The Possibility of Increasing Oocyte Capacity to Repair Sperm DNA Fragmentation 2 1 2 During spermatogenesis, the sperm loses most of its cytoplasm and acquires a shape commensurate with its main function for transferring the paternal genome to offspring through fertilization of oocyte and formation of early embryo. The different shape of mammalian sperm compared to somatic cells enables the sperm to pass through difficult pathways to reach and fertilize oocyte. Although the facilitating role of sperm’s structure and morphology in reaching the oocyte is significant, sperm chromatin condensation is consequential for protection of the paternal genome from harmful external and internal factors. Despite specific structure of sperm chromatin and changes in its nuclear proteins for greater compaction and protection against harmful agents, sperm is vulnerable to harmful agents such as free radicals since it loses most of its cytoplasm. However, seminal fluid plays a major role for protection of sperm when it has close contact with the seminal fluid. Normal semen contains sperm (Motile, immobile, and dead) and non-sperm cells, including epithelial cells, immature germ cells at different stages, leukocytes (WBC), and cell debris suspended in a mixture fluid from the testes, epididymis, and accessory sex glands. Increased defective sperm, dead cells, leukocytes, bacteria, and cell debris or removal of seminal fluid during sperm processing can disrupt protective effects of semen culminating in damages to sperm chromatin and its membrane. In addition, other factors such as infections, smoking, drug addiction, exposure to environmental and occupational pollutants, aging, varicocele, high fever, increased testis temperature (Using laptop, hot baths, and tight clothes), paralysis by spinal cord injuries, chronic diseases such as diabetes and kidney failure are associated with increased sperm DNA fragmentation (1). Therefore, the assessment of sperm chromatin damage and DNA fragmentation index (DFI) has received special attention by researchers and clinicians for diagnosis and treatment of male infertility, so it is often requested as a complementary test with semen analysis in IVF clinics. The DFI more than 30% is associated with reduced natural male fertility, risk of miscarriage, longer time to conceive, fertilization failure, low quality embryos, implantation failure, and repeated IVF cycles (2). However, the importance and role of DFI in assessment of male infertility and its measurement methods were widely criticized by the time of its introduction. However, extensive research gradually established its role in diagnosis and treatment of male infertility and also its measurement methods were somewhat improved and standardized. Therefore, based on the DFI results, a variety of new pharmacological and surgical therapeutic strategies have been recommended and evaluated for improvement of male fertility (1). A number of clinical and surgical interventions including changes in lifestyle, ejaculatory abstinence, oral antioxidant therapy, hormone therapy, varicocele repair and testicular sperm extraction (TESE) are suggested to improve sperm quality and reduce DFI. Oral antioxidant supplementation in infertile men with oxidative DNA damage could reduce the sperm DFI and significantly enhance sperm chromatin integrity. Recent studies have shown that shorter intervals between ejaculations improve DFI and sperm quality, which is associated with better intrauterine insemination (IUI) and assisted reproductive technology (ART) outcomes compared to recommended time for semen analysis. The sperm retrieved through testicular sperm extraction (TESE) in men with high levels of DFI has better DNA quality and lower DFI than ejaculated sperm following repeated IVF failure. The use of testicular sperm leads to an increase in fertilization rate, good quality embryos, implantation rate, and pregnancy rate compared to ejaculation in ICSI cycles among men with high level of DFI and resistance to drug interventions (3). In addition to mentioned interventions, an issue that has raised more attention is the role of the oocyte in repairing sperm DNA fragmentation. Oocytes are able to repair fragmented DNA following fertilization. The repairing system depends on the cytoplasmic and genomic mechanism and the quality of oocytes. The qualified early embryos also have the ability to repair remaining DNA breakage during pre-implantation at cleavage stage. However, the repairing ability of the oocyte has a limited capacity and vast amount of damage in two strands of DNA can not be properly repaired which may lead to fertilization failure, poor quality of embryo, and finally IVF failure (4, 5). The goal of above clinical and surgical interventions was to reduce the incidence of sperm DNA fragmentation. However, the main question is how to increas the oocyte to maximize its repairing capacity. In other words, the chief concern is whether our current practices in IVF cycles including type of ovarian stimulation protocol, denudation, micro-manipulation, length and quality of in vitro culture before and after fertilization are effective in changing the potential of oocyte in repairing sperm DNA fragmentation. In fact, further research needs to be conducted to evaluate whether special adjuvant therapies in induction of ovulation cycles would increase the repairing potential of oocytes. It seems that the possibility of adding specific ingredients or components to embryo and oocyte culture media to increase the repairing potential of oocyte can be a tentative hypothesis. Answer to these questions necessitates considering some ethical aspects that limit the possibility of study and research on the human oocyte and embryo; therefore, designing a series of animal studies for future research in the field of male fertility, with a focus on andrology, is an appropriate strategy which may be conducive to success of infertility treatment cycles. In fact, changes in lifestyle, increased occupational and environmental pollutants, and other harmful factors have led to threats to male fertility and subsequently sperm DNA fragmentation. It seems that along with all protocols for reduction of sperm DNA damages in various aspects of life and occupational and environmental practice, different solutions must be devised for in vitro and in vivo repairing of sperm DNA damages in future. 075 77 Mohammad Reza MR Sadeghi محمدرضا صادقی Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran sadeghi@ari.ir No Keyword 120111.pdf Evgeni E, Charalabopoulos K, Asimakopoulos B. Human sperm DNA fragmentation and its correlation with conventional semen parameters. J Reprod Infertil. 2014;15(1):2-14.##Kim GY. What should be done for men with sperm DNA fragmentation? Clin Exp Reprod Med. 2018;45(3):101-9.##Esteves SC, Santi D, Simoni M. An update on clinical and surgical interventions to reduce sperm DNA fragmentation in infertile men. Andrology. 2020;8(1):53-81.##Ménézo Y, Dale B, Cohen M. DNA damage and repair in human oocytes and embryos: a review Zygote. 2010;18(4):357-65.##Ješeta M, Myšková M, Žáková J, Crha I, Crha K, Chmelikova E, et al. Can oocytes repair fragmented DNA of spermatozoa? Med J Cell Biol. 2020;8(2):73-7.##

The Effect of Post warming Culture Period Between Thawing and Transfer of Cryopreserved Embryos on Reproductive Outcomes After In Vitro Fertilization (IVF): A Systematic Review and Meta-analysis 2 1 2 Background: The purpose of this study was to evaluate the effect of post warming culture period between thawing and transfer of cryopreserved embryos on reproduc-tive outcomes after in vitro fertilization (IVF). Methods: An extensive literature search was performed using PubMed, EmBase, and the Cochrane library from January 2000 to August 2019. A systematic review and meta–analysis of clinical trials was performed in this manuscript. The trials represented patients with embryo transfers of at least one previously cryopreserved good quality embryo. Main outcome measures of the study included clinical preg-nancy rate, live birth rate, miscarriage rate, and ectopic pregnancy rate. Results: A total of 5338 trial/abstracts were identified through a literature search. Totally, five studies were included in the systematic review, and three in the final meta–analysis. The studies included 1717 embryo transfers, 605 after short culture, and 1112 after long culture. The clinical pregnancy rate (CPR) was the most con-sistent outcome reported. The CPR was slightly better after short time culture with a RR of 1.09 (0.95–1.26, 95%CI) but this difference was not statistically significant. The great heterogenicity in the results reported in the included studies made it impossible to compare all planned outcomes. Conclusion: There are no differences in reproductive outcomes if cryopreserved embryos are transferred after overnight culture or after two hours of culture follow-ing thawing. Due to small number and the poor quality of trials reported on this topic, the results of this review should be treated with caution. 077 85 Luis H LH Sordia-Hernandez Department of Gynecology and Obstetrics, University Hospital “Dr. Jose Eleuterio González” Universidad Autónoma de Nuevo León Department of Gynecology and Obstetrics, University Hospital “Dr. Jose Eleuterio González” Universidad Autónoma de Nuevo León Mexico drsordia@gmail.com Felipe A FA Martinez Department of Gynecology and Obstetrics, University Hospital “Dr. Jose Eleuterio González” Universidad Autónoma de Nuevo León Department of Gynecology and Obstetrics, University Hospital “Dr. Jose Eleuterio González” Universidad Autónoma de Nuevo León Mexico Eduardo EG Orozco Department of Gynecology and Obstetrics, University Hospital “Dr. Jose Eleuterio González” Universidad Autónoma de Nuevo León Department of Gynecology and Obstetrics, University Hospital “Dr. Jose Eleuterio González” Universidad Autónoma de Nuevo León Mexico Andrea A Flores-Rodriguez Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Mexico Paloma C PC Leyva-Camacho Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Mexico Neri NA Alvarez-Villalobos Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Mexico Jorge JA Zuñiga-Hernandez Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Plataforma INVEST Medicina UANL–KER Unit Mayo Clinic (KER Unit México), Universidad Autonoma de Nuevo Leon Mexico CryopreservationEmbryo transferIn vitro fertilizationIntracytoplasmic sperm injection 120109.pdf Rienzi L, Gracia C, Maggiulli R, LaBarbera AR, Kaser DJ, Ubaldi FM, et al. Oocyte. embryo and blastocyst cryopreservation in ART: systematic review an meta analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance. Hum Reprod Update. 2017;23(2):139-55.##Shi Q, Xie Y, Wang Y, Li S. Vitrification versus slow freezing for human ovarian tissue cryopreservation: a systematic review and meta-anlaysis. Sci Rep. 2017;7(1):8538.##Rato ML, Gouveia-Oliveira A, Plancha CE. Influence of post-thaw culture on the developmental potential of human frozen embryos. J Assist Reprod Genet. 2012;29(8):789-95.##Hartshorne GM, Elder K, Crow J, Dyson H, Edwards RG. The influence of in-vitro development upon post-thaw survival and implantation of cryopreserved human blastocysts. Hum Reprod. 1991;6(1):136-41.##Fang C, Yue CM, Huang R, Wei LN, Jia L. Pregnancy outcomes of blas-tocysts cultured overnight after thawing. Arch Gynecol Obstet. 2016;293(6):1347-56.##Wang H, Ou Z, Chen Z, Yang L, Sun L. Influence of different post-thaw culture time on the clinical outcomes of different quality embryos. Adv Clin Exp Med. 2019;28(4):523-7.##Moher D, Liberati A, Tetzlaff J, Altman DG, PRISMA Group. Preferred reporting items for systematic reviews and meta-analyses: the PRISMA statement. PloS Med. 2009;6(7):e1000097.##Higgins JPT, Thomas J, Chandler J, Cumpston M, Li T, Page MJ, et al. Cochrane handbook for systematic reviews of interventions. 2nd ed. Chichester (UK): John Wiley & Sons; 2019. 27 p.##Guyatt GH, Oxman AD, Vist GE, Kunz R, Falck-Ytter Y, Alonso-Coello P, et al. GRADE: an emerging consensus on rating quality of evidence and strength of recommendations. BMJ. 2008;336(7650):924-6.##Herbemont C, Chekroune S, Bonan S, Cedrin-Durnerin I, Vivot A, Sonigo C, et al. Impact of post-warming culture duration on clinical outcomes of vitrified good-quality blastocyst transfers: a prospective randomized study. Fertil Steril. 2018;110(7):1290-7.##Agha-Rahimi A, Omidi M, Akyash F, Faramarzi A, Farshchi FA. Does overnight culture of cleaved embryos improve pregnancy rate in vitrified-warmed embryo transfer programme? Malays J Med Sci. 2019;26(2):52-8.##Jin R, Tong X, Wu L, Luan H, Zhou G, Johansson L, et al. Extended culture of vitrified–warmed embryos in day-3 embryo transfer cycles: a randomized controlled pilot study. Reprod Biomed Online. 2013;26(4):384-92.##Joshi BV, Manish R, Banker MR, Patel PM, Preeti B, Shah PB. Transfer of human frozen-thawed embryos with further cleavage during culture increases pregnancy rates. J Hum Reprod Sci. 2010;3(2):76-9.##Griesinger G. Beware of the ‘implantation rate’! Why the outcome parameter ‘implantation rate’ should be abandoned from infertility research. Hum Reprod. 2016;31(2):249-51.##Munne S, Alikani M, Tomkin G, Grifo J, Cohen J. Embryo morphology, developmental rates, and maternal age are correlated with chromo-some abnormalities. 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Characterization of DDX4 Gene Expression in Human Cases with Non-Obstructive Azoospermia and in Sterile and Fertile Mice 2 1 2 Background: In mammals, spermatogenesis is the main process for male fertility that is initiated by spermatogonial stem cells (SSCs) proliferation. SSCs are unipotent progenitor cells accountable for transferring the genetic information to the following generation by differentiating to haploid cells during spermato-and spermiogenesis. DEAD-box helicase 4 (DDX4) is a specific germ cell marker and its expression pattern is localized to, spermatocytes, and spermatids. The expression in the SSCs on the basement membrane of the seminiferous tubules is low. Methods: Immunohistochemistry (IHC) and Fluidigm reverse transcriptase-polymerase chain reaction (RT-PCR) were used to analyze the expression of DDX4 in testis tissue of fertile and sterile mice and human cases with non-obstructive azoospermia. Results: Our immunohistochemical findings of fertile and busulfan-treated mice showed expression of DDX4 in the basal and luminal compartment of seminiferous tubules of fertile mice whereas no expression was detected in busulfan-treated mice. The immunohistochemical analysis of two human cases with different levels of non-obstructive azoospermia revealed more luminal DDX4 positive cells. Conclusion: Our findings indicate that DDX4 might be a valuable germ cell marker for analyzing the pathology of germ cell tumors and infertility as global urological problems. 085 92 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 Amirreza AR NiaziTabar 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 Atiyeh A Mohammadi 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 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 DDX4 proteinSeminiferous tubulesSpermatogonial stem cellTesticles 120102.pdf Huang P, Wang T. Spermatogonial stem cell and TGF-\b {eta} involved regulation of proliferation and differentiation. arXiv preprint arXiv:1706.03892. 2017 Jun 13.##Yoshida S. Open niche regulation of mouse spermatogenic stem cells. Dev Growth Differ. 2018;60(9):542-52.##Meinhardt A, Wang M, Schulz C, Bhushan S. Microenvironmental signals govern the cellular identity of testicular macrophages. J leukoc Biol. 2018;104(4):757-66.##Wang F, Han D. Sertoli cell phagocytosis: an essential event for spermatogenesis. In: Wu W, Ziglioli F, Maestroni U, editors. Male reproductive health. UK: IntechOpen; 2019. p. 69-84.##Chen H, Wang Y, Ge R, Zirkin BR. Leydig cell stem cells: Identification, proliferation and differentiation. Mol Cell Endocrinol. 2017;445:65-73.##De Rooij DG. The nature and dynamics of spermatogonial stem cells. Development. 2017;144(17):3022-30.##Ishikura Y, Yabuta Y, Ohta H, Hayashi K, Nakamura T, Okamoto I, et al. In vitro derivation and propagation of spermatogonial stem cell activity from mouse pluripotent stem cells. Cell Rep. 2016;17(10):2789-804.##Zhou H, Zeng Z, Koentgen F, Khan M, Mombaerts P. The testicular soma of Tsc22d3 knockout mice supports spermatogenesis and germline transmission from spermatogonial stem cell lines upon transplantation. Genesis. 2019;57(6):e23295.##Chapman KM, Medrano GA, Jaichander P, Chaudhary J, Waits AE, Nobrega MA, et al. Targeted germline modifications in rats using CRISPR/Cas9 and spermatogonial stem cells. Cell Rep. 2015;10(11):1828-35.##Griswold MD. Spermatogenesis: the commitment to meiosis. Physiol Rev. 2016;96(1):1-17.##Robles V, Herraez P, Labbé C, Cabrita E, Pšenička M, Valcarce DG, et al. Molecular basis of spermatogenesis and sperm quality. Gen Comp Endocrinol. 2017;245:5-9.##Toocheck C, Clister T, Shupe J, Crum C, Ravindranathan P, Lee TK, et al. Mouse spermatogenesis requires classical and nonclassical testosterone signaling. Biol Reprod. 2016;94(1):11.##Silber S. Hormonal regulation of spermatogenesis. In: Fundamentals of male infertility. USA: Springer; 2018. p. 39-41.##Miller SR, Morales MN, Frost KL, Cherrington NJ, Wright SH. Cell model for studying nucleoside transporters, a key component of the blood-testis barrier. FASEB J. 2019;33(21):507-12.##Hui L, Nie Y, Li S, Guo M, Yang W, Huang R, et al. Matrix metalloproteinase 9 facilitates Zika virus invasion of the testis by modulating the integrity of the blood-testis barrier. PLoS Pathog. 2020;16(4):e1008509.##Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, et al. Derivation of male germ cells from bone marrow stem cells. Lab Invest. 2006;86(7):654-63.##Nayernia K, Nolte J, Michelmann HW, Lee JH, Rathsack K, Drusenheimer N, et al. In vitro-differentiated embryonic stem cells give rise to male gametes that can generate offspring mice. Dev Cell. 2006;11(1):125-32.##Cardoso HJ, Figueira MI, Socorro S. The stem cell factor (SCF)/c-KIT signalling in testis and prostate cancer. J Cell Commun Signal. 2017;11(4):297-307.##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.##Li R, Vannitamby A, Yue SS, Handelsman D, Hutson J. Mouse minipuberty coincides with gonocyte transformation into spermatogonial stem cells: a model for human minipuberty. Reprod Fertil Dev. 2017;29(12):2430-6.##Azizi H, Asgari B, Skutella T. Pluripotency potential of embryonic stem cell-like cells derived from mouse testis. Cell J. 2019;21(3):281-9.##Patel H, Bhartiya D. Direct action of FSH on testicular stem cells. Stem Cell Res Ther. 2019;10(1):261.##Ng CL, Yue Q, Cordula S. Notch signaling is required for survival of the germline stem cell lineage in testes of drosophila melanogaster. bioRxiv. 2019 Jan 1:682773.##Boellaard WP, Stoop H, Gillis AJ, Oosterhuis JW, Looijenga LH. 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Association Between Promoter Polymorphisms of IL-1Β, IL-4 and IL-6 Genes and a Viral Load Infected Women with Human Papillomavirus 2 1 2 Background: The purpose of this study was to investigate the association between IL-4 -589C>T, IL-6 -174G>C, IL-1β -31T>C and IL-1β -511C>T genes polymorphism with high concentrations of human papillomavirus (HPV), and the influence of gene-gene interactions on persistent human papillomavirus infection. Methods: In this study, 101 infected women with high HPV viral load and 93 healthy women were involved in a case-control study. Genotyping of SNPs for IL-4 -589C>T, IL-6 -174G>C, IL-1β -31T>C and IL-1β -511C>T genes was carried out by allele-specific PCR. Quantitative analysis of HPV-DNA was performed by Amplisens HPV HCR genotype-titer software. Gene-gene interactions were analyzed using multifactor dimensionality reduction (MDR) algorithm. Haplotype interactions were analyzed by HaploView 4.2 tool. Results: The study of single individual SNPs in promoters of IL-4 -589C>T, IL-6-174G>C and IL-1β -31T>C genes did not reveal statistically significant difference in genotypes and allele frequencies among women with high HPV viral load and control group. The frequency of -511T allele and TT genotype of the IL-1β gene in case group was significantly higher than the one in control group (OR=1.71, p=0.012 and OR=2.02, p=0.046, respectively). Haplotype analysis revealed that -511C/-31T haplotype for IL-1β gene is significantly less common among women with high HPV viral load (p=0.018). Conclusion: The haplotype -511C/-31T for IL-1β gene is associated with a protective effect against increasing HPV viral load. The frequencies of -511T allele and -511TT genotype of the IL-1β -511C>T were significantly higher among women with HPV in comparison to control group. 092 103 Abbas AH Albosale Department of Medical Laboratory Techniques, Al-Dour Technical Institute, Northern Technical University Department of Medical Laboratory Techniques, Al-Dour Technical Institute, Northern Technical University Iraq abbashammadi4@gmail.com Elena EV Mashkina Genetics Department of Academy of Biology and Biotechnology, Southern Federal University-Russia Genetics Department of Academy of Biology and Biotechnology, Southern Federal University-Russia Russia CytokinesHuman papillomavirus (HPV)InflammationPolymorphismViral load 120103.pdf Zur Hausen H. Papillomaviruses in the causation of human cancers-a brief historical account. Virology. 2009;384(2):260-5.##Choi YJ, Park JS. Clinical significance of human papillomavirus genotyping. J Gynecol Oncol. 2016;27(2):e21.##Gravitt PE, Winer RL. 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Genotype at a promoter polymorphism of the interleukin-6 gene is associated with baseline levels of plasma C-reactive protein. Cardiovasc Res. 2002;53(4):1029-34.##Hall SK, Perregaux DG, Gabel CA, Woodworth T, Durham LK, Huizinga TWF, et al. Correlation of polymorphic variation in the promoter region of the interleukin‐1β gene with secretion of interleukin‐1β protein. Arthritis Rheum. 2004;50(6):1976-83.##Federal Budget Institute of Science. Central research institute for epidemiology. DNA-sorb-AM nucleic acid extraction kit instruction manual. Moscow, Russia. Available from: https://interlabservice.ru/upload/iblock/bb8/DNA-sorb-AM 060318.pdf##Federal Budget Institute of Science. Central research institute for epidemiology. AmpliSens HPV HCR screen-titre-FRT PCR kit instruction manual. Moscow, Russia. Available from: https://interlabservice.ru/upload/iblock/614/HPV HCR screen-titre-FRT 261118.pdf.##Federal Budget Institute of Science. Central research institute for epidemiology. 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Evaluation of Microdissection Testicular Sperm Extraction (mTESE), Outcomes and Predictive Factors in Ireland: The Gold Standard for Men with Non-Obstructive Azoospermia 2 1 2 Background: Microdissection testicular sperm extraction (mTESE) is the gold standard approach in sperm retrieval in men with non-obstructive azoospermia (NOA). The purpose of the study was to assess the outcomes for Irish men who have undergone mTESE with a single surgeon. Methods: This is a retrospective, single cohort study. Thirty-four patients underwent mTESE between September 2015 and June 2019. A p<0.05 was considered statistically significant. Results: In this study, sperm retrieval rate (SRR) was 47.06%. (16/34). The mean age in those who had retrieved sperm at mTESE was 37.9±2.6 years. Johnson Score (JS) and FSH were statistically different between successful and unsuccessful mTESE groups (p=0.017*10-5 and p=0.004, respectively). Optimal cutoff values for FSH, T and JS were 15 IU/L, 13 nmol/L and 5, respectively. The pregnancy rate was 63.64% (7/11) among men who went on to use mTESE sperm in an ICSI cycle. Conclusion: The combination of mTESE/Intracytoplasmic sperm injection (ICSI) is the best option available for men with NOA who prefer to achieve paternity using their own DNA. Given the overall SRRs in mTESE, it is imperative to continue research for a predictive model to better counsel azoospermic men regarding the use of mTESE. For this purpose, large, multicenter, randomized controlled trials are needed. 103 110 Pat P Rohan Department of Urology, University Hospital Waterford Department of Urology, University Hospital Waterford Ireland pat.rohan@ucdconnect.ie Niamh N Daly Waterstone Clinic Waterstone Clinic Ireland Aoife A O’Kelly Department of Urology, University Hospital Waterford Department of Urology, University Hospital Waterford Ireland Martin M O’Leary College of Science, University College Dublin College of Science, University College Dublin Ireland Tim T Dineen Waterstone Clinic Waterstone Clinic Ireland Nigam N Shah Department of Pathology, University Hospital Waterford Department of Pathology, University Hospital Waterford Ireland Padraig P Daly Department of Urology, University Hospital Waterford Department of Urology, University Hospital Waterford Ireland John J Waterstone Waterstone Clinic Waterstone Clinic Ireland Ivor I Cullen Department of Urology, University Hospital Waterford Department of Urology, University Hospital Waterford Ireland Male infertilityMicrodissection testicular sperm extractionNon-obstructive azoospermiaPredictive markersSperm retrieval 120107.pdf Dabaja AA, Schlegel PN. Microdissection testicular sperm extraction: an update. Asian J Androl. 2013;15(1):35-9.##Bernie AM, Mata DA, Ramasamy R, Schlegel PN. Comparison of microdissection testicular sperm extraction, conventional testicular sperm extraction, and testicular sperm aspiration for nonobstructive azoospermia: a systematic review and meta-analysis. Fertil Steril. 2015;104(5):1099-103.e1-3.##Abdel Raheem A, Garaffa G, Rushwan N, De Luca F, Zacharakis E, Abdel Raheem T, et al. Testicular histopathology as a predictor of a positive sperm retrieval in men with non-obstructive azoospermia. BJU Int. 2013;111(3):492-9.##Aziz N, Agarwal A. The diagnosis and treatment of male infertility: a case-based guide for clinicians. 1st ed. USA: Springer; 2017. 318 p.##Donoso P, Tournaye H, Devroey P. Which is the best sperm retrieval technique for non-obstructive azoospermia? a systematic review. Hum Reprod Update. 2007;13(6):539-49.##Esteves SC, Miyaoka R, Agarwal A. An update on the clinical assessment of the infertile male. Clinics (Sao Paulo). 2011;66(4):691-700.##Johnson L. Evaluation of the human testis and its age-related dysfunction. Prog Clin Biol Res. 1989;302:35-60.##Schlegel PN. Testicular sperm extraction: microdissection improves sperm yield with minimal tissue excision. 1999;14(1):131-5.##Griswold MD. The central role of sertoli cells in spermatogenesis. Semin Cell Dev Biol. 1998;9(4):411-6.##Johnsen SG. Testicular biopsy score count--a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Hormes. 1970;1(1):2-25.##Devroey P, Liu J, Nagy Z, Tournaye H, Silber SJ, Van Steirteghem AC. Normal fertilization of human oocytes after testicular sperm extraction and intracytoplasmic sperm injection. Fertil Steril. 1994;62(3):639-41.##Flannigan R, Bach PV, Schlegel PN. Microdissection testicular sperm extraction. Transl Androl Urol. 2017;6(4):745-52.##Waterstone Clinic. First Pregnancy following Micro-TESE in Ireland [Internet]. Waterstone Clinic: Ireland; 1 Jun 2016. Available from: https://waterstoneclinic.ie/first-micro-tese-pregnancy-ireland/.##Tsujimura A, Matsumiya K, Miyagawa Y, Takao T, Fujita K, Koga M, et al. Prediction of successful outcome of microdissection testicular sperm extraction in men with idiopathic nonobstructive azoospermia. J Urol. 2004;172(5 Pt 1):1944-7.##Tsujimura A. Microdissection testicular sperm extraction: Prediction, outcome, and complications. Int J Urol. 2007;14(10):883-9. ##Schlegel PN, Tanrikut C, Li PS. Microdissection testicular sperm extraction (TESE) in non-obstructive azoospermia. Fertil Steril. 2006;86(3):S519.##Agarwal A, Deepinder F, Cocuzza M, Agarwal R, Short RA, Sabanegh E, et al. Efficacy of varicocelectomy in improving semen parameters: new meta-analytical approach. Urology. 2007;70(3):532-8.##Dadhich P, Ramasamy R, Scovell J, Wilken N, Lipshultz L. Testosterone versus clomiphene citrate in managing symptoms of hypogonadism in men. Indian J Urol. 2017;33(3):236-40.##Katz DJ, Nabulsi O, Tal R, Mulhall JP. Outcomes of clomiphene citrate treatment in young hypogonadal men. BJU Int. 2012;110(4):573-8.##Ishikawa T. Surgical recovery of sperm in non-obstructive azoospermia. Asian J Androl. 2012;14(1):109-15.##Crabbe E, Verheyen G, Silber S, Tournaye H, Van de Velde A, Goossens A, et al. Enzymatic digestion of testicular tissue may rescue the intracytoplasmic sperm injection cycle in some patients with non-obstructive azoospermia. Hum Reprod. 1998;13(10):2791-6.##Tournaye H, Verheyen G, Nagy P, Ubaldi F, Goossens A, Silber S, et al. Are there any predictive factors for successful testicular sperm recovery in azoospermic patients? Hum Reprod. 1997;12(1):80-6.##Eken A, Gulec F. Microdissection testicular sperm extraction (micro-TESE): Predictive value of preoperative hormonal levels and pathology in non-obstructive azoospermia. Kaohsiung J Med Sci. 2018;34(2):103-8.##Cetinkaya M, Onem K, Zorba OU, Ozkara H, Alici B. Evaluation of microdissection testicular sperm extraction results in patients with non-obstructive azoospermia: independent predictive factors and best cutoff values for sperm retrieval. Urol J. 2015;12(6):2436-43.##Deruyver Y, Vanderschueren D, Van der Aa F. Outcome of microdissection TESE compared with conventional TESE in non-obstructive azoospermia: a systematic review. Andrology. 2014;2(1):20-4.##Ziaee SA, Ezzatnegad M, Nowroozi M, Jamshidian H, Abdi H, Moghaddam SMMH. Prediction of successful sperm retrieval in patients with nonobstructive azoospermia. Urol J. 2006;3(2):92-6.##Ramasamy R, Lin K, Gosden LV, Rosenwaks Z, Palermo GD, Schlegel PN. High serum FSH levels in men with nonobstructive azoospermia does not affect success of microdissection testicular sperm extraction. Fertil Steril. 2009;92(2):590-3.##Ramasamy R, Yagan N, Schlegel PN. Structural and functional changes to the testis after conventional versus microdissection testicular sperm extraction. Urology. 2005;65(6):1190-4.##Tunc L, Kirac M, Gurocak S, Yucel A, Kupeli B, Alkibay T, et al. Can serum Inhibin B and FSH levels, testicular histology and volume predict the outcome of testicular sperm extraction in patients with non-obstructive azoospermia? Int Urol Nephrol. 2006;38(3-4):629-35.##Jensen CFS, Ohl DA, Hiner MR, Fode M, Shah T, Smith GD, et al. Multiple needle-pass percutaneous testicular sperm aspiration as first-line treatment in azoospermic men. Andrology. 2016;4(2):257-62.##Vernaeve V, Tournaye H, Schiettecatte J, Verheyen G, Steirteghem AV, Devroey P. Serum inhibin B cannot predict testicular sperm retrieval in patients with non-obstructive azoospermia. Hum Reprod. 2002;17(4):971-6.##Meachem SJ, Nieschlag E, Simoni M. Inhibin B in male reproduction: pathophysiology and clinical relevance. Eur J Endocrinol. 2001;145(5):561-71.##Katz MJ, Huland DM, Ramasamy R. Multiphoton microscopy: applications in Urology and Andrology. Transl Androl Urol. 2014;3(1):77–83.##Ramasamy R, Sterling J, Fisher ES, Li PS, Jain M, Robinson BD, et al. Identification of Spermatogenesis With Multiphoton Microscopy: An Evaluation in a Rodent Model. J Urol. 2011;86(6):2487-92.##Najari BB, Ramasamy R, Sterling J, Aggarwal A, Sheth S, Li PS, et al. Pilot study of the correlation of multiphoton tomography of ex vivo human testis with histology. J Urol. 2012;188(2):538-43.##

The Comparison of Resistance Index of Testicular Artery Using Color Doppler Ultrasound in Infertile Men Undergoing Varicocelectomy 2 1 2 Background: Varicocele is one of the leading causes of infertility in men. Resistance index (RI) in testis is a parameter indicating parenchymal perfusion and microvascular functions. Increased RI in the testis of patients with varicocele might be a sign of impairments in microvascularization and a significant decrease in testicular perfusion. In the present study, RI in capsular and intraparenchymal testicular arteries was evaluated in patients with varicocele who underwent varicocelectomy. Methods: This prospective cohort study was performed in 2019-2020 in Guilan, Iran. Sixty-six patients were included. Semen analysis was also done before surgeries. Patients with at least one disorder in semen analysis entered the study. RI in testicular arteries was measured by an experienced radiologist before surgeries. Six months after varicocelectomy, all patients underwent the same semen analysis and ultrasound imaging. Data were analyzed using SPSS software. The tests for analysis included McNemar Test and Wilcoxon and p<0.005 was considered as the significance level. Results: According to the results, 42 patients (63.6%) had positive changes in sperm analysis after surgeries. Sperm analysis showed a significant increase in number, concentration, morphology, and motility of sperm after surgeries (p<0.001). Further measurements of capsular and intratesticular RI in all patients also indicated a significant decrease (p<0.001). Conclusion: Increased RI might be associated with impaired microperfusion in testis followed by impairments in semen. Moreover, mean capsular and intratesticular RI in patients decreased after surgeries and this decrease was significantly more in patients who had improvement in their semen parameters. 110 116 Amir Reza AR Dalili Department of Radiology, Razi Hospital, Guilan University of Medical Sciences Department of Radiology, Razi Hospital, Guilan University of Medical Sciences Iran Ali A Hamidi Madani Urology Research Center, Razi Hospital, Guilan University of Medical Sciences Urology Research Center, Razi Hospital, Guilan University of Medical Sciences Iran Saeid S Sadeghi Joni Department of Radiology, Razi Hospital, Guilan University of Medical Sciences Department of Radiology, Razi Hospital, Guilan University of Medical Sciences Iran saeedsadeghi69@gmail.com Color Doppler ultrasoundResistance indexSemen analysisVaricoceleVaricocelectomy 120100.pdf Alsaikhan B, Alrabeeah K, Delouya G, Zini A. Epidemiology of varicocele. Asian J Androl. 2016;18(2):179-81.##Jensen CFS, Østergren P, Dupree JM, Ohl DA, Sønksen J, Fode M. Varicocele and male infertility. Nat Rev Urol. 2017;14(9):523-33.##Damsgaard J, Joensen UN, Carlsen E, Erenpreiss J, Jensen MB, Matulevicius V, et al. Varicocele is associated with impaired semen quality and reproductive hormone levels: a study of 7035 healthy young men from six European countries. Eur Urol. 2016;70(6):1019-29.##Pallotti F, Paoli D, Carlini T, Vestri A, Martino G, Lenzi A, et al. Varicocele and semen quality: a retrospective case–control study of 4230 patients from a single centre. J Endocrinol Invest. 2018;41(2):185-92.##Cho CL, Esteves SC, Agarwal A. Novel insights into the pathophysiology of varicocele and its association with reactive oxygen species and sperm DNA fragmentation. Asian J Androl. 2016;18(2):186-93.##Agarwal A, Sharma R, Harlev A, Esteves SC. Effect of varicocele on semen characteristics according to the new 2010 World Health Organization criteria: a systematic review and meta-analysis. Asian J Androl. 2016;18(2):163-70.##Gruhot TR, Rempel LA, White BR, Mote BE. The effect of varicocele on semen quality in boars exposed to heat stress. Transl Anim Sci. 2020;4(1):293-8.##Cocuzza MS, Tiseo BC, Srougi V, Wood GJ, Cardoso JP, Esteves SC, et al. Diagnostic accuracy of physical examination compared with color Doppler ultrasound in the determination of varicocele diagnosis and grading: impact of urologists’ experience. Andrology. 2020;8(5):1160-6.##Rocher L, Gennisson JL, Barranger J, Rachas A, Criton A, Izard V, et al. Ultrasensitive Doppler as a tool for the diagnosis of testicular ischemia during the Valsalva maneuver: a new way to explore varicoceles? Acta Radiol. 2019;60(8):1048-56.##Alsyouf MM, Stokes PK, Ko EY. Imaging and other diagnostic modalities in varicocele diagnosis. In: Esteves SC, Cho CL, Majzoub A, Agarwal A, editors. Varicocele and male infertility. USA: Springer; 2019. p. 123-35.##Hamada A, Esteves SC, Agarwal A. Varicocele classification. In: Esteves SC, Cho CL, Majzoub A, Agarwal A, editors. Varicocele and male infertility. USA: Springer; 2016. p. 37-43.##Rocher L, Gennisson JL, Ferlicot S, Criton A, Albiges L, Izard V, et al. Testicular ultrasensitive Doppler preliminary experience: a feasibility study. Acta Radiol. 2018;59(3):346-54.##Aguilar-García J, Cano-González H, Martínez-Jiménez M, de la Rosa-Zapata F, Sánchez-Aguilar M. Unilateral Lichtenstein tension-free mesh hernia repair and testicular perfusion: a prospective control study. Hernia. 2018;22(3):479-82.##Jedrzejewski G, Osemlak P, Wieczorek AP, Nachulewicz P. Testicular sonographic color Doppler dynamic tissue perfusion measurements in adolescents with varicocele. Urol Int. 2019;103(1):55-61.##Kohn TP, Kohn JR, Pastuszak AW. Varicocelectomy before assisted reproductive technology: are outcomes improved? Fertil Steril. 2017;108(3):385-91.##Kohn TP, Ohlander SJ, Jacob JS, Griffin TM, Lipshultz LI, Pastuszak AW. The effect of subclinical varicocele on pregnancy rates and semen parameters: a systematic review and meta-analysis. Curr Urol Rep. 2018;19(7):53.##Unsal A, Turgut AT, Taşkin F, Koşar U, Karaman CZ. Resistance and pulsatility index increase in capsular branches of testicular artery: indicator of impaired testicular microcirculation in varicocele? J Clin Ultrasound. 2007;35(4):191-5.##Akcar N, Turgut M, Adapınar B, Ozkan IR. Intratesticular arterial resistance and testicular volume in infertile men with subclinical varicocele. J Clin Ultrasound. 2004;32(8):389-93.##Tarhan S, Gümüs B, Gündüz İ, Ayyildiz V, Göktan C. Effect of varicocele on testicular artery blood flow in men color doppler investigation. Scand J Urol Nephrol. 2003;37(1):38-42.##Hassan A, Gad HM, Mostafa T. Radiologically assessed testicular changes in infertile males with varicocele. Andrologia. 2011;43(50:307-11.##Zhang M, Du L, Liu Z, Qi H, Chu Q. The effects of varicocelectomy on testicular arterial blood flow: laparoscopic surgery versus microsurgery. Urol J. 2014;11(5):1900-6.##Ener K, Üçgül YE, Okulu E, Aldemir M, Işik E, Kayigil Ö, et al. Comparison of arterial blood supply to the testicles in the preoperative and early postoperative period in patients undergoing subinguinal varicocelectomy. Scand J Urol. 2015;49(2):169-73.##Balci A, Karazincir S, Gorur S, Sumbas H, Egilmez E, Inandi T. Long‐term effect of varicocele repair on intratesticular arterial resistance index. J Clin Ultrasound. 2008;36(3):148-52.##Akand M, Koplay M, Islamoglu N, Altintas E, Kilic O, Gul M, et al. Color Doppler ultrasound characteristics after subinguinal microscopic varicocelectomy. Med Ultrason. 2017;19(1):59-65.##Semiz I, Tokgöz Ö, Tokgoz H, Voyvoda N, Serifoglu I, Erdem Z. The investigation of correlation between semen analysis parameters and intraparenchymal testicular spectral Doppler indices in patients with clinical varicocele. Ultrasound Q. 2014;30(1):33-40.##

Clinical Efficacy of Follitropin Alfa in GnRH-Antagonist Protocols: A Prospective Observational Phase IV Study on the Use of Biosimilar Follitropin Alfa r-hFSH in Assisted Reproductive Technology in a Routine Care Setting 2 1 2 Background: This phase IV routine care study evaluated ovarian responses when using a biosimilar follitropin alfa r-hFSH (Bemfola®) for controlled ovarian stimulation (COS) in women undergoing assisted reproductive technology (ART) treatment who were pituitary-suppressed with a gonadotrophin-releasing hormone (GnRH) antagonist. Methods: This multicenter, prospective, non-comparative, non-interventional study (Germany/Austria) was conducted with 885 women (Mean age of 34.0±4.4 years) for whom COS with Bemfola® and GnRH-antagonist for pituitary suppression were applied as part of in vitro fertilization (IVF) treatment with/without intracytoplasmic sperm injection (ICSI) observing routine clinical-practice protocols. Primary endpoint was the number of retrieved cumulus-oocyte-complexes (COCs). Results: Among 986 ART cycles, COS was given for 9.9±1.8 days (First-day r-hFSH dose of 220.7±68.9 IU; mean total dose of 2184.3±837.5 IU). It was revealed that 99.1% of cycles resulted in follicular puncture, with mean of 10.7±6.6 oocytes retrieved. Successful fertilization took place after IVF/ICSI in 93.8% of follicular punctures. Freeze-all was performed in 14.2% of cycles. Fresh embryo transfer was performed in 76.9% of cycles with follicular puncture; mean day of transfer was 3.5±1.3 and average number of transferred embryos was 1.76±0.50. Clinical pregnancy rate was 30.2% of embryo-transfer cycles and 23.4% of started cycles. Sixty-nine reports of ovarian hyperstimulation syndrome (7.0% of started cycles) were documented. Conclusion: COS with Bemfola® in GnRH-antagonist IVF/ICSI protocols in a routine care setting led to an appropriate ovarian response allowing oocyte retrieval in 99.1% of initiated cases. 116 125 Georg G Griesinger Sektion Für Gynaekologische Endokrinologie und Reproduktionsmedizin, Klinik Für Frauenheilkunde & Geburtshilfe (Gynaekologie), Universitaetsklinikum Schleswig-Holstein-Campus Luebeck Sektion Für Gynaekologische Endokrinologie und Reproduktionsmedizin, Klinik Für Frauenheilkunde & Geburtshilfe (Gynaekologie), Universitaetsklinikum Schleswig-Holstein-Campus Luebeck Germany georg.griesinger@uni-luebeck.de Thilo T Schill MVZ Kinderwunschzentrum Langenhagen-Wolfsburg MVZ Kinderwunschzentrum Langenhagen-Wolfsburg Germany Michael M Sator Kinderwunsch im Zentrum GmbH Kinderwunsch im Zentrum GmbH Australia Michael M Schenk Das Kinderwunsch Institut Schenk GmbH Das Kinderwunsch Institut Schenk GmbH Australia Jan-Steffen JS Krüssel Universitaeres interdisziplinäres Kinderwunschzentrum Duesseldorf (UniKiD), Klinik für Frauenheilkunde und Geburtshilfe, Universitätsklinikum Duesseldorf Universitaeres interdisziplinäres Kinderwunschzentrum Duesseldorf (UniKiD), Klinik für Frauenheilkunde und Geburtshilfe, Universitätsklinikum Duesseldorf Germany Assisted reproductive technologyBiosimilar pharmaceuticalsFollitropin alfaGonadotropin-releasing hormone antagonist 120104.pdf Wolzt M, Gouya G, Sator M, Hemetsberger T, Irps C, Rettenbacher M, et al. Comparison of pharmacokinetic and safety profiles between Bemfola(®) and Gonal-f(®) after subcutaneous application. Eur J Drug Metab Pharmacokinet. 2016;41(3):259-65.##Rettenbacher M, Andersen AN, Garcia-Velasco JA, Sator M, Barri P, Lindenberg S, et al. A multi-centre phase 3 study comparing efficacy and safety of Bemfola(®) versus Gonal-f(®) in women undergoing ovarian stimulation for IVF. Reprod Biomed Online. 2015;30(5):504-13.##Niederberger C, Pellicer A, Cohen J, Gardner DK, Palermo GD, O’Neill CL, et al. Forty years of IVF. Fertil Steril. 2018;110(2):185-324.e5.##Imthurn B, McVeigh E, Stiller R, Arnold M, Pringle F, Irps C, et al. Evaluation of the use and handling of three different pen systems considered for in vitro fertilization treatment. Expert Opin Drug Deliv. 2014;11(12):1859-64.##Quintero LA, Verdú Merino V, Olivares Vela R, Rayward J, Torres Afonso A, Báez Quintana D, et al. An evaluation by potential IVF/donor oocyte patients of the use and handling of the Bemfola® Pen compared with the Gonal-f® Pen and Puregon Pen®. Revista Iberoamericana de Fertilidad y Reproducción Humana. 2017; 34:64-73.##Saunders H, de la Fuente Bitaine L, Eftekhar C, Howles CM, Glaser J, Hoja T, et al. Functionality of a novel follitropin alfa pen injector: results from human factor interactions by patients and nurses. Expert Opin Drug Deliv. 2018;15(6):549-58. ##Blumenauer V, Czeromin U, Fehr D, Fiedler K, Gnoth C, Krüssel JS, et al. D.I.R annual 2017 - the German IVF-registry. J Reproduktionsmed Endokrinol. 2018;15:217-50.##Van Tilborg TC, Oudshoorn SC, Eijkemans MJ, Mochtar MH, Van Golde RJ, Hoek A, et al. Individualized FSH dosing based on ovarian reserve testing in women starting IVF/ICSI: a multicentre trial and cost-effectiveness analysis. Hum Reprod. 2017;32(12):2485-95.##Nastri CO, Teixeira DM, Moroni RM, Leitão VM, Martins WP. Ovarian hyperstimulation syndrome: pathophysiology, staging, prediction and prevention. Ultrasound Obstet Gynecol. 2015;45(4):377-93.##Roque M, Valle M, Kostolias A, Sampaio M, Geber S. Freeze-all cycle in reproductive medicine: current perspectives. JBRA Assist Reprod. 2017;21(1):49-53.##Griesinger G, Von Otte S, Schroer A, Ludwig AK, Diedrich K, Al-Hasani S, et al. Elective cryopreservation of all pronuclear oocytes after GnRH agonist triggering of final oocyte maturation in patients at risk of developing OHSS: a prospective, observational proof-of-concept study. Hum Reprod. 2007;22(5):1348-52.##

The Impact of COVID-19 Pandemic on Stress and Anxiety of Non-infected Pregnant Mothers 2 1 2 Background: The newly emerging COVID-19 has caused severe anxiety around the world and it is infecting more people each day since there is no preventive measure or definite therapy for the diseases. The present study aimed to evaluate its effect on anxiety and stress of pregnant mothers during perinatal care. Methods: Three–hundred pregnant mothers without COVID–19 infection who were referred to the hospitals affiliated to Iran University of Medical Sciences for delivery during April 2020, based on negative clinical symptoms and the results of polymerase chain reaction (rt-PCR) for COVID–19, were recruited by census method and asked to complete the Persian version of the perceived stress scale (PSS); participants views about their anxiety level and the role of COVID–19 as the source of their stress and worries were recorded. Women who refused to continue the study were excluded. The frequency of variables and mean scores were calculated using SPSS v. 21. Results: Mean age of mothers was 30.20±16.19 years; 31.3% were primigravida and mean gestational age was 38.00±4.14 weeks. Moreover, 16.3% asked for earlier pregnancy termination and 39% requested Cesarean section (C/S). Assessing the mothers’ anxiety revealed a high/very high level of anxiety in 51.3%. The majority felt worried and frustrated because of COVID–19 (86.4%). Social media had a great impact on the level of stress among these mothers (60.3%). Conclusion: COVID-19 pandemic is an important source for the increased anxiety and stress among healthy pregnant mothers. 125 133 Abolfazl A Mehdizadehkashi Abolfazl Mehdizadehkashi Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran Shahla Sh Chaichian Shahla Chaichian Pars Advanced and Minimally Invasive Medical Manners Research Center, Pars Hospital, Iran University of Medical Sciences Pars Advanced and Minimally Invasive Medical Manners Research Center, Pars Hospital, Iran University of Medical Sciences Iran chaichian.sh@iums.ac.ir, shchaichian@gmail.com Ladan L Haghighi Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran Nooshin N Eshraghi Department of Perinatology, Shahid Akbarabadi Hospital, Iran University of Medical Sciences Department of Perinatology, Shahid Akbarabadi Hospital, Iran University of Medical Sciences Iran Arash A Bordbar Shahid Akbarabadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences (IUMS) Shahid Akbarabadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences (IUMS) Iran Neda N Hashemi Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran Roya R Derakhshan Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran Shahla Sh Mirgalobayat Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran Samaneh S Rokhgireh Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran Kobra K Tahermanesh Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Endometriosis Research Center, Iran University of Medical Sciences (IUMS) Iran No Keyword 120108.pdf Thoits PA. Stress and health: Major findings and policy implications. J Health Soc Behav. 2010;51 Suppl:S41-S53.##Keller A, Litzelman K, Wisk LE, Maddox T, Cheng ER, Creswell PD, et al. Does the perception that stress affects health matter? the association with health and mortality. Health Psychol. 2012;31(5):677-84.##Schneiderman N, Ironson G, Siegel SD. Stress and health: psychological, behavioral, and biological determinants. Annu Rev Clin Psychol. 2005;1:607-28.##Goodman JH, Chenausky KL, Freeman MP. Anxiety disorders during pregnancy: a systematic review. J Clin Psychiatry. 2014;75(10):e1153-84.##Schetter CD, Tanner L. Anxiety, depression and stress in pregnancy: implications for mothers, children, research, and practice. Curr Opin Psychiatry. 2012;25(2):141-8.##Grote NK, Bridge JA, Gavin AR, Melville JL, Iyengar S, Katon WJ. A meta-analysis of depression during pregnancy and the risk of preterm birth, low birth weight, and intrauterine growth restriction. Arch Gen Psychiatry. 2010;67(10):1012-24.##Lima SAM, El Dib RP, Rodrigues MRK, Ferraz GAR, Molina AC, Neto CAP, et al. Is the risk of low birth weight or preterm labor greater when maternal stress is experienced during pregnancy? a systematic review and meta-analysis of cohort studies. PloS One. 2018;13(7):e0200594.##Biaggi A, Conroy S, Pawlby S, Pariante CM. Identifying the women at risk of antenatal anxiety and depression: a systematic review. J Affect Disord. 2016;191:62-77.##Satyanarayana VA, Lukose A, Srinivasan K. Maternal mental health in pregnancy and child behavior. Indian J Psychiatry. 2011;53(4):351-61.##Singhal T. A review of coronavirus disease-2019 (COVID-19). Indian J Pediatr. 2020;87(4):281-6.##Hu Y, Sun J, Dai Z, Deng H, Li X, Huang Q, et al. Prevalence and severity of corona virus disease 2019 (COVID-19): a systematic review and meta-analysis. J Clin Virol. 2020;127:104371.##Hellewell J, Abbott S, Gimma A, Bosse NI, Jarvis CI, Russell TW, et al. Feasibility of controlling COVID-19 outbreaks by isolation of cases and contacts. Lancet Glob Health. 2020;8(4):e488-e96.##World Health Organization. Coronavirus disease 2019 (COVID-19). Geneva: World Health Organization; 2020 Mar. 10 p. Report No. 70.##Gao J, Zheng P, Jia Y, Chen H, Mao Y, Chen S, et al. Mental health problems and social media exposure during COVID-19 outbreak. PloS One. 2020;15(4):e0231924.##Wu YC, Chen CS, Chan YJ. The outbreak of COVID-19: an overview. J Chin Med Assoc. 2020;83(3):217-20.##Rajkumar RP. COVID-19 and mental health: a review of the existing literature. Asian J Psychiatr. 2020;52:102066.##Lancet T. COVID-19: protecting health-care workers. Lancet. 2020;395(10228):922.##Akkerman D, Cleland L, Croft G, Eskuchen K, Heim C, Levine A, et al. Routine prenatal care. 5th ed. Bloomington (MN): Institute for clinical systems improvement (ICSI); 2012. 116 p.##Alzamora MC, Paredes T, Caceres D, Webb CM, Valdez LM, La Rosa M. Severe COVID-19 during pregnancy and possible vertical transmission. Am J Perinatol. 2020;37(8):861-5.##Corbett GA, Milne SJ, Hehir MP, Lindow SW, O’connell MP. Health anxiety and behavioural changes of pregnant women during the COVID-19 pandemic. Eur J Obstet Gynecol Reprod Biol. 2020;249:96-7.##Loomans EM, Van Dijk AE, Vrijkotte TG, Van Eijsden M, Stronks K, Gemke RJ, et al. Psychosocial stress during pregnancy is related to adverse birth outcomes: results from a large multi-ethnic community-based birth cohort. Eur J Public Health. 2013;23(3):485-91.##Dashraath P, Jeslyn WJL, Karen LMX, Min LL, Sarah L, Biswas A, et al. Coronavirus disease 2019 (COVID-19) pandemic and pregnancy. Am J Obstet Gynecol. 2020;222(6):521-31.##Cohen S, Kessler RC, Gordon LU. Measuring stress: a guide for health and social scientists. 1st ed. New York: Oxford University Press; 1997. 235 p.##Lee EH. Review of the psychometric evidence of the perceived stress scale. 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A Novel Balanced Chromosomal Translocation in an Azoospermic Male: A Case Report 2 1 2 Background: Balanced translocation and azoospermia as two main reasons for recurrent pregnancy loss are known to be the leading causes of infertility across the world. Balanced translocations in azoospermic males are very rare and extensive studies need to be performed to elucidate the translocation status of the affected individuals. Case Presentaion: The cytogenetic characterization of a 28 year old male and his female partner is reported in this study. The male partner was diagnosed with non-obstructive azoospermia (NOA) and the couple was unable to conceive. Cytogenetic analysis by karyotyping through Giemsa-trypsin-giemsa banding technique (GTG) showed a novel balanced translocation, 46,XY,t(19;22)(19q13.4;22q11.2), 13ps+ in the male and the female karyotype was found to be 46,XX. Multicolor fluorescence in situ hybridization (mFISH) analysis on paternal chromosomal preparations confirmed both the region and origin of balanced translocation. The status of Y chromosome microdeletion (YMD) was analyzed and no notable microdeletion was observed. Furthermore, protein-protein interaction (PPI) network analysis was performed for breakpoint regions to explore the possible functional genetic associations. Conclusion: The azoospermic condition of the male patient along with novel balanced chromosomal translocation was responsible for infertility irrespective of its YMD status. Therefore, cytogenetic screening of azoospermic patients should be performed in addition to routine semen analysis to rule out or to confirm presence of any numerical or structural anomaly in the patient. 133 138 Abhik A Chakraborty Molecular Stress and Stem Cell Biology Group, School of Biotechnology, KIIT University, Bhubaneswar Molecular Stress and Stem Cell Biology Group, School of Biotechnology, KIIT University, Bhubaneswar India Indira I Palo Department of Obstetrics and Gynecology, Amit Hospital Department of Obstetrics and Gynecology, Amit Hospital India Souvick S Roy Molecular Stress and Stem Cell Biology Group, School of Biotechnology, KIIT University, Bhubaneswar Molecular Stress and Stem Cell Biology Group, School of Biotechnology, KIIT University, Bhubaneswar India Shu SW Koh Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore Singapore Manoor MP Hande Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore Singapore Birendranath B Banerjee Molecular Stress and Stem Cell Biology Group, School of Biotechnology, KIIT University, Bhubaneswar Molecular Stress and Stem Cell Biology Group, School of Biotechnology, KIIT University, Bhubaneswar India bnbanerjee@kiitbiotech.ac. in AzoospermiaBalanced translocationInfertilityMulticolor fluorescence in situ hybridization 120105.pdf Esteves SC, Miyaoka R, Agarwal A. An update on the clinical assessment of the infertile male. [corrected]. Clinics (Sao Paulo). 2011;66(4):691-700.##Esteves SC. Novel concepts in male factor infertility: clinical and laboratory perspectives. J Assist Reprod Genet. 2016;33(10):1319-35.##Frydman N, Romana S, Le Lorc'h M, Vekemans M, Frydman R, Tachdjian G. Assisting reproduction of infertile men carrying a robertsonian translocation. Hum Reprod. 2001;16(11):2274-7.##Wang H, Jia Z, Mao A, Xu B, Wang S, Wang L, et al. Analysis of balanced reciprocal translocations in patients with subfertility using single-molecule optical mapping. J Assist Reprod Genet. 2020;37(3):509-16.##Foresta C, Garolla A, Bartoloni L, Bettella A, Ferlin A. Genetic abnormalities among severely oligospermic men who are candidates for intracytoplasmic sperm injection. J Clin Endocrinol Metab. 2005;90(1):152-6.##Arafa MM, Majzoub A, AlSaid SS, El Ansari W, Al Ansari A, Elbardisi Y, et al. Chromosomal abnormalities in infertile men with azoospermia and severe oligozoospermia in Qatar and their association with sperm retrieval intracytoplasmic sperm injection outcomes. Arab J Urol. 2017;16(1):132-9.##Sheth FJ, Liehr T, Kumari P, Akinde R, Sheth HJ, Sheth JJ. Chromosomal abnormalities in couples with repeated fetal loss: An Indian retrospective study. Indian J Hum Genet. 2013;19(4):415-22.##Hande MP, Azizova TV, Burak LE, Khokhryakov VF, Geard CR, Brenner DJ. Complex chromosome aberrations persist in individuals many years after occupational exposure to densely ionizing radiation: an mFISH study. Genes Chromosomes Cancer. 2005;44(1):1-9.##Agarwal S, Agarwal A, Khanna A, Singh K. Microdeletion of Y chromosome as a cause of recurrent pregnancy loss. J Hum Reprod Sci. 2015;8(3):159-64.##Krausz C, Hoefsloot L, Simoni M, Tüttelmann F, European Academy of A, European Molecular Genetics Quality N. EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: state-of-the-art 2013. Andrology. 2014;2(1):5-19.##Simoni M, Bakker E, Krausz C. EAA/EMQN best practice guidelines for molecular diagnosis of y-chromosomal microdeletions. State of the art 2004. Int J Androl. 2004;27(4):240-9.##Arumugam B, Samuel CR, Thyagarajan SS. Balanced autosomal translocations in two women reporting recurrent miscarriage. J Clin Diagn Res. 2016;10(12):Gd01-GD03.##Dutta UR, Ponnala R, Dalal A. A novel de novo balanced reciprocal translocation t(18;22) associated with recurrent miscarriages: a case report. J Reprod Infertil. 2014;15(2):113-6.##Ogilvie CM, Scriven PN. Meiotic outcomes in reciprocal translocation carriers ascertained in 3-day human embryos. Eur J Hum Genet. 2002;10(12):801-6.##Harton GL, Tempest HG. Chromosomal disorders and male infertility. Asian J Androl. 2012;14(1):32-9.##Ray PF, Toure A, Metzler-Guillemain C, Mitchell MJ, Arnoult C, Coutton C. Genetic abnormalities leading to qualitative defects of sperm morphology or function. Clin Genet. 2017;91(2):217-32.##Kfir S, Basavaraja R, Wigoda N, Ben-Dor S, Orr I, Meidan R. Genomic profiling of bovine corpus luteum maturation. PloS One. 2018;13(3):e0194456.##Zuazua-Villar P, Rodriguez R, Gagou ME, Eyers PA, Meuth M. DNA replication stress in CHK1-depleted tumour cells triggers premature (S-phase) mitosis through inappropriate activation of Aurora kinase B. Cell Death Dis. 2014;5(5):e1253.##Lin M, Pedrosa E, Hrabovsky A, Chen J, Puliafito BR, Gilbert SR, et al. Integrative transcriptome network analysis of iPSC-derived neurons from schizophrenia and schizoaffective disorder patients with 22q11.2 deletion. BMC Syst Biol. 2016;10(1):105.##Devaraju P, Yu J, Eddins D, Mellado-Lagarde MM, Earls LR, Westmoreland JJ, et al. Haploinsufficiency of the 22q11.2 microdeletion gene Mrpl40 disrupts short-term synaptic plasticity and working memory through dysregulation of mitochondrial calcium. Mol Psychiatry. 2017;22(9):1313-26.##Guna A, Butcher NJ, Bassett AS. Comparative mapping of the 22q11.2 deletion region and the potential of simple model organisms. J Neurodeve Disord. 2015;7(1):18.##

Chromosomal Analysis of Cumulus Cells as a Future Predictor for Oocyte Aneuploidy: A Case Report 2 1 2 Background: Within the ovary, the optimal growth of the follicle, oocyte maturation and ovulation are highly conditioned by the two-way cross talk and interactions between the oocyte and the immediate somatic cells, known as cumulus cells (CCs). This biological communication between cell lines triggered the interest in the study of CCs as a biomarker of oocyte competence. Case Presentation: The findings of a 45,X mosaic pattern on CCs from a female patient with unremarkable medical history are reported in this study. The patient came to the Centre for Reproductive and Genetic Health, London on 14th August 2019 for her first visit and the follow up procedures were done for her to determine underlying genetic status. For this purpose, four sources of DNA including CCs, blood lymphocytes, buccal cells and immature oocytes were analyzed in the present report. Conclusion: In the present case study, the hypothesis of the female patient being mosaic 45,X was confirmed although the degree of mosaicism and whether this was affecting the germinal line could not be determined. In the event of the discovery of a cell line with an apparently abnormal genetic makeup, genetic counselling is important in order to understand the implications from somatic to germinal cells for patients exploring fertility journeys. 138 144 Xavier XV Gonzalez Department of Embryology, The Centre for Reproductive and Genetic Health Department of Embryology, The Centre for Reproductive and Genetic Health UK xavier.vinalsgonzalez@crgh.co.uk Rabi R Ahmed-Odia Department of Embryology, The Centre for Reproductive and Genetic Health Department of Embryology, The Centre for Reproductive and Genetic Health UK Himanshu H Gupta Himanshu Gupta Institute for Women’s Health, University College London Institute for Women’s Health, University College London UK Roy RP Naja Institute for Women’s Health, University College London Institute for Women’s Health, University College London UK Falak F Arshad School of Medicine, Cardiff University School of Medicine, Cardiff University UK Paul P Serhal Department of Embryology, The Centre for Reproductive and Genetic Health Department of Embryology, The Centre for Reproductive and Genetic Health UK Wael W Saab Department of Embryology, The Centre for Reproductive and Genetic Health Department of Embryology, The Centre for Reproductive and Genetic Health UK Srividya S Seshadri Department of Embryology, The Centre for Reproductive and Genetic Health Department of Embryology, The Centre for Reproductive and Genetic Health UK BiomarkerCounsellingDNAGeneticsMosaicismOocyte 120110.pdf Son WY, Das M, Shalom-Paz E, Holzer H. Mechanisms of follicle selection and development. Minerva Ginecol. 2011;63(2):89-102. ##Assou S, Haouzi D, Mahmoud K, Aouacheria A, Guillemin Y, Pantesco V, et al. A non-invasive test for assessing embryo potential by gene expression profiles of human cumulus cells: a proof of concept study. Mol Hum Reprod. 2008;14(12):711-9. ##Feuerstein P, Puard V, Chevalier C, Teusan R, Cadoret V, Guerif F, et al. Genomic assessment of human cumulus cell marker genes as predictors of oocyte developmental competence: impact of various experimental factors. PLoS One. 2012;7(7):e40449. ##Fragouli E, Wells D, Iager AE, Kayisli UA, Patrizio P. Alteration of gene expression in human cumulus cells as a potential indicator of oocyte aneuploidy. Hum Reprod. 2012;27(8):2559-68.##Russell LM, Strike P, Browne CE, Jacobs PA. X chromosome loss and ageing. Cytogenet Genome Res. 2007;116(3):181-5.##Sugawara N, Kimura Y, Araki Y. Case report: a successful pregnancy outcome in a patient with non-mosaic Turner syndrome (45, X) via in vitro fertilization. Hum Cell. 2013;26(1):41-3. ##Peek R, Schleedoorn M, Smeets D, van de Zande G, Groenman F, Braat D, et al. Ovarian follicles of young patients with Turner’s syndrome contain normal oocytes but monosomic 45,X granulosa cells. Hum Reprod. 2019;34(9):1686-96. ## Gilbert SF. Developmental Biology. 6th ed. Sunderland (MA): Sinauer Associates; 2000. ##Peek R, Schleedoorn M, Smeets D, van de Zande G, Groenman F, Braat D, et al. Ovarian follicles of young patients with Turner's syndrome contain normal oocytes but monosomic 45,X granulosa cells. Hum Reprod. 2019;34(9):1686-96.##Turner JMA. Meiotic silencing in mammals. Annu Rev Genet. 2015;49:395-412. ##Machiela MJ, Zhou W, Karlins E, Sampson JN, Freedman ND, Yang Q, et al. Female chromosome X mosaicism is age-related and preferentially affects the inactivated X chromosome. Nat Commun. 2016;7:11843.##Hook EB. Exclusion of chromosomal mosaicism: tables of 90%, 95% and 99% confidence limits and comments on use. Am J Hum Genet. 1977;29(1):94-7. ##Deng X, Berletch JB, Nguyen DK, Disteche CM. X chromosome regulation: diverse patterns in development, tissues and disease. Nat Rev Genet. 2014;15(6):367-78. ##Aller V, Gargallo M, Abrisqueta JA. Familial transmission of a duplication-deficiency X chromosome associated with partial Turner syndrome. Clin Genet. 1995;48(6):317-20.##Held KR, Kerber S, Kaminsky E, Singh S, Goetz P, Seemanova E, et al. Mosaicism in 45,X Turner syndrome: does survival in early pregnancy depend on the presence of two sex chromosomes? Hum Genet. 1992;8(3):288-94.##Varela M, Shapira E, Hyman DB. Ullrich-Turner syndrome in mother and daughter: prenatal diagnosis of a 46,X,del(X)(p21) offspring from a 45,X mother with low-level mosaicism for the del(X)(p21) in one ovary. Am J Med Genet. 1991;39(4):411-2.##Goldman RH, Racowsky C, Farland LV, Munné S, Ribustello L, Fox JH. Predicting the likelihood of live birth for elective oocyte cryopreservation: a counseling tool for physicians and patients. Hum Reprod. 2017;32(4):853-9. ##Cil AP, Bang H, Oktay K. Age-specific probability of live birth with oocyte cryopreservation: an individual patient data meta-analysis. Fertil Steril. 2013;100(2):492-9.e3.##Verpoest W, Staessen C, Bossuyt PM, Goossens V, Altarescu G, Bonduelle M, et al. Preimplantation genetic testing for aneuploidy by microarray analysis of polar bodies in advanced maternal age: a randomized clinical trial. Hum Reprod. 2018;33(9):1767-76.##

Pregnancy Consequences and Vertical Transmission Potential of SARS- CoV-2 Infection: Seeking Answers From a Preliminary Observation 2 1 2 Dear Editor, The pandemic of COVID-19 has transpired as the stiffest public health problem in 2020. For pregnant females infected with SARS-CoV-2, breach of placental barrier, i.e., vertical transmission and successive fetal complications are significant concerns (1). Pregnant females are susceptible to the invasion of respiratory pathogens causing pneumonia. During this period, physiological changes in the immune and cardiopulmonary system occur (2). Maternal pneumonia is associated with adverse obstetrical outcomes such as premature rupture of membranes, preterm labor, stillbirth, and neonatal deaths (3). Since the emergence of SARS-CoV-2, which was not previously seen in humans, little knowledge about COVID-19 and its adverse maternal and fetal outcomes exists. Earlier in 2012, researchers in Taiwan, in their population-based study, showed that mothers suffering from known pneumonia (Non-COVID-19) were more likely to have low birth weight, preterm, small for gestational age, and low Apgar score babies than healthy mothers. These mothers also have more chances of caesarean section and pre-eclampsia/eclampsia (4). The objective of the present study was to assess the effect of COVID-19 on maternal and perinatal outcomes and the potential for vertical transmission of SARS-CoV-2. The present study was a cross-sectional observational study conducted between April 2020 and August 2020 period. All 28 women admitted for delivery in dedicated COVID-19 rural tertiary care hospital during the study period were included. All 29 neonates of the index pregnancy of above females (Including one stillbirth and a twin delivery) were included in the present study. Nasopharyngeal swabs were obtained for SARS-CoV-2 RT-PCR test from all pregnant females. Samples of all neonate’s nasopharyngeal swabs were collected immediately after birth for the evidence of vertical transmission of COVID-19. The mean±SD of maternal age was 27.43±4.94 years (Minimum age of 20 years and maximum age of 38 years). As there was one twin pregnancy, therefore, the number of newborns that were included in the present study was 29. The mean birth weight of newborns in the present study was 2.57±0.67 kgs (Minimum weight was 700 grams, and the maximum reported weight was 4.0 kilograms). The mean period of gestation was 37.90± 3.75 weeks. Maternal characteristics and outcomes: All the mothers in the study had COVID-19 (By RT-PCR test), but none of them had any COVID-19 symptoms (A pneumonia-like fever, cough, fatigue, sore throat, shortness of breath, and diarrhea). Twenty females (71.4%) out of 28 came to the hospital with no complications. One female (3.6%) came with twin pregnancy, two females (7.1%) came with the transverse lie, and two (7.1%) with leaking per vaginum. Also, one female (3.6%) presented with placenta previa, one (3.6%) with "pregnancy with an IUD" and one (3.6%) with antepartum hemorrhage. Also, there was no complication reported in any women during the delivery and after delivery. Neonatal characteristics and outcomes: All the neonates, including stillborn and preterm, were negative cases of SARS-CoV-2 during the observation period and the rest (Excluding one stillbirth and two newborns which could not be revived probably due to pre-maturity) were normal and healthy without any COVID-19 symptoms. Out of 29 newborns, 26 (89.7%) were healthy, one (3.4%) was a stillbirth, and two newborns (6.9%) could not be revived (The premature baby of 700 grams at 27 weeks and 1000 grams baby at 28 weeks). Nasopharyngeal swabs for SARS-CoV-2 RT-PCR tests were repeated after the first sampling before the discharge of all cases. Twenty cases (69.0%) were delivered by LSCS for common obstetric factors such as the previous history of cesarean, pre-eclampsia, or fetal distress. At the same time, 9 cases (31.0%) had a vaginal delivery. The gestational age of six newborns was between "more than 28 weeks and less than 37 weeks". About 10.3% of pregnancies (3 newborns) were post-dated, i.e., more than 42 completed weeks of gestational age. The birth weight in 72.4% of babies was ≥2.5 kg, six babies (20.7%) had a birth weight between "1.5 to 2.4 kg", while 6.9% of babies had a birth weight of less than 1.5 kgs. Tang et al. observed that mothers having pneumonia (Non-COVID-19) during pregnancy had more chances of preterm births and caesarean sections than the healthy mothers (5). In a retrospective study, Chen et al. investigated the probability of intrauterine vertical transmission of SARS-CoV-2 virus in pregnant women with laboratory-confirmed RT-PCR test for COVID-19 in late pregnancy; amniotic fluid, cord blood, and neonatal pharyngeal swab samples from six neonates were tested for SARS-CoV-2 using quantitative RT-PCR. All were tested negative for the virus, implying that the vertical transmission of the virus did not occur during the third trimester of pregnancy (6). There was no evidence of vertical transmission in nine pregnant women with COVID-19 in the third trimester of pregnancy, and vaginal secretion samples were also tested negative for SARS-CoV-2 RNA in a study done by Lei et al. (7). Also, Chen et al. analyzed placental tissue from three pregnant women with confirmed COVID-19 infection and neonatal pharyngeal swab samples to assess the possible risk of vertical transmission, and all were tested negative for SARS-CoV-2 RNA (8). Liu et al. conducted a study of three pregnant females who acquired SARS-CoV-2 infection in their late pregnancy. They found that two of the cases had an uneventful caesarean delivery and healthy newborns. None of the three infants became infected by vertical transmission. Li et al. published a case report of a pregnant female who was COVID-19 positive in her 35th week of pregnancy and delivered an infant by caesarean section. This infant was found negative for SARS-CoV-2 infection. Yang et al. had done a study on seven females who got infected in their late pregnancy. All the females were delivered by caesarean section. They took samples of six newborns out of seven, and all the samples were found negative for SARS-CoV-2 infection (9-11). These results are in accordance with the present research where most of the deliveries (69%) were by LSCS, and no newborn was found COVID-19 positive. Chen et al. collected data on five COVID-9 positive pregnant females between the ages of 25 and 31 years. The gestational age at delivery among them was between 38 weeks to 40 weeks. Three of them were gravida one, and two were gravida three. Four had zero parity, while one female had parity of one. Caesarean section was done among two females out of five (40%). There were no complications after delivery. All the neonates were negative for SARS-CoV-2 RT-PCR, and none of them had any symptoms of COVID-19 (12). The age group, gravida, and parity of the above study were similar to the current study and only the rate of caesarean section was slightly higher (69%) in the present study. Similarly, in the present study, there were no complications after delivery, and all the neonates (100.0%) were negative for SARS-CoV-2 RT-PCR, and none of them had any symptoms of COVID-19. Similar to the results of the present study, Lu et al. showed that there was no maternal to child vertical transmission in a 22-year-old pregnant female asymptomatically infected with SARS-CoV-2 in her late pregnancy (13). In the present study, neonatal pharyngeal swabs were taken for the SARS-CoV-2 RT-PCR test, and all tested negative. Hence, our study results are consistent with the above-reported studies. In contrast, some studies showed RT-PCR positive results in neonates born to COVID-19 positive mothers. Hu et al. conducted a study on seven RT-PCR confirmed COVID-19 positive pregnant females who were between the ages of 30 to 34 years. Six out of seven females experienced caesarean section while one had vaginal delivery. One female had a liver complication, but the other six had no complications during their pregnancy. They found that one infant out of seven was positive for SARS-CoV-2 infection (14). Alzamora et al. published a study on a 41-year gravida 3 para 2 pregnant woman female (with a history of previous caesarean delivery and diabetes mellitus) who had severe symptoms of COVID-19 and required mechanical ventilation on day 5 of disease onset. Caesarean delivery was conducted, and isolation of newborn was done immediately after birth, with no delay in cord clamping or any skin-to-skin contact. The nasopharyngeal swab specimen for SARS- CoV-2 was found positive 16 hr after delivery, but immunoglobulin M (IgM) and IgG for SARS-CoV-2 were negative. The mother’s IgM and IgG were found positive on day 4 postpartum (15). Nie et al. conducted research on 33 pregnant females who were positive for COVID-19. The perinatal transmission rate for COVID-19 in their study was 3.6% (16). Recently, two studies exhibited the possible risk of vertical transmission of SARS-CoV-2. Dong et al. and Zeng et al. with a combined total of seven COVID-19 infected pregnancies noted a virus-specific IgM antibody in neonatal blood tested immediately after birth, with a negative SARS-CoV-2 RT-PCR test result by nasopharyngeal swab. Cord blood and placental tissue sample testing was not done; thus, no direct evidence of infection was found. Furthermore, these antibodies (IgM) analyses are likely to provide false-positive results (17, 18). The possibility of postpartum neonatal infection cannot be completely excluded because of many factors such as the delay in testing and high viral load of severely symptomatic mothers. The present results showed no evidence of vertical transmission of SARS-CoV-2 in pregnant females suggesting the function of placenta as a barrier to this viral disease. There was no complication encountered during the delivery of any females in this study. Conflict of Interest Authors declare no conflict of interest. 144 148 Kalpana K Kumari Department of Obstetrics and Gynecology, Uttar Pradesh University of Medical Sciences Department of Obstetrics and Gynecology, Uttar Pradesh University of Medical Sciences India Ramakant R Yadav Department of Neurology, Uttar Pradesh University of Medical Sciences Department of Neurology, Uttar Pradesh University of Medical Sciences India rkyadav_2003@yahoo.com Sangh S Mittra Veerangana Avantibai Mahila Hospital Veerangana Avantibai Mahila Hospital India Arushi A Kumar Department of Community Medicine, Uttar Pradesh University of Medical Sciences Department of Community Medicine, Uttar Pradesh University of Medical Sciences India Jigyasa J Singh Department of Community Medicine, Uttar Pradesh University of Medical Sciences Department of Community Medicine, Uttar Pradesh University of Medical Sciences India Prashant PK Bajpai Department of Community Medicine, Uttar Pradesh University of Medical Sciences Department of Community Medicine, Uttar Pradesh University of Medical Sciences India Dhiraj DK Srivastava Department of Community Medicine, Uttar Pradesh University of Medical Sciences Department of Community Medicine, Uttar Pradesh University of Medical Sciences India Raj R Kumar Uttar Pradesh University of Medical Sciences Uttar Pradesh University of Medical Sciences India No Keyword 120099.pdf Sutton D, Bertozzi-Villa C, Lasky J, Fuchs K, Friedman A. 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