Ejaculate Parameters Research Articles

A view of immunologic male infertility from the perspective of preserving seminal fluid homeostasis

The basis of the pathogenesis of immunologic male infertility is the production of antisperm antibodies against sperm antigens by plasmocytes. Extracellular nucleic acids, among other antigens, are perceived as DAMPs. The clearance system for biological fluids is complex. Endonucleases are responsible for the utilization of extracellular DNA. Among endonucleases, DNase 1 is the most studied. Its concentration in biological fluids is higher than others. For the first time, we determined the concentration of DNase 1 in the seminal fluid of men of reproductive age. DNase 1 concentration values were compared with the level of antisperm antibodies in the ejaculate, after which the relationship between the enzyme level and standard spermogram parameters was determined. The material for the study was the ejaculate of 44 relatively healthy men of reproductive age. During spermiological analysis, macroscopic and microscopic parameters of seminal fluid were determined, the concentration of deoxyribonuclease 1 (DNase 1) in the material was determined by enzyme immunoassay, and the presence of antisperm antibodies was assessed using a direct diagnostic method – the MAR test. Statistical processing of the obtained data was carried out using analytical software PAST v. 4.06. Reference intervals for DNase 1 concentrations were calculated using the MedCalc v. 17.4 program. The relationship between spermogram parameters and DNase 1 concentration was assessed by calculating Spearman’s linear rank correlation coefficients. As a result, a negative correlation was found between the DNase 1 ratio and such ejaculate parameters as liquefaction time (rS = -0.37; p = 0.013) and viscosity (rS = -0.37; p = 0.013), as well as the level of antisperm antibodies A (rS = -0.43; p = 0.003) and G (rS = -0.33; p = 0.027), the mucous component of the ejaculate (rS = -0.37; p = 0.012) and agglutination of male germ cells (rS = -0.33; p = 0.029). According to the data obtained, the optimal level of DNase 1 is directly related to the quality of the ejaculate. Determination of DNase 1 concentration can serve as an additional diagnostic criterion for male immunological infertility. These data will expand our understanding of a man’s fertile potential.

Direct and indirect connections of androgen status with ejaculate parameters in men from infertile couples

In men from infertile couples the serum level of total testosterone (tT) has been shown to vary widely. Is it possible to expect that there is an association of tT content with spermogram disorders in men from infertile couples? Aim of the study was to investigate the patterns of changes in the spermiological status of men from infertile couples depending on tT level in blood serum. Material and methods. Design – observational, retrospective, one-stage study. The analysis of medical histories of 358 men with infertility in marriage was carried out. The sample was divided into comparison groups according to tT level: group 1 – less than 12.1, group 2 – from 12.1 to 20.9, group 3 – 21.0 nmol/l or more. Results. From group 1 to group 3, tT content increases more than twice, as well as concentration of indicators related to the level of T – sex hormone binding globulin (SHBG) and free testosterone (fT). There are no significant differences in luteinizing hormone (LH) and follicle stimulating hormone (FSH) level, although there is a tendency to its increase from group 1 to group 3. From group 1 of men with androgen deficiency to group 3, not only body weight and body mass index (BMI), but also waist circumference (WC) and hip circumference (HC), as well as the WC/HC index, characterizing the degree of abdominal obesity, decrease. The groups examined did not differ in the values of all studied ejaculate parameters. In group 1, a pronounced correlation between the content of tT and fT was found, in groups 2 and 3 – statistically significant inverse relationships between the level of Tob and the values of anthropometric indicators (body weight, BMI, WC and HC), as well as direct ones - with the concentration of SHBG, tT, LH and estradiol, in group 3 – with FSH levels. There were no correlations between tT content and spermogram indicators in any group of examined men. Conclusions. The results obtained suggest that only at high-normal level of testosterone in the blood it can have a stimulating effect on spermatogenesis. As a result of the accumulation of cases of androgenic deficiency in the population, the direct positive effect of serum testosterone on spermatogenesis is becomes insufficient for normal regulation, and the negative effect of testosterone deficiency on spermatogenesis, mediated through the accumulation of overweight and obesity comes to the fore.

Influence of surgical treatment of varicocele on the pathospermia and the level of sperm DNA fragmentation

Introduction. Varicocele is a varicose vein in the testicle caused by pathological venous reflux. Clinically, varicocele is found in 15% of the male population, with 35% of men with primary infertility and 70-81% of men with secondary infertility. Uncontrolled, excessive amounts of reactive oxygen species (ROS) can cause single- and/ or double-strand breaks in sperm DNA, commonly referred to as sperm DNA fragmentation. The purpose of the study was to determine the effect of surgical treatment of varicocele on the level of DNA fragmentation and to develop prognostic criteria for restoring the fertility of patients.Materials and methods. The study is based on a comparative analysis of the results of examination and treatment of 93 patients without concomitant pathology, with pathospermia, increased levels of DNA fragmentation and varicocele, who received surgical treatment. All patients were followed up for 3–9 months after surgery. The control group did not receive postoperative therapy – 34 people, the main group (59 patients) received drug rehabilitation – a complex of spermatoprotectors and antioxidants – for 3 months after surgical treatment.Results of the study and their discussion. In patients who received surgical treatment, there was a normalization of the level of DNA fragmentation after 3 months in the postoperative period, on average the positive dynamics was 21% (from an average fragmentation level of 22.82% to 15.84%, p <0.05), the prognosis for pregnancy in the couple became favorable. An improvement in DNA fragmentation indicators was observed in parallel with an improvement in spermogram indicators in the postoperative period. Progression of left testicular hypotrophy in the postoperative period was not detected. No relapses or other complications were noted.Conclusions. Treatment of varicocele improves fertility in patients with pathospermia, increased levels of DNA fragmentation, which is consistent with research data from other authors, which confirmed the fact of a significant improvement in ejaculate parameters (sperm concentration, motility and morphology) However, the dynamics of normalization of sperm DNA fragmentation occurs, according to our data, more dynamically (within 3 months in the postoperative period) than the improvement of pathospermia (OAT syndrome) and does not depend on the presence of postoperative drug rehabilitation.

Dynamics of ejaculate parameters from 2017 to 2023: a retrospective single-center analysis

Introduction: One of the most important tests to evaluate male fertility is a semen analysis. Assessment of ejaculate parameters allows to assume possible causes of male infertility and helps to determine further treatment. According to modern literature there is a slight but constant deterioration of ejaculate parameters with time.Purpose of the study: to evaluate semen analysis parameters in patients examined in one medical center.Materials and Methods: retrospective evaluation of semen analyses performed in the same laboratory from January 1, 2017 to October 31, 2023 was performed with a total of 47902 patients included. The ejaculate volume, concentration and number of spermatozoa, the proportion of spermatozoa of different categories of motility, sand the proportion of morphologically normal spermatozoa were evaluated. Statistical processing of data was performed using MS Office and SPSS v26 software packages.Results: the mean semen volume from 2017 to 2023 was 3.5 ± 1.5 mL, sperm concentration was 51.9 ± 39.3 million/mL, progressively motile spermatozoa was 48.2 ± 16.5%, number of progressively motile spermatozoa in the sample was 93.7 ± 94.3 million, motile spermatozoa 55.6 ± 17.4%, total motile spermatozoa was 107.5 ± 105.6 million, mean number of morphologically normal forms was 2.8 ± 1.4% in 2017, 2.2 ± 0.9% in 2023, and 3.5 ± 1.5% during the observation period. Mean pH of ejaculate was 7.6 ± 0.3 with noted tendency to increase (7.38 ± 0.33 in 2017, 7.6 ± 0.38 in 2023). The proportion of samples with azoospermia increased from 2.8% in 2017 to 4.5% in 2023. Mean number of patients with cryptozoospermia was 1.5%.Conclusion: there is a trend of decreasing proportion of morphologically normal spermatozoa, increasing ejaculate pH and increasing frequency of azoospermia from 2017 to 2023.

Identification of two hidden clinical subgroups among men with idiopathic cryptozoospermia.

Are there subgroups among patients with cryptozoospermia pointing to distinct etiologies? We reveal two distinct subgroups of cryptozoospermic (Crypto) patients based on testicular tissue composition, testicular volume, and FSH levels. Cryptozoospermic patients present with a sperm concentration below 0.1 million/ml. While the etiology of the severely impaired spermatogenesis remains largely unknown, alterations of the spermatogonial compartment have been reported including a reduction of the reserve stem cells in these patients. To assess whether there are distinct subgroups among cryptozoospermic patients, we applied the statistical method of cluster analysis. For this, we retrospectively selected 132 cryptozoospermic patients from a clinical database who underwent a testicular biopsy in the frame of fertility treatment at a university hospital. As controls (Control), we selected 160 patients with obstructive azoospermia and full spermatogenesis. All 292 patients underwent routine evaluation for endocrine, semen, and histological parameters (i.e. the percentage of tubules with elongated spermatids). Moreover, outcome of medically assisted reproduction (MAR) was assessed for cryptozoospermic (n = 73) and Control patients (n = 87), respectively. For in-depth immunohistochemical and histomorphometrical analyses, representative tissue samples from cryptozoospermic (n = 27) and Control patients (n = 12) were selected based on cluster analysis results and histological parameters. This study included two parts: firstly using clinical parameters of the entire cohort of 292 patients, we performed principal component analysis (PCA) followed by hierarchical clustering on principal components (i.e. considering hormonal values, ejaculate parameters, and histological information). Secondly, for histological analyses seminiferous tubules were categorized according to the most advanced germ cell type present in sections stained with Periodic acid Schif. On the selected cohort of 39 patients (12 Control, 27 cryptozoospermic), we performed immunohistochemistry for spermatogonial markers melanoma-associated antigen 4 (MAGEA4) and piwi like RNA-mediated gene silencing 4 (PIWIL4) followed by quantitative analyses. Moreover, the morphologically defined Adark spermatogonia, which are considered to be the reserve stem cells, were quantified. The PCA and hierarchical clustering revealed three different clusters, one of them containing all Control samples. The main factors driving the sorting of patients to the clusters were the percentage of tubules with elongated spermatids (Cluster 1, all Control patients and two cryptozoospermic patients), the percentage of tubules with spermatocytes (Cluster 2, cryptozoospermic patients), and tubules showing a Sertoli cells only phenotype (Cluster 3, cryptozoospermic patients). Importantly, the percentage of tubules containing elongated spermatids was comparable between Clusters 2 and 3. Additional differences were higher FSH levels (P < 0.001) and lower testicular volumes (P < 0.001) in Cluster 3 compared to Cluster 2. In the spermatogonial compartment of both cryptozoospermic Clusters, we found lower numbers of MAGEA4+ and Adark spermatogonia but higher proportions of PIWIL4+ spermatogonia, which were significantly correlated with a lower percentage of tubules containing elongated spermatids. In line with this common alteration, the outcome of MAR was comparable between Controls as well as both cryptozoospermic Clusters. While we have uncovered the existence of subgroups within the cohort of cryptozoospermic patients, comprehensive genetic analyses remain to be performed to unravel potentially distinct etiologies. The novel insight that cryptozoospermic patients can be divided into two subgroups will facilitate the strategic search for underlying genetic etiologies. Moreover, the shared alterations of the spermatogonial stem cell compartment between the two cryptozoospermic subgroups could represent a general response mechanism to the reduced output of sperm, which may be associated with a progressive phenotype. This study therefore offers novel approaches towards the understanding of the etiology underlying the reduced sperm formation in cryptozoospermic patients. German research foundation CRU 326 (grants to: SDP, NN). Moreover, we thank the Faculty of Medicine of the University of Münster for the financial support of Lena Charlotte Schülke through the MedK-program. We acknowledge support from the Open Access Publication Fund of the University of Münster. The authors have no potential conflicts of interest. N/A.

Monitoring of taurine dietary supplementation effect on parameters of Duroc boar ejaculate in summer season.

The aim of this experiment was to find out whether the taurine supplementation in daily ration had an effect on quantity or quality of Duroc boar ejaculate. The experiment duration was from June to August, when it could assumed the possible occurrence of heat stress. For the study was chosen 12 Duroc boars of approximately the same age and condition. The control group of 6 Duroc boars was fed only by basic diet and the experimental group of 6 Duroc boars was fed by the same basic diet with supplementation of 15 g taurine/boar/day. Ejaculate was collected once a week by hand glowed technique. From ejaculate parameters were monitored volume of ejaculate, sperm concentration, total amount of sperm, morphologically abnormal sperm, taurine concentration and GSH/GSSH concentration. From microscopic analysis, results were statistically significant in motility in June and July (P<0.05). In biochemical results, a significant difference (P<0.05) has been found between the experimental groups in the concentrations of taurine as well as GSH/GSSG in ejaculate which indicates the effect of heat stress on boars during the experimental period.

Влияние хирургического лечения варикоцеле на патоспермию и уровень фрагментации ДНК сперматозоидов

Introduction Varicocele is a varicose vein in the testicle caused by pathological venous reflux. Clinically, varicocele is found in 15% of the male population, with 35% of men with primary infertility and 70-81% of men with secondary infertility. Uncontrolled, excessive amounts of reactive oxy-gen species (ROS) can cause single- and/or double-strand breaks in sperm DNA, commonly referred to as sperm DNA fragmentation. The purpose of the study was to determine the effect of surgical treatment of varicocele on the level of DNA fragmentation and to develop prognostic criteria for restor-ing the fertility of patients. Materials and methods. The study is based on a comparative analysis of the results of examination and treatment of 93 patients without concomi-tant pathology, with pathospermia, increased levels of DNA fragmentation and varicocele, who received surgical treatment. All patients were followed up for 3-9 months after surgery. The control group did not receive postop-erative therapy - 34 people, the main group (59 patients) received drug re-habilitation - a complex of spermatoprotectors and antioxidants - for 3 months after surgical treatment. Results of the study and their discussion. In patients who received surgical treatment, there was a normalization of the level of DNA fragmenta-tion after 3 months in the postoperative period, on average the positive dy-namics was 21% (from an average fragmentation level of 22.82% to 15.84%, p &lt;0.05), the prognosis for pregnancy in the couple became favorable. An improvement in DNA fragmentation indicators was observed in parallel with an improvement in spermogram indicators in the postoperative period. Progression of left testicular hypotrophy in the postoperative period was not detected. No relapses or other complications were noted. Conclusions. Treatment of varicocele improves fertility in patients with pathospermia, increased levels of DNA fragmentation, which is con-sistent with research data from other authors, which confirmed the fact of a significant improvement in ejaculate parameters (sperm concentration, mo-tility and morphology) However, the dynamics of normalization of sperm DNA fragmentation oc-curs, according to our data, more dynamically (within 3 months in the post-operative period) than the improvement of pathospermia (OAT syndrome) and does not depend on the presence of postoperative drug rehabilitation.

Динамика показателей эякулята за период с 2017 по 2023 год: ретроспективный одноцентровой анализ

Introduction: One of the most important tests to evaluate male fertility is a semen analysis. Assessment of ejaculate parameters allows to assume possible causes of male infertility and helps to determine further treatment. According to modern literature there is a slight but constant deterioration of ejaculate parameters with time. Purpose of the study: to evaluate semen analysis parameters in patients examined in one medical center. Materials and Methods: retrospective evaluation of semen analyses performed in the same laboratory from January 1, 2017 to October 31, 2023 was performed with a total of 47902 patients included. The ejaculate volume, concentration and number of spermatozoa, the proportion of spermatozoa of different categories of motility, sand the proportion of morphologically normal spermatozoa were evaluated. Statistical processing of data was performed using MS Office and SPSS v26 software packages. Results: the mean semen volume from 2017 to 2023 was 3.5 В± 1.5 mL, sperm concentration was 51.9 В± 39.3 million/mL, progressively motile spermatozoa was 48.2 В± 16.5%, number of progressively motile spermatozoa in the sample was 93.7 В± 94.3 million, motile spermatozoa 55.6 В± 17.4%, total motile spermatozoa was 107.5 В± 105.6 million, mean number of morphologically normal forms was 2.8 В± 1.4% in 2017, 2.2 В± 0.9% in 2023, and 3.5 В± 1.5% during the observation period. Mean pH of ejaculate was 7.6 В± 0.3 with noted tendency to increase (7.38 В± 0.33 in 2017, 7.6 В± 0.38 in 2023). The proportion of samples with azoospermia increased from 2.8% in 2017 to 4.5% in 2023. Mean number of patients with cryptozoospermia was 1.5%. Conclusion: there is a trend of decreasing proportion of morphologically normal spermatozoa, increasing ejaculate pH and increasing frequency of azoospermia from 2017 to 2023.

Effects of heat stress on oxidative balance and sperm quality in dogs.

Heat stress negatively affects both animal reproductive performance and their overall wellbeing and welfare. When temperatures soar, the body responds to maintain balance, resulting in heat stress. This triggers various responses, including the buildup of reactive oxygen species (ROS), which can harm sperm vitality through lipid peroxidation. Oxidative damage can result in sperm dysfunction. This study aimed to evaluate the effects of environmental heat stress on canine quantitative and qualitative ejaculate parameters. Thirty-six male crossbred dogs were involved in the trial. This study was performed in 2022, precisely from May to August. Dogs were subdivided in two groups, one heat stressed (HS) and one in thermoneutrality (TN). Thermo Hygrometric Index (THI) was hourly measured and ranged from 60 to 71 in TN dogs and from 60 to 83 for HS dogs. Semen and blood samples were collected at 30-day intervals, starting from May (0 days), and then at 30 days, 60 days, and 90 days and analysis for evaluating biochemical profile, semen oxidative status, and semen quality were performed. In HS dogs, serum total protein, albumin, and urea concentrations showed a significant decrease after 60 days (P < 0.01), with values lower than those observed in TN dogs (P < 0.01). Both catalase and glutathione peroxidase concentrations were reduced after 60 days in HS dogs, showing lower levels than the TN group (P < 0.01 and P < 0.05, respectively). Antioxidant potential increased over time in HS dogs, reaching higher values at 60 days (P < 0.05) and 90 days (P < 0.01). On the other hand, ROS in the sperm of HS animals rose by day 90, surpassing the values recorded at previous time points and in TN dogs (P < 0.01). Semen concentration (P > 0.01) and total sperm count (P < 0.05) declined after 30 days in the HS group and remained lower than the TN group throughout the trial. The study demonstrates that heat stress negatively affects the oxidative status and sperm quality of male dogs, reducing reproductive performance. However, further research is needed due to the lack of complete breed homogeneity in the study groups.

Dynamics of sperm DNA fragmentation in patients with clinically significant varicocele after Marmar operation and laparoscopic varicocelectomy

BACKGROUND: Infertility is detected in 15% of sexually active couples who engage in unprotected sexual intercourse. At the same time, the male factor of infertility accounts from 30 to 50%. It is known that in 15–25% the cause of frozen pregnancy and miscarriage is the male factor of infertility associated with violations of the integrity of sexual chromatin – pathological fragmentation of sperm DNA (FDNAS). One of the main pathophysiological mechanisms of the occurrence of pathozoospermia and FDNAS in secretory male infertility is oxidative stress, which occurs in patients with varicocele. It has been proven that varicocelectomy increases the frequency of spontaneous pregnancy and improves the results of IVF-procedures. However, there are currently insufficient studies devoted to the comparative evaluation of the effectiveness of various methods of varicocelectomy in patients with pathological FDNAS.&#x0D; AIM: To conduct a comparative assessment of the dynamics of sperm DNA fragmentation after subinguinal and laparoscopic varicocelectomy in male infertility.&#x0D; MATERIALS AND METHODS: The study included the results of examination and treatment of 87 men who complained of infertility in marriage for 1–9 years (on average, 3.8 ± 2.7 years). The age of the patients was 24–42 years (the average, 28.6 ± 11.3 years). All patients were diagnosed with grade II left-sided varicocele. Other pathological conditions potentially affecting ejaculate parameters were excluded. All patients underwent ejaculate analysis — sperm analysis, determined the degree of FDNAS by TUNEL method and the level of IgG class antisperm antibodies in the ejaculate by direct Mar-test. The patients were divided into 2 groups by randomization. The group 1 included 41 patients who underwent subinguinal varicocelectomy (Marmar operation), the group 2 included 46 patients who underwent laparoscopic varicocelectomy. Studies of the ejaculate — a sperm-analysis (according to WHO criteria, 2010), a MAR test and FDNAS — were carried out before, 3 and 6 months after surgery.&#x0D; RESULTS: In all patients of both groups in six months after surgical correction of varicocele the concentration of spermatozoa, the number of progressively mobile and normal forms of spermatozoa (according to the strict Kruger criteria) were significantly increased. The MAR test value in all patients of both groups before surgery did not exceed 10%. There were no statistically significant changes in this indicator in the postoperative period. FDNAS values in patients of group 1 before and 3 months after surgical treatment no statistically significant differences were found (27.6 ± 6.7% and 22.4 ± 8.3%, respectively, p 0.1). In patients in group 2, 3 months after surgical treatment a statistically significant decrease in the FDNAS value was noted compared with that before treatment (from 26.1 ± 8.9% to 13.3 ± 7.7%, p 0.001). 6 months after surgery, patients in both groups showed a significant (p 0.001) decrease in FDNAS values compared to the value before treatment: up to 14.6 ± 7.8% in patients of group 1 and up to 12.1 ± 8.0% — in patients of group 2.&#x0D; CONCLUSIONS: 1. Performing subinguinal and laparoscopic varicocelectomy contributes to the significant improvement of the main parameters of sperm in six months after the surgical operation. 2. The DNA fragmentation level significantly decrease already in 3 months after laparoscopic varicocelectomy, and just in 6 months after Marmar operation.

Reproductive system status in men after coronavirus infection and vaccination

Introduction The COVID-19 pandemic is a serious problem for the health care system in Russia and worldwide, as the pathogenic influence of the pathogen can cause a systematic range of complications in the body, especially in the reproductive system.The aim of the work was to determine the state of the reproductive system in men after coronavirus infection and vaccination based on the analysis of literature data.Materials and methods The search for papers on the state of the reproductive system in men after coronavirus infection and vaccination was conducted in PubMed, Medline, EMBASE and eLibrary.ru databases.Results The reproductive capacity of male patients who have undergone COVID-19 is affected by numerous factors: general health status, activity of the renin-angiotensin-aldosterone system, development of systemic inflammatory response, activation of lipoperoxidation processes, etc. This changes cause inflammatory response in the tissues of the reproductive system, secretion of pro-inflammatory cytokines, imbalance of the antioxidant system, formation of oxidative stress, damage to Leydig and Sertoli cells, formation of reproductive insufficiency and infertility.Discussion The pathogenetic role of SARS-CoV-2 in the development of reproductive depression in men with coronavirus infection, which has a prolonged, negative and persistent effect on the state of testicular hemodynamics, the level of total testosterone in blood and ejaculate parameters, was confirmed.Conclusion The results of the study suggest a deterioration of the reproductive system in men following coronavirus infection.

The prevalence of social risk factors for the development of male infertility: smoking, alcohol and narcotic use in men from infertile couples, the influence of the “northern type” of alcohol consumption on ejaculate indicators

Introduction. Lifestyle factors, including smoking, alcohol (AU) and drug use (DU), can affect male fertility. The aim of the study was to investigate the frequency and characteristics of smoking, AU and DU in men from infertile couples, to identify the most significant associations of disorders of spermatogenesis and social risk factors.Material and methods. At the 1st stage of the study, the case histories of 1198 men from infertile couples were analyzed to determine the frequency of smoking, AU and DU, at the 2nd stage, a more detailed questionnaire of 239 patients from the general sample was conducted for detailed characteristics of smoking and/or AU and/or DU, at the 3rd stage, a comprehensive analysis of ejaculate from men who consumed strong alcohol, but did not smoke or use DU (n = 46) was performed in comparison with men without bad habits taken into account (n = 60).Results. In the general sample, the frequency of AU was 73 %, smoking – 41 %, DU –17 % (mostly in anamnesis). 47.9 % of AU men consumed beer (1.5 (1–2.5) liters per week, 25.6 % – strong alcohol (250 (100–500) ml per week), 7.6 % – champagne/wine (500 (250–725) ml per week); 92.7 % of smokers used cigarettes, 7.3 % – electronic cigarettes (smoking experience was 15 (10–20) years, the number of cigarettes per day was 15 (10–20) pieces); more than 90 % of DU men have a history of non-injection DU. Men who consumed only strong alcohol, but did not smoke and did not use drugs, increased sperm DNA fragmentation compared to men without bad habits taken into account: 16.0 (13.5–19.6) and 12.8 (8.8–19.4) %, respectively, p = 0.018.Conclusions. Among men from infertile couples, a high frequency of AU was revealed with the predominant use of beer or strong alcohol, the use of the latter increases sperm DNA fragmentation; smoking is characterized by a long experience; active DU is rare. Given the specifics of the requirements for the state of reproductive health of men applying to reproductive medicine centers, it is important to have knowledge about the frequency and severity of smoking, AU and DU among them, as well as about the relationship of risk factors of infertility with the parameters of ejaculate.

O-015 Spontaneous alterations in semen parameters are associated with age, accessory gland function and the FSHB c.-211G&amp;gt;T variant

Abstract Study question Which parameters are associated with spontaneous alterations in semen analysis in infertile men? Summary answer Changes in semen parameters are modulated by abstinence time, accessory gland function and fluctuations in FSH serum concentration, which is bound to FSHB c.-211G&amp;gt;T variant. What is known already There is a strong within-subject alteration of semen parameters in men with infertility. The WHO therefore recommends two ejaculate analyses. However, it remains unknown in which subgroup variations are likely to occur and which semen parameters are affected. We assume that in some men with idiopathic infertility, ejaculate parameters may spontaneously change without interventional medication apart from regression to the mean effects. The study aims to identify factors in conventional semen analysis based on the patients’ clinical data, including reproductive hormones like follicle stimulating hormone (FSH), and factors impacting endocrine activity, like the FSHB-211 promoter gene. Study design, size, duration In this retrospective longitudinal study we selected 3456 deeply phenotyped men with idiopathic infertility who had visited our centre for fertility workup from 2010-2020. The patients were selected from our well curated database Androbase© for analysis of repeated ejaculate samples. Participants/materials, setting, methods Men with idiopathic infertility and at least two ejaculate analyses (70-90 days distance) were included. Exclusion criteria comprised: sperm concentration &amp;lt;1 million/ml, abnormal FSH or low testosterone, bitesticular volume (bTV) &amp;lt;10ml, orchiectomy, cryptorchidism, varicocele testis, oncological disease, genetic findings (i.e. abnormal karyotype, AZF deletions), medication affecting spermatogenesis. Grouped linear 2-level nested mixed-effect models were applied. Analyzed parameters included: somatic, hormonal (including FSH and FSHB c.-211 variants), and semen parameters (including abstinence time, accessory gland markers). Main results and the role of chance Groups were built according to sperm concentration: A (n = 397): ≥1.0 to &amp;lt; 5.0 mill/ml, B (n = 708): ≥5.0 to &amp;lt; 15.0 mill/ml, C (n = 2351): ≥15.0 mill/ml. A, B and C: changes in ejaculate volume were associated with alterations in total sperm count and motility (p &amp;lt; 0.003). Changes were, controlled for abstinence time (p &amp;lt; 0.001), related to alpha-glucosidase, fructose or zinc (p = 0.005-0.02). A+B: fluctuations in FSH concentration influenced sperm concentration/count (p = 0.004-0.02), albeit only in men with FSHB c.-211 GG (p = 0.007-0.02). T-allele carriers did not show changes in FSH concentration (p &amp;gt; 0.1). B: age &amp;lt;50 years (p = 0.007-0.01) and normal bTV (p = 0.008-0.02) were associated with spontaneous increases of sperm concentration, count and motility. Semen parameters exhibit intra-individual alterations associated with organic, hormonal and genetic variables. Changes are pronounced in younger men with normal bTV and oligo- to almost normozoospermia. The effect is modulated by abstinence time, accessory gland function and fluctuations in FSH concentrations which, in turn, are bound to the FSHB-211 promotor gene polymorphism. Limitations, reasons for caution A limitation of our approach is the retrospective nature of the study, which cannot exclude the possibility of bias within the data set. Wider implications of the findings Judgement of semen analysis should be based on two semen samples, with abstinence time between 4-5 days. In future clinical trials, abstinence time should be kept constant. It might be investigated if younger men with normal bTV and FSHB c.-211 GT/TT benefit from improving accessory gland function and increasing FSH. Trial registration number not applicable

Correlation of resistive index of prostatic capsular artery to clinical symptoms in men with chronic prostatitis/chronic pelvic pain syndrome.

To evaluate the relation between resistive index (RI) of prostatic capsular arteries by transrectal Doppler ultrasonography in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and its correlation with lower urinary tract symptoms, erectile dysfunction and premature ejaculation parameters of CP/CPPS. In total, we included 68 patients presenting with chronic prostatitis/chronic pelvic pain syndrome. We formed two groups, as Group 1 of 35 patients with a RI ≥ 0.7 and Group 2 of 33 patients with RI < 0.7. All patients were assessed with International Prostate Symptom Score (IPSS), International Index of Erectile Function (IIEF-5), premature ejaculation diagnostic tool (PEDT) and National Institutes of Health Chronic Prostatitis Symptom Index (CPSI). In addition, the RI of the prostate capsular artery was measured in all patients using Doppler ultrasound. Statistical analyses were performed with SPSS version 18. A p value < .05 was considered significant. Demographic characteristics were similar in the two groups. IPSS was 11.3 ± 6 in Group 1 and 9.7 ± 5.3 in Group 2. IIEF-5 was 18.6 ± 2 in Group 1 and 20.4 ± 2.3 in Group 2. PEDT was 12.4 ± 5.6 in Group 1 and 11.2 ± 4 in Group 2. CPSI (total) was 19.3 ± 12.3 in Group 1 and 10.6 ± 7.7 in Group 2. There was significant statistical difference in IPSS, IIEF-5, and CPSI between the two groups (p < .001, p < .001, p < .001 respectively). However, we found no significant difference in PEDT between the two groups (p = .19). There is a significant correlation between the lower urinary tract symptoms and erectile dysfunction parameters and the RI of the prostatic capsular artery in CP/CPPS and RI is an effective and noninvasive method to assess the severity of the disease.

Peculiarities of the formation of sperm production and the course of peroxide oxidation in boars

The intensive use of boars and receiving biologically complete valuable ejaculates from them is possible due to early assessment of the quality of sperm production and timely determination of their genetic potential. The research aimed to determine the peculiarities of the formation of sperm production and the course of peroxide oxidation in boars during puberty. For the experiment, 5–10 month-old boars of Poltava Meat, the Large White, and Myrhorod breeds were used. The quality of the obtained ejaculates was evaluated according to standard indicators according to the instructions for artificial insemination of pigs. The research results indicate a significant increase in the quantitative indexes of sperm production in young boars in the age range with different features for each breed. By the eighth month of life, a general tendency to increase the parameters in the received ejaculates is noted. In contrast, the highest increase in ejaculate volume and sperm concentration was found in young boars of the sebaceous direction of productivity on the 240th day of the development (4.6 and 1.9 times). The minimum motility and survival of gametes were observed at five months, regardless of the breed, with a gradual increase to maximum values at the seventh and eighth months of age, depending on the direction of animal productivity. The use of moderate sexual load for young boars after they reach the age of 9 months ensures the functional activity of sperm suitable for artificial insemination (motility above 80 % and survival above 70 %). The significant changes in the pro-oxidant-antioxidant homeostasis in the sperm of boars of 7–8 months occur in the direction of the acceleration of the peroxygen oxidation of lipids, which is evidenced by an increase in the content of diene conjugates and TBC-active compounds and an increase in the activity of enzymes of the antioxidant link. An earlier formation of pro-oxidant-antioxidant homeostasis in this secretion is noted in representatives of the meat and fat direction of productivity. Thus, the obtained results indicate significant age-related changes in the ejaculate parameters and peculiarities of the course of peroxide oxidation processes depending on the breed of boars.

Low frequency neuromuscular electrical stimulation applied to the bulbospongiosus muscle prolongs the ejaculation latency in a rat model.

Premature ejaculation (PE) is common, but its true pathophysiology is not clear, and treatments are limited. We aimed to investigate the effect of neuromuscular electrical stimulation applied in different modes and frequencies to the bulbospongiosus muscle on ejaculation parameters. In this study, 24 male Wistar albino rats were used. The rats were equally divided into three groups: control, high-frequency burst (HFB) and low-frequency (LF) (n = 8 each). Neuromuscular electrical stimulation was applied to the rats for 30 min. In the HFB group, this stimulation was applied in the burst mode at 80 Hz frequency using 200 microseconds (µs) transition time. In the LF group, manual stimulation was applied using a 2 Hz frequency and 200 µs transition time. Following the intraperitoneal administration of para-chloroamphetamine at a dose of 5 mg/kg, ejaculation time, increase in basal seminal vesicle pressure, seminal vesicle maximum pressure, number and interval time of seminal vesicle contractions and bulbospongiosus muscle electromyography activities were evaluated over 30 min. There was a significant difference between the groups in terms of ejaculation time (p = 0.002). The ejaculation times of the LF, HFB and control groups were 1344.71 ± 105.9, 908 ± 62 and 672 ± 149.7 s, respectively. The post hoc analysis revealed that the ejaculation time of the LF group was significantly longer than that of the HFB and control groups (p = 0.033 and p = 0.001, respectively). The remaining parameters did not differ significantly between the groups. The results showed that low-frequency (2 Hz) electrical stimulation applied to the male rats significantly prolonged the ejaculation time. It is thus considered that applying neuromuscular electrical stimulation before planned sexual activity can prevent the rhythmic contractions necessary for completing the ejaculatory process by maintaining subtetanic continuous contraction and prolonging the ejaculation time in patients with premature ejaculation complaints.

Bilateral orchiectomy deteriorates the structure and function of seminal vesicles in a rat model.

The effects of hormone levels on ejaculation are known. In addition to thyroid hormone levels, testosterone levels are also associated with ejaculation, but no consensus has been reached on this issue. Thus, we investigated the effect of decreased testosterone levels due to bilateral orchiectomy on the chemical stimulation-induced ejaculation phases in rats. Twenty-one male Wistar rats were randomized into the orchiectomy, sham, and control groups, with seven rats in each group. Bilateral orchiectomy was performed. The ejaculation parameters were evaluated 5 days after the sham and bilateral orchiectomy operations and the waiting period in the control group. The seminal vesicle (SV) phasic contraction number and increase in basal pressure amplitude were significantly lower in the orchiectomy group (6.9 ± 3.3 and 0.6 ± 0.3 mmHg) than in the sham and control groups (11.2 ± 1.7 and 1.0 ± 0.4 mmHg, and 14.5 ± 6.6 and 1.1 ± 0.2 mmHg, respectively; p = 0.016 and p = 0.03, respectively). The interval between the SV contractions was significantly longer in the orchiectomy group (166.2 ± 104.3 s) than in the sham and control groups (76.0 ± 15.5 s and 63.1 ± 31.1 s, respectively; p = 0.014 (between groups), orchiectomy vs sham p = 0.040 and orchiectomy vs control p = 0.018). The SV weights of the rats were significantly lower in the orchiectomy group (0.14 ± 0.01 g) than in the sham and control groups (0.37 ± 0.05 g and 0.48 ± 0.03 g respectively; p < 0.0001 (between groups), orchiectomy vs sham p < 0.0001 and orchiectomy vs control p < 0.0001). The groups showed no significant differences in ejaculation time, SV basal pressure, SV maximum amplitude, and bulbospongiosus muscle contraction electromyographic activity. Our results partially clarified the relationship between decreased testosterone levels and ejaculation. Decreased testosterone levels caused statistically significant changes in SV functions and affected the ejaculation emission phase.

The efficacy of novel centrifugation-free sperm selection (Io-Lix) on sperm parameters and ICSI reproductive outcomes

Research questionWhat is the effect of a novel non-centrifugation method (Io-Lix) of sperm selection on sperm parameters and intracytoplasmic sperm injection (ICSI) reproductive outcomes? DesignThis pilot study elevated the capacity of the Io-Lix sperm selection protocol to improve sperm parameters (concentration, motility and sperm DNA fragmentation) of the neat ejaculate. Once established, the reproductive outcomes of Io-Lix selected spermatozoa were used for autologous and donor oocyte ICSI programmes and their efficacy compared with those using conventional swim-up. ResultsIo-Lix sperm selection resulted in lower sperm concentration yield (P < 0.001) and sperm DNA fragmentation (P < 0.001) but higher sperm motility (P < 0.001) when compared with spermatozoa in the neat ejaculate. When compared with swim-up sperm selection the Io-Lix protocol resulted in a 14.7% (P = 0.028) increase in pregnancy rate and 16.3% (P = 0.047) reduction in miscarriages in the autologous ICSI programme. A similar comparison of sperm selection procedures employed for a donor oocyte ICSI programme showed no difference in terms of their respective reproductive outcomes. ConclusionsThe Io-Lix sperm selection protocol resulted in improved pregnancy rate and reduction in miscarriage when applied to autologous ICSI, which was attributed to a reduction in the proportion of spermatozoa with DNA damage post-selection. A similar finding was not apparent in the donor oocyte programme, which may be associated with the capacity of the donor oocyte to repair sperm DNA post-syngamy.

Effects of virtual reality erotica on ejaculate quality of sperm donors: a balanced and randomized controlled cross-over within-subjects trial

BackgroundPrevious research has shown that the type and duration of erotic material that men have access to during masturbation can influence semen parameters. To our knowledge, the use of virtual reality (VR) headsets to present erotica has not previously been studied. We reasoned that, because VR can provide a more immersive experience to the user, semen parameters of masturbatory ejaculates may be altered.MethodsThis study had a balanced and randomized controlled cross-over within-subjects design. 504 ejaculates were collected from 63 sperm donors at 4 locations in Denmark. During masturbation each donor was instructed to observe erotic material either on a touch screen monitor or using a VR headset. The order of each pair of within-subject treatments was randomized by the throw of a dice. Anonymized data were analysed with linear mixed and piecewise structural equation models.ResultsBoth abstinence period and VR-use influenced the total number of motile spermatozoa ejaculated. For short abstinence periods, VR-use increased the number of motile sperm in the ejaculate. However, the difference between VR and non-VR ejaculates decreased as abstinence period increased such that there was no difference at the mean abstinence period of 58 h. For longer abstinence periods, total motile sperm counts were lower, on average, when men used VR compared to those that did not.ConclusionThe use of VR headsets to view erotica had a strong positive effect on the number of motile sperm in an ejaculate when the donor’s abstinence time was short (< 24 h). VR-use could improve the ejaculate quality of men who are asked to provide samples after a short period of abstinence, such as men in infertile partnerships producing samples for ART or cancer patients depositing sperm before treatment.Trial registrationTrial retrospectively registered on 13 July 2022 at ClinicalTrials.gov. Identifier: NCT05457764.

Biometric analysis spermograms of men of different age groups in normal and pathology in Lviv region, Ukraine

Background. Analysis of the causes of infertility revealed an increase in the proportion of male factors over the past 20 years by an average of 10–12 %. World Health Organization data, which summarize the observations of many authors in recent years, show a decrease in male reproductive potential. The ejaculate parameters of the modern average man are steadily declining. For example, the concentration of germ cells in the seminal fluid has decreased three times, and this trend continues: the concentration of sperm decreases annually by about 2 %, and their motility – by 1.5 %. Not only is infertility a pressing problem in modern medicine, but also an important social issue, as it is associated with the mental state of men and the preservation of the family. Materials and Methods. Spermograms were obtained during the study of patients in the clinic of reproductive medicine “Alternative Clinic” (Lviv, Ukraine). A total of 50 men were surveyed, including 16 men aged 20 to 29 (junior age group), 17 men aged 30 to 39 (middle age group) and 17 men 40 to 49 years old (senior age group), respectively. We found that out of the 16 men in the junior group, 4 spermograms corresponded to asthenozoospermia, and 12 – normozoospermia, of the 17 middle-aged group, 7 spermograms corresponded to asthenozoospermia, and 10 – normozoospermia and 10 men of the senior group to 15 spermograms asthenozoospermia, 2 – normospermia. Spermograms were evaluated by the following indicators: ejaculate count, viscosity, sperm count per 1 mL of ejaculate, sperm motility in categories “A” and “B”, the number of morphologically normal and morphologically degenerative sperm, Farris index. For control we took spermograms of patients with normozoospermia. Results. Analyzing spermograms according to the studied indicators, we found a decrease in the amount of ejaculate in older men relative to the norm, which may indicate a lack of function of the seminal vesicles, prostate. According to the criteria for the number of sperm in 1 ml of ejaculate and the total number of sperm in the whole ejaculate, it was shown that in the spermograms of patients with asthenozoospermia of different age, these figures are much lower than normal. This indicates a decrease in male fertility in these diseases. Examining sperm motility according to the following criteria: “A” – fast translational movements and “B” – slow, sluggish translational movements, we found a decrease in these indicators compared with the norm in men of all ages with asthenozoospermia. This indicates a decrease in the quality of ejaculate in men, and, consequently, reduce the likelihood of fertilization. Examining sperm morphology, we assessed the number of normal and degenerative sperm. We have shown that in men of all ages with asthenozoospermia, the number of morphologically normal sperm is lower than in the control, and the number of morphologically degenerative sperm is increasing. Probably, such pathology can lead to a decrease in the possibility of fertilization and increases the likelihood of malformations in the fetus, if fertilization has occurred. Evaluating the spermograms of men in the study groups according to the Farris index, we found a significant reduction in this number in men with asthenozoospermia. This indicates a low probability of fertilization. Conclusions. Male infertility (in particular, asthenozoospermia) can be caused by many different diseases in which there are deviations from the norms of qualitative and quantitative indicators of ejaculate. According to the data obtained from the observation of men, there is a tendency for the reduction of ejaculate volume, concentration and motility of sperm, as well as their morphological status.