Nowadays, the search for etiopathogenetic factors of infertility is an urgent medical and social task. 23 men with chronic abacterial prostatitis in a barren marriage underwent the follow-up. Herpesviruses of types IV–VI were detected in urogenital samples (urethra, ejaculate, prostate secretions) in 8 of 23 men who were treated with valacyclovir (500 mg 2 times a day for 3 months) and interferon α-2b in combination with antioxidants — vitamins E and C (rectal suppositories). Based on the personal experience of the authors, the patients were prescribed the following drug use regimen: 3 million IU twice a day — 10 days, then 3 million IU twice a day — 3 times per week for the next 3 weeks. The remaining 15 patients without any detected viruses were prescribed the following: levofloxacin 500 mg — 30 days, tamsulosin 0.4 mg — 90 days, diclofenac 50 mg — 10 days. By the end of therapy, electron microscopy and virology showed a decrease in the concentration and number of virus-positive urogenital samples, as well as a decrease in the number of spermatozoa with viral capsids. Within 6 months from the therapy initiation, the onset of pregnancy was observed in the married couples of 6 out of 8 men with herpesviruses vs. in 4 out of 11 without viruses (p=0.026). Thus, we obtained the data on the role of herpesviruses of types IV–VI in male infertility. A good reproductive effect was achieved with valacyclovir and interferon α-2b in combination with antioxidants in most patients. KEYWORDS: herpesviruses, Epstein-Barr virus, cytomegalovirus, human herpes virus type VI, male infertility. FOR CITATION: Kovalyk V.P., Gomberg M.A., Bragina E.E. et al. Herpesvirus role in male infertility. Russian Medical Inquiry. 2021;5(3):123– 129. DOI: 10.32364/2587-6821-2021-5-3-123-129.

From our point of view, the problem of infertility in a pair of appropriately addressed jointly by the two related disciplines. This process has been actively helping set newsupplements Spematon and Pregnoton. The study included 50 couples planning to conceive: 25 couples with male factor infertility secretory type (group 1), 25 couples with infertility caused by varicocele in the postoperative period (group 2). The first group included patients with hyperprolactinemia relative to a second group – 25 somatically healthy patients. All groups patients takes Spematon and Pregnoton during 3 months. Terms of infertility in couples ranged from 1 to 4,5 years and an average reached 2,8 ± 1,6 years. According to the results of the comparative analysis the reproductive value of sperm had been enhanced, functional status of the female reproductive system had been improved, the menstrual cycle had been normalized and relatively elevated levels of prolactin had been decreased after co-application of these supplements. Five woman became pregnant.

Mechnikov’s hypothesis that the key to prolongation of life lies in the introduction of useful microflora to the gut was not proved. Any microflora needs nutrition and perceives the human body only as a nutrient substrate. Destruction of the basement membranes, that delimit the epithelium contacting with aggressive microbiological environment from the deeper parts of the body, can lead to chronic inflammatory diseases and aging of the skin as a consequence of invasion of microorganisms. At the ultrastructural level this has been shown by the example of prostatitis and skin aging changes. Coupled with the penetration of germs, a flux of immune cells may cause autoimmune reactions due to abrupt changes in the molecular design during intermembrane transport. Thus, the physiological process of macroorganism aging can be viewed as a consequence of its microbiological destruction.

The dynamics of string growth was studied in model homochiral solutions of biomimetics, trifluoroacetylated amino alcohols (TFAAA) in heptane, water, and inverted heptane-water emulsion. In heptane and water, a thick (~1 μ in diameter) string had a crown of thin strings on its growing terminal and these thin strings effectively adsorbed dissolved TFAAA. In emulsion, the strings grew inside the water droplets, in which this TFAAA cannot be solved, presumably due to transport of TFAAA molecules from heptane into water in the surface layer surrounding the string. Applications of these phenomena to in vivo cell commutation were discussed.

Prostatic inflammation is associated with infections penetrating through the urethra. This inflammation is treated by long courses of wide-spectrum antibiotics. However, the most frequent cause of prostatitis is Escherichia coli and other enteric flora. Electron microscopy of biopsy specimens from the prostate detected gaps in the prostatic epithelium basement membrane, their size explaining the penetration of enteric flora into the prostate. These data suggest another view on the pathogenesis of prostatitis and approaches to improvement of therapy for this disease.

Changes in chromatin structure at different stages of differentiation of human spermatids were studied. It was shown that, in nuclei of early spermatids, chromatin is loosely packed and its structural element is an 8-nm fiber. This “elementary” fiber is predominant at the initial stages of differentiation; in the course of maturation, it is replaced by globular elements approximately 60 nm in diameter. In intermediate spermatids, these globules start to condense into fibrillar aggregates and reduce their diameter to 30–40 nm. At all stages of spermatid maturation, except the final stages, these globules are convergence centers for elementary fibers. This remodelling process is vectored and directed from the apical (acrosomal) to the basal pole of the nucleus. In mature spermatids, the elementary 8-nm fibers are almost absent and the major components are 40-nm fibrillar aggregates. The nuclei of mature spermatids are structurally identical with the nuclei of spermatozoa with the so-called “immature chromatin,” which are commonly found in a low proportion in sperm samples from healthy donors and may prevail over the normal cells in spermiogenetic disorders. The cause of this differentiation blockade remains unknown. Possibly, the formation of intermolecular bonds between protamines, which are required for the final stages of chromatin condensation, is blocked in a part of spermatids. The results of this study are discussed in comparison with the known models of nucleoprotamine chromatin organization in human spermatozoa.

Processes of cell commutation via anisometric supramolecular structures (strings) are considered in vitro and using physicochemical models of lipids and trifluoroacetylated amino alcohols. The biological effectiveness of commutation via strings is demonstrated to be higher compared to the diffusion mechanism of transfer of biologically active molecules over distances between objects typical of populations of microorganisms. A kinetic model of strings is developed, and the rate of signal transmission in such systems is estimated. It is shown that the condition of the chirality of lipids forming the biomembranes of cells follows from the experimentally observed cell commutation by means of supramolecular anisometric structures.

В работе изучена динамика изменений структуры хроматина на различных стадиях дифференцировки сперматид человека. Показано, что в ядрах ранних сперматид хроматин полностью декомпактизован, его структурной единицей является фибрилла толщиной около 8 нм. На начальных стадиях дифференцировки эта “элементарная” фибрилла является преобладающей структурой в ядрах, а на последующих этапах она постепенно замещается крупными глобулярными комплексами диаметром около 60 нм. В дальнейшем глобулы ассоциируют в фибриллярные структуры, толщина которых уменьшается до 30–40 нм. На всех стадиях дифференцировки, кроме конечных, глобулярно-фибриллярные комплексы представляют собой центры ассоциации элементарных фибрилл. Процесс конденсации хроматина имеет векторный характер – он направлен от апикальной зоны ядра к базальной. В зрелых сперматидах преобладающим компонентом являются фибриллы толщиной около 40 нм, тонкие фибриллы практически отсутствуют. По своей структурной организации ядра зрелых сперматид полностью соответствуют ядрам сперматозоидов с “незрелым” хроматином, которые в небольшом количестве обнаруживаются в эякуляте здоровых доноров и могут преобладать над нормальными клетками при паталогиях спермиогенеза. Причина такой остановки дифференцировки остается непонятной. Возможно, в части сперматид блокируется формирование межмолекулярных дисульфидных связей, которые необходимы для завершающей фазы компактизации генома. Полученные результаты обсуждаются в связи с известными моделями организации нуклеопротаминового хроматина в сперматозоидах человека.

Most of the researchers attribute amniotic fluid stem cells (AF SCs) to mesenchymal stem cells (MSCs). However, AF SCs express both mesenchymal and epithelial markers, which distinguishes them from postnatal MSCs. Cultivation in the three-dimensional (3D) matrix provides a different look at the nature of the cells. We showed that in 3D collagen gel AF SCs form epithelial structures (tubules and cysts). The active contraction of the gel during the first days of cultivation, which is characteristic of mesenchymal cells, does not occur. Electron microscopic study showed that adherent junctions typical to epithelial cells are formed between AF SCs. On the other hand, during culturing in the gel AF SCs continue to express MSCs markers. Thus, AF SCs may be not true mesenchymal cells because they can display properties of epithelial cells. Perhaps these cells undergo epithelial-mesenchymal transition, a process which actively takes place during embryogenesis.

The relation between the activity of the sperm-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDS) and the motility of sperms was investigated. It was found that the mean value of GAPDS activity in sperm samples with low motility is 2.5–3-fold lower than that in samples with high motility. Sperm motility was shown to diminish in the presence of superoxide anion, hydroxyl radical, and hydrogen peroxide. The decrease in sperm motility in the presence of hydrogen peroxide was proportional to the concentration of the oxidant and correlated with the decrease in GAPDS activity (r = 0.96). Based on the literature data on the importance of GAPDS for the motility of sperms together with the presented observations, it was concluded that the decrease in the sperm motility in the presence of reactive oxygen species is due to the oxidation of GAPDS and inhibition of glycolysis.

The results of an electron microscopy study of the ultrastructural characteristics of stallion spermatozoa in native sperm and after freezing-thawing are given. Changes occurring in spermatozoa under the effect of cryopreservation are shown. Maximum damages are found in acrosomes.

WARM 2010 was lower than nonsmoker men (p < 0.01).Therefore, it appears that cigarette smoking is associated.

Cytological and molecular genetics methods were used to study sperm from patients with sperm infected with herpes simplex virus (HSV) as indicated by virological and immunocytochemical tests. The following methods were used: (1) sperm analysis to evaluate the morphology and functional properties of sperm; (2) fluorescence in situ hybridization (FISH) with DNA probes specific for chromosomes 1, X, and Y to evaluate nondisjunction frequencies of these chromosomes in sperm; and (3) quantitative analysis of immature germ cells in the ejaculate to identify spermatogenic abnormalities. The total sperm count and the count of sperm with normal motility proved similar to the norm. FISH analysis demonstrated no difference in the nondisjunction frequency of chromosomes 1, X, and Y between infertile patients with HSV-infected sperm and fertile donors. Comparative quantitative analysis of immature germ cells from the ejaculate has demonstrated a significant and considerable (threefold) increase in the number of spermatocytes I in pachytene and diplotene, at the prepachytene stages of prophase I (preleptotene, leptotene, and zygotene) in HSV patients compared to normal donors. At the same time, HSV patients demonstrated a significant decrease in the number of spermatocytes I, in pachytene and diplotene, decrease in the proportion of spermatocytes II and spermatids, and a twofold increase in the number of unidentifiable immature germ cells. The data obtained indicate a partial spermatogenic arrest at the early stages of meiotic prophase I in HSV patients, which prompts further research into the cellular mechanisms of abnormal spermatogenesis after viral infection in humans.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDs) was purified from human sperms and properties of the enzyme were investigated. After sonication of sperms, the most part of GAPDs is associated with the insoluble cell fraction. Trypsin treatment results in the cleavage of part of the N-terminal domain of the enzyme yielding a soluble fragment that was purified by hydrophobic chromatography on Phenyl-Sepharose. The isolated fragment was shown to be a tetramer with molecular weight of approximately 150 kD (according to Blue Native PAGE) and composed of subunits of 40 kD (according to SDS-PAGE). The specific activity of the isolated fragment reached 374 U/mg. It is supposed that GAPDs exists in sperms as the tetrameric molecule bound to the fibrous sheath of the flagellum through the N-terminus of one or two subunits. Comparative study of the amino acid sequences of mammalian GAPDs revealed conservative cysteine residues (C21, C94, and C150) that are specific for the sperm isoenzyme and absent in the somatic isoenzyme. Residue C21 can be involved in the formation of the disulfide bond between the N-terminal domain of GAPDs and fibrous sheath proteins.

We developed a model of herpetic orchitis in guinea pigs. Intratesticular inoculation of type 2 herpes simplex virus suspension results in infection of the testicular spermatocytes and spermatides. The possibility of viral infection dissemination from infected into intact testis is proven.