye M, Qin X, Li L, Cheng S, et al. In vitro differentiation of human embryonic stem cells into ovarian follicle-like cells. Nat Commun. 2017;8:15680.##Law NC, Oatley MJ, Oatley JM. Developmental kinetics and transcriptome dynamics of stem cell specification in the spermatogenic lineage. Nat Commun. 2019;10(1):2787.##Mardanpour P, Nayernia K, Khodayari S, Khodayari H, Molcanyi M, Hescheler J. Application of stem cell technologies to regenerate injured myocardium and improve cardiac function. Cell Physiol Biochem. 2019;53(1):101-20.##Kurimoto K, Saitou M. Germ cell reprogramming. Curr Top Dev Biol. 2019;135:91-125.##Hoang SN, Ho CNQ, Nguyen TTP, Doan CC, Tran DH, Le LT. Evaluation of stemness marker expression in bovine ovarian granulosa cells. Anim Reprod. 2019;16(2):277-81.##Kuzmin AA, Ermakova VV, Sinenko SA, Ponomartsev SV, Starkova TY, Skvortsova EV, et al. Genetic tool for fate mapping of Oct4 (Pou5f1)-expressing cells and their progeny past the pluripotency stage. Stem Cell Res Ther. 2019;10(1):391.##Mercurio S, Serra L, Nicolis SK. More than just stem cells: functional roles of the transcription factor Sox2 in differentiated glia and neurons. Int J Mol Sci. 2019;20(18):4540.##Ryu MS, Zhang D, Protchenko O, Shakoury-Elizeh M, Philpott CC. PCBP1 and NCOA4 regulate erythroid iron storage and heme biosynthesis. J Clin Invest. 2017;127(5):1786-97.##Hashemi Karoii D, Azizi H. OCT4 protein and gene expression analysis in the differentiation of spermatogonia stem cells into neurons by immunohistochemistry, immunocytochemistry, and bioinformatics analysis. Stem Cell Rev Rep. 2023 Apr 29. Ahead of Print.##Reza E, Azizi H. Comparing the expression levels of Alkaline Phosphatase, Gfra1, Lin28, and Sall4 Genes in embryonic stem cells, spermatogonial stem cells, and embryonic stem-like cells in mice. J Mazandaran Univ Med Sci. 2022;32(210):13-25.##Karagiannis P, Takahashi K, Saito M, Yoshida Y, Okita K, Watanabe A, et al. Induced pluripotent stem cells and their use in human models of disease and development. Physiol Rev. 2019;99(1):79-114.##Sojoudi K, Azizi H, Skutella T. A fundamental research in in vitro spermatogonial stem cell culturing: what are clump cells? Cell Reprogram. 2023;25(2):65-72.##Cannarella R, Condorelli RA, Mongioì LM, La Vignera S, Calogero AE. Molecular biology of spermatogenesis: novel targets of apparently idiopathic male infertility. Int J Mol Sci. 2020;21(5):1728.##Conrad S, Azizi H, Hatami M, Kubista M, Bonin M, Hennenlotter J, et al. Differential gene expression profiling of enriched human spermatogonia after short-and long-term culture. Biomed Res Int. 2014;2014:138350.##Wang Z, Kang L, Zhang H, Huang Y, Fang L, Li M, et al. AKT drives SOX2 overexpression and cancer cell stemness in esophageal cancer by protecting SOX2 from UBR5-mediated degradation. Oncogene. 2019;38(26):5250-64.##Xiao W, Zheng S, Xie X, Li X, Zhang L, Yang A, et al. SOX2 promotes brain metastasis of breast cancer by upregulating the expression of FSCN1 and HBEGF. Mol Ther Oncolytics. 2020;17:118-29.##Alatzoglou KS, Andoniadou CL, Kelberman D, Buchanan CR, Crolla J, Arriazu MC, et al. SOX2 haploinsufficiency is associated with slow progressing hypothalamo‐pituitary tumours. Hum Mutat. 2011;32(12):1376-80.##Gregory LC, Dattani MT. The molecular basis of congenital hypopituitarism and related disorders. J Clin Endocrinol Metab. 2020;105(6):dgz184.##Huangfu D, Osafune K, Maehr R, Guo W, Eijkelenboom A, Chen S, et al. Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol. 2008;26(11):1269-75.##Driessens G, Blanpain C. Long live sox2: sox2 lasts a lifetime. Cell Stem Cell. 2011;9(4):283-4.##Alonso MM, Diez-Valle R, Manterola L, Rubio A, Liu D, Cortes-Santiago N, et al. Genetic and epigenetic modifications of Sox2 contribute to the invasive phenotype of malignant gliomas. PloS One. 2011;6(11):e26740.##Xia P, Wang S, Ye B, Du Y, Huang G, Zhu P, et al. Sox2 functions as a sequence-specific DNA sensor in neutrophils to initiate innate immunity against microbial infection. Nat Immunol. 2015;16(4):366-75.##Campolo F, Gori M, Favaro R, Nicolis S, Pellegrini M, Botti F, et al. Essential role of Sox2 for the establishment and maintenance of the germ cell line. Stem Cells. 2013;31(7):1408-21.##Azizi H, Hamidabadi HG, Skutella T. Differential proliferation effects after short-term cultivation of mouse spermatogonial stem cells on different feeder layers. Cell J. 2019;21(2):186-93.##Azizi H, Conrad S, Hinz U, Asgari B, Nanus D, Peterziel H, et al. Derivation of pluripotent cells from mouse SSCs seems to be age dependent. Stem Cells Int. 2016;2016:8216312.##Azizi H, Niazi Tabar A, Skutella T. Successful transplantation of spermatogonial stem cells into the seminiferous tubules of busulfan-treated mice. Reprod Health. 2021;18(1):189.##Steevens AR, Sookiasian DL, Glatzer JC, Kiernan AE. SOX2 is required for inner ear neurogenesis. Sci Rep. 2017;7(1):4086.##Som P, Grapin-Botton A. The current embryology of the foregut and its derivatives. Neurographics. 2016;6(1):43-63.##Varela-Nallar L, Inestrosa NC. Wnt signaling in the regulation of adult hippocampal neurogenesis. Front Cell Neurosci. 2013;7:100.##Feng R, Wen J. Overview of the roles of Sox2 in stem cell and development. Biol Chem. 2015;396(8):883-91.##Dodonova SO, Zhu F, Dienemann C, Taipale J, Cramer P. Nucleosome-bound SOX2 and SOX11 structures elucidate pioneer factor function. Nature. 2020;580(7805):669-72.##Kim DK, Lee JS, Lee EY, Jang H, Han S, Kim HY, et al. O-GlcNAcylation of Sox2 at threonine 258 regulates the self-renewal and early cell fate of embryonic stem cells. Exp Mol Med. 2021;53(11):1759-68.##Myers SA, Peddada S, Chatterjee N, Friedrich T, Tomoda K, Krings G, et al. SOX2 O-GlcNAcylation alters its protein-protein interactions and genomic occupancy to modulate gene expression in pluripotent cells. Elife. 2016;5:e10647.##Jullien J, Astrand C, Halley-Stott RP, Garrett N, Gurdon JB. Characterization of somatic cell nuclear reprogramming by oocytes in which a linker histone is required for pluripotency gene reactivation. Proc Natl Acad Sci USA. 2010;107(12):5483-8.##Muhr J. Genomic occupancy in various cellular contexts and potential pioneer factor function of SOX2. In: Kondoh H, Lovell-Badge R, editors. Sox2: biology and role in development and disease. Academic Press; 2016. p. 145-59.##Chocu S, Calvel P, Rolland AD, Pineau C. Spermatogenesis in mammals: proteomic insights. Syst Biol Reprod Med. 2012;58(4):179-90.##Darzi MB, Nemati F, Azizi H, Jouybari AD. Immunohistochemistry and immunocytochemistry analysis of PLZF and VASA in mice testis during spermatogenesis. Zygote. 2023;31(3):273-80.##Bunina D, Abazova N, Diaz N, Noh KM, Krijgsveld J, Zaugg JB. Genomic rewiring of SOX2 chromatin interaction network during differentiation of ESCs to postmitotic neurons. Cell Syst. 2020;10(6):480-94.e8.##Niknejad P, Aziz H, Sojoudi K. POU5F1 protein and gene expression analysis in neonate and adult mouse testicular germ cells by immunohistochemistry and immunocytochemistry. Cell Reprogram. 2021;23(6):349-58.##Uysal F, Akkoyunlu G, Ozturk S. DNA methyltransferases exhibit dynamic expression during spermatogenesis. Reprod Biomed Online. 2016;33(6):690-702.##Wang F, Chen R, Han D. Innate immune defense in the male reproductive system and male fertility. In: Saxena ShK, Prakash H, editors. Innate immunity in health and disease: UK: IntechOpen; 2019. 229-318.##Kubo N, Toh H, Shirane K, Shirakawa T, Kobayashi H, Sato T, et al. DNA methylation and gene expression dynamics during spermatogonial stem cell differentiation in the early postnatal mouse testis. BMC Genomics. 2015;16(1):624.##Yu J, Zheng Q, Li Z, Wu Y, Fu Y, Wu X, et al. CG6015 controls spermatogonia transit-amplifying divisions by epidermal growth factor receptor signaling in Drosophila testes. Cell Death Dis. 2021;12(5):491.##Wu H, Coskun V, Tao J, Xie W, Ge W, Yoshikawa K, et al. Dnmt3a-dependent nonpromoter DNA methylation facilitates transcription of neurogenic genes. Science. 2010;329(5990):444-8.##Fan W, Xu Y, Liu Y, Zhang Z, Lu L, Ding Z. Obesity or overweight, a chronic inflammatory status in male reproductive system, leads to mice and human subfertility. Front Physiol. 2018;8:1117.##Singh V, Bala R, Chakraborty A, Rajender S, Trivedi S, Singh K. Duplications in 19p13. 3 are associated with male infertility. J Assist Reprod Genet. 2019;36(10):2171-9.##Yu Z, Guo R, Ge Y, Ma J, Guan J, Li S, et al. Gene expression profiles in different stages of mouse spermatogenic cells during spermatogenesis. Biol Reprod. 2003;69(1):37-47.##Yang C, Bae H, Song G, Lim W. Quercetin affects spermatogenesis-related genes of mouse exposed to high-cholesterol diet. J Anim Reprod Biotechnol. 2020;35(1):73-85.##Sharma M, Braun RE. Cyclical expression of GDNF is required for spermatogonial stem cell homeostasis. Development. 2018;145(5):dev151555.##Ivanski F, de Oliveira VM, de Oliveira IM, de Araújo Ramos AT, de Oliveira Tonete ST, de Oliveira Hykavei G, et al. Prepubertal acrylamide exposure causes dose-response decreases in spermatic production and functionality with modulation of genes involved in the spermatogenesis in rats. Toxicology. 2020;436:152428.##Ernst C, Eling N, Martinez-Jimenez CP, Marioni JC, Odom DT. Staged developmental mapping and X chromosome transcriptional dynamics during mouse spermatogenesis. Nat Commun. 2019;10(1):1251.##Endo T, Freinkman E, de Rooij DG, Page DC. Periodic production of retinoic acid by meiotic and somatic cells coordinates four transitions in mouse spermatogenesis. Proc Natl Acad Sci USA. 2017;114(47):E10132-41.##Ni FD, Hao SL, Yang WX. Multiple signaling pathways in Sertoli cells: recent findings in spermatogenesis. Cell Death Dis. 2019;10(8):541.##Wu FJ, Lin TY, Sung LY, Chang WF, Wu PC, Luo CW. BMP8A sustains spermatogenesis by activating both SMAD1/5/8 and SMAD2/3 in spermatogonia. Sci Signal. 2017;10(477):eaal1910.##Hadziselimovic F, Verkauskas G, Vincel B, Stadler MB. Testicular expression of long non–coding RNAs is affected by curative GnRHa treatment of cryptorchidism. Basic Clin Androl. 2019;29:18.##Bo D, Jiang X, Liu G, Xu F, Hu R, Wassie T, et al. Multipathway synergy promotes testicular transition from growth to spermatogenesis in early-puberty goats. BMC Genomics. 2020;21(1):372.##Arzate-Mejía RG, Recillas-Targa F, Corces VG. Developing in 3D: the role of CTCF in cell differentiation. Development. 2018;145(6):dev137729.##Sarakul M, Elzo MA, Koonawootrittriron S, Suwanasopee T, Jattawa D, Laodim T. Characterization of biological pathways associated with semen traits in the Thai multibreed dairy population. Anim Reprod Sci. 2018;197:324-34.##Wang J, Gao WJ, Deng SL, Liu X, Jia H, Ma WZ. High temperature suppressed SSC self-renewal through S phase cell cycle arrest but not apoptosis. Stem Cell Res Ther. 2019;10(1):227.##

Follicle Stimulating Hormone (FSH) as a Predictor of Decreased Oocyte Yield in Patients with Normal Anti-Müllerian Hormone (AMH) and Antral Follicle Count (AFC) 2 1 2 Background: The purpose of the current study was to determine the utility of early follicular phase follicle-stimulating hormone (FSH) testing in patients undergoing in vitro fertilization (IVF). Methods: This was a retrospective review of patients from 2012 to 2015 at Mayo Clinic in Rochester, Minnesota, USA. Included subjects had a normal anti-Müllerian hormone (AMH) of 1 to 9 ng/ml and antral follicle count (AFC) of 10 to 29. Patients were stratified by FSH level when associated estradiol was less than 50 ng/ml. In total, 225 patients were categorized into three groups: high FSH (FSH ≥10 IU/L; n= 36), normal FSH (>5 IU/L and <10 IU/L; n=170), and low FSH (FSH ≤5 IU/L; n= 19). ANOVA and multiple logistic regression were used for statistical comparisons and for evaluation of the relationships between variables; significance level was set at <0.05. Results: There were no significant differences in demographics, IVF cycle type, or peak estradiol level between the groups. Patients with a high basal FSH level had a similar clinical pregnancy rate and live birth rate compared to controls and patients with low FSH. High FSH level was associated with decreased follicular development (17 versus 22; p<0.01), oocyte yield (15 versus 18; p=0.02), and embryo yield (8 versus 10; p=0.04) despite higher total doses of gonadotropins. Conclusion: Patients with normal AMH and AFC levels could be further stratified into lower responders and starting doses of medications can be adjusted based on high basal FSH levels. Therefore, it is suggested to counsel patients on pregnancy outcomes which seem to be quite similar regardless of the FSH level. 181 188 Colleen CM Miller Division of Reproductive Endocrinology and Infertility, Mayo Clinic Division of Reproductive Endocrinology and Infertility, Mayo Clinic USA Ryan REM Melikian School of Medicine, Wayne State University School of Medicine, Wayne State University USA Tiffanny TL Jones Conceive Fertility Center Conceive Fertility Center USA Mackenzie MP Purdy Kindbody - St. Louis Kindbody - St. Louis USA Zaraq Z Khan Division of Reproductive Endocrinology and Infertility, Mayo Clinic Division of Reproductive Endocrinology and Infertility, Mayo Clinic USA Jessica JL Bleess Division of Reproductive Endocrinology and Infertility, Mayo Clinic Division of Reproductive Endocrinology and Infertility, Mayo Clinic USA Elizabeth EA Stewart Division of Reproductive Endocrinology and Infertility, Mayo Clinic Division of Reproductive Endocrinology and Infertility, Mayo Clinic USA Charles CC Coddington Division of Reproductive Endocrinology and Infertility, Mayo Clinic Division of Reproductive Endocrinology and Infertility, Mayo Clinic USA Chandra CC Shenoy Division of Reproductive Endocrinology and Infertility, Mayo Clinic Division of Reproductive Endocrinology and Infertility, Mayo Clinic USA shenoy.chandra@mayo.edu Anti- mullerian hormoneFollicle stimulating hormoneIn vitro fertilizationOvarian reserve 140196.pdf Ubaldi FM, Cimadomo D, Vaiarelli A, Fabozzi G, Venturella R, Maggiulli R, et al. Advanced maternal age in IVF: still a challenge? the present and the future of its treatment. Front Endocrinol (Lausanne). 2019;10:94.##Gougeon A, Ecochard R, Thalabard JC. Age-related changes of the population of human ovarian follicles: increase in the disappearance rate of non-growing and early-growing follicles in aging women. Biol Reprod. 1994;50(3):653-63.##te Velde ER, Pearson PL. The variability of female reproductive ageing. Hum Reprod Update. 2002;8(2):141-54.##4. van Tilborg TC, Broekmans FJ, Dolleman M, Eijkemans MJ, Mol BW, Laven JS, et al. Individualized follicle-stimulating hormone dosing and in vitro fertilization outcome in agonist downregulated cycles: a systematic review. Acta Obstet Gynecol Scand. 2016;95(12):1333-44.##Kelly J, Hughes CM, Harrison RF. The hidden costs of IVF. Ir Med J. 2006;99(5):142-3.##Tal R, Seifer DB. Ovarian reserve testing: a user's guide. Am J Obstet Gynecol. 2017;217(2):129-40.##Broekmans FJ, Soules MR, Fauser BC. Ovarian aging: mechanisms and clinical consequences. Endocr Rev. 2009;30(5):465-93.##Roudebush WE, Kivens WJ, Mattke JM. Biomarkers of Ovarian Reserve. Biomark Insights. 2008;3:259-68.##Jirge PR. Ovarian reserve tests. J Hum Reprod Sci. 2011;4(3):108-13.##Practice committee of the american society for reproductive medicine. Testing and interpreting measures of ovarian reserve: a committee opinion. Fertility and Sterility. 2020;114(6):1151-7.##Abdalla H, Thum MY. An elevated basal FSH reflects a quantitative rather than qualitative decline of the ovarian reserve. Hum Reprod. 2004;19(4):893-8.##Verhagen TE, Hendriks DJ, Bancsi LF, Mol BW, Broekmans FJ. The accuracy of multivariate models predicting ovarian reserve and pregnancy after in vitro fertilization: a meta-analysis. Hum Reprod Update. 2008;14(2):95-100.##Rosenwaks Z, Reichman DE. Use of antimullerian hormone: the risks of interpreting ovarian reserve markers in isolation. Fertil Steril. 2013;99(7):1850.##La Marca A, Ferraretti AP, Palermo R, Ubaldi FM. The use of ovarian reserve markers in IVF clinical practice: a national consensus. Gynecol Endocrinol. 2016;32(1):1-5.##Broer SL, van Disseldorp J, Broeze KA, Dolleman M, Opmeer BC, Bossuyt P, et al. Added value of ovarian reserve testing on patient characteristics in the prediction of ovarian response and ongoing pregnancy: an individual patient data approach. Hum Reprod Update. 2013;19(1):26-36.##van Rooij IA, Bancsi LF, Broekmans FJ, Looman CW, Habbema JD, te Velde ER. Women older than 40 years of age and those with elevated follicle-stimulating hormone levels differ in poor response rate and embryo quality in in vitro fertilization. Fertil Steril. 2003;79(3):482-8.##Chuang CC, Chen CD, Chao KH, Chen SU, Ho HN, Yang YS. Age is a better predictor of pregnancy potential than basal follicle-stimulating hormone levels in women undergoing in vitro fertilization. Fertil Steril. 2003;79(1):63-8.##Daney de Marcillac F, Pinton A, Guillaume A, Sagot P, Pirrello O, Rongieres C. What are the likely IVF/ICSI outcomes if there is a discrepancy between serum AMH and FSH levels? A multicenter retrospective study. J Gynecol Obstet Hum Reprod. 2017;46(8):629-35.##Metello JL, Tomas C, Ferreira P. Can we predict the IVF/ICSI live birth rate? JBRA Assist Reprod. 2019;23(4):402-7.##Dhillon RK, McLernon DJ, Smith PP, Fishel S, Dowell K, Deeks JJ, et al. Predicting the chance of live birth for women undergoing IVF: a novel pretreatment counselling tool. Hum Reprod. 2016;31(1):84-92.##Buratini J, Dellaqua TT, Dal Canto M, La Marca A, Carone D, Mignini Renzini M, et al. The putative roles of FSH and AMH in the regulation of oocyte developmental competence: from fertility prognosis to mechanisms underlying age-related subfertility. Hum Reprod Update. 2022;28(2):232-54.##

Increased Risk of Infertility in Women Infected with Human Papillomavirus 2 1 2 Background: Among several causes of infertility, urogenital infections seem to be influencing factors. The effect of bacterial or viral sexually transmitted infections (STIs) on human fertility is not well understood. The aim of this study was to determine the frequency of STIs in cervical samples of infertile and fertile women and study the relationship between these agents and infertility. Methods: In this case-control study, cytobrush was used for collecting of cervical sample from each infertile and fertile woman (n=95) who attended Research and Clinical Centers for Infertility in Kerman, Iran. PCR and real-time PCR methods were used to detect the presence of bacterial (genital Ureaplasma species, genital Mycoplasma species, Chlamydia trachomatis (C. trachomatis), and Gardnerella vaginalis) and viral (herpes simplex virus, human papillomavirus (HPV), and Epstein-Barr virus) agents, respectively. Fisher's exact test and the logistic regression with the significance level of ≤5% were used for statistical analyses. Results: In general, 78.94% and 14.73% of specimens were positive for one or more studied microorganisms, respectively. Among studied agents, only the infection with HPV was significantly different between infertile and fertile groups (p=0.005) which may enhance the likelihood of female infertility (OR=5.30, 95% CI:1.47-19.11, p<0.05). After adjusting for age, irregular menstrual cycle, abnormal vaginal discharge, and ectopic pregnancy, the odds ratio of infertility in HPV-infected women increased (OR=7.02, 95% CI:1.52-32.3, p<0.05). Conclusion: Since HPV infection is asymptomatic, periodic screening of women in reproductive age especially infertile couples is recommended for early diagnosis and prevention of infection progression and cross contamination. 188 198 Parastou P Heidari Pebdeni Student Research Committee, Kerman University of Medical Sciences Student Research Committee, Kerman University of Medical Sciences Iran Fereshteh F Saffari Department of Medical Microbiology (Bacteriology & Virology), Afzalipour Faculty of Medicine, Kerman University of Medical Sciences Department of Medical Microbiology (Bacteriology & Virology), Afzalipour Faculty of Medicine, Kerman University of Medical Sciences Iran Hamid Reza HR Mollaei Department of Medical Microbiology (Bacteriology & Virology), Afzalipour Faculty of Medicine, Kerman University of Medical Sciences Department of Medical Microbiology (Bacteriology & Virology), Afzalipour Faculty of Medicine, Kerman University of Medical Sciences Iran Toraj Reza TR Mirshekari Afzalipour Clinical Center for Infertility, Kerman University of Medical Sciences Afzalipour Clinical Center for Infertility, Kerman University of Medical Sciences Iran Robabeh R Hosseini Sadat Department of Obstetrics and Gynecology, School of Medicine, Kerman University of Medical Sciences Department of Obstetrics and Gynecology, School of Medicine, Kerman University of Medical Sciences Iran Victoria V Habibzadeh Department of Obstetrics and Gynecology, School of Medicine, Kerman University of Medical Sciences Department of Obstetrics and Gynecology, School of Medicine, Kerman University of Medical Sciences Iran Lida L Saeed Department of Obstetrics and Gynecology, School of Medicine, Kerman University of Medical Sciences Department of Obstetrics and Gynecology, School of Medicine, Kerman University of Medical Sciences Iran Moslem M Taheri Soodejani Center for Healthcare Data Modeling, Department of Biostatistics and Epidemiology, School of Public Health, Shahid Sadoughi University of Medical Sciences Center for Healthcare Data Modeling, Department of Biostatistics and Epidemiology, School of Public Health, Shahid Sadoughi University of Medical Sciences Iran Roya R Ahmadrajabi Medical Mycology and Bacteriology Research Center, Kerman University of Medical Sciences Medical Mycology and Bacteriology Research Center, Kerman University of Medical Sciences Iran ahmadrajabi3@ yahoo.com, r.ahmadi@kmu.ac.ir FemaleInfertilityIranSexually transmitted infections 140189.pdf Ruggeri M, Cannas S, Cubeddu M, Molicotti P, Piras GL, Dessole S, etal. Bacterial agents as a cause of infertility in humans. New Microbiol. 2016;39(3):206-9.##Naz MS, Ozgoli G, Sayehmiri K. Prevalence of infertility in Iran: a systematic review and meta-analysis. Urol J. 2020;17(4):338-45.##Janghorban R, Azarkish F. An overview on sexually transmitted infections in Iran. Int J Reprod Contracept Obstet Gynecol. 2016;5(3):585-95.##Mwatelah R, McKinnon LR, Baxter C, Abdool Karim Q, Abdool Karim SS. Mechanisms of sexually transmitted infection‐induced inflammation in women: implications for HIV risk. J Int AIDS Soc. 2019;22(Suppl 6):e25346.##Tsevat DG, Wiesenfeld HC, Parks C, Peipert JF. Sexually transmitted diseases and infertility. Am J Obstet Gynecol. 2017;216(1):1-9.##Sweet RL. Treatment of acute pelvic inflammatory disease. Infect Dis Obstet Gynecol. 2011;2011:561909.##Piscopo RC, Guimarães RV, Ueno J, Ikeda F, Jarmy-Di Bella ZI, Girão MJ, et al. Increased prevalence of endocervical Mycoplasma and Ureaplasma colonization in infertile women with tubal factor. JBRA Assist Reprod. 2020;24(2):152-7.##Schwebke JR, Muzny CA, Josey WE. Role of Gardnerella vaginalis in the pathogenesis of bacterial vaginosis: a conceptual model. J Infect Dis. 2014;210(3):338-43.##Ravel J, Moreno I, Simón C. Bacterial vaginosis and its association with infertility, endometritis, and pelvic inflammatory disease. Am J Obstet Gynecol. 2021;224(3):251-7.##Jaworek H, Zborilova B, Koudelakova V, Vrbkova J, Oborna I, Hajduch M. Prevalence of human papillomavirus infection in oocyte donors and women treated for infertility: an observational laboratory-based study. Eur J Obstet Gynecol Reprod Biol X. 2019;4:100068.##Jersovienė V, Gudlevičienė Z, Rimienė J, Butkauskas D. Human papillomavirus and infertility. Medicina (Kaunas). 2019;15;55(7):377.##Prabha V, Aanam TD, Kaur S. Bacteriological study of the cervix of females suffering from unexplained infertility. Am J Biomed Sci. 2011;3(2):84-9.##Pebdeni PH, Saffari F, Mirshekari TR, Ashourzadeh S, Soodejani MT, Ahmadrajabi R. Bacteriospermia and its association with seminal fluid parameters and infertility in infertile men, Kerman, Iran: a cross-sectional study. Int J Reprod Biomed. 2022;20(3):202-12.##Adamson PC, Krupp K, Freeman AH, Klausner JD, Reingold AL, Madhivanan P. Prevalence and correlates of primary infertility among young women in Mysore, India. Indian J Med Res. 2011;134(4):440-6.##Moghadam NM, Kheirkhah B, Mirshekari TR, Harandi MF, Tafsiri E. Isolation and molecular identification of mycoplasma genitalium from the secretion of genital tract in infertile male and female. Iran J Reprod Med. 2014;12(9):601-8.##Grześko J, Elias M, Mączyńska B, Kasprzykowska U, Tłaczała M, Goluda M. Occurrence of Mycoplasma genitalium in fertile and infertile women. Fertil Steril. 2009;91(6):2376-80.##Malaguti N, Bahls LD, Uchimura NS, Gimenes F, Consolaro ME. Sensitive detection of thirteen bacterial vaginosis-associated agents using multiplex polymerase chain reaction. Biomed Res Int. 2015;2015:645853.##Tavakoli A, Monavari SH, Bokharaei-Salim F, Mollaei H, Abedi-Kiasari B, Fallah FH, et al. Asymptomatic herpes simplex virus infection in Iranian mothers and their newborns. Fetal Pediatr Pathol. 2017;36(1):27-32.##Ghorbani S, Mollaei H, Mirshekari T, Ahmadipour H, Arabi Mianroodi AA, Iranmanesh K, et al. Frequency of human papillomavirus genotypes in head and neck epithelial cancers in the ear, throat, and nose department of Shafa hospital, Kerman, Iran in 2017. J Kerman Univ Med Sci. 2021;28(4):382-90.##Zaruni L, Arabzadeh SA, Afshar RM, Afshar AA, Mollaei HR. Detection of Epstein-Barr virus and cytomegalovirus in gastric cancers in Kerman, Iran. Asian Pac J Cancer Prev. 2016;17(5):2423-8.##Yuan S, Qiu Y, Xu Y, Wang H. Human papillomavirus infection and female infertility: a systematic review and meta-analysis. Reprod Biomed Online. 2020;40(2):229-37.##Hsu LC, Tsui KH, Wei JC, Yip HT, Hung YM, Chang R. Female human papillomavirus infection associated with increased risk of infertility: a nationwide population-based cohort study. Int J Environ Res Public Health. 2020;17(18):6505.##Rocha RM, Souza RP, Gimenes F, Consolaro ME. The high-risk human papillomavirus continuum along the female reproductive tract and its relationship to infertility and endometriosis. Reprod Biomed Online. 2019;38(6):926-37.##Lundqvist M, Westin C, Lundkvist O, Simberg N, Strand A, Andersson S, et al. Cytologic screening and human papilloma virus test in women undergoing artificial fertilization. Acta Obstet Gynecol Scand. 2002;81(10):949-53.##Perino A, Giovannelli L, Schillaci R, Ruvolo G, Fiorentino FP, Alimondi P, et al. Human papillomavirus infection in couples undergoing in vitro fertilization procedures: impact on reproductive outcomes. Fertil Steril. 2011;95(5):1845-48.##Zhang LD, Zhang HM, Pei J, He GR, Sun XF, Li B. [Investigation on HPV viral load and high risk HPV types infection among patients with infertility]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2007;21(2):159-61. Chinese.##Strehler E, Sterzik K, Malthaner D, Hoyer H, Nindl I, Schneider A. Influence of ovarian stimulation on the detection of human papillomavirus DNA in cervical scrapes obtained from patients undergoing assisted reproductive techniques. Fertil Steril. 1999;71(5):815-20.##Berntsson M, Dubicanac L, Tunbäck P, Ellström A, Löwhagen GB, Bergström T. Frequent detection of cytomegalovirus and Epstein–Barr virus in cervical secretions from healthy young women. Acta Obstet Gynecol Scand. 2013;92(6):706-10.##Cameron JE, Rositch AF, Vielot NA, Mugo NR, Kwatampora JK, Waweru W, et al. Epstein-Barr virus, high-risk human papillomavirus and abnormal cervical cytology in a prospective cohort of African female sex workers. Sex Transm Dis. 2018;45(10):666-72.##Virant-Klun I, Vogler A. In vitro maturation of oocytes from excised ovarian tissue in a patient with autoimmune ovarian insufficiency possibly associated with Epstein-Barr virus infection. Reprod Biol Endocrinol. 2018;16(1):33.##Li D, Huang T, Zhang Z. [The relationship between herpes simplex virus II, human papillomavirus infection and infertility after artificial abortion]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 1998;12(2):155-7. Chinese.##Menon S, Timms P, Allan JA, Alexander K, Rombauts L, Horner P, et al. Human and pathogen factors associated with Chlamydia trachomatis-related infertility in women. Clin Microbiol Rev. 2015;28(4):969-85.##Hasanabad MH, Mohammadzadeh M, Bahador A, Fazel N, Rakhshani H, Majnooni A. Prevalence of Chlamydia trachomatis and Mycoplasma genitalium in pregnant women of Sabzevar-Iran. Iran J Microbiol. 2011;3(3):123-8.##Rawre J, Dhawan B, Malhotra N, Sreenivas V, Broor S, Chaudhry R. Prevalence and distribution of Chlamydia trachomatis genovars in Indian infertile patients: a pilot study. APMIS. 2016;124(12):1109-15.##Ramadhani MY, Mirambo MM, Mbena H, Kihunrwa A, Mshana SE. High prevalence of Chlamydia trachomatis infection among infertile women in Mwanza city, Tanzania: a need to introduce screening and treatment programme. Sex Transm Infect. 2017;93(2):111.##Haghdoost M, Mousavi S, Gol MK, Montazer M. Frequency of Chlamydia trachomatis infection in spontaneous abortion of infertile women during first pregnancy referred to tabriz university of medical sciences by nested PCR method in 2015. Int J Women's Health Reprod Sci. 2019;7(4):526-30.##Frej-Mądrzak M, Gryboś A, Gryboś M, Teryks-Wołyniec D, Jama-Kmiecik A, Sarowska J, et al. PCR diagnostics of Chlamydia trachomatis in asymptomatic infection by women. Ginekol Pol. 2018;89(3):115-9.##Afrasiabi Sh, Moniri R, Samimi M, Khorshidi A, Mousavi SG. The prevalence of endocervical Chlamydia trachomatis infection among young females in Kashan, Iran. Jundishapur J Microbiol. 2015;8(4):e15576.##Joolayi F, Navidifar T, Jaafari RM, Amin M. Comparison of Chlamydia trachomatis infection among infertile and fertile women in Ahvaz, Iran: a case-control study. Int J Reprod Biomed. 2017;15(11):713-18.##Passos LG, Terraciano P, Wolf N, de Oliveira FD, de Almeida I, Passos EP. The correlation between Chlamydia trachomatis and female infertility: a systematic review. Rev Bras Ginecol Obstet. 2022;44(6):614-20.##Dehghan Marvast L, Aflatoonian A, Talebi AR, Eley A, Pacey AA. Relationship between Chlamydia trachomatis and Mycoplasma genitalium infection and pregnancy rate and outcome in Iranian infertile couples. Andrologia. 2017;49(9).##Hoenderboom BM, van Bergen JEAM, Dukers-Muijrers NHTM, Götz HM, Hoebe CJPA, de Vries HJC, et al. Pregnancies and time to pregnancy in women with and without a previous Chlamydia trachomatis infection. Sex Transm Dis. 2020;47(11):739-47.##Ahmadi MH, Mirsalehian A, Bahador A. Prevalence of urogenital mycoplasmas in Iran and their effects on fertility potential: a systematic review and meta-analysis. Iran J Public Health. 2016;45(4):409-22.##Bahaabadi SJ, Moghadam NM, Kheirkhah B, Farsinejad A, Habibzadeh V. Isolation and molecular identification of Mycoplasma hominis in infertile female and male reproductive system. Nephrourol Mon. 2014;6(6):e22390.##Seifoleslami M, Safari A, Khameneie MK. Prevalence of Ureaplasma urealyticum and Mycoplasma hominis in high vaginal swab samples of infertile females. Iran Red Crescent Med J. 2015;17(12):e16823.##Casari E, Ferrario A, Morenghi E, Montanelli A. Gardnerella, Trichomonas vaginalis, Candida, Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum in the genital discharge of symptomatic fertile and asymptomatic infertile women. New Microbiol. 2010;33(1):69-76.##

The Child's Right to Know Versus the Parents’ Right Not to Tell: The Attitudes of Couples Undergoing Fertility Treatments Towards Identity-Release Gamete Donation 2 1 2 Background: In Brazil, donor anonymity is mandatory; however, the tendency of Brazilians towards the practice is unknown. In this study, an attempt was made to investigate whether couples undergoing assisted reproductive technology (ART) have a different perception of anonymous versus identity-release gamete donation than a target population in Brazil. Methods: This cross-sectional study was performed from September 1, 2020 to December 15, 2020. For that purpose, surveys through online platforms were conducted, including either patients undergoing ART (ART-group, n=400) or subjects interested in the theme (interested-group, n=100) randomized by age at a 1:4 ratio. The survey collected information on the participants’ attitudes towards anonymity of gamete donors, and answers were compared between the groups. Results: Most participants stated that the relationship between children and their parents would be affected by the child's knowledge of the origin of its conception. Most participants in the ART-group believed that the gamete donor’s identity should not be revealed to the child, while only half of the interested-group stated the same. Most of the participants stated that "the donor's identity should be revealed if the child questions its biological origin". "From birth" was the second most common response, while "when the child turns 18 years old" and "sometime during teenage years" were less common answers. Conclusion: The attitudes of ART patients about anonymity are conservative, with most participants believing that family relationships may be affected if the child is aware of the origin of his/her conception. These patients also believe that the identity of the gamete donor should not be revealed to the child. 198 206 Douglas DR Rocha Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Brazil Rose RMM Melamed Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Brazil Daniela DPAF Braga Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Brazil Amanada AS Setti Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Brazil Assumpto JrA Iaconelli Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Brazil Edson E Borges Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Associação Instituto Sapientiae-Centro de Estudos e Pesquisa em Reprodução Assistida Brazil edson@fertility.com.br Donor conceptionDonorSpermSurvey 140191.pdf Wang TK, Yuen KY, Wong SS. Asymptomatic Penicillium marneffei fungemia in an HIV-infected patient. Int J Infect Dis. 2007;11(3):280-1.##Ombelet W, Van Robays J. Artificial insemination history: hurdles and milestones. Facts Views Vis Obgyn. 2015;7(2):137-43.##Gerkowicz SA, Crawford SB, Hipp HS, Boulet SL, Kissin DM, Kawwass JF. Assisted reproductive technology with donor sperm: national trends and perinatal outcomes. Am J Obstet Gynecol. 2018;218(4):421.e1-10.##Bustillo M, Buster JE, Cohen SW, Hamilton F, Thorneycroft IH, Simon JA, et al. Delivery of a healthy infant following nonsurgical ovum transfer. JAMA. 1984;251(7):889.##Cobo A, Remohí J, Chang CC, Nagy ZP. Oocyte cryopreservation for donor egg banking. Reprod Biomed Online. 2011;23(3):341-6.##De Geyter Ch, Calhaz-Jorge C, Kupka MS, Wyns C, Mocanu E, Motrenko T, et al. ART in Europe, 2014: results generated from European registries by ESHRE: the European IVF-monitoring Consortium (EIM) for the European Society of Human Reproduction and Embryology (ESHRE). Hum Reprod. 2018;33(9):1586-601.##Skoog Svanberg A, Sydsjö G, Lampic C. Psychosocial aspects of identity-release gamete donation–perspectives of donors, recipients, and offspring. Ups J Med Sci. 2020;125(2):175-82.##Hibino Y, Allan S. Absence of laws regarding sperm and oocyte donation in Japan and the impacts on donors, parents, and the people born as a result. Reprod Med Biol. 2020;19(3):295-8.##Blyth E, Frith L. Donor-conceived people's access to genetic and biographical history: an analysis of provisions in different jurisdictions permitting disclosure of donor identity. Int J Law Policy Fam. 2009;23(2):174-91.##Lampic C, Skoog Svanberg A, Gudmundsson J, Leandersson P, Solensten NG, Thurin-Kjellberg A, et al. National survey of donor-conceived individuals who requested information about their sperm donor-experiences from 17 years of identity releases in Sweden. Hum Reprod. 2022;37(3):510-21.##Miettinen A, Rotkirch A, Suikkari AM, Söderström-Anttila V. Attitudes of anonymous and identity-release oocyte donors towards future contact with donor offspring. Hum Reprod. 2019;34(4):672-8.##Ethics committee of the American society for reproductive medicine. Informing offspring of their conception by gamete or embryo donation: an ethics committee opinion. Fertil Steril. 2018;109(4):601-5.##Scheib JE, Ruby A, Benward J. Who requests their sperm donor's identity? the first ten years of information releases to adults with open-identity donors. Fertil Steril. 2017;107(2):483-93.##Anvisa. Resolution of the Collegiate Board - RDC No. 771. 2022 Jul [cited 2022 Dec 28]; 1 (244) p 306-313. Available: http://antigo.anvisa.gov.br/ legislacao/?inheritRedirect=true#/visualizar/500343.##Cheib JE, Riordan M, Shaver PR. Choosing between anonymous and identity-release sperm donors: recipient and donor characteristics. Reprod Technol. 2000;10(1):50.##Brewaeys A, De Bruyn JK, Louwe LA, Helmerhorst FM. Anonymous or identity-registered sperm donors? a study of Dutch recipients' choices. Hum Reprod. 2005;20(3):820-4.##Daniels K. Donor gametes: anonymous or identified? Best Pract Res Clin Obstet Gynaecol. 2007;21(1):113-28.##Nordqvist P, Smart C. Relative strangers. Relative strangers and the paradoxes of genetic Kinship. In: Nordqvist P, Smart C, editors. Family life, genes and donor conception. London: Springer; 2014. p. 144-66.##Indekeu A, Dierickx K, Schotsmans P, Daniels K, Rober P, D'Hooghe T. Factors contributing to parental decision-making in disclosing donor conception: a systematic review. Hum Reprod Update. 2013;19(6):714-33.##Kovacs GT, Wise S, Finch S. Keeping a child's donor sperm conception secret is not linked to family and child functioning during middle childhood: an Australian comparative study. Aust N Z J Obstet Gynaecol. 2015;55(4):390-6.##Golombok S, Brewaeys A, Giavazzi M, Guerra D, MacCallum F, Rust J. The European study of assisted reproduction families: the transition to adolescence. Hum Reprod. 2002;17(3):830-40.##Golombok S, Blake L, Casey P, Roman G, Jadva V. Children born through reproductive donation: a longitudinal study of psychological adjustment. J Child Psychol Psychiatry. 2013;54(6):653-60.##Freeman T, Golombok S. Donor insemination: a follow-up study of disclosure decisions, family relationships and child adjustment at adolescence. Reprod Biomed Online. 2012;25(2):193-203.##Ilioi E, Blake L, Jadva V, Roman G, Golombok S. The role of age of disclosure of biological origins in the psychological wellbeing of adolescents conceived by reproductive donation: a longitudinal study from age 1 to age 14. J Child Psychol Psychiatry. 2017;58(3):315-24.##Isaksson S, Sydsjo G, Skoog Svanberg A, Lampic C. Disclosure behaviour and intentions among 111 couples following treatment with oocytes or sperm from identity-release donors: follow-up at offspring age 1–4 years. Hum Reprod. 2012;27(10):2998-3007.##Greenfeld DA, Klock SC. Disclosure decisions among known and anonymous oocyte donation recipients. Fertil Steril. 2004;81(6):1565-71.##Kalampalikis N, Haas V, Fieulaine N, Doumergue M, Deschamps G. Giving or giving back: new psychosocial insights from sperm donors in France. Psychol Health Med. 2013;18(1):1-9.##Salevaara M, Suikkari AM, Söderström-Anttila V. Attitudes and disclosure decisions of Finnish parents with children conceived using donor sperm. Hum Reprod. 2013;28(10):2746-54.##Freeman T, Zadeh S, Smith V, Golombok S. Disclosure of sperm donation: a comparison between solo mother and two-parent families with identifiable donors. Reprod Biomed Online. 2016;33(5):592-600.##Lycett E, Daniels K, Curson R, Golombok S. School-aged children of donor insemination: a study of parents' disclosure patterns. Hum Reprod. 2005;20(3):810-9.##Lalos A, Gottlieb C, Lalos O. Legislated right for donor-insemination children to know their genetic origin: a study of parental thinking. Hum Reprod. 2007;22(6):1759-68.##Golombok S, Murray C, Jadva V, Lycett E, MacCallum F, Rust J. Non-genetic and non-gestational parenthood: consequences for parent–child relationships and the psychological well-being of mothers, fathers and children at age 3. Hum Reprod. 2006;21(7):1918-24.##Söderström-Anttila V, Sälevaara M, Suikkari A. Increasing openness in oocyte donation families regarding disclosure over 15 years. Hum Reprod. 2010;25(10):2535-42.##Ethics ommittee of the American society for reproductive medicine. Informing offspring of their conception by gamete donation. Fertil Steril. 2004;81(3):527-31.##Indekeu A, Maas AJB, McCormick E, Benward J, Scheib JE. Factors associated with searching for people related through donor conception among donor-conceived people, parents, and donors: a systematic review. F&S Rev. 2021;2(2):93-119.##Practice committee of the American society for reproductive medicine. The clinical utility of sperm DNA integrity testing: a guideline. Fertil Steril. 2013;99(3):673-7.##Isaksson S, Skoog Svanberg A, Sydsjö G, Thurin-Kjellberg A, Karlström PO, Solensten NG, et al. Two decades after legislation on identifiable donors in Sweden: are recipient couples ready to be open about using gamete donation? Hum Reprod. 2011;26(4):853-60.##Gebhardt AJ, Sydsjö G, Skoog Svanberg A, Indekeu A, Lampic C. Parenting stress and its association with perceived agreement about the disclosure decision in parents following donor conception. Acta Obstet Gynecol Scand. 2017;96(8):968-75.##Daniels KR, Grace VM, Gillett WR. Factors associated with parents’ decisions to tell their adult offspring about the offspring's donor conception. Hum Reprod. 2011;26(10):2783-90.##Pennings G. Disclosure of donor conception, age of disclosure and the well-being of donor offspring. Hum Reprod. 2017;32(5):969-73.##Gottlieb C, Lalos O, Lindblad F. Disclosure of donor insemination to the child: the impact of Swedish legislation on couples' attitudes. Hum Reprod. 2000;15(9):2052-6.##Rumball A, Adair V. Telling the story: parents' scripts for donor offspring. Hum Reprod. 1999;14(5):1392-9.##Mac Dougall K, Becker G, Scheib JE, Nachtigall RD. Strategies for disclosure: how parents approach telling their children that they were conceived with donor gametes. Fertil Steril. 2007;87(3):524-33.##Turner AJ, Coyle A. What does it mean to be a donor offspring? The identity experiences of adults conceived by donor insemination and the implications for counselling and therapy. Hum Reprod. 2000;15(9):2041-51.##Jadva V, Freeman T, Kramer W, Golombok S. The experiences of adolescents and adults conceived by sperm donation: comparisons by age of disclosure and family type. Hum Reprod. 2009;24(8):1909-19.##Beeson DR, Jennings PK, Kramer W. Offspring searching for their sperm donors: how family type shapes the process. Hum Reprod. 2011;26(9):2415-24.##Hershberger PE, Driessnack M, Kavanaugh K, Klock SC. Oocyte donation disclosure decisions: a longitudinal follow-up at middle childhood. Hum Fertil (Camb). 2021;24(1):31-45.##Applegarth LD, Kaufman NL, Josephs-Sohan M, Christos PJ, Rosenwaks Z. Parental disclosure to offspring created with oocyte donation: intentions versus reality. Hum Reprod. 2016;31(8):1809-15.##Daniels KR, Grace VM, Gillett WR. Factors associated with parents' decisions to tell their adult offspring about the offspring's donor conception. Hum Reprod. 2011;26(10):2783-90.##Blyth E, Farrand A. Anonymity in donor-assisted conception and the UN Convention on the Rights of the Child. Int J Children's Rights. 2004;12(2):89-104.##

Fumarase Deficiency and Its Effect on Infertility: A Case Series 2 1 2 Background: Fumarase deficiency is an autosomal recessive condition characterized by severe neurologic abnormalities due to homozygous mutations in the fumarate hydratase (FH) gene. Heterozygous carriers of FH mutations have increased risk of developing uterine fibroids that can be associated with hereditary leiomyomatosis and renal cell cancer (HLRCC). The association between FH mutations and infertility remains uncertain. The objective of our study was to characterize the infertility diagnoses, treatments, and outcomes in women presenting to a fertility center who were found to be carriers of fumarase deficiency based on the presence of heterozygous FH mutations. Case Presentation: A retrospective case series was conducted including 10 women presenting to an academic fertility center who were found to be FH carriers based on genetic carrier screening. Of the 9 women who were engaged in further workup, 2 had imaging results consistent with uterine fibroids. One woman underwent hysteroscopic myomectomy prior to two courses of ovulation induction with timed intercourse (OI/TIC) followed by one successful cycle of IVF. Of the remaining patients, only 1 woman successfully delivered after a cycle of ovulation induction with intrauterine insemination (OI/IUI). Other patients pursuing OI/IUI, OI/TIC, or monitored natural cycles had unsuccessful experiences. Conclusion: Patients with infertility who are offered genetic testing should be screened for FH mutations, as the carriers are at risk of developing HLRCC-associated uterine fibroids, which can influence fertility and pregnancy. Additional research is needed to investigate the impacts of FH mutations on infertility. 206 212 Jessica JS Schwartz Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Hofstra University Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Hofstra University USA jessicaws737@gmail.com Alexandra A Peyser Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Northwell Health Fertility, North Shore University Hospital Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Northwell Health Fertility, North Shore University Hospital USA Miriam M Tarrash Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Northwell Health Fertility, North Shore University Hospital Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Northwell Health Fertility, North Shore University Hospital USA Randi RH Goldman Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Hofstra University Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Hofstra University USA FibroidsFumarate hydrataseGenetic counselingHereditary leiomyomatosis and renal cell cancer (HLRCC)InfertilityLeiomyoma 140195.pdf Coman D, Kranc KR, Christodoulou J. Fumarate Hydratase Deficiency. 2006 Jul 5 [Updated 2020 Apr 23]. In: Adam MP, Mirzaa GM, Pagon RA, et al., editors. GeneReviews® [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2023. Available from: https://www.ncbi.nlm.nih.gov/books/NBK1506/##Patel VM, Handler MZ, Schwartz RA, Lambert WC. Hereditary leiomyomatosis and renal cell cancer syndrome: an update and review. J Am Acad Dermatol. 2017;77(1):149-58.##Shuch B, Li S, Risch H, Bindra RS, McGillivray PD, Gerstein M. Estimation of the carrier frequency of fumarate hydratase alterations and implications for kidney cancer risk in hereditary leiomyomatosis and renal cancer. Cancer. 2020;126(16):3657-66. ##Marieke P, Susan G, Martijn B, Monique W, Jaap B, Ed J, et al. Fumarase deficiency: a case with a new pathogenic mutation and a review of the literature. J Child Neurol. 2021;36(4):310-23. ##Lehtonen HJ, Kiuru M, Ylisaukko‐oja SK, Salovaara R, Herva R, Koivisto PA, et al. Increased risk of cancer in patients with fumarate hydratase germline mutation. J Med Genet. 2006;43(6):523-6.##Toro JR, Nickerson ML, Wei MH, Warren MB, Glenn GM, Turner ML, et al. Mutations in the fumarate hydratase gene cause hereditary leiomyomatosis and renal cell cancer in families in north America. Am J Hum Genet. 2003;73(1):95-106.##Whynott RM, Vaught KC, Segars JH. The effect of uterine fibroids on infertility: a systematic review. Semin Reprod Med. 2017;35(6):523-32.##Khaund A, Lumsden MA. Impact of fibroids on reproductive function. Best Pract Res Clin Obstet Gynaecol. 2008;22(4):749-60.##Giuliani E, As-Sanie S, Marsh EE. Epidemiology and management of uterine fibroids. Int J Gynecol Obstet. 2020;149(1):3-9. ##Freytag D, Günther V, Maass N, Alkatout I. Uterine fibroids and infertility. Diagnostics (Basel). 2021;11(8):1455.##Zepiridis LI, Grimbizis GF, Tarlatzis BC. Infertility and uterine fibroids. Best Pract Res Clin Obstet Gynaecol. 2016;34:66-73.##Stewart L, Glenn GM, Stratton P, Goldstein AM, Merino MJ, Tucker MA, et al. Association of germline mutations in the fumarate hydratase gene and uterine fibroids in women with hereditary leiomyomatosis and renal cell cancer. Arch Dermatol. 2008;144(12):1584-92.##Harrison WJ, Andrici J, Maclean F, Madadi-Ghahan R, Farzin M, Sioson L, et al. Fumarate hydratase–deficient uterine leiomyomas occur in both the syndromic and sporadic Settings. Am J Surg Pathol. 2016;40(5):599-607.##Schmidt LS, Linehan WM. Hereditary leiomyomatosis and renal cell carcinoma. Int J Nephrol Renovasc Dis. 2014;7:253-60.##Smit DL, Mensenkamp AR, Badeloe S, Breuning MH, Simon MEH, van Spaendonck KY, et al. Hereditary leiomyomatosis and renal cell cancer in families referred for fumarate hydratase germline mutation analysis. Clin Genet. 2011;79(1):49-59. ##Chan E, Rabban JT, Mak J, Zaloudek C, Garg K. Detailed morphologic and immunohistochemical characterization of myomectomy and hysterectomy specimens from women with hereditary leiomyomatosis and renal cell carcinoma syndrome (HLRCC). Am J Surg Pathol. 2019;43(9):1170-9.##Chayed Z, Kristensen LK, Ousager LB, Rønlund K, Bygum A. Hereditary leiomyomatosis and renal cell carcinoma: a case series and literature review. Orphanet J Rare Dis. 2021;16(1):34.##Bayley JP, Launonen V, Tomlinson IP. The FH mutation database: an online database of fumarate hydratase mutations involved in the MCUL (HLRCC) tumor syndrome and congenital fumarase deficiency. BMC Med Genet. 2008;9:20.##Rivera-Cruz G, Boyraz B, Petrozza JC. How a woman’s myomectomy saved her father’s life: evidence of fumarate hydratase deficient uterine leiomyoma and early detection of germline variants in fumarate hydratase. F S Rep. 2021;3(1):26-31. ##Zhang L, Walsh MF, Jairam S, Mandelker D, Zhong Y, Kemel Y, et al. Fumarate hydratase FH c.1431_1433dupAAA (p.Lys477dup) variant is not associated with cancer including renal cell carcinoma. Hum Mutat. 2020;41(1):103-9. ##American Society for Reproductive Medicine. Age and Fertility [Internet]. Birmingham: American Society for Reproductive Medicine; 2012 [cited 2021 Sep 10]. Available from: https://www.reproductivefacts.org/news-and-publications/patient-fact-sheets-and-booklets/documents/fact-sheets-and-info-booklets/age-and-fertility/.##

Treatment of Immature Teratoma Associated with Ovarian Endometrioma: A Case Report 2 1 2 Background: Mature teratoma is a benign neoplasm, mostly composed of well-differentiated derivations of almost two or three germ cell layers, while immature teratoma is a malignant neoplasm composed of immature neural and embryonic tissue. Immature teratoma in the context of ovarian endometrioma has not been reported yet. Case Presentation: A 34-year-old woman with primary infertility is reported in this study who suffered from immature teratoma associated with ovarian endometrioma. After several rounds of fertility treatment, the patient was referred for severe abdominal pain and underwent emergency surgery for the rupture of ovarian endometrioma. To preserve the ovary, the cyst was not resected in areas attached to the ovary. Some months later, the patient noticed a progressive abdominal enlargement. The sonographic evaluation revealed multiple solid-cystic lobulated masses on the abdominal wall and throughout the pelvic cavity. The histologic diagnosis was consistent with immature teratoma. The patient was treated with high-dose neoadjuvant chemotherapy and fertility-sparing surgery (FSS). The histologic evaluation of the extracted masses revealed teratoma maturation. Conclusion: This study reveals the importance of complete removal of endometrioma and highlights the role of neoadjuvant chemotherapy in fertility-sparing surgery and potentiating teratoma maturation. 212 216 Soheila S Amini Moghadam Department of Obstetrics and Gynecology, Firouzgar Hospital, Iran University of Medical Sciences Department of Obstetrics and Gynecology, Firouzgar Hospital, Iran University of Medical Sciences Iran Niloufar N Sarchami Department of Obstetrics and Gynecology, Firouzgar Hospital, Iran University of Medical Sciences Department of Obstetrics and Gynecology, Firouzgar Hospital, Iran University of Medical Sciences Iran Niloo.sarchami@gmail.com Ali A Rahbari Department of Pathology, Jam Hospital Department of Pathology, Jam Hospital Iran EndometriomaFertility preservationImmature teratomaNeoadjuvant chemotherapy 140194.pdf Matalliotakis M, Matalliotaki C, Zervou MI, Krithinakis K, Goulielmos GN, Kalogiannidis I, et al. Retrospective evaluation of pathological results among women with ovarian endometriomas versus teratomas. Mol Clin Oncol. 2019;10(6):592-6.##Schmidt D, Kommoss F. [Teratoma of the ovary. clinical and pathological differences between mature and immature teratomas]. Pathologe. 2007;28(3):203-8. Germany.##Donadio AC, Motzer RJ, Bajorin DF, Kantoff PW, Sheinfeld J, Houldsworth J, et al. Chemotherapy for teratoma with malignant transformation. J Clin Oncol. 2003;21(23):4285-91.##Imran H, Siddiqui AH, Wilson F, Pashankar F. Growing teratoma syndrome After chemotherapy for ovarian immature teratoma. J Pediatr Hematol Oncol. 2020;42(7):e630-3.##Vasta FM, Dellino M, Bergamini A, Gargano G, Paradiso A, Loizzi V, et al. Reproductive outcomes and fertility preservation strategies in women with malignant ovarian germ cell tumors after fertility sparing surgery. Biomedicines. 2020;8(12):544.##Der EM, Seidu S. Case report of three immature cystic teratomas in northern Ghana. Case Rep Obstet Gynecol. 2019;2019:1210509.##Matalliotakis M, Matalliotaki C, Zervou MI, Krithinakis K, Goulielmos GN, Kalogiannidis I, et al. Retrospective evaluation of pathological results among women with ovarian endometriomas versus teratomas. Mol Clin Oncol. 2019;10(6):592-6.##Takagi H, Takata E, Sakamoto J, Fujita S, Takakura M, Sasagawa T. Malignant transformation of an ovarian endometrioma during endometriosis treatment: a case report. Case Rep Obstet Gynecol. 2018;2018:6210172.##Li J, Wu X. Current strategy for the treatment of ovarian germ cell tumors: role of extensive surgery. Curr Treat Options Oncol. 2016;17(8):44.##Pashankar F, Hale JP, Dang H, Krailo M, Brady WE, Rodriguez-Galindo C, et al. Is adjuvant chemotherapy indicated in ovarian immature teratomas? a combined data analysis from the malignant germ cell tumor international collaborative. Cancer. 2016;122(2):230-7.##Talukdar S, Kumar S, Bhatla N, Mathur S, Thulkar S, Kumar L. Neo-adjuvant chemotherapy in the treatment of advanced malignant germ cell tumors of ovary. Gynecol Oncol. 2014;132(1):28-32.##Gorbatiy V, Spiess PE, Pisters LL. The growing teratoma syndrome: current review of the literature. Indian J Urol. 2009;25(2):186-9.##Sato S, Itamochi H. Neoadjuvant chemotherapy in advanced ovarian cancer: latest results and place in therapy. The Adv Med Oncol. 2014;6(6):293-304.##Culine S, Lhomme C, Kattan J, Michel G, Duvillard P, Droz J. Pure malignant immature teratoma of the ovary: the role of chemotherapy and second-look surgery. Int J Gynecol Cancer. 1995;5(6):432-7.##