Normal Semen Parameters Research Articles

Comparative analysis of short versus long co-incubation of gametes on post-insemination outcomes and embryo morphokinetics: a sibling oocyte randomized study

Research questionDoes a short co-incubation of gametes in conventional IVF impact post-insemination outcomes and embryo morphokinetics? DesignSibling oocyte randomized pilot study performed between December 2020 and March 2023. Eligible couples (n=55) were female aged 18-43 years with body mass index (BMI) ≤35 and male normal semen parameters. Cumulus oocyte complexes (COCs) (min 6 - max 12) were randomized in a 1:1:2 proportion in long (16-18 hrs), or short (2 hrs) co-incubation IVF exposure, and ICSI, respectively. All oocytes inseminated via IVF were monitored by time-lapse (TL). Blastocysts of ≥BL3CC were subjected to trophectoderm (TE) biopsy on day (D)5, D6 or D7 by next generation sequencing (NGS). ResultsA total of 954 COCs were distributed to long co-incubation (n=235, 24.6%), short co-incubation (n=235, 24.6%), and ICSI (n=484, 50.3%). In total, 202 were MII after long co-incubation, 209 after short co-incubation and 394 at ICSI. Regression analyses showed both short co-incubation groups (OR: 0.72, 95% CI: 0.46–1.11, P=0.139) and ICSI (OR: 1.43, 05% CI: 0.95–2.15, P=0.090) did not significantly impact normal fertilization rates (2PN) when compared to long co-incubation. Usable blastocysts per MII, per 2PN, and euploid blastocysts per MII, per 2PN and per tested blastocysts were similar among the groups. The rate of embryo arrest (HR: HR: 0.79, 95% CI: 0.57-1.11, P=0.183) and timings to reach morphokinetic milestones among embryos reaching blastulation were similar between IVF groups (P>0.05, for all). ConclusionA short 2-hour co-incubation of gametes in IVF yields comparable fertilization, blastocyst, and euploidy rates without adverse embryo morphokinetic events, compared to standard overnight co-incubation.

Attitudes and preferences towards erectile dysfunction treatment among men with fertility needs: insights from a clinical study.

Erectile dysfunction (ED) is prevalent among males experiencing fertility challenges, yet attitudes towards actively treating ED in this group are under-researched. From a cohort of 1256 men with reproductive needs, 303 were identified with ED. The survey encompassed 296 respondents who correctly completed the second questionnaire, revealing that 50.3% sought ED treatment, with higher ED severity increasing the likelihood of seeking treatment. Infertile men were more likely to seek treatment than those with pregnancy loss (OR 3.18, 95% CI 1.74-5.83). Men with normal semen parameters were more open to ED therapy (OR 3.02, 95% CI 1.69-5.36), whereas those undergoing Assisted Reproduction Treatment were less inclined (OR 0.32, 95% CI 0.18-0.58). PDE-5 inhibitors (PDE-5Is) were preferred by 51.0% of those seeking treatment, with 29.7% of men with pregnancy loss and 60.1% of infertile men choosing PDE-5Is as their first option. Concerns included potential adverse effects of PDE-5Is on fetal health (78.7% of men with pregnancy loss) and on sperm quality (44.2% of infertile men). In conclusion, the different fertility requirements, semen parameters, and whether received ART are significant factors influencing the acceptance of treatment, PDE-5Is utilization among individuals in men with couple pregnancy loss is notably limited.

Correlation of mediterranean diet pattern and lifestyle factors with semen quality of men attending fertility clinics: A cross-sectional study

BackgroundWhile numerous studies have examined the impact of individual or combined nutrients on semen quality, research on the correlation between overall dietary patterns and semen quality remains limited. This cross-sectional study investigates the relationship between adherence to the alternative Mediterranean diet (aMED) and semen quality. MethodsA total of 274 men, presenting with both normal and abnormal semen parameters, participated in this study. Dietary data were collected using a 147-item semi-quantitative food frequency questionnaire (FFQ). Participants were divided into three groups based on their adherence to the Mediterranean diet (T1, T2, and T3). Statistical analyses, including ANOVA for numerical data and Chi-square tests for categorical data, were conducted. Multivariable logistic regression models were employed to estimate the association between aMED scores and abnormal semen parameters. ResultsParticipants in the highest adherence group (T3) were younger and had lower BMI compared to those in the other tertiles. Significant differences in sperm parameters were observed across the tertiles. T3 recorded the highest levels of sperm concentration, total sperm count, motility, progressive motility, and normal morphology (57.53 ± 36.16, 213.8 ± 158.9, 73.4 ± 25.9, 61.2 ± 24.6, and 6.42 ± 1.51, respectively), whereas T1 had the lowest values (11.92 ± 22.29, 43.3 ± 73.8, 36.7 ± 33.8, 4.6 ± 5.7, respectively). No significant differences were found in semen volume and viscosity. The regression analysis revealed a significant positive correlation between aMED scores and sperm concentration (B = 1.32, P = 0.001), total sperm count (B = 1.12, P = 0.001), and total motility (B = 0.71, P = 0.001). ConclusionAdherence to a high-quality Mediterranean diet is positively associated with improved semen quality and increased male fertility potential. Promoting healthy dietary patterns may be an effective strategy to enhance sperm motility and overall male reproductive health.

A Retrospective Study of Leparoscopic Evaluation of Tubal Factors in Case of Infertility

Background: Tubal factor infertility accounts for a large portion of female factor infertility. The most prevalent cause of tubal factor infertility is pelvic inflammatory disease and acute salpingitis. Tubal factor infertility accounts for approximately 25-35% of cases of female infertility. The aim of this study was to determine the role of leparoscopic evaluation of Tubal factors in case of infertility. Methods: A retrospective study was carried out at Department of Gynae and Obstetrics UHC, Dhamrai, Dhaka, Bangladesh from January to December 2023. 55 patients were selected for laparoscopic evaluation. Inclusion criteria were patients with no prior pelvic surgery, normal bimanual pelvic examination, normal semen parameters of partner and no ovulatory dysfunction. After obtaining thorough history & detailed examination, patients were informed about the procedure and written informed consent was taken. Results: 55 patients underwent laparoscopy during the study period. Most patients 10 were in the age group of ˂20 yr, 15 were between 21-30 yr & 20 were 31- 40 yr of age and 10 were between 36 – 40 yrs of age. Among these 55 patients 24 patients had primary infertility while 31 had presented with secondary infertility. Diagnostic laparoscopy revealed normal pelvic organs in 9 (16.3%) cases, Chronic pelvic inflammatory disease in 3 (5.4%), endometriosis in 13 (23.6%) and congenital anomalies of uterus in 1(1.8%) cases. 7.2% cases had bilateral tubal block, 27.3% had unilateral block and in 7.2% cases though the tubes were found patent significant peritubal adhesions were noted. Tubal pathology was detected in 55.5% cases of primary infertility and 42.1% cases of secondary infertility. Among those who had tubal pathology, nonspecific pelvic inflammatory disease was noted in 33.3% and genital tuberculosis was diagnosed in 2.7% cases. The most common complications were pyrexia, shoulder tip pain, nausea and vomiting. Conclusion: Laparoscopy is a safe and effective diagnostic tool in evaluating tubal status of infertile patients. Laparoscopy and chromopertubation test should be recommended as a first step in the investigation of infertile women with tubal factor.

Current landscape of fertility induction in males with congenital hypogonadotropic hypogonadism.

Congenital hypogonadotropic hypogonadism (CHH) is a rare reproductive disorder caused by deficient secretion or action of gonadotropin-releasing hormone (GnRH) and is a hormonally treatable form of male infertility. Both pulsatile GnRH treatment and combined gonadotropin therapy effectively induce spermatogenesis in 75%-80% of males with CHH, albeit the ejaculate does not usually approach normal semen parameters by WHO criteria. This is in some contrast to the cumulative fertility outcomes in females with CHH on gonadotropin treatment that are indistinguishable from those of reproductively normal females. Emerging data provide insights into early life determinants of male fertility (i.e., minipuberty), and research has identified key predictors of outcomes for fertility-inducing treatment in men with CHH. Such developments provide mounting evidence for tailoring approaches to maximize fertility potential in CHH, although there is no clear consensus to date on the optimal approach to fertility-inducing treatment. This review provides an up-to-date review on the current evidence underpinning therapeutic approaches for inducing spermatogenesis in males with CHH. In the absence of evidence-based clinical guidelines, this synthesis of current evidence provides guidance for clinicians working with males with CHH seeking fertility.

Analysis of 12 kinds of cytokines in seminal plasma by flow cytometry and their correlations with routine semen parameters

ObjectiveTo investigate the levels of 12 kinds of cytokines in seminal plasma and their correlations with routine semen parameters. MethodsThe remaining seminal plasma samples of 134 patients undergoing routine semen examination were collected for detecting cytokines. The parameters for sperm concentration, percentage of progressively motile sperm (PR), and motility were analyzed by a computer-assisted sperm analysis (CASA) system. According to the results of sperm concentration, PR and motility, 134 patients were divided into the normal routine semen parameters group, oligoasthenospermia group and azoospermia group. The levels of 12 kinds of cytokines in seminal plasma, including interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17, interferin (IFN)-α, IFN-γ, and tumor necrosis factor (TNF)-α, were detected by flow cytometry. Two seminal plasma samples were detected for 10 times, respectively, to calculate the coefficients of variation (CV) of each cytokine. The linear range of each cytokine was measured using the standard, and the correlation coefficient (r) was calculated. ResultsThe r2 of 12 kinds of cytokines detected by flow cytometry were all greater than 0.99. The reproducibility of 2 seminal plasma samples showed that the CVs of all cytokines were lower than 15 % except for TNF-α in sample 1 (15.15 %). Seminal plasma IL-6 levels were negatively correlated with semen volume (P < 0.01). Seminal plasma IL-5 levels were positively correlated with sperm concentration (P < 0.01). Seminal plasma IL-8 levels were negatively correlated with sperm motility (P < 0.01). Seminal plasma IL-8, IL-17 and IL-12P70 levels were negatively correlated with sperm PR (P < 0.05). In addition to the significant negative correlation between IL-5 and IL-17 (P < 0.05), there was a significant positive correlation between the majority of other cytokines. The levels of seminal plasma IL-17 and IL-12P70 in the oligoasthenospermia group and IL-1β and IL-12P70 in the azoospermia group were significantly higher than those in the normal routine semen parameters group (P ≤ 0.05), while the levels of IL-10 in the azoospermia group were significantly lower than that in the normal routine semen parameters group (P < 0.05). ConclusionThere are certain correlations between seminal plasma cytokines and routine semen parameters and strong correlations between different seminal plasma cytokines, suggesting that the imbalance between seminal plasma cytokines may affect sperm quality. However, it still needs to be further confirmed by large samples and multi-center clinical studies and related basic researches.

P-106 A novel sperm selection technique to select the most motile spermatozoa with superior genomic integrity for intrauterine insemination

Abstract Study question Does the utilization of a novel sperm selection method via microfluidics(MFSS) confer advantages over the conventional density gradient(DG) technique for couples undergoing intrauterine insemination(IUI)? Summary answer MFSS efficiently selected spermatozoa with increased motility and genomic integrity in an expedited manner, leading to an enhanced IUI clinical outcome. What is known already The first-line reproductive treatments for infertile couples are timed intercourse and IUI. In IUI, DG is commonly utilized to isolate motile spermatozoa. However, this method requires technical skills and processing time. Furthermore, DG exposes spermatozoa to silica gel particles and reactive oxygen species during centrifugation. MFSS represents a safer and more efficient sperm processing method to minimize human intervention. It has been reported by several investigators that MFSS selects spermatozoa with higher progressive motility and lower DNA fragmentation in ART. We have decided to investigate the efficacy of MFSS as an alternative method to select spermatozoa for IUI. Study design, size, duration Since 2017, post-processing semen parameters and Sperm Chromatin Fragmentation (SCF) was measured in the raw, DG-, and MFSS-processed specimens. Clinical outcomes were compared between IUI cycles using DG versus MFSS processing. Paired analyses were performed on patients who underwent history IUI-DG and subsequent IUI-MFSS, and cycles were split into natural (NC) or clomid or letrozole superovulation (CLO) cohorts. Participants/materials, setting, methods A total of 855 couples underwent 922 IUI-MFSS. Post-processing semen parameters and clinical outcomes were compared with those of 2325 age-matched couples who underwent 8322 IUI-DG during the same period. All male partners included in this study had normal semen parameters. DG was performed according to WHO, while MFSS as per manufacturer protocol(ZyMōt® Multi 850µL). SCF was assessed by TUNEL(normal threshold,≤15%). For all comparisons, +bHCG and clinical pregnancy(CPR,+FHB) were compared. Main results and the role of chance IUI-DG(maternal age:32.7±2;paternal age:37.9±6) resulted in a final concentration of 48.4±33x106/mL and motility of 88.1±4%. IUI-MFSS resulted in a decrease in a final concentration at 36.4±17x106/mL with a remarkable increase in motility at 98.6±1%(P&amp;lt;0.01). SCF was 6.0±4% in raw specimen, became 4.7±2% after DG, and 2.5±2% after MFSS(P&amp;lt;0.01). Overall, IUI-DG(n = 2325) resulted in a +bHCG of 12.7%(1065/8322) and a CPR(+FHB) of 11.0%(919/8322). IUI-MFSS(n = 855) resulted in a higher +bHCG of 16.1%(149/922,P&amp;lt;0.01) and a CPR of 12.5%(116/922). Next, we divided cycles into NC or CLO cohorts. In couples with NC, IUI-DG(n = 420) resulted in a +bHCG of 9.3%(132/1416) and a CPR of 8.8%(124/1416). IUI-MFSS(n = 107) resulted in a +bHCG of 11.4%(15/131) and a CPR of 11.4%(15/131). In CLO cycles, IUI-DG(n = 1990) resulted in a +bHCG of 13.0%(788/6048) and CPR of 10.9% (659/6048), while IUI-MFSS(n = 496) resulted in a significantly higher +bHCG at 17.4%(122/698) and CPR at 14.3%(100/698,P&amp;lt;0.01). In the paired sub-analysis, NC couples (n = 30) underwent IUI-DG and subsequently IUI-MFSS. IUI-DG resulted in a 9.4% +bHCG(7/74) and 8.1% CPR(6/74). IUI-MFSS yielded a 10.2% +bHCG(4/39) and 10.2% CPR(4/39). CLO couples (n = 132) underwent IUI-DG and resulted in a 13.6% +bHCG(85/623) and 11.2% CPR(70/623), followed by IUI-MFSS yielding a 15.2% +bHCG(27/178) and 12.4% CPR(22/178). Limitations, reasons for caution IUI-MFSS cycles resulted in an increased proportion of progressively motile spermatozoa, with a superior genomic integrity and higher clinical outcome. The benefit of MFSS may be somewhat limited because of the low SCF of this particular cohort and only a larger case series would be able to confirm this finding. Wider implications of the findings DG is labor intensive, technician-dependent, and unable to meaningfully ameliorate SCF. MFSS, while reducing exposure to reactive oxygen species, grants the highest sperm progressive motility with superior genomic integrity that may be responsible for an improved pregnancy outcome. Trial registration number not applicable

O-120 The role of CatSper in male infertility - from bench to bedsite and back

Abstract Infertility is on the rise, but the underlying causes still remain mostly unknown. In fact, for about one third of infertile men, semen parameters are within reference limits. This so called ‘unexplained male infertility’ seems to rest on elusive functional defects of the sperm undetectable by semen analysis, precluding early diagnosis of the infertility and evidence-based treatment by medically assisted reproduction (MAR). As a result, affected couples often experience failed MAR attempts, subjecting the ‘healthy’ female to recurring medical risks. We set out to gain insight into the pathomechanisms underlying unexplained male infertility. To this end, we systematically assessed the function of the sperm-specific multisubunit CatSper-channel complex in sperm of 2,300 men undergoing a fertility workup at our center, using a simple motility-based ‘CatSper-Test’ combined with [Ca2+]i fluorimetric and electrophysiological recordings. Thereby, we identified a group of men with normal semen parameters, but defective CatSper function. These men or couples failed to conceive naturally and upon MAR via intrauterine insemination and in vitro fertilization. Intracytoplasmic sperm injection (ICSI) was, ultimately, required to conceive a child. We revealed that the defective CatSper function was caused by variations in CATSPER genes. Moreover, we unveiled that CatSper-deficient human sperm were unable to undergo hyperactive motility and, therefore, failed to penetrate the egg coat. Thus, we provide the experimental evidence that sperm hyperactivation is required for human fertilization, explaining the infertility of CatSper-deficient men and need of ICSI for medically assisted reproduction. Finally, our study also revealed that defective CatSper function and ensuing failure to hyperactivate represents the most common cause of unexplained male infertility known thus far and that this sperm channelopathy can readily be diagnosed, enabling future evidence-based treatment of affected couples.

P-006 Fertility outcomes in men diagnosed with oncohematological diseases who stored their semen for fertility preservation

Abstract Study question What is the prevalence of conceptions among patients who underwent semen cryopreservation before receiving treatment for oncohematological disease? Summary answer Natural live birth rate after oncohematological treatments is around 38%(95%CI:31-45%), but can rise to 61%(95%CI:54-67%) with the use of frozen semen during ART cycles. What is known already Due to advancements in cancer treatments, there is a growing population of male adult cancer survivors. These individuals are at risk of experiencing treatment-induced infertility, and it is generally advised for them to preserve their semen before starting therapies. However, the details regarding the long-term utilization and effectiveness of frozen sperm samples in this particular population remain unclear. Study design, size, duration The study included males with oncohematological disease who stored semen (1987-2018) at Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico’s Assisted Reproductive Technology Unit in Milan. Data, gathered from records or proactive inquiry, assessed natural and ART conceptions post-treatment. The study also examined semen analysis, parenthood attempts, and the usage rates of stored spermatozoa as primary outcomes. Participants/materials, setting, methods The study is limited to men who meet the following eligibility criteria: i) semen cryopreserved for fertility preservation between 1987 and 2018; ii) diagnosis of oncohematological disease; iii) survival of at least 5 years from cryopreservation. Main results and the role of chance A total of 424 patients who underwent semen cryopreservation for oncohematological diseases were included in the study. The median [IQR] age at the time of sperm banking was 27 [23-33]. The most common indication was lymphoma (86%), and 102 subjects (24%) underwent hematopoietic stem cell transplantation. The median sperm concentration at the time of storage was 54 [20-98] million/ml. Among the participants, 242 (57%) men underwent post-treatment sperm analysis, revealing azoospermia in 87 of them (36%, 95%CI: 30-42%). Of the remaining, only 93 (38%) exhibited normal semen parameters. A total of 188 out of the 424 men included (44%) expressed a desire for childbearing and seventy-one achieved a natural live birth (38%, 95%CI: 31-45%). Eighty-nine individuals used their cryopreserved sperm samples, constituting 21% (95%CI: 17-25%) of the entire cohort. Among these 89 men, 45 experienced at least one live birth (51%, 95%CI: 40-61%). Notably, thirty-three men, despite actively seeking pregnancy, did not utilize their frozen semen. In summary, out of the 188 men interested in fatherhood, a total of 114 achieved a live birth (61%, 95%CI: 54-67%). Limitations, reasons for caution Our study is constrained by the lack of detailed information on the illness status, comorbidities, and specific cancer therapies administered. Furthermore, the data concern a single-center population, potentially limiting the generalizability of the results. Wider implications of the findings Oncohematological disease often require systemic therapies or even hematopoietic stem cell transplantation resulting in a very high percentage of infertile patients These findings support the need to prioritize sperm preservation before therapies in these patients, even in cases who are not candidates for stem cell transplantation in the first instance Trial registration number NA

O-196 Men who fathered embryos with poor pre-implantation development rates exhibit an increased frequency of sperm with high DNA accessibility

Abstract Study question Do differing levels of DNA accessibility in sperm affect the ability of embryos generated by ART to undergo pre-implantation development? Summary answer Individuals whose sperm generated embryos that failed to efficiently form blastocysts tend to have more sperm with high levels of DNA accessibility. What is known already Chromatin is a complex composed of nucleic acids and proteins that serves to condense and regulate the genome in all eukaryotic cells. Sperm chromatin is packaged into a structure with higher density than that found in precursor germ cells or somatic cells. Samples from individuals with decreased normal semen parameters have previously been shown to possess an increased frequency of sperm with aberrant chromatin packaging. However, existing methodologies for measuring sperm chromatin packaging are indirect and non-quantitative. Study design, size, duration Prospective cohort study, registered January 2020. We adapted an enzymatic methodology for measuring DNA accessibility known as NicE-view (Nicking Endonuclease-mediated visualization), for use in human sperm. DNA accessibility was quantified in three millions of individual sperm from semen obtained from 60 individuals. Both total sperm and sperm selected for motility via swim-up were analyzed for each participant. Participants/materials, setting, methods Participants sought treatment for infertility at Universitätsspital Basel and possessed normospermic levels in conventional semen analysis. Participant partners had normal endocrine parameters and ovulation response to hormonal stimulation. 20 couples obtained pregnancies without the need for ART, while 40 individuals who were treated using ART and divided into two groups differing in pre-implantation development rates: (1) 20 with blastocyst development of &amp;gt; 50% of all zygotes and (2) 20 produced 0-1 blastocyst following embryo culture. Main results and the role of chance We quantified DNA accessibility and the presence of single stranded DNA nicks in more than 3 million sperm derived from 117 samples. We found that while DNA nicks were only present in a small fraction of the sperm analyzed, DNA accessibility levels varied dramatically with the data indicating a bimodal distribution. At a single sperm level, DNA accessibility and the presence of single stranded DNA damage both correlated with nuclear size. We found that DNA nicking was more abundant in total sperm samples compared to sperm selected for motility via swim-up preparation. In contrast, sperm with high levels of DNA accessibility were more abundant following swim-up, perhaps resulting from capacitation. We observed that individuals with a history of low blastocyst development were more likely to have high frequencies of sperm harboring DNA nicks and high DNA accessibility. We also measured increased frequencies of sperm with high DNA accessibility to negatively correlate with conventional semen parameter values. Limitations, reasons for caution Our study cohort consists of only 60 individuals. Variation is also observed between individuals within each participant category, so further analysis with increased participant numbers is required to strengthen these conclusions. Wider implications of the findings Our results suggest measuring DNA accessibility in sperm from normospermic individuals provide additional information about the efficiency of pre-implantation development following ART. Future work examining the sources of increased DNA accessibility in sperm will also be interesting to identify possible treatments for such individuals. Trial registration number SNCTP000003654 | NCT04256668

Genetic Insights into Azoospermia and Severe Oligozoospermia: Discovering Seven SNPs through GWAS and In Silico Analysis.

Azoospermia and severe oligozoospermia represent the most extreme forms of male infertility. Despite their prevalence, the genetic foundations of these conditions are not well understood, with only a limited number of genetic factors identified so far. This study aimed to identify single-nucleotide polymorphisms (SNPs) linked to both azoospermia and severe oligozoospermia. We conducted a genome-wide association study (GWAS) involving 280 Greek males with normal semen parameters and 85 Greek males diagnosed with either azoospermia or severe oligozoospermia. Following rigorous quality control measures, our analysis identified seven SNPs associated with azoospermia/severe oligozoospermia. An in silico functional annotation was subsequently used to further investigate their role. These SNPs, found in regions not previously associated with male reproductive disorders, suggest novel genetic pathways that may contribute to these forms of infertility and pave the way for future studies. Additionally, this study sheds light on the significant role of noncoding RNAs in the pathogenesis of male infertility, with three of the identified SNPs situated in long intergenic non-coding RNAs (lincRNAs). Our findings highlight the intricate genetic landscape of azoospermia and severe oligozoospermia, underlining the necessity for more detailed studies to fully grasp the underlying mechanisms and their potential for informing diagnostic and therapeutic strategies.

Comparison of Testicular Sonography and Elastography Findings With Semen Parameters in Cases Investigated for Infertility.

This study aimed to investigate the correlation between testicular shear wave elastography (SWE) values and semen analysis results in men with infertility. This was a retrospective case-control study. Patients were categorized as normal, abnormal, or azoospermic based on sperm analysis results. Testicular volume was measured using B-mode ultrasonography using the Lambert formula. Subsequently, 40-80 regions of interest measuring 1.5 × 1.5 mm were manually positioned in both testicles based on their size, and two-dimensional SWE was applied through virtual touch imaging quantification software. The patients had a mean age of 33.79 ± 6.3 years, with semen analysis revealing normal results in 15 patients (22.4%), pathological findings in 35 patients (52.2%), and azoospermia in 17 patients (25.4%). Right, left, total, and mean testicular volumes were significantly lower in patients with azoospermia compared to those in both normal and impaired semen parameters (P < .05). Conversely, testicular elastography scores were higher in patients with azoospermia than in the other groups (P < .05). The significant negative correlation between volume and elastographic findings remained independent of age (r = 0.4, P < .001). The accuracy rates for detecting impaired semen parameters and azoospermia were 94.3% and 94.1%, respectively, after considering factors such as age, testicular volume (right/left/total), and elastography (right/left/total). Notably, the total mean elastography score ranked first, with 100% in the independent normalized importance distribution of these variables. SWE can be used effectively alone or in combination with other diagnostic tools to evaluate histopathological changes in the testicles of male patients with infertility.

Characterization of seminal microbiome associated with semen parameters using next-generation sequencing

BackgroundTo characterize the seminal microbiome associated with normal and abnormal semen parameters, towards the prediction of reproductive health and sperm quality. Despite the association between bacteria and infertility, few studies have looked at the beneficial effects of the seminal microbiome on infertility.The study comprised semen samples from 69 men with normal spermiograms and 166 men with at least 1 abnormal spermiogram parameter from the Institutional IVF Center between October 2019 and October 2022.We hypothesized that the composition of the microbiota may affect semen parameters. To determine the composition of uncultured bacteria, the 16S ribosomal RNA (rRNA) gene was amplified using Oxford Nanopore Technology.ResultsDifferent groups of bacteria were present in the semen samples of patients with normal semen parameters, such as female factor infertility and abnormal sperm parameters. Bacterial communities differed between samples. However, the relative distribution of Lactobacillus and Prevotella in the normal and abnormal semen groups differed (p = 0.05) and was statistically significant.In the abnormal semen group, the incidence of Lactobacillus probiotics was lower and the frequency of Prevotella was higher. Additionally, principal component analysis (PCA) revealed differences in the microbial composition of normal and abnormal semen.ConclusionsIn our study, NGS analysis revealed the increased presence of harmful bacteria Prevotella in groups with abnormal semen raises the possibility that certain microbiota may be associated with semen quality and male infertility.

MTHFRC 677T Gene Polymorphism and Homocysteine in North China Patients with Varicocele

To investigate the correlation between methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism, homocysteine, and male infertility with varicocele in North China. One Hundred infertile males with varicocele (VC; grade II-III, VC group) and 100 healthy males with normal semen parameters and no varicocele (NC group) were recruited for PCR microarray, blood and semen testing. Compared with the CC genotype in the NC group, the TT genotype in the NC group and the CC genotype in the VC group showed no significant changes in sperm motility (P=0.191; P=0.130), sperm density (P=0.591, P=0.643), plasma homocysteine level (P=0.511; P=0.677), and seminal plasma MDA (P=0.752; P=0.451). In contrast, VC patients with the TT genotype had higher plasma homocysteine level and seminal plasma MDA levels (P &lt;0.001), lower partial pressure of oxygen in seminal pulse (PO2; P &lt;0.001) and poorer sperm quality (P &lt;0.001), as compared with the CC genotype. This suggests that MTHFR C677C&gt;T may not be a risk factor for male Varicocele in North China. However, this may affect the oxidative stress associated with homocysteine expression, which in turn affects semen parameters in VC patients. Larger studies are needed to validate our findings.

Paternal aging impacts expression and epigenetic markers as early as the first embryonic tissue lineage differentiation

BackgroundAdvanced paternal age (APA) is associated with adverse outcomes to offspring health, including increased risk for neurodevelopmental disorders. The aim of this study was to investigate the methylome and transcriptome of the first two early embryonic tissue lineages, the inner cell mass (ICM) and the trophectoderm (TE), from human blastocysts in association with paternal age and disease risk. High quality human blastocysts were donated with patient consent from donor oocyte IVF cycles from either APA (≥ 50 years) or young fathers. Blastocysts were mechanically separated into ICM and TE lineage samples for both methylome and transcriptome analyses.ResultsSignificant differential methylation and transcription was observed concurrently in ICM and TE lineages of APA-derived blastocysts compared to those from young fathers. The methylome revealed significant enrichment for neuronal signaling pathways, as well as an association with neurodevelopmental disorders and imprinted genes, largely overlapping within both the ICM and TE lineages. Significant enrichment of neurodevelopmental signaling pathways was also observed for differentially expressed genes, but only in the ICM. In stark contrast, no significant signaling pathways or gene ontology terms were identified in the trophectoderm. Despite normal semen parameters in aged fathers, these significant molecular alterations can adversely contribute to downstream impacts on offspring health, in particular neurodevelopmental disorders like autism spectrum disorder and schizophrenia.ConclusionsAn increased risk for neurodevelopmental disorders is well described in children conceived by aged fathers. Using blastocysts derived from donor oocyte IVF cycles to strategically control for maternal age, our data reveals evidence of methylation dysregulation in both tissue lineages, as well as transcription dysregulation in neurodevelopmental signaling pathways associated with APA fathers. This data also reveals that embryos derived from APA fathers do not appear to be compromised for initial implantation potential with no significant pathway signaling disruption in trophectoderm transcription. Collectively, our work provides insights into the complex molecular mechanisms that occur upon paternal aging during the first lineage differentiation in the preimplantation embryo. Early expression and epigenetic markers of APA-derived preimplantation embryos highlight the susceptibility of the future fetus to adverse health outcomes.

Comparative analysis of conventional in vitro fertilization and intracytoplasmic sperm injection in patients with polycystic ovarian syndrome, tubal factor infertility, and unexplained infertility whose partners exhibit normal semen parameters: A retrospective study of sibling oocytes.

This study compared the outcomes of conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in patients with polycystic ovarian syndrome (PCOS), tubal factor (TF) infertility, and unexplained infertility whose partners had normal semen parameters. This retrospective study included 360 couples diagnosed with infertility involving PCOS (n=157), unexplained infertility (n=140), and TF infertility (n=63). Sibling oocytes were randomly assigned to undergo ICSI or conventional IVF insemination. The fertilization rate and embryo morphology were evaluated as outcomes. Retrieved cumulus-oocyte complexes from patients with PCOS (2,974), unexplained infertility (1,843), and TF infertility (844) were split and inseminated by conventional IVF and ICSI respectively. In comparison to the ICSI method, the conventional IVF approach was linked to a significantly higher fertilization rate in groups with PCOS (68.81% vs. 77.49%), unexplained infertility (67.62% vs. 78.84%), and TF issues (69.23% vs. 78.63%) (p<0.05). The proportion of embryos with grade A produced by the conventional IVF method was significantly higher than that produced using the ICSI method in the PCOS and unexplained infertility groups (p<0.05). Additionally, the percentage of grade B embryos produced with the ICSI method was significantly higher than that produced with the conventional IVF method in PCOS patients (p=0.002). Our results indicated that the conventional IVF method was associated with higher zygote production and a higher proportion of grade A embryos when all infertile groups were evaluated together. Thus, ICSI is not suggested for patients with these causes of infertility if their partner has normal semen parameters.

Impact of semen microbiota on the composition of seminal plasma.

Several studies have found associations between specific bacterial genera and semen parameters. Bacteria are known to influence the composition of their niche and, consequently, could affect the composition of the seminal plasma. This study integrated microbiota profiling and metabolomics to explore the influence of seminal bacteria on semen metabolite composition in infertile couples, revealing associations between specific bacterial genera and metabolite profiles. Amino acids and acylcarnitines were the predominant metabolite groups identified in seminal plasma. Different microbiota profiles did not result in globally diverse metabolite compositions in seminal plasma. Nevertheless, levels of specific metabolites increased in the presence of a dysbiotic microbiota. Urocanate was significantly increased in abnormal semen samples (adjusted P-value < 0.001) and enriched in samples dominated by Prevotella spp. (P-value < 0.05), which was previously linked to a negative impact on semen. Therefore, varying microbiota profiles can influence the abundance of certain metabolites, potentially having an immunomodulatory effect, as seen with urocanate.IMPORTANCEMale infertility is often considered idiopathic since the specific cause of infertility often remains unidentified. Recently, variations in the seminal microbiota composition have been associated with normal and abnormal semen parameters and may, therefore, influence male infertility. Bacteria are known to alter the metabolite composition of their ecological niches, and thus, seminal bacteria might affect the composition of the seminal fluid, crucial in the fertilization process. Our research indicates that distinct seminal microbiota profiles are not associated with widespread changes in the metabolite composition of the seminal fluid. Instead, the presence of particular metabolites with immunomodulatory functions, such as urocanate, could shed light on the interplay between seminal microbiota and variations in semen parameters.

Human fertilization in vivo and in vitro requires the CatSper channel to initiate sperm hyperactivation.

The infertility of many couples rests on an enigmatic dysfunction of the man's sperm. To gain insight into the underlying pathomechanisms, we assessed the function of the sperm-specific multisubunit CatSper-channel complex in the sperm of almost 2,300 men undergoing a fertility workup, using a simple motility-based test. We identified a group of men with normal semen parameters but defective CatSper function. These men or couples failed to conceive naturally and upon medically assisted reproduction via intrauterine insemination and in vitro fertilization. Intracytoplasmic sperm injection (ICSI) was, ultimately, required to conceive a child. We revealed that the defective CatSper function was caused by variations in CATSPER genes. Moreover, we unveiled that CatSper-deficient human sperm were unable to undergo hyperactive motility and, therefore, failed to penetrate the egg coat. Thus, our study provides the experimental evidence that sperm hyperactivation is required for human fertilization, explaining the infertility of CatSper-deficient men and the need of ICSI for medically assisted reproduction. Finally, our study also revealed that defective CatSper function and ensuing failure to hyperactivate represents the most common cause of unexplained male infertility known thus far and that this sperm channelopathy can readily be diagnosed, enabling future evidence-based treatment of affected couples.

Investigation of sperm chromatin condensation by aniline blue staining in infertile men with normal and abnormal semen parameters

Investigation of sperm chromatin condensation by aniline blue staining in infertile men with normal and abnormal semen parameters

#116 : Prevalence of Aneuplodies in Chromosomes 13, 18, 21, X and Y in Embryos Generated by ICSI of Fertile Couples

Background and Aims: It has previously been reported that up to 70% of the embryos generated in an ART lab are chromosomally abnormal. Abnormalities in chromosomes 13, 18, 21, X and Y have been observed to be as common as 63% in embryos of sub-fertile couples. The aim of this study was to find if these abnormalities are just as common in embryos of fertile couples who have no history of an affected child. Method: A retrospective cohort study was conducted on 409 embryos obtained from 113 couples undergoing preimplantation genetic testing at an ART center in Pakistan, between 1 January 2021 and 31 December 2022. The inclusion criteria was that fertile couples who had come for gender selection for family balancing. Age range of female partners was 28 years to 38 years. Male partners had normal semen parameters as per the World Health Organization (WHO) guidelines. Results: A total of 70.7% (289/409) were unaffected by aneuploides in the five chromosomes. There was a significant difference in the ratio of normal to abnormal embryos. Out of the normal embryos, there was no significant difference in the male (34.5%) and female (36.1%) embryos. There was no significant difference in the number of abnormal female and male embryos (p=0.127). Among the aneuploidies in the five chromosomes, trisomy 21 was the most common (n=27; 6.6%). Conclusion: Approximately 20% of sub-fertile couples have an unknown etiology. Our results indicate that the five commonly known aneuploidies found in embryos of sub-fertile patients have a low prevalence in fertile patients. It is worth noting that this study is limited by the fact that abnormalities in sub-fertile couple were not analyzed and statistics from other already published studies was relied upon. Nonetheless, this data has provided perspectives for counselling couples of unexplained sub-fertility.