Viral infection of sperm. Part 2. Human herpes viruses, human immunodeficiency virus, hepatitis C virus, Zika virus (review) [Вирусное инфицирование сперматозоидов. Часть 2. Герпесвирусы человека, вирус иммунодефицита человека, вирус гепатита С, вирус Зика (обзор литературы)]

Intragametal viral infection of spermatozoa can cause true vertical transmission of viruses through germ cells. Currently, human immunodeficiency virus, hepatitis C viruses, herpes simplex virus, cytomegalovirus, Zika virus have been detected in spermatozoa. The possibility of vertical transmission of human immunodeficiency virus, cytomegalovirus, herpes simplex virus and Zika virus has been proven. Intragametal infection of spermatozoa with viruses of the herpes group leads to abnormalities in the development of the embryo and can cause spontaneous abortions both during natural conception and when using assisted reproductive technologies. The development of adequate methods for diagnosing an intragametal spermatozoa virus infection will make it possible to find out, at least in some patients, the cause of infertility and pregnancy abnormalities and apply appropriate antiviral therapy in preparation for natural conception or the use of assisted reproductive technologies. © 2020 ABC-press Publishing House. All rights reserved

Фенотип сперматозоидов при варикоцеле

The study objective is to microscopically evaluate the morphology of sperm in patients with varicocele after surgical treatment and therapy with Prostatilen® AC.Materials and methods. The treatment group included 20 males between 26 and 45 years of age (mean age 31.6 ± 6.1 years) with various stages of varicocele and fertility problems; among them, 10 patients with subclinical stages of varicocele received conservative treatment with Prostatilen® AC (group 1) and were examined before and after the treatment; 10 males with infertility caused in part by varicocele of the spermatic cord veins were examined 6 months to 3 years after surgical treatment (group 2). Standard clinical and lab tests, sperm analysis, electron microscopy of the ejaculate were performed. The control group included 65 fertile males whose sperm samples were obtained from a bank of reproductive cells and tissues and used for comparison in microscopic examination.Results. In patients who received conservative treatment the number of sperm with immature chromatin decreased (p = 0.045) compared to the control group. This characteristic differed in patients after varicocelectomy and patients after conservative treatment (p = 0.037). Compared to control, the number of sperm with excess residual cytoplasm in the head and neck was higher in patients after varicocelectomy (p = 0.011). After conservative treatment, the number of sperm with excess residual cytoplasm was close to the control number and lower than in patients after varicocelectomy (р = 0.028).Conclusion. In patients with subclinical varicocele, conservative treatment with Prostatilen® AC leads to significant improvement in sperm ultrastructure compared to patients who underwent surgery to treat this pathology.Цель исследования - оценить морфологию сперматозоидов у пациентов с варикоцеле после его хирургического лечения и после терапии «Простатиленом® АЦ» по данным электронно-микроскопического исследования.Материалы и методы. В основную группу были включены 20 мужчин в возрасте 26-45 лет (средний возраст 31,6 ± 6,1 года) с различными стадиями варикоцеле и нарушением фертильности; из них 10 больных с субклинической формой варикоцеле проходили консервативное лечение «Простатиленом® АЦ» (1-я группа) и были обследованы до и после него, 10 мужчин с бесплодием, обусловленным в том числе варикозным расширением вен семенного канатика, обследованы в сроки от 6 мес до 3 лет после хирургического лечения (2-я группа). Проведены стандартные клинико-лабораторные тесты, спермиологическое исследование, электронное микроскопическое исследование эякулята. В контрольную группу вошли 65 фертильных мужчин, образцы спермы которых были получены из банка репродуктивных клеток и тканей и использованы при сравнении результатов микроскопического исследования.Результаты. У пациентов, прошедших консервативную терапию, содержание сперматозоидов с незрелым хроматином было меньше, чем у мужчин контрольной группы (p = 0,045). Различались значения этого показателя и у пациентов 1-й и 2-й групп (после консервативной терапии и после варикоцелэктомии) (p = 0,037). По сравнению с контролем было выше содержание сперматозоидов с избыточной остаточной цитоплазмой на головке и на шейке у пациентов, перенесших варикоцелэктомию (p = 0,011). После консервативной терапии содержание сперматозоидов с избыточной остаточной цитоплазмой было практически равным таковому в контроле и было меньше, чем у пациентов после варикоцелэктомии (р = 0,028).Заключение. У пациентов с субклинической формой варикоцеле после консервативной терапии «Простатиленом® АЦ» происходит статистически значимое улучшение ультраструктуры сперматозоидов по сравнению с таковой у пациентов, оперированных по поводу данной патологии

Sperm phenotypes in varicocele [Фенотип сперматозоидов при варикоцеле]

The study objective is to microscopically evaluate the morphology of sperm in patients with varicocele after surgical treatment and therapy with Prostatilen® AC. Materials and methods. The treatment group included 20 males between 26 and 45 years of age (mean age 31.6 ± 6.1 years) with various stages of varicocele and fertility problems; among them, 10 patients with subclinical stages of varicocele received conservative treatment with Prostatilen® AC (group 1) and were examined before and after the treatment; 10 males with infertility caused in part by varicocele of the spermatic cord veins were examined 6 months to 3 years after surgical treatment (group 2). Standard clinical and lab tests, sperm analysis, electron microscopy of the ejaculate were performed. The control group included 65 fertile males whose sperm samples were obtained from a bank of reproductive cells and tissues and used for comparison in microscopic examination. Results. In patients who received conservative treatment the number of sperm with immature chromatin decreased (p = 0.045) compared to the control group. This characteristic differed in patients after varicocelectomy and patients after conservative treatment (p = 0.037). Compared to control, the number of sperm with excess residual cytoplasm in the head and neck was higher in patients after varicocelectomy (p = 0.011). After conservative treatment, the number of sperm with excess residual cytoplasm was close to the control number and lower than in patients after varicocelectomy (р = 0.028). Conclusion. In patients with subclinical varicocele, conservative treatment with Prostatilen® AC leads to significant improvement in sperm ultrastructure compared to patients who underwent surgery to treat this pathology. © 2019 Andrologia i Genital'naa Hirurgia. All rights reserved

High-throughput analysis of TUNEL-stained sperm using image cytometry

Sperm DNA integrity is an essential factor for successful fertilization and proper pregnancy progression. The terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay is commonly used for the estimation of the DNA fragmentation index. Analysis of TUNEL-stained sperm is often performed by flow cytometry, an approach that allows high-throughput analysis but in which any morphological information is lost. In this study, results of an automated image cytometry estimation of TUNEL-stained sperms were presented. The results of visual counting and automatic analysis were closely correlated, indicating that image cytometry is suitable for such analysis and may be applied in a clinical setting. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometr

High-throughput analysis of TUNEL-stained sperm using image cytometry

Sperm DNA integrity is an essential factor for successful fertilization and proper pregnancy progression. The terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay is commonly used for the estimation of the DNA fragmentation index. Analysis of TUNEL-stained sperm is often performed by flow cytometry, an approach that allows high-throughput analysis but in which any morphological information is lost. In this study, results of an automated image cytometry estimation of TUNEL-stained sperms were presented. The results of visual counting and automatic analysis were closely correlated, indicating that image cytometry is suitable for such analysis and may be applied in a clinical setting. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometr

Evaluation of the Rotaprost mineral-herbal complex effectiveness for improvement of functional condition of the male urogenital system [Оценка эффективности минерально-растительного комплекса Ротапрост для улучшения функционального состояния мочеполовой системы мужчины]

Introduction. Modern studies indicate a close relationship between the deterioration of the quality of the ejaculate and the increase in the symptoms of the lower urinary tract (LUTS) with the increase in the age of the man. Taking into account the common pathogenesis of reproductive dysfunction, LUTS and the development of possible preventive measures, we conducted an independent study. The study objective was to evaluate the effectiveness of the combined dietary supplement Rotaprost to improve the functional state of the male genitourinary system. Materials and methods. The study included 30 patients with clinical and laboratory signs of infertility and LUTS aged 23-65 years (mean age 44.91 ± 4.5). Patients were divided into two groups of 15 patients. Group 1 with chronic pelvic pain syndrome (CPPS) of IIIb category (US National Institutes of Health, 1995) with secretory type of infertility and LUTS, group 2 of 15 patients with stage 1 benign prostatic hyperplasia (BPH), pathozoospermia and LUTS. During the study, clinical and laboratory parameters were registered, NIH-CPSI, IPSS-QoL, IIEF-5, NRS questionnaires were used, spermogram data, MAR test, oxidative stress, DNA fragmentation, and electron microscopy of spermatozoa were evaluated. All patients before and after treatment underwent triplex ultrasound of the scrotum and transrectal ultrasound of the prostate. Expert-class devices E-CUBE 15 (Alpinion) were used. Patients included in the design of the study took Rotaprost capsules No. 30, 1-2 times a day for 30 days at the place with meals, drinking a small amount of water. After the course of treatment, patients were examined again. Results. The results of the study showed a more significant effect on the parameter of progressive mobility after treatment in the group with CPPS IIIb (group 1) by 23.5 % compared to the BPH group, where the total increase was 10.7 %. Also, an interesting finding was the trend towards a decrease in the number of leukocytes in the semen while taking Rotaprost in patients with CPPS IIIb (from 1.22 million/ml to 0.43 million/ml) compared with the BPH group, where changes are less significant (from 0.68 million/ml to 0.36 million/ml). Comparing the level of reactive oxygen species in the ejaculate and assessing the DNA fragmentation of spermatozoa revealed a statistically significant decrease during treatment in both groups, reducing the amount of residual urine. The drug had a significant effect on the level of pain, assessed by the NRS scale and improved the quality of life, including IPSS-QoL and NIH-CPSI data. Conclusions. Rotaprost mineral-herbal complex allows to correct moderate pathozoospermia and indicators of impaired urination in the vast majority of patients with CPPS IIIb and men with stage 1 BPH. The use of the Rotaprost mineral-herbal complex for a month leads to an improvement in the quality of life of patients with CPPS IIIb, reducing their pain by 25 %, restoring the quality of urination, and reduce the prostate volume of men with stage 1 BPH. High safety and adherence to therapy in the studied groups of patients is explained by the general pathogenetic mechanisms of the development of these diseases in men over 40 years of age and the possibility of a complex effect on the key pathophysiological cellular mechanisms of age-related changes in the genitourinary system of men by multifactorial components of modern dietary supplements. © 2021 Cyrillo-Methodian Research Centre of BAS. Anales de Investigación,. All rights reserved

Peptide bioregulator amprost® in the correction of pathospermia in various types of secretory infertility [Пептидный биорегулятор Сампрост® в коррекции патоспермии при различных видах секреторного бесплодия]

The aim of the study was to evaluate the clinical and laboratory effectiveness of the peptide bioregulator Samprost® in the correction of pathospermia in men with secretory type of infertility caused by chronic nonbacterial prostatitis (CNP) and varicocele in the postoperative period. Materials and methods. The multicenter study included 30 patients between 23 and 51 years of age: 15 patients with CNP (1st group) and 15 men with infertility due to varicocele in the postoperative period (2nd group). Patients of all groups received Samprost® for 20 days in the form of intramuscular injections of 5 mg daily. The study design provided for a traditional clinical, laboratory and specialized examination with the study of the level of antisperm antibodies, the study of sperm DNA fragmentation by the TUNEL method, the assessment of oxidative stress, a cryotolerance test, and an electron microscopic examination of the ejaculate. Statistical processing of the material was carried out using the paired Student’s t test for dependent samples, and the Wilcoxon nonparametric T test was also used. The calculations were performed using the SigmaPlot 12.5 program. Results. The average age of the study participants was 36.91 ± 4.5 years (23–51 years). Among the concomitant and past diseases, the most common were patients with recurrent varicocele, May–Turner syndrome, varicose veins of the pelvis. When evaluating the effectiveness, the results of the study showed a more significant effect of the drug Samprost® in the group of patients with CNP. This concerned an increase in ejaculate volume by 11.5 %, an increase in sperm concentration by 19.4 %. Progressive sperm motility increased more in the group of patients with varicocele by 12.2 %, compared with the CNP group – 11.4 %. Similar data were obtained during cryotest. After 20-day use of the Samprost® a significant decrease in the level of free radicals in the ejaculate was noted: by 2.5 times in the group with CNP and by 2.3 in the group of patients with varicocele. Analysis of these indicators may indicate an antioxidant effect on the effect on the sperm cell, which is under serious stress in the fight against free radicals. This assumption is also confirmed by the EMIS data, which showed a positive dynamics in the number of intact heads of normal form with a normal structure of chromatin and acrosome, the number of spermatozoa with insufficiently condensed immature chromatin. Conclusion. Samprost® is an effective and safe drug for patients with secretory type of male infertility. Based on the methods of statistical analysis, including statistically significant differences in the study groups, Samprost® is more effective in patients with secretory type of male infertility in the case of CNP than in patients with varicocele in the postoperative period. More significant changes relate to the volume, concentration of sperm viability and a decrease in the level of free radicals The 20-day course of Samprost® increases progressive sperm motility during cryotest in patients with varicocele and CNP and can be used for substantiated preparation of patients for gamete cryopreservation and its use in preparatory assisted reproductive technologies programs using intra cytoplasmic sperm injection. A decrease in DNA fragmentation and the received electron microscopic examination of the sperm data indicate a possible influence of Samprost® on sperm morphology and require further research in this area. © 2021 ABC-press Publishing House. All right reserved

Evaluation and comparative analysis of sperm characteristics in men before and after vaccination with Sputnik V (Gam-COVID-Vac) [Оценка и сравнительный анализ спермиологических показателей у мужчин до и после вакцинации препаратом «Спутник V» (Гам-КОВИД-Вак)]

The study objective is to evaluate reproductive function in men who underwent vaccination with Sputnik V (Gam-COVID-Vac) based on quantitative and qualitative sperm characteristics. Materials and methods. Mean ejaculate characteristics were analyzed in 3 independent groups formed from patients examined at the Laboratory of Genetics of Impaired Reproduction of the Research Center for Medical Genetics in 2021: the comparison group consisted of 759 unvaccinated men; the group of 73 men who were vaccinated with Sputnik V and for whom the time between the full vaccination (2 components of the vaccine) and semen analysis was less than 75 days; and the group of 58 men who underwent vaccination more than 75 days prior to semen analysis. The results of semen analysis of 53 patients before and after full vaccination procedure were compared. In 32 patients, less than 75 days passed since vaccination; in 21 patients, more than 75 days; for 5 patients data prior to 75 days since vaccination and after are available. The study group included patients without history of COVID-19. Standard semen analysis was performed in accordance with the World Health Organization Guidelines. For statistical analysis of the differences between the groups Student's t-test was used with significance level of p <0.05. Results. Comparison of semen analysis of the same patients before and after vaccination procedure showed no statis- tically significant differences. Small decrease in progressive motility was observed (from 19.0 ± 0.4 to 13.8 ± 1.1 %), as well as decline in sperm morphology (from 4.53 ± 0.12 to 3.55 ± 0.31 %) in the group who underwent vaccination less than 75 days prior to the analysis. In the group who underwent vaccination more than 75 days before semen analy- sis, sperm characteristics do not differ from the unvaccinated group. Conclusion. There were no statistically significant differences in the ejaculate indices of the same patients before and after vaccination with Sputnik V (Gam-COVID-Vac). In the group of patients examined in a short time after vaccination, there is a decrease in the quality of sperm compared to the comparison group (unvaccinated). In the group of patients examined in a more distant period, there are no significant differences in the quality of sperm from the group of unvac- cinated, which indicates the absence of long-term consequences of vaccination for the quality of ejaculate. © 2021 ABC-press Publishing House. All Rights Reserved

Effect of COVID-19 infection on characteristics of sperm in men with impaired fertility [Влияние перенесенной инфекции COVID-19 на спермиологические показатели мужчин с нарушением фертильности]

Introduction. The risk of infection with SARS-CoV-2 virus and potential cross-infection in an in vitro fertilization laboratory remains unclear. SARS-CoV-2 enters the body through angiotensin-converting enzyme 2 receptor. There is a possibility that high expression of angiotensin-converting enzyme 2 on testicular cells can lead to spermatogenesis disfunction and decreased quality of the ejaculate. The study objective is to compare quantitative and qualitative sperm characteristics in patients who sought medical help at the Laboratory of Genetics of Reproductive Disorders at the Research Centre for Medical Genetics with reproduction problems between 2017 and 2020 with a focus on the period of COVID-19 pandemic. Materials and methods. Results of semen analysis of 4403 patients (mean age 34 years) were considered. Taking into account histories of patients examined in 2020, a separate group of 428 men was formed with 42 men with a history of coronavirus infection confirmed by lab tests (40 cases of mild disease, 2 cases of moderate disease). In 14 patients, sperm analysis was performed twice: prior to COVID-19 and after. Standard sperm analysis was performed in accordance with the World Health Organization Guidelines. For statistical analysis Student's t-test was used with significance level p <0.05. Results. Results of 2020 semen analyses were compared to the control group including 3300 patients examined between 2017 and 2019 prior to emergence of COVID-19 virus in Russia. Decreased mean sperm count was observed in patients examined in 2020 compared to the control group: 302 ± 6 million versus 250 ± 8 million with concurrent increased sperm motility (14.7 ± 0.2 % versus 16.5 ± 0.4 %) and improved sperm morphological (3.07 ± 0.05 % versus 3.92 ± 0.11 %). The differences for these three sperm parametres are statistically significant. In 2020, normospermia was observed in 10.3 % of cases which does not significantly differ from 2017 and 2019 levels. Comparison of semen analyses in 2020 showed that the percentage of normospermia in patients after COVID-19 infection was almost 2-fold lower than in the control group of patients without history of the infection (7.1 % vs 13 %) and percentage of azpoospermia was more 2.5-fold higher (7.1 % vs 2.5 %). In 14 patients, sperm analyses before and after COVID-19 were compared. Mean sperm count after the disease decreased from 244 ± 74 million to 178 ± 34 million. Mean motility characteristics (12.7 and 15.8 %) and normal sperm morphology (2.6 and 3.2 %), on the other hand, had an upward trend after the disease. However, these differences were not statistically significant. Conclusion. In conclusion, SARS-CoV-2 virus and condition of COVID-19 pandemic did not lead to serious decrease in sperm quality in the studied group of Russian patients in 2020. This can be associated with small number of patients after the infection included in the sample, as well as general trend toward improvement in sperm quality observed in recent years. © 2021 Andrologia i Genital'naa Hirurgia. All rights reserved