The Association of HLA-G Gene Polymorphism and Its Soluble Form With Male Infertility

Successful reproduction depends on many factors. Male factors contribute to infertility in approximately 50% of couples who fail to conceive. Seminal plasma consists of secretions from different accessory glands containing a mixture of various cytokines, chemokines, and growth factors, which together can induce a local immune response that might impact on a male’s as well as a female’s fertility. Human leukocyte antigen (HLA)-G expression has been suggested as an immunomodulatory molecule that influences pregnancy outcome. The HLA-G gene encodes either membrane-bound or/and soluble proteins. The aim of this study was the evaluation of HLA-G polymorphisms and their impact on soluble HLA-G (sHLA-G) production. We tested the HLA-G polymorphism in three positions: rs1632947: c.-964G>A; rs1233334: c.-725G>C/T in the promoter region; rs371194629: c.∗65_∗66insATTTGTTCATGCCT in the 3′ untranslated region. We tested two cohorts of men: 663 who participated in in vitro fertilization (test material was blood or sperm), and 320 fertile controls who possessed children born after natural conception (test material was blood). Since 50% of men visiting assisted reproductive clinics have abnormal semen parameters, we wondered if men with normal sperm parameters differ from those with abnormal parameters in terms of HLA-G polymorphism and secretion of sHLA-G into semen. We found that certain rs1632947-rs1233334-rs371194629 HLA-G haplotypes and diplotypes were associated with male infertility, while others were protective. Normozoospermic men with the A-C-del haplotype and A-C-del/A-C-del diplotype secreted the most sHLA-G into semen (574.1 IU/mL and 1047.0 IU/mL, respectively), while those with the G-C-ins haplotype and G-C-ins/G-C-ins diplotype – the least (80.8 IU/mL and 75.7 IU/mL, respectively). Men with the remaining haplotypes/diplotypes secreted sHLA-G at an intermediate level. However, only in one haplotype, namely G-C-ins, did we observe strong significant differences in the concentration of sHLA-G in the semen of men with teratozoospermia compared to men with normal sperm parameters (p = 0.009). In conclusion, fertile men differ in the profile of HLA-G polymorphism from men participating in IVF. Among all HLA-G haplotypes, the most unfavorable for male fertility is the G-C-ins haplotype, which determines the secretion of the lowest concentration of the soluble HLA-G molecule. This haplotype may reduce sperm parameters.

SEMEN ANALYSIS; CORNER STONE IN EVALUATING INFERTILITY

Objective: To study the patterns and distribution of various abnormal semen parameters in infertile males. Study Design: Cross sectional study. Place and Duration of Study: Department of Pathology, Combined Military Hospital, Karachi, from Nov 2019 to Oct 2020. Methodology: The study included 364 patients who presented with primary and secondary infertility. Consecutive convenient sampling was done. Semen analysis was performed using World Health Organization latest guidelines. Samples were categorized as normospermia, azoospermia, oligospermia, asthenozoospermia and necrospermia. Results: The study comprised of 364 samples of infertile males. Normal sperm count was observed in 317 (87%) males, azoospermia in 28 (7.6%) and oligospermia in 19 (5.2%) males. Low ejaculated volume and higher non-motile sperms were noted in oligospermia samples in comparison with normospermia samples. Asthenozoospermia was observed in 102 (28%) and oligoasthenospermia was noted in 15 (4.1%) samples. Conclusion: Good quality semen analysis is a corner stone to diagnose the cause of male infertility.Sperm concentration and motility are the important markers of normal male reproductive system and are related to each other.

Comparative Study of Abnormalities of the Spermocytogram between Manual Techniques versus Automated SCA (Sperm Class Analyzer)

It is already well established that the percentage of normal sperm and specific sperm abnormalities has diagnostic value in vivo. The spermocytogram is an important part of the analysis of human semen, this kind of analysis, simple at first sight, brings real difficulties because the results from one laboratory to another are very relatively reliable. When analyzing abnormal sperm morphology, the observer has to estimate the respective size of the different spermatozoa or their components, the length of all the spermatozoa (too small or too big) and to identify the shape (sperm with a coiled flagellum, or no flagellum as an example). The objective of our work is first of all to make a comparative study of the results of the morphological evaluation of 50 semen samples between the manual technique versus the automated one (SCA) in order to find a possible correlation between these two techniques. According to the analytical stage of the manual and automatic results, the two latter led us to obtain significantly different percentages.

Establishment of semen collection technique using electroejaculator and semen cryopreservation of Javan leopard (Panthera pardus melas Cuvier, 1809)

Background and Aim: The Javan leopard (Panthera pardus melas Cuvier, 1809) is a subspecies of Panthera pardus spp., spread across the African and Asian regions. Information on reproductive aspects is crucial for wild animals, including the Javan leopard. In this study, we aimed to develop electroejaculator (EE) techniques and evaluate cryopreservation success in Javan leopard semen. Materials and Methods: The semen of four adult Javan leopards was collected once a week using EE. Placement of the EE probe in the rectum was performed after ultrasound imaging (ultrasonography) to determine the prostate body location. The semen obtained was then evaluated macroscopically and microscopically. Three Javan leopards were used for cryopreservation. The ejaculate was divided into two parts [i.e., one part diluted with AndroMed® (Minitüb, Tiefenbach, Germany) and the other part with Steridyl® (Minitüb, Tiefenbach, Germany)] at a 1:1 ratio immediately after collection and evaluation. The semen was then packed in a 0.25 mL MiniStraw® (Minitüb, Tiefenbach, Germany) then equilibrated at 4°C for 2 h. After equilibration, the straw was then frozen in liquid nitrogen vapor. Frozen semen was then stored in containers until further evaluation. Results: The results showed that ejaculation response occurred at all levels of stimulation, while erections did not always occur. The fastest ejaculation and erection occurred at the fourth voltage. The macroscopic evaluation showed that the semen volume was 0.80±0.26 mL, cloudy white, pH 7.44±0.14, and with watery semen consistency. The microscopic evaluation showed that the sperm motility was 66.98±0.39%, with sperm viability of 75.6±1.79%. Sperm concentration was 62.17±46.95×106 mL–1 with a total concentration of 42.14±23.51×106 cells. Normal sperm morphology is only 40.72±6.26%. Conclusion: This study concluded that the development of a semen collection technique using an EE preceded by imaging of the EE probe location using ultrasound was effective for the ejaculation of Javan leopards. The characteristics of the semen of the Javan leopard showed moderate semen volume, sperm motility, and viability. Javan leopard showed low sperm concentration and normal sperm morphology.

Sheng Jing Decoction Can Promote Spermatogenesis and Increase Sperm Motility of the Oligozoospermia Mouse Model

Sheng Jing Decoction (SJD), as a traditional Chinese medicine prescription, is mainly be used to treat male infertility. However, the pharmacological functions and molecular mechanisms of SJD are poorly understood. In this study, we investigated the functions of SJD on spermatogenesis and sperm motility and explored the potential mechanisms involved. Here, we demonstrated that high, medium, and low doses of SJD are effective in restoring the impairments of the whole body and testicular tissue by cyclophosphamide inducing and to rescue the damage of testicular tissue cells including Sertoli cells and germ cells. SJD can partly restore the decrease in sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate in oligozoospermic mouse models. Ki67 staining analyses confirm SJD can promote testicular tissue cell proliferation. Real-time RT-PCR analyses also reveal that SJD can upregulate the expression of proliferation-associated gene Lin28a and differentiation-associated genes Kit, Sohlh2, and Stra8. SJD can also reduce the impairment of mitochondrial membrane potential (MMP) and sperm plasma membrane integrity by cyclophosphamide inducing. Our results reveal that SJD is effective in improving both sperm quantity and quality by increasing the sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate. SJD can promote spermatogenesis by upregulating the expression of the proliferation-associated gene Lin28a and the differentiation-associated genes (Kit, Sohlh2, and Stra8). SJD can sustain MMP and sperm plasma membrane integrity to increase sperm motility.

An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section I

As the use of assisted reproductive technologies (ART) and in vitro embryo production (IVP) expand in the equine industry, it has become necessary to further our understanding of semen physiology as it applies to overall fertility. This segment of our two-section review will focus on normal sperm parameters, beginning with development and extending through the basic morphology of mature spermatozoa, as well as common issues with male factor infertility in IVP. Ultimately, the relevance of sperm parameters to overall male factor fertility in equine IVP will be assessed.

Kisspeptin and Spexin: lipid profile and sperm parameters in infertile and fertile male subjects

BACKGROUND & OBJECTIVE: Kisspeptin (KP) and spexin (SPX) are neuropeptides that play a substantial role in metabolism and sexual function. Our objective is to explore the association of spexin and kisspeptin with sperm parameters and lipid profile in the infertile male population. METHODOLOGY: The cross-sectional study was conducted from March 2017 till July 2018 in Aga Khan University Hospital, Karachi, Pakistan, after acquiring ethical approval (4813-BBS-ERC-17). Convenient sampling was applied to recruit 44 normozoospermic subjects as controls and 44 patients with abnormal sperm parameters”. Serum levels of kisspeptin and spexin were measured by enzyme linked immunosorbent assay and Cobas c111 analyzer was used for estimation of high and low density-lipoprotein, triglyceride and serum cholesterol levels. Mann-Whitney U tests and spearman’s rank correlation test were used to analyze quantitative variables, and data was presented by means, standard error (SE) and correlations coefficient, with a statistical significance set at p≤0.05. RESULTS: The mean kisspeptin values of ‘altered sperm parameters’ males 13.23 ± 3.34 were lower than normal sperm parameters 31.89 ± 3.25 (p<0.001). The mean spexin values of altered sperm parameters 383.57 ± 21.09 were less than compared to normal sperm parameters 502.76 ± 19.77 (p<0.001). Correlation of KP and SPX levels was (r-value = 0.332, p= 0.002). There was no significant correlation was found between KP and SPX with lipid profile parameters. CONCLUSION: Spexin and kisspeptin levels observed were higher in male subjects with normal sperm parameters and had a significant positive correlation with each other. There was no impact of spexin on lipid profile and obesity.

Clinical significance of sperm morphology in male infertility treatment

Introduction. Sperm morphology is one of the most common tests in fertility practice. The interlaboratory variability is the main drawback of the method. The clinical significance of sperm morphology in assisted reproductive technologies is controversial. The aim of the review is to address this question. Materials and methods. The search of relevant publications was carried out in PubMed and e-Library databases using the keywords «male infertility», «sperm morphology», «teratozoospermia», «IUI», «IVF», «ICSI». Conference abstracts and dissertations were excluded from analysis and 56 publications were included in this literature review. Results. The small numbers of studies were evaluated to the effect of teratozoospermia on the likelihood of natural pregnancy. The pregnancy rate was higher in the group of couples with normozoospermia. In the couples with severe teratozoospermia pregnancies rate was also detected. The most studies did not reveal a statistically significant effect of tertozoospermia on the frequency of pregnancy during intrauterine insemination. The data about the influence of sperm morphology on in vitro fertilization are contradictory. Early studies showed a positive correlation between normal sperm morphology and frequency of conception, but these results were not confirmed in further studies. The most studies have not been revealed the correlation between normal sperm morphology male fertility status, clinical and live birth rate. Discussion. To date the spermatozoa mofophology studying remains the «starting point» ofa man's examination for infertility. However, the data available do not confirm its role in choosing the method ofassisted reproductive technologies or predicting their results. Conclusions. To date the role of sperm morphology on conceive and pregnancy frequencies in ART use is controversal. The data available does not confirm the value of this test as a proxy of higher pregnancy and birth of healthy child probability. The further studies are required to address this question.

Using Data Mining to Assist in Predicting Reproductive Outcomes Following Varicocele Embolization

We carried out a retrospective analysis of infertile couple data using several methodologies and data analysis techniques, including the application of a novel data mining approach for analyzing varicocele treatment outcomes. The aim of this work was to characterize embolized varicocele patients by ascertaining the improvement of some of their clinical features, predicting the success of treatment via pregnancy outcomes, and identifying data patterns that can contribute to both ongoing varicocele research and the more effective management of patients treated for varicocele. We retrospectively surveyed the data of 293 consenting couples undergoing infertility treatment with male varicocele embolization over a 10-year period, and sperm samples were collected before and at 3, 6, and 12 months after varicocele embolization treatment and analyzed with World Health Organization parameters—varicocele severity grades were assessed with medical assessment and scrotal ultrasound, patient personal information (e.g., age, lifestyle, and embolization complications) was collected with clinical inquiries, and varicocele embolization success was measured through pregnancy outcomes. Varicocele embolization significantly improved sperm concentration, motility, and morphology mean values, as well as sperm chromatin integrity. Following this study, we can predict that a male patient without a high varicocele severity grade (with grade I or II) has a 70.83% chance of conceiving after embolization treatment if his partners’ age is between 24 and 33 with an accuracy of 70.59%. Furthermore, male patients successful in achieving pregnancy following embolization are mostly characterized by having a normal sperm progressive motility before treatment, a normal sperm concentration after treatment, a moderate to low varicocele severity grade, and not working in a putatively hazardous environment.

Scrotal thermographic profile and seminal characteristics of Mangalarga Marchador stallions bred in the Atlantic Rainforest biome

ABSTRACT The objective of this study was to determine the scrotal thermographic profile and to verify the influence of temperature and humidity of the humid tropical climate on testicular temperature and seminal quality of Mangalarga Marchador stallions. The thermal profiles of the proximal, middle, and distal zones of the testicles and total surface temperature (TSTT) were recorded using an FLIR E60bx thermal imager. The average air temperature (°C) and relative humidity (%) were obtained 1, 5, 9, 33, and 66 days before semen collection and showed a mean value of 26.5±2.4 and 80.4±6.0 respectively. The scrotal surface temperature was close to 34°C and there was no variation with the age of the stallion, reproductive activity, and characteristics of the ejaculate (P>0.05). The only significant correlations obtained were between TSTT and minor defects (R = 0.41; P<0.05), between TSTT and total defects (R = 0.46; P<0.01), and between TSTT and percentage of morphologically normal sperm (R = -0.46; P<0.05). It was concluded that the Mangalarga Marchador stallions maintained the testicular temperature within favorable conditions for spermatogenesis, demonstrating the efficiency of testicular thermoregulation mechanisms in the Atlantic Forest biome.