[Study of the resistome of human microbial communities using a targeted panel of antibiotic resistance genes in COVID-19 patients].

AIM: To study overall drug resistance genes (resistome) in the human gut microbiome and the changes in these genes during COVID-19 in-hospital therapy. MATERIALS AND METHODS: A single-center retrospective cohort study was conducted. Only cases with laboratory-confirmed SARS-CoV-2 RNA using polymerase chain reaction in oro-/nasopharyngeal swab samples were subject to analysis. The patients with a documented history of or current comorbidities of the hepatobiliary system, malignant neoplasms of any localization, systemic and autoimmune diseases, as well as pregnant women were excluded. Feces were collected from all study subjects for subsequent metagenomic sequencing. The final cohort was divided into two groups depending on the disease severity: mild (group 1) and severe (group 2). Within group 2, five subgroups were formed, depending on the use of antibacterial drugs (ABD): group 2A (receiving ABD), group 2AC (receiving ABD before hospitalization), group 2AD (receiving ABD during hospitalization), group 2AE (receiving ABD during and before hospitalization), group 2B (not receiving ABD). RESULTS: The median number of antibiotic resistance (ABR) genes (cumulative at all time points) was significantly higher in the group of patients treated with ABD: 81.0 (95% CI 73.8-84.5) vs. 51.0 (95% CI 31.1-68.4). In the group of patients treated with ABD (2A), the average number of multidrug resistance genes (efflux systems) was significantly higher than in controls (group 2B): 47.0 (95% CI 46.0-51.2) vs. 21.5 (95% CI 7.0-43.9). Patients with severe coronavirus infection tended to have a higher median number of ABR genes but without statistical significance. Patients in the severe COVID-19 group who did not receive ABD before and during hospitalization also had more resistance genes than the patients in the comparison group. CONCLUSION: This study demonstrated that fewer ABR genes were identified in the group with a milder disease than in the group with a more severe disease associated with more ABR genes, with the following five being the most common: SULI, MSRC, ACRE, EFMA, SAT.

[Composition of oropharyngeal microbiota in patients with COVID-19 of different pneumonia severity].

AIM: To identify features of the taxonomic composition of the oropharyngeal microbiota of COVID-19 patients with different disease severity. MATERIALS AND METHODS: The study group included 156 patients hospitalized with confirmed diagnosis of COVID-19 in the clinical medical center of Yevdokimov Moscow State University of Medicine and Dentistry between April and June 2021. There were 77 patients with mild pneumonia according to CT (CT1) and 79 patients with moderate to severe pneumonia (CT2 and CT3). Oropharyngeal swabs were taken when the patient was admitted to the hospital. Total DNA was isolated from the samples, then V3V4 regions of the 16s rRNA gene were amplified, followed by sequencing using Illumina HiSeq 2500 platform. DADA2 algorithm was used to obtain amplicon sequence variants (ASV). RESULTS: When comparing the microbial composition of the oropharynx of the patients with different forms of pneumonia, we have identified ASVs associated with the development of both mild and severe pneumonia outside hospital treatment. Based on the results obtained, ASVs associated with a lower degree of lung damage belong predominantly to the class of Gram-negative Firmicutes (Negativicutes), to various classes of Proteobacteria, as well as to the order Fusobacteria. In turn, ASVs associated with a greater degree of lung damage belong predominantly to Gram-positive classes of Firmicutes Bacilli and Clostridia. While being hospitalized, patients with severe pneumonia demonstrated negative disease dynamics during treatment significantly more often. CONCLUSION: We have observed differences in the taxonomic composition of the oropharyngeal microbiota in patients with different forms of pneumonia developed outside hospital treatment against COVID-19. Such differences might be due to the presumed barrier function of the oropharyngeal microbiota, which reduces the risk of virus titer increase.

[Polyamines of urogenital tract men as factors of apoptosis regulation spermatozoids].

The aim of the study was to compare the content of polyamines (spermine and spermidine) in the seminal plasma of men of different fertility and to reveal the relationship between their level in sperm and the presence of markers of apoptosis in gametes. MATERIALS AND METHODS: ejaculates of 34 fertile men and 40 infertile men with various forms of subfertility between the ages of 22 and 40 were examined. The determination of polyamines was carried out by electrophoretic fractionation in an agar gel. As markers of apoptosis, externalization of phosphatidylserine (PS) to the external side of the spermatozoon membrane was determined and the receptor for the initiation of apoptosis CD95 in the gametes was detected. RESULTS: In sperm of infertile men, more spermatozoa with markers of apoptosis were revealed in comparison with fertile donors (p0,001). Against the background of a general decrease in the concentration of polyamines in ejaculates of infertile patients, the ratio between polyamines with a predominance of sperm in the seminal fluid is observed, which is the initiating factor for the initiation of apoptosis in gametes. This was proved by the relationship between the externalization of FS in gametes and the concentration of sperm in the spermoplasm in accordance with the coefficient of positive correlation (r=0.5, p<0.01). CONCLUSIONS: Thus, a change in the concentration of polyamines in the seminal fluid of men is one of the factors in the regulation of apoptosis of the sex cells. Determination of the content of polyamines in seminal plasma can be recommended to increase the information content of the study of the causes of impaired fertility of the ejaculate, and the results obtained for the development of the algorithm for examining infertile patients.

[Diagnosis and treatment of abacterial category III CP/CPPS, associated with herpes viruses].

INTRODUCTION: The etiology of abacterial CP/CPPS (category III) has not been studied enough. Currently, there is no gold standard of diagnostic study and optimal treatment algorithm. AIM: The aim of our study was to study three human herpes viruses (HHV) in clinical samples from patients with inflammatory diseases of urogenital tract and to evaluate the efficiency of proposed treatment algorithm for abacterial CP/CPPS. MATERIALS AND METHODS: The biological samples from the urogenital tract (urethral swab, ejaculate and expressed prostatic secretions) from 101 patients with category III CP/CPPS were studied. Quantitative analysis of HHV DNA (CMV, EBV and HHV-6) was performed by PCR. RESULTS: HHV DNA was detected in 38/101 patients (37.6%) in Group 1. Among the detected viral types, HHV-6 was the most common (52%). Analysis of biological samples form the three sources revealed that viral DNA was determined in urethral swab in concentration of 3,703,900 copies/ml. In Group 2, viral DNA was not detected in 63 patients. Evaluation of results of the standard treatment in HHV-negative patients (n=63) and antibiotic-free scheme, including the immunoregulatory drug Viferon, in HHV-positive patients (n=38) showed that the number of HHV-positive samples after treatment decreased by 54.3%. In addition, severity of all symptoms according to NIH-CPSI scale significantly decreased in both groups (p<0.0001). There was an improvement in all clinical symptoms in Group 1 by 47.9%, especially for pain + urination (52%). It should be noted that a positive response to treatment, which was confirmed by the changes in total score of NIH-CPSI scale, was noted in all patients in Group 1. CONCLUSION: Detection of herpes viruses in the urogenital tract of patients with abacterial CP/CPPS suggests possible role of viral infections in its etiology. The comparative analysis of the results of standard treatment including antiviral, immunomodulatory and antioxidant drugs showed that the use of complex therapy without antibiotics allowed to eliminate or significantly reduce the concentration of viruses in urogenital tract, as well as significantly reduce the clinical manifestations of abacterial CP/CPPS.

[QUANTITATIVE DNA EVALUATION OF THE HIGH CARCINOGENIC RISK OF HUMAN PAPILLOMA VIRUSES AND HUMAN HERPES VIRUSES IN MALES WITH FERTILITY DISORDERS].

Infertility is an actual medical and social problem. In 50% of couples it is associated with the male factor and in more than 50% of cases the etiology of the infertility remains insufficiently understood. The goal of this work was to study the prevalence and to perform quantitative analysis of the human herpes viruses (HHV) and high carcinogenic risk papilloma viruses (HR HPV) in males with infertility, as well as to assess the impact of these infections on sperm parameters. Ejaculate samples obtained from 196 males fall into 3 groups. Group 1 included men with the infertility of unknown etiology (n = 112); group 2, patients who had female partners with the history of spontaneous abortion (n = 63); group 3 (control), healthy men (n = 21). HHV and HR HPV DNA in the ejaculates were detected in a total of 42/196 (21.4%) males: in 31 and 11 patients in groups 1 and 2, respectively (p > 0.05) and in none of healthy males. HHV were detected in 24/42; HR HPV, in 18/42 males (p > 0.05) without significant difference between the groups. Among HR HPV genotypes of the clade A9 in ejaculate were more frequent (14/18, p = 0.04). Comparative analysis of the sperm parameters showed that in the ejaculates of the infected patients sperm motility as well as the number of morphologically normal cells were significantly reduced compared with the healthy men. The quantification of the viral DNA revealed that in 31% of the male ejaculates the viral load was high: > 3 Ig10/100000 cells. Conclusion. The detection of HHV and HR HPV in the ejaculate is associated with male infertility. Quantification of the viral DNA in the ejaculate is a useful indicator for monitoring viral infections in infertility and for decision to start therapy.

[Desadaptive Reaction and Development of Arterial Hypertension in Conditions of Professional Stress].

PURPOSE: to compare prevalence of maladaptive responses and risk of hypertension and cardiovascular remodeling. MATERIAL AND METHODS: We examined using clinical and instrumental methods 1321 workers of locomotive crews; 472 of those examined were followed-up for 12 years. RESULTS: Machinists with maladaptive responses compared with the group of individuals with normal tension of adaptation processes had greater left ventricular myocardial mass and carotid artery intima-media thickness regardless of blood pressure (BP) level. During follow up 380/472 persons maintained normal BP while 87 developed hypertension (5 patients with symptomatic hypertension were excluded from analysis). According to modified MMPI test persons with hypertension had higher scores on scales 2, 8, and 6. Personality profile was characterized by presence of mixed type of response with combination of high need for self-realization and tendency to curb behavioral reactions.

[CLINICAL EFFICACY OF THE RECEPTOR BETA AND ALPHA BLOCKERS - CARVEDILOL AND ORNITHINE-ASPARTATE IN PATIENTS WITH CHD AND CHF COMBINED WITH ALCOHOLIC LIVER DISEASE].

Aim: The aim of the paper is to study the clinical efficacy of carvedilol and ornithine-aspartate in the complex therapy (ACE inhibitors, diuretics, cardiac glycosides, nitrates indication), to assess their impact on quality of life, intracardiac hemodynamics, remodeling of the left (LV) and right ventricular (RV), indicators of the inflammatory enzyme activity in blood serum of patients with coronary heart disease with CHF II-III FC and alcoholic liver disease (ALD). Materials and Methods: 95 patients were studied 45-75 years (mean age - 58,2 ± 1,2) with CHF II-III FC and postinfarction cardiosclerosis, LVEF less than 45%. ALD was diagnosed in 58 patients. In 23 (39%) patients among them had steatosis, 18 (30.5%) - chronic hepatitis (CH), 17 (30.5%) - liver cirrhosis (LC). Patients were divided into 3 groups. Patients in the first group (37 people) with coronary artery disease and heart failure without a UPS received an average dose of carvedilol in - 32,8 ± 4,7 mg / day. Patients in the 2nd group (32 persons) suffering from coronary artery disease, heart failure, and UPS received carvedilol in an average dose of 25,4 ± 1,6- mg / day and L-ornithine-L-aspartate in a dose of 10 g granulate per day for 2 weeks, and then by 5g a day for 4 months. Patients in the third control group (26 people) with CHD and CHF and CHF ABP received basic therapy (without ß-blocker) and ademetionine at a dose of 800 mg / day for 2 weeks, followed by 400 mg / day for 4 months. Results: After 4 months of observation, it was noted that CHF patients with IHD in combination with BPO flows less favorably. In all groups, the clinical status of patients was improved on the background of the therapy, however, the clinical status was more pronounced while using carvedilol and ornithine-aspartate (Group 2): summary measure of quality of life has improved by 38 points, the speed of the test increased with a digital sequence up to 54.4, decreased shortness of breath, edema, ascites, portal hypertension effects, hepatocellular insufficiency and hepatic encephalopathy. In general, was shown the normalization of sleep rhythm, reducing sleepiness, improved memory, attention, reduced asterixis and sweeping hand tremor, asthenia. Conclusions: The use of carvedilol and ornithine-aspartate in the treatment of patients with CHF FC II-III with CHD and BPO improves the clinical condition of patients, quality of life, hemodynamics, reduces the severity of pulmonary hypertension and normalizes serum biochemical parameters.

[Interaction of herpesviruses with mature human spermatozoa in the model system in vitro].

The DNA of human herpesviruses (HHV), including the herpes simplex virus (HSV) and cytomegalovirus (CMV), is often identified in ejaculates of patients with urogenital diseases and infertility. At least a part of viral DNA is associated with cell fraction of ejaculate. However, it remains unclear how the semen is infected by the virus. It can be located in gametes or be capable of infecting mature germ cells, including motile sperm cells. In order to resolve this issue, interactions of the CMV and HSV with human sperm cells were studied using an original optimized model of the herpesviral infection of male gametes in vitro. The analysis of the immunofluorescent staining of gametes for viral antigens has shown that CMV infected 2% gametes, while HSV infected 17.26 ± 2.58% gametes. The fraction of progressively motile sperm cells contained 13.99 ± 4.64% infected cells. Localization of HSV was studied by the confocal microscopy. Sometimes, viral gB protein was found on sperm cell membrane. In addition, optical scanning of other cells has shown the intracellular localization of the viral proteins. In the majority of spermatozoa, the viral proteins were observed in the head and neck. In some cells, they were located in the middle piece or, rarely, in the equatorial segment. In general, after in vitro infection HSV antigens were located in the same areas of the sperm cells as in ejaculates from infected patients. According to DNA-DNA hybridization in situ, gametes containing HSV DNA accounted for 16.94 ± 5.28%, which is consistent with the results obtained in the immunofluorescence assay. It can be concluded that mature male gametes are infected by HHV in the genital tract, where the virus binds to the sperm cell membrane and enters the cell. Interaction of HHV with progressively motile sperm cells implies a vertical viral transmission upon fertilization and points to the necessity of testing ejaculate for herpesviruses infections.

Low Concentrations of Hydrogen Peroxide Activate the Antioxidant Defense System in Human Sperm Cells.

The effect of low concentrations of hydrogen peroxide (10-100 µM) on sperm motility and on the activity of the sperm enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDS) was investigated. Incubation of semen samples with 10 and 100 µM hydrogen peroxide increased the content of spermatozoa with progressive motility by 20 and 18%, respectively, and enhanced the activity of GAPDS in the sperm cells by 27 and 20% compared to a semen sample incubated without additions. It was also found that incubation with 10 µM hydrogen peroxide increased the content of reduced glutathione (GSH) in sperm cells by 50% on average compared to that in the control samples. It is supposed that low concentrations of hydrogen peroxide activate the pentose phosphate pathway, resulting in NADPH synthesis and the reduction of the oxidized glutathione by glutathione reductase yielding GSH. The formed GSH reduces the oxidized cysteine residues of the GAPDS active site, increasing the activity of the enzyme, which in turn enhances the content of sperm cells with progressive motility. Thus, the increase in motile spermatozoa in the presence of low concentrations of hydrogen peroxide can serve as an indicator of normal functioning of the antioxidant defense system in sperm cells.

[New approaches to therapy of Helicobacterpylori infection (by the materials of the Maastricht Consensus-IV, Florence, 2010)].

European recommendations for eradication of Helicobacter pylori are presented. The increase of the resistance to clarithromycin requires the necessity of rational use of quadritherapy as the first line treatment and sequential therapy including drugs active against the pathogen with low resistance. New treatment regiments for patients with allergy to penicillin are recommended.

[Optimization of treatment of patients with chronic cardiac insufficiency of the ischemic genesis and hobnail liver].

Clinical efficacy of combined therapy including the use of rifaximin and L-ornithin-l-aspartate, as well as the dynamics of the biochemical indices, the manifestation levels of portal-systemic-encephalopathy and intestinal microbiocynosis were investigated in patients with chronic cardiac insufficiency of ischemic genesis and hobnail liver. The combined therapy resulted in improvement of the patients clinical state, lower levels of the portal-systemic encephalopathy manifectation by decreasing hyperammonium, normalization of the large intestine microflora, and blood serum biochemical parameters.

[Selzink plus study in patients with chronic non-infectious prostatitis and abnormal fertility].

Correction of subnormal spermatogenesis caused by chronic prostatitis is made with application of different therapeutic schemes. Our aim was to investigate efficacy of selzink medicine which corrects deficiency of trace elements Se and Zn in ejaculate and thus improves spermatogenesis. One-month course of therapy produced no side effects, had a positive effect on low fertility of ejaculate. Therefore, selzink can be recommended in combined treatment of male infertility.

[Seasonal characteristics of gametogenesis of some fishery hydrobionts].

The data received on histological organization and cell composition of gonads in some hydrobionts complete the information about their reproductive biology and allow giving concrete expression to spawning period. The influence of some important ecological factors on reproduction of these hydrobionts has been analysed.

[Characteristics of monoclonal antibody binding with the C domain of human angiotensin converting enzyme].

Binding of a panel of eight monoclonal antibodies (mAbs) with the C domain of angiotensin converting enzyme (ACE) to human testicular ACE (tACE) (corresponding to the C domain of the somatic enzyme) was studied and the inhibition of the enzyme by the mAb 4E3 was found. The dissociation constants of complexes of two mAbs, IB8 and 2H9, with tACE were 2.3 +/- 0.4 and 2.5 +/- 0.4 nM, respectively, for recombinant tACE and 1.6 +/- 0.3 nM for spermatozoid tACE. Competition parameters of mAb binding with tACE were obtained and analyzed. As a result, the eight mAbs were divided into three groups, whose binding epitopes did not overlap: (1) 1E10, 2B11, 2H9, 3F11, and 4E3; (2) 1B8 and 3F10; and (3) IB3. A diagram demonstrating mAb competitive binding with tACE was proposed. Comparative analysis of mAb binding to human and chimpanzee ACE was carried out, which resulted in revealing of two amino acid residues, Lys677 and Pro730, responsible for binding of three antibodies, 1E10, 1B8, and 3F10. It was found by mutation of Asp616 located close to Lys677 that the mAb binding epitope 1E10 contains Asp616 and Lys677, whereas mAbs 1B8 and 3F10 contain Pro730.

[Effect of mineral water on metabolic processes].

The paper comparatively analyzes the effect of the mineral water "Penta" on the biochemical parameters of blood and urine, which characterize the functional activity of renal metabolic processes. A group comprising 10 examinees without renal disease took water by the routine mineral water scheme for 4 days. Comparison of the biochemical parameters before and after water taking revealed a significant reduction in azotemia and uric acid levels with its simultaneously enhanced excretion, as well as an increasing tendency for the excretion of oxalates, i.e. the most important parameters determining the formation of urate and oxalate calculi. These findings allow use "Penta", to a certain degree, in the treatment of renal disease, urolithiasis in particular, and in the prevention of stone formation.

[Seasonal characteristics of gametogenesis in the Corbicula japonica].

A morphological study of gonads in Corbicula japonica made it possible to distinguish between five stages of gonad maturity. It has been established that stages of active gametogenesis, prespawning and of spawning go almost in parallel and in the shortest time. In July, with the beginning of spawning, gametogenesis does not stop, and oogenesis proceeds up to the end of August. Dynamics of changes of some chemical substances in oocytes was followed. High concentrations of, respectively, RNA, albumens, lipids, glycoproteins and proteoglycans were revealed in oocyte nuclei and nucleoli both at the beginning of the development, and at the stage of active gametogenesis. This testifies in favor of a synthetic activity of these cells.

[Reproductive function in chronic abacterial prostatitis patients].

The results of examination of 325 patients with chronic abacterial prostatitis, varicocele showed marked changes in ejaculate parameters of these patients. Prostatitis of different duration in the same degree causes pathozoospermia, reduces ejaculate fertility. Such condition of spermatogenesis indicates involvement of other sex organs suggested by the literature data and previous experience. This necessitates extended diagnostic examination of patients with chronic prostatitis.