Ya.V. Prokopyev

Introduction. The problem of male infertility remains a pressing issue in the field of reproductive medicine, as male infertility progresses year after year. The male reproductive microbiome is a complex community of microorganisms that inhabit various parts of the male body, including the urethra, prostate, and seminal vesicles. Research shows that the microbiome plays an important role in maintaining reproductive health and may influence fertility. Objective. To identify differences in the composition of the testicular and urethral microbiota between groups of patients with positive and negative results of assisted reproductive technologies (ART). Materials and methods. A comparative analysis of the taxonomic structure of the testicular and urethral microbiota was conducted in 24 infertile men with obstructive azoospermia (OA) who underwent ART procedures. Pregnancy occurred in 13 cases (ART+ group), while pregnancy was not achieved in 11 cases (ARTgroup). To analyze the bacterial composition of the urogenital tract, biological material was collected from the urethra of all study participants, as well as a testicular biopsy. Analysis of bacterial 16S rRNA gene amplicons was performed using high-throughput next-generation sequencing (NGS). The obtained data were processed using QIIME software (version 1.9.1), which allowed for taxonomic analysis and assessment of microbiota diversity. Results. Our study revealed significant differences in the microbiota composition between testicular and urethral tissue in infertile men with OA, as well as between groups of patients with positive and negative ART results. In testicular tissue, the main microbiota families were Enterobacteriaceae and Burkholderiaceae, while in the urethra, the dominant families were Mycobacteriaceae, Staphylococcaceae, Lactobacillaceae, and Peptoniphilaceae. Despite differences in microbiota composition, no significant differences in alpha diversity indices of testicular and urethral microbiota were found between the ARTand ART+ groups. However, in the ARTgroup, a significantly increased relative representation of 19 bacterial families was observed in the testicular microbiota compared to patients in the ART+ group, including increased representation of two families: Mycobacteriales and Legionellaceae. This may indicate a possible link between microbiota composition and ART success. Correlation analysis revealed a positive association between inhibin levels and testicular microbiota composition. This suggests that testicular microbiota may influence inhibin levels, which are an important marker of testicular function and may play a role in fertility. Conclusions. The results demonstrate that OA-associated disorders lead to changes in the relative representation of individual families of the urethral and testicular microbiota, which has a significant impact on ART outcomes. The practical significance of these data is that microbiota correction may be a potential strategy for improving ART outcomes in patients with OA, opening up new approaches to treatment and enhancing fertility, and suggests the theory of «testicular fertility microbiota» and may subsequently play an important role in pre-conception cryopreservation in ART programs and the creation of new tests for sperm selection in artificial insemination programs.

Introduction. Male infertility is a complex condition with many potential causes, including hormonal imbalances, anatomical problems, genetic factors, lifestyle factors and more. But today, there is a fairly large group of infertile men with unknown causes of the disease.Objective. To analyze the taxonomic microbial diversity of testicular tissue and the urogenital tract of infertile men and to identify correlations between the microbiome and mastocytes in the testicular parenchyma.Materials & methods. The study was performed on testicular tissue samples from infertile patients with azoospermia (n = 33). All patients were divided into two groups based on the form of azoospermia: group 1 — infertile patients with non-obstructive azoospermia (NOA) (n=21); and group 2 — infertile with obstructive azoospermia (OA) patients (n=12). The bacterial diversity of testicular tissue was studied by the method of high-performance new generation sequencing (NGS). Immunohistochemical staining with anti-MCT (Anti-Mast Cell Tryptase) was used to determine the IHC expression of mastocyte markers.Results. The microbiome of patients with NOA differs markedly from the microbiome of patients with OA (p < 0.05). In group 1, representatives of the Enterobacteriaceae and Xanthomonadaceae families, the genera Finegoldia, Bifidobacterium, Porphiromonas, Prevotella, Peptoniphilus and Pseudomonas are significantly more often found. A distinctive feature of group 2 is the rare occurrence of the genus Prevotella. Histochemical analysis revealed mastocytosis in the in-between-canalicular stroma approximately in 83% of azoospermia cases. Mastocytes are found in tubule structures in 68% of cases and correlate with the microbiome of testicular tissue.Conclusions. Injuries caused by mastocytes in the stroma and tubular structures are interrelated with the taxonomic diversity of testicular tissue. Moreover, the testicles of NOA-patients have a qualitatively and quantitatively more diverse spectrum both at the level of families and genera, unlike OA-patients.

Aim. To determine the role of taxonomic bacterial diversity of the urethral and testicular microbiota in the development of male infertility using high-throughput next-generation sequencing (NGS). Material and methods. In the period between 2018 and 2022 we examined 53 patients. Three study groups were formed: Infertile patients with nonobstructive azoospermia and concomitant varicocele (n = 13) – group 1; Infertile patients with obstructive azoospermia (n = 29) – group 2; Fertile patients with confirmed paternity and testicular pathology requiring histological verification to exclude testicular oncopathology (n = 11) – group 3. Each patient underwent TESE with testicular both and urethral tissue sampling in order to compare the bacterial landscape and control the purity of the method. The obtained material was sequenced using a high-throughput method (NGS). Results. According to our findings, the testicular tissue of obstructive azoospermia patients had significantly depleted bacterial diversity in comparison to nonobstructive azoospermia and concomitant varicocele ones. Also, the fertile group turned out to be the most diverse in its taxonomic community. These results may suggest the bacterial microbiome’s influence on men’s reproductive health. Discussion. Our urogenital bacterial diversity analysis showed that human testicular tissue is not a microbiologically sterile environment and also presented new data associated with testicular tissue and its possible relations with male infertility. Conclusion. A focus on the qualitative and quantitative composition of the testicular microbial community may form the new basis in the diagnosis and treatment of male infertility associated with various types of azoospermia

Introduction. The study of the human microbiome has become of increasing scientific interest. A balanced and diverse microbiota of the male re- productive system is essential for optimal reproductive health. Today, there are no diagnostic tests to determine the effect of specific microorganisms on the quality of seminal fluid, however, the use of next-generation sequencing (NGS), using the 16S rRNA gene sequence as a molecular marker, provides us with the opportunity to understand the complexity of the relationship between the microbiome and its host. The aim of the study. To conduct a comparative analysis of the taxonomic structure of the microbiota between patients with obstructive and nonobstructive azoospermia and the fertility group (patients with a history of having children). Materials and methods. The study included (n=57) patients (infertile patients with obstructive azoospermia (OA), non-obstructive azoospermia (NOA) and a control group (FERT). All patients underwent percutaneous aspiration of sperm from the testicle (testicular sperm aspiration, MESA), with subsequent in vitro fertilization using ICSI technology, or cryopreservation of biological material. The amplicons of the bacterial 16S rRNA gene were analyzed using high-performance next-generation sequencing (NGS). For this research of bacterial composition of the urogenital tract and control the purity of the method, samples has been taken from the urethra of each patient who underwent testicular biopsy. The data was processed using the QUICKTIME program (version 1.9.1). Results. No significant differences were found in the comparison groups during the study of the urethral bacterial composition. The species diversity, as well as the total number of detected taxonomic units (OTE) in the NOA and OA groups are significantly lower than in the FERT group. The presentation of the phylum like Proteobacteria, Firmicutes, Bacteroidetes, Bifidobacterium adolescentis in the studied groups is different. Three microorganisms were identified with the same frequency in the urethra and testicular tissue (the exact Fisher criterion was used, adjusted for multiple comparison by the Benjamin-Hochberg method). The representation of the phylum Prevotella, Comamonadaceae, Veillonella dispa, Comamonadaceae in the studied groups differs. Conclusion. Testicular tissue is not sterile and has its own unique microbial landscape for each comparison group. Bacterial communities of testicular tissue in patients with azoospermia are characterized by a reduced diversity and a specific composition that differs from the urethral microbiota. These results may be useful in further to study the role of microbiota in the pathology of spermatogenesis and developing new approaches to the treatment and diagnosis of male infertility.

Aim. To determine the role of taxonomic bacterial diversity of the urethral and testicular microbiota in the development of male infertility using high-throughput next-generation sequencing (NGS). Material and methods. In the period between 2018 and 2022 we examined 53 patients. Three study groups were formed: Infertile patients with nonobstructive azoospermia and concomitant varicocele (n = 13) – group 1; Infertile patients with obstructive azoospermia (n = 29) – group 2; Fertile patients with confirmed paternity and testicular pathology requiring histological verification to exclude testicular oncopathology (n = 11) – group 3. Each patient underwent TESE with testicular both and urethral tissue sampling in order to compare the bacterial landscape and control the purity of the method. The obtained material was sequenced using a high-throughput method (NGS). Results. According to our findings, the testicular tissue of obstructive azoospermia patients had significantly depleted bacterial diversity in comparison to nonobstructive azoospermia and concomitant varicocele ones. Also, the fertile group turned out to be the most diverse in its taxonomic community. These results may suggest the bacterial microbiome’s influence on men’s reproductive health. Discussion. Our urogenital bacterial diversity analysis showed that human testicular tissue is not a microbiologically sterile environment and also presented new data associated with testicular tissue and its possible relations with male infertility. Conclusion. A focus on the qualitative and quantitative composition of the testicular microbial community may form the new basis in the diagnosis and treatment of male infertility associated with various types of azoospermia

Introduction. Everywhere, about 15% of couples of reproductive age have the problem of infertility, with the male factor being the main cause in 50% of cases. The lack of effective treatment for male

Introduction. The concept of microbiome is a collective term for many microorganisms, such as bacteria, archaea, fungi, protists, and viruses, inhabiting various anatomical areas of the human body. Th

Геном и функционирование митохондрий играют центральную роль в развитии, течении и исходе многочисленных заболеваний человека. Однако оценка функции митохондрий в клинической медицине затруднена, в первую очередь из-за инвазивного получения жизнеспособной ткани для биоэнергетического анализа и отсутствия адаптированных лабораторных технологий. В исследованиях последних лет сформулирована гипотеза, в соответствии с которой митохондрии легкодоступных циркулирующих клеток крови могут быть использованы в качестве биомаркера системной и/или клеточно-специфической биоэнергетической функции. В настоящем обзоре будут освещены методы оценки биоэнергетики клеток крови и клинические исследования, в которых эти методы применялись. Суммированы результаты оценки эффективности анализа биоэнергетического статуса у здоровых индивидуумов, а также динамики биоэнергетики клеток крови при заболеваниях и корреляции биоэнергетического статуса с клиническими параметрами. Обсуждаются пути внедрения биоэнергетического анализа клеток крови в поле трансляционных исследований и персонализированной медицины.