Infertile Group Research Articles

Women may not benefit from repeated frozen embryo transfers: A retrospective analysis of the Cumulative live birth rate of 43,972 women

Abstract STUDY QUESTION Which specific groups of women would not benefit from repeated frozen embryo transfers (FETs)? SUMMARY ANSWER Women over 45 years of age should stop treatment after three FET attempts due to the absence of further benefits, while women aged 40-45 years and those with a diminished ovarian reserve and other causes of infertility have a lower chance of improving their cumulative live birth rate (CLBR) within five FET cycles and experience fewer advantages from repeated transfers. WHAT IS KNOWN ALREADY In real-life scenarios of ART, women who fail to achieve a live birth often choose to undergo repeated FETs via the freeze-all strategy. STUDY DESIGN, SIZE, DURATION This retrospective study included 43,972 women who underwent 86,496 oocyte retrieval cycles and 82,022 FET cycles between January 2010 and March 2023 under the freeze-all strategy. PARTICIPANTS/MATERIALS, SETTING, METHODS We categorized the population based on the female’s age at the first oocyte pick-up (OPU) cycle (Groups 1-6: <30, 30-34, 35-39, 40-42, 43-44, and ≥45 years of age), number of retrieved oocytes at the first OPU cycle (Groups 1-5: 1-5, 6-10, 11-15, 16-20, and >20 oocytes), and causes of infertility (Groups 1-9: tubal factor, male factor, polycystic ovary syndrome, diminished ovarian reserve, endometriosis, other uterine factors, combined factors, unexplained infertility, and other infertility) to analyse their CLBRs within different FET cycles via Kaplan–Meier analysis (optimistic method) and the competing risk method (conservative method). We utilized multivariate Cox and Fine-Gray models to examine the associations between the CLBR and age, the number of retrieved oocytes, and nine causes of infertility. MAIN RESULTS AND THE ROLE OF CHANCE The CLBR decreased with increasing female age over five FET cycles (Groups 1-6: optimistic method: 96.4%, 94.2%, 86.0%, 50.2%, 23.1% and 10.1%; conservative method: 87.1%, 82.0%, 67.8%, 33.9%, 13.8% and 3.5%, respectively). Moreover, there was an increasing trend in the number of retrieved oocytes (Groups 1-5: optimistic method: 82.5%, 91.7%, 93.6%, 94.1%, and 96.2%; conservative method: 58.6%, 76.7%, 84.8%, 88.0%, and 92.5%, respectively). Furthermore, the CLBR varied across different causes of infertility (Groups 1-9: optimistic method: 91.7%, 93.1%, 96.6%, 79.2%, 89.9%, 76.1%, 90.0%, 92.9% and 35.4%; conservative method: 77.3%, 79.4%, 88.9%, 46.7%, 72.7%, 62.1%, 74.4%, 78.8% and 20.1%, respectively). LIMITATIONS, REASONS FOR CAUTION Calculating the actual CLBR for each person is difficult because some patients have remaining embryos that have not been transferred; additionally, the current statistical methodology uses both optimistic and conservative methods to calculate the CLBR, and in real life, the CLBR falls between the optimistic and conservative curves. WIDER IMPLICATIONS OF THE FINDINGS Our study is the first to identify specific subgroups of women who fail to benefit from repeated FETs and who require rational discontinuation of treatment following unsuccessful transfer. STUDY FUNDING/COMPETING INTEREST(S) This study was financially supported by grants from the National Natural Science Foundation of China (grant numbers: 82271732 to YP Kuang, 82071603 to L.W., 82001502 to Y. Liu., and 82201888 to X. Shen). The authors declare that they have no conflicts of interest in the present study. TRIAL REGISTRATION NUMBER N/A

The Study of Dermatoglyphic Patterns in Females with Primary Infertility: A Case-Control Study in the Maharashtra Population

Introduction: Dermatoglyphics is a widely known technique applied in genetic and medical research for the examination of finger, palm, and sole patterns. Dermatoglyphic characters are formed in the fetal stage and remain unaltered throughout their life. Some previous studies have indicated that dermatoglyphic patterns are linked with some reproductive disorders, like primary infertility. This study focuses on the evaluation and comparison of the dermatoglyphic patterns of females with primary infertility and healthy controls in the Maharashtra population. Methods: It involves a case-control study on two hundred females diagnosed with primary infertility and two hundred age-matched healthy controls. Several dermatoglyphic patterns were found, including total ridge count (TRC), atd angles, loops, whorls, and arches, which were assessed and compared among the groups. Results: When compared to the control group, the dermatoglyphic patterns of the infertility group were significantly different. Whorls and arches were more common in this group, and loops were less common. Furthermore, the infertility group exhibited a higher mean atd angle value and a lower total ridge count in comparison to the control group. Conclusion: Certain dermatoglyphic features correlated with female primary infertility in Maharashtra have been identified. These results can be taken as a precursor to further research work on the genetic basis of infertility and also with regard to whether dermatoglyphics can be used as a diagnostic tool.

The association between serum HDL levels and infertility among American women aged 20-44 years: A retrospective cross-sectional study of NHANES, 2013-2020.

Infertility is a significant national public health concern, and the World Health Organization (WHO) predicts that it will rank as the third most prevalent disease following tumors, cardiovascular and cerebrovascular diseases. The impact of dysfunctional lipoproteins on female infertility remains relatively understudied; therefore, the research focuses on exploring the relationship between serum high-density lipoprotein (HDL) concentration and infertility. This is a retrospective cross-sectional study where we employed multivariate logistic regression analysis to examine the association between serum HDL concentrations and female infertility. The strength of association was quantified using odds ratios (OR) along with their corresponding 95% confidence intervals and statistical significance was evaluated at a level of P < 0.05 (two-tailed). The study found that there was a significant correlation between serum HDL and infertility without adjusting the model (OR = 0.62, 95%CI 0.48-0.82, P<0.001). After adjusting for covariates, a weak correlation between HDL and infertility remained (OR = 0.70, 95%CI 0.49-1.00). When HDL concentrations were divided into quartiles, there was a trend of strengthened correlation between HDL and infertility risk with the increase in HDL concentrations. Specifically, individuals in the highest concentration quartile exhibited a 44.0% lower risk of infertility compared to those in the lowest concentration quartile (95% CI 0.38-0.84). In the age-stratified analysis, after adjusting for covariates, the correlation between HDL and infertility was statistically insignificant across all age groups. Furthermore, after categorizing HDL levels into quartiles, we observed a dose-dependent trend between HDL and the reduction of female infertility risk in the adjusted models of the secondary infertility group. Specifically, in the adjusted model, the high-concentration group exhibited a 67.0% lower risk of infertility compared to the low-concentration group (OR = 0.33; 95% CI: 0.12-0.940, P = 0.04). Our research findings suggest weak negative correlation between serum HDL and female infertility. However, upon stratified analysis by age, the correlation between HDL and infertility did not attain statistical significance. In cases of secondary infertility, a subtle dose-dependent trend was observed between serum HDL and infertility.

High levels of bisphenol A among infertile men can impair spermatogenesis by oxidative stress and elevated levels of microRNA-337

BackgroundStudies have shown that Bisphenol A may interfere with the process of spermatogenesis and result in a decrease in the quality of semen. Nevertheless, the fundamental processes remain unclear. This study was done to investigate the connections between exposure to Bisphenol A, spermatogenesis with microRNA-337, and malondialdehyde in infertile men.MethodsThis study was a case–control study on 73 participants. Infertile group (1a): azoospermia (n = 16), infertile group (1b): oligozoospermia (n = 22), and control group (2): normospermic (n = 35) were enrolled in this study. Full history, local examination, semen analysis, and urine and blood samples were taken from all participants. Urinary Bisphenol A, malondialdehyde, and serum microRNA-337 were measured.ResultsThe mean Bisphenol A level in azoospermia group shows statistically significant increase comparing to fertile control group. The mean microRNA-337 level in oligozoospermia and azoospermia group shows statistically significant increase comparing to fertile controls. The mean malondialdehyde level in infertility groups shows statistically significant increase comparing to fertile control group. No linear correlations were recorded between Bisphenol A levels with semen quality parameters, hormonal profile, and microRNA-337.ConclusionWhile there is no significant change in the levels of Bisphenol A between normal fertile males and infertile males with oligozospermia, a significant elevation in the BPA level was observed in infertile males with azoospermia. A significant upregulation of the miRNA-337 gene expression in infertile males either oligozospermia or azoospermia was also observed. In addition, lipid peroxidation as evident by the significant elevation of MDA levels was marked among infertile patients.

Investigation of endoplasmic reticulum stress-regulated chaperones as biomarkers in idiopathic nonobstructive azoospermia.

Azoospermia is a condition in which sperm cells are completely absent in a male's ejaculate. Typically, sperm production occurs in the testes and is regulated by a complex series of cellular and molecular interactions. Endoplasmic reticulum (ER) stress arises when there is a deviation from or damage to the normal functions of the ER within cells. In response to this stress, a cascade of response mechanisms is activated to regulate ER stress within cells. This study aims to investigate the role of ERstress-regulated chaperones as potential biomarkers in male infertility. ER stress associated with azoospermia can manifest in cells such as spermatogonia in the testes and can impact sperm production. As a result of ER stress, the expression and activity of a variety of proteins within cells can be altered. Among these proteins are chaperone proteins that regulate the ER stress response. The sample size was calculated to be a minimum of 36 patients ineach group. In this preliminary study, we measured and compared serum levels of protein disulfide-isomerase A1, protein disulfide-isomerase A3 (PDIA3), mesencephalic astrocyte-derived neurotrophic factor (MANF), glucose regulatory protein 78 (GRP78), clusterin (CLU), calreticulin (CRT), and calnexin (CNX) between male subjects with idiopathic nonobstructive azoospermia and a control group of noninfertile males. Serum PDIA1 (P=0.0004), MANF (P=0.018), PDIA3 (P<0.0001), GRP78 (P=0.0027), andCRT (P=0.0009) levels were higher in the infertile group compared to the control. In summary, this study presents novel findings in a cohort of male infertile patients, emphasizing the significance of incorporating diverse biomarkers. It underscores the promising role of ER stress-regulated proteins as potential serum indicators for male infertility. By elucidating the impact of ER stress on spermatogenic cells, the research illuminates the maintenance or disruption of cellular health. A deeper understanding of these results could open the door to novel treatment approaches for reproductive conditions, including azoospermia.

Common mental disorders and risk of female infertility: a two-sample Mendelian randomization study.

Female infertility is a global issue that impacts on public health seriously and many mental disorders are observed in infertility groups. To investigate the casual relationship between those, genome-wide association studies summary data of anxiety disorder (n=9,897), broad depression (n=322,580), major depressive disorder (n=480,359 and n=500,199), bipolar disorder (n=51,710), insomnia (n= 462,341), and female infertility (n=126,342) were extracted from the existing datasets and was analyzed through the two-sample mendelian randomization study. The following heterogeneity and sensitivity test were applied to ensure the robustness of results. Based on inverse variance weighted results, major depressive disorder was associated with female infertility (P = 0.0001, odds ratio 1.396, 95 % confidence interval 1.175-1.658). No causal relationship was identified between the other four mental disorders and infertility. was found. Additionally, reverse mendelian randomization did not indicate a causal relationship among these disorders. The early identification and management of anxiety symptoms in women of reproductive age, in conjunction with the effective treatment of major depressive disorder, may be crucial for preserving female fertility.

Estimation Of Serum Ionized Calcium In Relation With Anti Mullerian Hormone In Women With Infertility

This study has discussed how the condition of calcium equilibrium associated with AMH levels affects the ability to reproduce in some pre-menopausal women. Forty-five females (20-35 age) selected with pre-infertility from infertility unit centers in AL-Saader Medical City from January 2023 to October 2023; also, the study involved group consisted of 45 health volunteers. They were nearly similar in age. The findings showed that the mean serum ions of calcium levels (4.25 ± 0.72 mg/dl) were considerably (p&lt;0.001) less in the infertility group than the non-infertile group (5.33 ± 0.31), accompanied by a statistical increase in PTH in patients (98.23±33.5) than in control(94.6± 28.2). Also, there is a significant decrease in AMH in the patient (2.35±1.57) than the control (4.53±0.95). Our findings indicated that the amount of ionized calcium might have to be balanced in some way for every physiological function, including reproduction.

Relationship Between Anti-Mullerian Hormone (AMH) and Other Influencing Factors with in Vitro Fertilization (IVF) Outcome of Iraqi Infertile Women

The clinical application of Anti-Mullerian Hormone (AMH) measurement has been proposed in the prediction of qualitative and quantitative aspects of assisted reproductive technologies (ART). The aim of this study is to investigate the role of AMH level and other influencing factors in the outcomes of in vitro fertilization (IVF) treatment. The study focused on AMH and its association with infertile women undergoing IVF. The population included in this study was 131 infertile women who sought therapy for infertility at Kamal Al-Samarra'ay Specialized Hospital. They were divided into three groups according to the level of AMH hormone; group A (0.5-1.5 ng/mL, n=52), group B (1.6-4 ng/mL, n=70), and group C (&gt; 4 ng/mL, n=9). Data collection was performed by means of a specially designed questionnaire, independent of demographic data. It included questions related to infertility. Statistical analysis was performed with SPSS 26. Clinical parameters of infertile women, infertility hormones and their relationship with AMH, as well as FMS-like Tyrosine kinase 1 levels, were all investigated. The study findings revealed a significant difference in the concentration of AMH between infertile women (p&lt;0.01). Our analysis further demonstrated that the level of AMH reduces with advancing age in infertile women. Additionally, a positive correlation was observed between the number of oocytes obtained and AMH levels in infertile women. Furthermore, the study found that AMH levels had an effect on clinical pregnancy outcomes. Specifically, the highest success rates for IVF were observed in women with normal AMH levels falling within the ranges of group B (6-4 ng/mL(. Additionally, the study demonstrated significant differences in AMH and TSH levels among infertile groups (p&lt;0.05). Our study concluded that there is a significant correlation between clinical parameters and IVF outcomes, which can provide information for future clinical interventions aimed at improving success rates. Our analysis highlighted the role of FMS-like Tyrosine kinase 1 in infertility, where higher FMS-like Tyrosine kinase 1 levels were associated with higher success rates in IVF. The results indicate that the FMS-like Tyrosine kinase 1 level is a valuable marker for confirming the success or failure of IVF and can help determine the appropriate treatment plan.

Pre- and Procedural Factors Influencing the Success of In Vitro Fertilization: Evaluating Embryo Quality and Clinical Pregnancy in Cases of Tubal Factor Infertility.

Introduction: While tubal occlusion is a prevalent cause of infertility, accounting for 11-35% of infertility cases among women, there remains a limited understanding of the factors influencing clinical pregnancy following in vitro fertilization (IVF). Methods: In our retrospective, cross-sectional cohort study conducted at a single tertiary center, medical records of women aged 19 to 43 years were analyzed. Logistic regression models were employed to identify the prognostic factors associated with clinical pregnancy after IVF in patients with tubal factor infertility, excluding cases with hydrosalpinx. Results: Data from 219 women diagnosed with tubal occlusion were compared to 1140 cases with non-tubal indication, covering a total of 1359 IVF cycles. A lower maternal age (adjusted odds ratio [AOR]: 0.89, p = 0.001) and a higher embryo quality (AOR: 1.26, p = 0.01) emerged as important factors in clinical pregnancy in the tubal infertility group. Moreover, a lower maternal (AOR:0.91, p < 0.01) and paternal age (p = 0.001), and favorable semen quality (AOR: 1.32, p = 0.03) were critical determinants in the non-tubal infertility group. BMI was generally higher in tubal infertility patients (p = 0.01). Furthermore, FSH level (AOR: 0.93, p = 0.004), AMH level (p < 0.04), number of embryos transferred (AOR: 2.04, p < 0.001), and embryo quality (AOR: 1.26, p < 0.001) came into prominence only in the non-tubal infertility group. The clinical pregnancy rate (34.2%) of women with tubal occlusion did not differ significantly from those in other forms of infertility undergoing IVF (35.4%). Conclusions: Although tubal infertility is typically anticipated to yield the highest clinical pregnancy rates following IVF, it is crucial to acknowledge that both maternal and paternal characteristics can also significantly impact the outcomes.

Fertility status and depression: A case-control study among women in Herat, Afghanistan.

On a global scale, an estimated 17.5% of the reproductive-aged population experiences clinical infertility. Beyond its categorization as a reproductive health concern, infertility emerges as a substantial independent risk factor for the development of various mental health disorders. The current study aims to investigate the prevalence of depression among women experiencing infertility compared to fertile women in Herat city, Afghanistan. This hospital-based case-control study was conducted in Herat city, Afghanistan, during the latter half of 2023 on women aged 15-49. Sociodemographic and infertility data were gathered by a gynecologist using a structured questionnaire with 14 items. Assessment of depression employed the Dari-translated version of the Patient Health Questionnaire. The data were scored and categorized following the instrument's guidelines. To examine the association between categorical variables, a chi-square test was conducted, with a significance level set at 0.05 for all analyses. These statistical procedures were performed using IBM Statistical Package for Social Sciences, version 27. Our investigation revealed a statistically significant association (p < 0.001) between infertility and depression. Infertility was associated with a markedly increased risk of depression, as evidenced by a prevalence of 79.0% in infertile women compared to 44.4% in fertile women. The study found a link between depression, health, and nutritional status in both infertile and fertile participants. Notably, within the infertile group, the longer a woman struggled with infertility, the more likely she was to experience depression. Given the substantial prevalence and important correlates of depression among infertile women in Herat, Afghanistan, it is imperative for policymakers, mental health professionals, and gynecologists to specifically address the mental well-being of this vulnerable population.

Do aluminum, boron, arsenic, cadmium, lipoperoxidation, and genetic polymorphism determine male fertility?

Male infertility is a world multifactorial problem modulated by environmental and genetic factors. Male aspects account for 20–50 % of infertility cases. Our results are unique because they treat the importance of components participating in the determination of male infertility (environmental and immunogenetic determinants, seminological analysis, lipoperoxidation, genetic determinants, role of aluminum, arsenic, cadmium and boron). We analyzed agents affecting male reproductive potential (aluminum, boron, cadmium, arsenic, lipid peroxidation, gene polymorphisms (MTHFRv.C677T (rs1801133) (chromosome-1) and IL-4v.C589T (rs2243250) (chromosome-5) in men with semen disorders (n=76) and with normozoospermia (n=87) from Central Poland. Polymorphisms of MTHFRv.C677T and IL-4v.C589T genes indirectly shape toxic metals concentration and lipoperoxidation but do not exert direct influence on male fertility disorders (monomorphism and lack of differences in genotypes frequency). Men with genotype TT or CC (IL-4v.C589T) show some differentiation in elements concentration and intensity of lipoperoxidation. Analysis of TT or CC (IL-4v.C589T) genotype brought correlations with B, Al, Cd, and lipoperoxidation (P<0.05) and suggesting that mentioned factors jointly shape male reproductive capability. Toxic metals may play an important role in shaping of men genetic polymorphisms, since Cd was identified as a factor increasing risk of qualification to infertile group, predisposing to fertility disorders. B, Al and Cd may be considered as important modulators of reproductive condition. However, lipoperoxidation as an isolated predictive parameter does not produce convincing results in male reproductive potential (higher MDA concentration in healthy men). Our results may be helpful in the diagnosis of male infertility, in the reduction of idiopathic cases of unknown origin and in implementation of targeted and more effective treatment (pharmacological, hormonal). Identification of environmental stressors and their correlations with fertility disorders can help to eliminate or reduce the impact of factors unfavorable to fertility. Our results highlight the importance of environmental and immunogenetic factors in shaping of defensive potential against destruction of spermatozoa and infer a role of oxidative stress in the induction of gene polymorphisms, affecting male fertility.

Analysis of genital chlamydia and gonococcal infections in infertility

To analyze the infection of chlamydia (CT) and gonorrhea (NG) in female infertility and male infertility population, and to explore the correlation between CT and NG infection and infertility. A case-control study was conducted to retrospectively analyze the specimens submitted by patients from the Third Xiangya Hospital of Central South University from January 2021 to December 2022. The results showed that a total of 32 184 specimens were collected, and the positive rates of CT were 4.41% (1 419/32 184), and positive rats of NG were 1.42% (457/32 184). In the infertility group (n=3 366), 2 987 were females and 379 were males. In the control group (n=3 366), 2 509 were females and 857 were males. The CT positive rate of the infertility group was 13.61% (458/3 366), which was significantly higher than that of the control group 3.30% (111/3 366), and the difference was statistically significant (χ2=4.245, P<0.05), and the NG positive rate of the infertility group was 6.36% (214/3 366), which was significantly higher than that of the control group 0.89% (30/3 366), and the difference was statistically significant (χ2=4.011, P<0.05). A total of 23 992 female genital tract swab specimens were collected, including 2 987 in the infertility group and 2 509 in the control group, and the positive rate of CT in the female infertility subgroup was 10.41% (311/2 987), which was significantly higher than that in the control group 3.75% (94/2 509), the difference was statistically significant (χ2=4.132, P<0.05), and the NG positive rate of 8.73% (261/2 987) in the female infertility subgroup was significantly higher than that in the control group 0.40% (10/2 509), and the difference was statistically significant (χ2=4.242, P<0.05). A total of 8 192 male urine samples were collected, including 379 in the infertility group and 857 in the control group, and the CT positive rate of the male infertility subgroup was 13.72% (52/379), which was significantly higher than that of the control group 3.38% (29/857), and the difference was statistically significant (χ2=5.267, P<0.05), and the positive rate of NG in the male infertility subgroup was 12.66% (48/379), which was significantly higher than that of the control group 0.93% (8/857), and the difference was statistically significant (χ2=4.166, P<0.05). Among the 2 987 female specimens in the infertility group, 1 034 were in the primary infertility subgroup and 1 953 were in the secondary infertility subgroup, and the positive rates of CT were 7.93% (82/1 034) and 15.72% (307/1 953), respectively, and the positive rates of NG were 3.87% (40/1 034) and 8.65% (169/1 953) respectively, and the difference was not statistically significant (χ2=0.185, P>0.05) and (χ2=0.002, P>0.05). In conclusion, the infection rate of genital tract CT and NG is high in the infertility population, CT and NG are recommended as routine examination indicators for eugenics and infertility screening.

Beyond the Incision: A Comparative Study of Suprainguinal and Inguinal Varicocele Surgeries.

Background Varicocele, characterised by the abnormal dilation of the pampiniform plexus of scrotal veins, is a prevalent and treatable cause contributing to male infertility, affecting 40% of men experiencing primary infertility and 80% of those with secondary infertility. Often asymptomatic, it can lead to chronic scrotal pain or a feeling of heaviness. Surgical interventions such as open, laparoscopic, or microsurgical varicocelectomy aim to eliminate venous reflux into the scrotum. This study sought to assess and evaluate the surgical outcomes and postoperative complications of the suprainguinal and inguinal approaches to varicocelectomy, offering evidence-based insights to improve varicocele management. Methodology A total of 60 males diagnosed with grade II or III unilateral or bilateral varicocele were included in the study. Patients with subclinical or grade I varicocele, recurrent varicocele, or concomitant inguinoscrotal pathology (hydrocele, epididymal cyst, and inguinal hernia) were excluded. Participants were randomly assigned to one of two groups based on the surgical technique: Group A (suprainguinal approach) and Group B (inguinal approach). Surgical outcomes were assessed by evaluating operating time, postoperative pain, wound hematoma, wound infection, hydrocele development, testicular atrophy, and semen analysis, both pre- and postoperatively. Results The study included 60 patients with a mean age of 29.05±5.96 years and an age range of 18-40 years. The suprainguinal approach offers a significantly shorter operating time than the inguinal approach (33.1 vs. 40.8 minutes). Both surgical techniques resulted in similar postoperative pain levels. The incidence of complications, such as wound hematoma, wound infection, and hydrocele development, showed no significant differences between the two approaches. In Group A (suprainguinal approach), the rates were 3.3%, 6.6%, and 3.3%, respectively, while in Group B (inguinal approach), they were 6.6%, 13.3%, and 6.6%. Additionally, 75% of patients in the infertility group showed improvements in semen parameters, with 80% in Group A and 71.4% in Group B, with no significant difference between the surgical approaches. Conclusion The suprainguinal and inguinal approaches to varicocelectomy effectively manage varicoceles, with the suprainguinal approach offering a shorter operating time. Postoperative complications and improvements in semen parameters were comparable between the two methods.

Male infertility is associated with differential DNA methylation signatures of the imprinted gene GNAS and the non-imprinted gene CEP41.

To investigate whether the DNA methylation profiles of GNAS(20q13.32), MEST(7q32.2), MESTIT1(7q32.2), IGF2(11p15.5), H19 (7q32.2), and CEP41(7q32.2) genes are related to the transcriptomic and epigenomic etiology of male infertility. The DNA methylation levels of spermatozoa were obtained from fertile (n = 30), oligozoospermic (n = 30), and men with normal sperm count (n = 30). The methylation status of each CpG site was categorized as hypermethylated or hypomethylated. Expression levels of target gene transcripts were determined using real-time PCR. The oligozoospermia showed a higher frequency of hypermethylation at GNASAS 1st, 3rd, and 5th CpG dinucleotides (66.7%, 73.3%, 73.3%) compared to the fertile group (33.3%, 33.3%, 40%, respectively). The normal sperm count exhibited a higher frequency of hypermethylation at the 3rd CpG of CEP41 (46.7%) than the fertile group (16.7%). Normal sperm count was predicted by CEP41 hypermethylation (OR = 1.750, 95%CI 1.038-2.950) and hypermethylation of both CEP41 and GNASAS (OR = 2.389, 95%CI 1.137-5.021). Oligozoospermia was predicted solely by GNASAS hypermethylation (OR = 2.460, 95%CI 1.315-4.603). In sperms with decreased IGF2 expression in the fertile group, we observed hypomethylation in the 2nd CpG of IGF2 antisense (IFG2AS), and hypermethylation in the 1st, 2nd, and 4th CpGs of H19. No significant relationship was found between IGF2 expression and methylation status of IGF2AS and H19 in infertile groups. The disappearance of the relationship between IGF2 expression and IGF2AS and H19 methylations in the infertile group provides new information regarding the disruption of epigenetic programming during spermatogenesis. A better understanding of sperm GNASAS and CEP41 hypermethylation could advance innovative diagnostic markers for male infertility.

An insight into the vaginal microbiome of infertile women in Bangladesh using metagenomic approach.

The dysbiosis of vaginal microbiota is recognized as a potential underlying factor contributing to infertility in women. This study aimed to compare the vaginal microbiomes of infertile and fertile women to investigate their relationship with infertility. Metagenomic analysis was conducted on samples from 5 infertile and 5 fertile individuals using both amplicon 16S and metagenomics shotgun sequencing methods. In the infertile group, the bacterial community was primarily represented by three major bacterial genera: Lactobacillus (79.42%), Gardnerella (12.56%) and Prevotella (3.33%), whereas, the fertile group exhibited a more diverse composition with over 8 major bacterial genera, accompanied by significantly reduced abundance of Lactobacillus (48.79%) and Gardnerella (6.98%). At the species level, higher abundances of L. iners, L. gasseri and G. vaginalis were observed in the infertile group. Regarding the microbiome composition, only one fertile and two infertile subjects exhibited the healthiest Community State Types, CST-1, while CST-3 was observed among two infertile and one fertile subject, and CST-4 in three other fertile and one infertile subject. Overall, alpha diversity metrics indicated greater diversity and lower species richness in the control (fertile) group, while the infertile group displayed the opposite trend. However, beta-diversity analysis did not show distinct clustering of samples associated with any specific group; instead, it demonstrated CST-type specific clustering. Shotgun metagenomics further confirmed the dominance of Firmicutes, with a greater abundance of Lactobacillus species in the infertile group. Specifically, L. iners and G. vaginalis were identified as the most dominant and highly abundant in the infertile group. Fungi were only identified in the control group, dominated by Penicillium citrinum (62.5%). Metagenome-assembled genomes (MAGs) corroborated read-based taxonomic profiling, with the taxon L. johnsonii identified exclusively in disease samples. MAG identities shared by both groups include Shamonda orthobunyavirus, L. crispatus, Human endogenous retrovirus K113, L. iners, and G. vaginalis. Interestingly, the healthy microbiomes sequenced in this study contained two clusters, Penicillium and Staphylococcus haemolyticus, not found in the public dataset. In conclusion, this study suggests that lower species diversity with a higher abundance of L. iners, L. gasseri and G. vaginalis, may contribute to female infertility in our study datasets. However, larger sample sizes are necessary to further evaluate such association.

O-016 To be or not to be capacitated after sperm preparation: tyrosine phosphorylation as predictive indicator for capacitation status and male fertility potential

Abstract Study question Can post-capacitation tyrosine phosphorylation (TP) levels distinguish “capacitated” from “non-capacitated” molecular profiles, and are these similarly distributed among infertile and fertile men? Summary answer TP levels after 3 hours of incubation post-capacitation robustly predicted “capacitated” and “non-capacitated” molecular profiles, with the latter being more common in the infertile group. What is known already TP is considered one of the most relevant intracellular signalling events in the capacitation process and in the regulation of sperm function. Despite the recognized general trend of increased TP post-capacitation, interindividual variability can be observed, with some samples showing phosphorylation levels like those before capacitation. Currently, no test is conducted on capacitated samples to verify whether they have indeed acquired a molecular pattern consistent with capacitation. Therefore, the objective of this study was to evaluate the predictive ability of post-capacitation TP levels measured at various time points in distinguishing molecularly “capacitated” patterns from “non-capacitated” ones. Study design, size, duration Prospective experimental study conducted from June 2021 to September 2023 with the aim of assessing TP levels in different populations before (B-cap) and after (A-cap) in vitro capacitation and after different incubation times (1h and 3h) under capacitating conditions. A total of 335 samples, including infertile patients (n = 111), tests for the inclusion in the sperm donation program (n = 57) and sperm donors (n = 167) were included. Participants/materials, setting, methods Semen samples were examined following WHO guidelines and in vitro capacitation was performed using density gradient centrifugation. Flow cytometry was used to assess global TP (%) (B-cap, A-cap, 1h and 3h). Receiver operating characteristic (ROC) curve analyses were performed to evaluate the discriminatory potential of post-capacitation TP levels (A-cap, 1h and 3h) against B-cap, aiming to distinguish between “capacitated” and “non-capacitated” molecular profiles. Chi-square test was used to compare proportions. P &amp;lt; 0.05 was considered statistically significant. Main results and the role of chance The dataset was randomly split into two subsets, one for model development and the other for subsequent performance evaluation. The areas under the curve (AUC) for A-cap, 1h and 3h were 73.8% (95% CI = 69.3%-78.3%), 84.5% (81.0%-88.0%), and 92.7% (90.4%-95.0%), respectively. Sensitivity and specificity values were calculated for the curve with the highest AUC, constructed with TP values from non-capacitated (B-cap) and 3-hours incubated (3h) samples. The selected cut-off point was 4.35% (sensitivity: 85.0%, specificity: 85.7%). The model exhibited robust overall performance in identifying molecularly “capacitated” and “non-capacitated” samples, with a sensitivity, specificity, false positive rate and false negative rate of 85.2%, 89.7%, 9.4% and 12.9%, respectively. Accuracy, positive predictive value and negative predictive value were 87.3%, 90.2%, and 84.5%, respectively. In infertile patients, using TP values at the third hour of incubation and the selected cut-off, the model identified 73.9% (82/111) of samples as molecularly “capacitated” and 26.1% (29/111) as “non-capacitated,” despite laboratory capacitation. In contrast, in the control group, the model identified 93.4% (156/167) of samples as “capacitated” and 6.6% (11/167) as “non-capacitated”. A significantly higher number of seminal samples were identified as “non-capacitated” in the infertile patient group compared to the proven fertile donor group (p &amp;lt; 0.001). Limitations, reasons for caution The model's performance demonstrates its utility and reliability in identifying samples with a molecular “non-capacitated” profile, even after in vitro capacitation. However, further refinement is possible by incorporating additional data. External validation is essential to confirm generalizability of the results. Wider implications of the findings The assumption that a molecular capacitation profile is acquired following in vitro sperm processing may be questioned. Our results suggest a potential link between the selected threshold and fertility, offering insights into male infertility aetiology and paving the way for personalized intervention strategies. Trial registration number NCT04962100

O-305 Mapping the entire functionally active seminal microbiota and its association with semen quality parameters

Abstract Study question Does semen harbour functionally active microorganisms, and whether the microbial composition associate with semen quality parameters? Summary answer Seminal fluid harbours functionally alive microorganisms including bacteria, viruses, archaea, viroids and fungi whose composition and metabolic functions associate with semen quality parameters. What is known already Seminal fluid has been shown to possess different microbes, where different bacteria have been associated with spermatogenesis, seminal parameters and infertility. Nevertheless, previous results are inconclusive and the core microorganismal composition in semen is not determined and whether the identified bacterial DNA sequences refer to alive/functionally active microbes is not clear. Further, there is limited knowledge of the potential host-microbe interactions. Study design, size, duration In total 78 men with age between 18 and 45 years donated semen for the study, 59 men with good quality semen parameters and 19 men with low semen quality and diagnosed asthenospermia, oligoasthenozoospermia or oligoasthenoteratozoospermia comprised control and case groups, respectively. Participants/materials, setting, methods Semen quality parameters were analysed following WHO, 2021 guidelines. Seminal fluid underwent NextSeq total RNA sequencing, separating human and non-human sequences with Hisat2 aligner. Taxonomic classification of microorganisms was obtained using Kraken2. Microbial species and human RNA-Seq analyses, comparing control and infertility groups, utilized metagenomeSeq and edgeR R packages. Human and microbial pathways were annotated using MetaCyc, followed by GSEA analysis. The integration of metabolic pathways between host and microbes was investigated. Main results and the role of chance With our total RNAseq analysis, we mapped the entire alive microbiota composing of 6453 microorganisms within the seminal fluid samples. Microbes such as bacteria, fungi, viruses, viroids and archea were identified. When analysing microbiota associations with seminal parameters, 404 microbial species (Lactobacillus, Gardnerella, Prevotella, Atopobium, Pseudomonas among others) were significantly differently present among infertile men vs. good quality semen group. The host transcriptomic analysis identified 2569 differentially expressed genes in infertile vs. good quality semen groups, involved in sperm differentiation, spermatid development, reproduction, cilium movement and meiosis. Metabolic pathway analysis detected possible metabolic activity in the host-microbiota crosstalk in FAO-PWY: fatty acid &amp; beta-oxidation, LIPASYN-PWY: phospholipases, PWY66-429: fatty acid biosynthesis, PWY-3781: aerobic respiration I pathways. Limitations, reasons for caution Bigger sample size of infertile men would be needed to validate the study findings. Wider implications of the findings We confirm the presence of active microbes in semen with implications in seminal quality parameters such as motility, sperm count and morphology. Our results contribute to better understanding of seminal microbiota dysregulation in infertility with broader implications for reproductive health, suggesting functional links between seminal microbiota and sperm parameters. Trial registration number NA

P-319 Chronic endometritis prevalence, associated factors and pregnancy outcome among infertile women

Abstract Study question What are the prevalence, associated factors and pregnancy outcome of chronic endometritis(CE) among infertile women? Summary answer The overal prevalence of chronic endometritis in infertile women was 24.7%. Diagnosis rate of CE was higher in primary infertility compared to other infertility groups. What is known already Chronic endometritis is defined as a chronic inflammation of uterine endometrium. The prevalence of chronic endometritis varies across different groups and diagnostic criteria. The pathophysiology of chronic endometritis remained unclear, therefore, associated factors of chronic endometritis are uncertain. Study design, size, duration We perfomed an single-center, retrospective study between May 2019 and July 2023. By doing so, we collected data on 486 infertile women. Participants/materials, setting, methods A total of 486 infertile women, who underwent hysteroscopy by single surgeon, were recruited. The diagnosis of chronic endometritis was confirmed by presence of plasma cell per section in hematoxylin-eosin stain or CD138 immunohistochemical staining. Preoperative information (including age, body mass index, smoking, alcohol consumption, clinical presentation of infertility, endometrial thickness at ovulatory phase, anti-mullerian hormone (AMH) level and infertility factors) and post-operative information (including endometrial histology and pregnancy outcome) were collected. Main results and the role of chance The prevalence of chronic endometritis in the 486 infertile women with hysteroscopy guided endometrial biopsy was 24.7%. Prevalence of chronic endometritis was 28.1%, 19%, 22.2%, 27.2% in women with primary infertility, secondary infertility, recurrent pregnancy loss (RPL) and repeated implantation failure (RIF), respectively. A multivariate analysis revealed that overweight was associated with lower risk of chronic endometritis than underweight (relative risk (RR) = 0.56, 95% confidence interval (CI): 0.32-0.98, p = 0.042). Primary infertility was associated with higher risk of chronic endometritis than non-primary infertility (RR = 1.44, 95% CI: 1.02-2.03, p = 0.039). The pregnancy rate (including spontaneous pregnancies) was comparable between women with treated chronic endometritis and without chronic endometritis; 54.7% versus 61.7% in clinical pregnancy rate and 82.8% vs 85.4% in ongoing pregnancy rate. The prevalence of chronic endometritis with coexisting endometrial lesion (endometrial polyps, submucous myoma, or both of them) was 61.7%. Limitations, reasons for caution Our retrospective study has a less powerful design compared to a prospective randomized controlled study. Wider implications of the findings In women with primary infertility, the diagnosis rate of chronic endometritis was higher compared to other infertility groups. Therefore, even in cases of primary infertility, chronic endometritis should be considered as a potential cause of infertility. Trial registration number not applicable

P-094 Genetic variations of antioxidant genes may coordinate with lifestyle to define male fertility: A cross-sectional study with control group

Abstract Study question Are polymorphisms of the oxidative stress-related genes different between infertile men and reproductively healthy men? Does lifestyle contribute to fertility outcomes? Summary answer A strong association between the polymorphisms of two antioxidant genes and sperm quality was identified. The infertile men had higher BMI and alcohol usage. What is known already Antioxidant genes including catalase (CAT), superoxide dismutase (SOD), and nitric oxide synthase (NOS) play a critical role in spermatogenesis and sperm functions, especially in stress-response cell signaling pathways. Polymorphisms of antioxidant genes have been reported to be associated with gene function, hence sperm quality, ultimately affecting male fertility in various cohorts. Body mass index (BMI) is universally considered a marker of health reflecting whole health, nutrition, and lifestyle. Smoking and alcohol consumption expose the body to harmful chemicals and oxidative stress. Here, we examined the polymorphisms of antioxidant genes in Vietnamese men and the link with lifestyle to regulate fertility. Study design, size, duration This cross-sectional study with control group recruited from Hanoi Medical University Hospital from 2018-2020. The participants were divided into two groups regarding semen analysis based on WHO 2010 criteria. The participants gave consent, then were clinically examined, interviewed for lifestyle, and genetically analyzed for antioxidant gene variations. An extra inclusion criterion for the control men was having at least one healthy baby. The exclusion criteria were chromosome aberrations and Y chromosome microdeletion. Participants/materials, setting, methods The control group (N = 85) and infertile group (N = 107) were recruited. Semen analysis was carried out by the CASA instrument based on WHO 2010 criteria. Lifestyle characteristics including exercise (per week), smoking (yes/no), alcohol consumption (the threshold for “yes” was five standard drinks/occasion, at least one occasion/month) were recorded. Four variants SOD1:7958G&amp;gt;A, SOD2:c.47T&amp;gt;C, CAT:-262C&amp;gt;T, and NOS3:-786C&amp;gt;T were analyzed by Sanger sequencing. Pearson’s, Chi-squared (χ2), Wilcoxon rank-sum, ANOVA test, and Bonferroni correction were applied where specifically required. Main results and the role of chance The average age of the two groups was not different (31.96 ± 4.87 vs 31.93 ± 6.3 years old, p = 0.92). The body mass index (BMI) of infertile men was significantly higher than the control (24.84 ± 2.31 vs 23.53 ± 2.55 kg/m2, p = 0.00026). No significant difference was identified regarding exercise and smoking. Notably, 96.26% of infertile men consumed alcohol in comparison with 88.23% of the control (p = 0,034). Genetically, SOD1:7958GA genotype and NOS3:-786CT genotype in the infertility group were significantly higher than in the control, OR = 2.191 (95% CI: 1.226–3.915, p = 0.004) and OR = 3.135 (95% CI: 1.591–6.180, p &amp;lt; 0.001), respectively. The frequency of the SOD2:c.47TC genotype was significantly higher in the male infertility group than in the control (OR = 1.941, 95% CI: 1.063–3.595, p = 0.029). Individuals carrying the SOD1:GA+AA, SOD2:TC+CC, and CAT:CT/TT genotype pattern had an increased risk of male infertility (OR = 7.614, p = 0.007), suggesting that gene-gene interaction plays an important role in male reproduction. To our knowledge, this is the first report worldwide on the association between the SOD1:7958G&amp;gt;A polymorphism and male infertility. Together, the data suggest that antioxidant gene polymorphisms and lifestyle, especially alcohol usage, coordinate to regulate male fertility. Limitations, reasons for caution Direct measurement of oxidative stress in the semen of men having different gene variations would be required to ascertain the link between the polymorphisms of antioxidant genes and fertility. The sample size was relatively small, a larger sample size would be required to accurately represent the Vietnamese population. Wider implications of the findings The findings can assist in infertility diagnosis focusing on the group of higher infertile risk genetic variations. Interventions such as antioxidant supplements would be developed and applied, lifestyle advice would be given specifically for a group having high-risk gene variations to accurately target fertility issues. Trial registration number Not applicable

O-187 Real-word data on effectiveness of ICSI over conventional IVF according to infertility factors based on human fertilisation and embryology authority dataset

Abstract Study question Can the analysis of Human Fertilisation and Embryology Authority (HFEA) dataset provide real-word data on effectiveness of ICSI over conventional IVF according to infertility factors? Summary answer A statistically significant reduction in live birth rate was shown to be associated with ICSI use in cycles with female infertility factors. What is known already The adoption of ICSI has seen a considerable increase over the years, even in cases without an evident male-factor infertility. Available studies including large retrospective studies and meta-analyses evaluating the effectiveness of the two strategies tend to support the idea that in the absence of a male factor, ICSI does not offer a significant advantage over conventional IVF(cIVF). However, some uncertainties still exist regarding live birth rates and in relation to various infertility factors. Study design, size, duration Retrospective study of 275.825 first cycles treated with cIVF or ICSI between 2005-2018, included in the HFEA database. Infertility indications were simplified into three groups: 1) female only (endometriosis, tubal, ovulatory and cervical factors); 2) male (male only or with male and female factors together); 3) unexplained infertility. The primary outcome was live birth rate in the first fresh cycle. Secondary outcomes included fertilization, implantation, miscarriage rates, cycle cancellation rate and neonatal outcomes. Participants/materials, setting, methods Outcomes were evaluated through binomial logistic regression including the following predictors: female age, inseminated oocytes, number/stage of transferred embryos (condensed with principal component analysis), subtype of female infertility, year of treatment. A paired-analysis was also performed matching couples in a 1:1 ratio between ICSI and conventional IVF based on female age, indication, inseminated oocytes, year of treatment. Results were reported according to infertility cause using crude and adjusted Odds Ratios (OR) with 95%CI. Main results and the role of chance The study revealed a consistent balance in treatment distribution between conventional IVF and ICSI, with a slight increase in ICSI cycles in recent years. Based on logistic regression analysis, a significant reduction in live birth rate was demonstrated using ICSI compared to conventional IVF in cycles with female factors (adjusted OR = 0.95, 95%CI: 0.91-0.99). No significant differences emerged in unexplained factor cycles. With the use of conventional IVF, the overall odds of having a total fertilization failure increased by a factor of 1.84 (95%CI: 1.75-1.94). No significant differences in implantation rates between conventional VF and ICSI were seen after adjustment. The OR for male sex in newborns compared to female sex from single pregnancies was significantly influenced by ICSI use across the three groups of infertility causes and ranged between 0.84 and 0.87. The matched dataset confirmed a reduced occurrence of live birth per cycle in case of female factors (aOR=0.91, 95%CI: 0.86-0.96) and an overall significant lower proportion of cycles ending in cancellation for absence of viable embryos using ICSI compared to conventional IVF. Limitations, reasons for caution Some biases are intrinsic in the availability and quality of retrospective data obtained from registries and unmeasured confounders may influence the generalizability of the results. The study does not provide information on cumulative live birth rates. Wider implications of the findings ICSI introduces an additional, invasive step in assisted reproduction with a potential reduction in live birth rates for specific patients’ groups. These findings emphasize the importance of tailoring treatments to specific patient profile.The routine use of ICSI for all cases should not be considered an appropriate clinical practice. Trial registration number not applicable