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Annually, data are accumulating on the involvement of opportunistic microorganisms in the development of inflammatory diseases in humans, maintaining a chronic inflammatory response and, thus, adapting to the conditions of existence in the biotopes of the human body. This review provides information on the interactions of microorganisms of medical importance, which affects the virulence of both opportunistic pathogens and classical pathogens, which probably underlie the chronicity of infection and inflammation. Often, opportunistic pathogenic species cannot fully realize their pathogenic potential, which is observed in numerous cases under conditions of microbial symbiosis. Thus, a revision of approaches to interpreting the results of microbiological methods is necessary, which takes into account the functional activity of the total microflora and the search for individual extrachromosomal genetic elements as a marker of the pathogenicity of microorganisms.

The intestinal microbiota is a marker of the organism state, capable of direct and indirect interacting. The main mechanisms underlying interactions are immunoregulation and energy metabolism. Metabolites, formed during the life of microbiota, realize their actions through these engagements. Some metabolites arouse negative effects on endothelial vessels, causing and maintaining a systemic inflammatory response, which stands behind major cardiac risk factors. Metabolites and molecules such as lipopolysaccharide or trimethylamine N-oxide initiate endothelial dysfunction, and thus trigger the processes of atherogenesis, insulin resistance and even increase blood pressure by activating inflammasomes and pro-inflammatory cytokines. Short-chain fatty acids including the main metabolites such as acetate, propionate and butyrate are antagonists of lipopolysaccharide and trimethylamine N-oxide. These substances are a source of energy for intestinal epithelial cells. They maintain homeostasis, stimulate the production of anti-inflammatory components and activating reparative processes. Another important factor influencing levels on blood pressure and systemic inflammation is intestinal barrier dysfunction, which is determined by the regulatory protein zonulin. Besides, there are proved receptor interactions, as well as qualitative and quantitative changes in the composition of the microbiota that can influence blood pressure and atherogenesis. Each cardiovascular disease is characterized by its own microbiological pattern and the predominance of specific metabolites. The article presents a review, summarizing experimental and clinical data on the role of microbiota in the development of atherosclerosis and cardiovascular diseases.

Introduction. Currently, there is an increasing importance of microbial associations in the pathogenesis of genital inflammatory diseases. However, the issues of deciphering the taxonomic affiliation and the diagnostic significance of the bacteria detected in this case remain unresolved.The aim of the study was to review the diagnostic significance of the quantitative approach in determining the etiological role of microorganisms in andrology.Materials and methods. For the study, samples of ejaculate and/or discharge from the urethra from 15 men who were in infertile marriages, 12 with a confirmed diagnosis of "acute genital gonococcal infection" were used. The testing included a classic bacteriological study and the metagenomic analysis of 16S ribosomal RNA samples carried out at the Department of Collection Cultures of the State Research Center for Applied Microbiology (Obolensk). To standardize the distribution of samples into groups based on the indicators of alpha diversity and the concentration of putrescine, the range of variation and the average linear deviation were used. Statistical analysis of the obtained data was carried out using Excel software (MS Office package).Results. The microbial landscape of the ejaculate was characterized by the predominance of representatives of the genera Enterococcus, Neisseria, Lactobacillus, Corynebacterium, Prevotella, Bacteroides. Stable associations of E. faecalis and M. osloensis were detected in the ejaculate. When using the cultural method, representatives of the genus Moraxella were not isolated in any sample. It was shown that discrepancies can affect not only quantitative indicators, but also reveal inconsistencies between the qualitative assessment of detected genetic markers and the results of identification in a bacteriological study of individual representatives of similar or phenotypically similar taxa.Discussion. The results of this study indicate that the lesser diversity of opportunistic pathogens gives them more opportunities to realize their pathogenic potential. On the other hand, in a complex community with greater alpha diversity, its realization is hindered by a complex intermicrobial relationships and the need to survive.Conclusion. As a result, it seems that the most promising approach is the integrated use of cultural methods and metagenomics with a comparative statistical analysis of the obtained qualitative and quantitative indicators.

Recently, there have been increasing an interest to study a role of polyamines in intercellular interactions, especially in the focus of inflammation, where accumulation of such polycations is observed. In this regard, products of microbial origin cadaverine and putrescine are of particular attention. The role of polyamines as scavengers of free radicals has been described, but no data of their effect on the leukocyte radical-producing activity have been obtained so far. The aim of the investigation was to study features of hydroxyl radical generation by human leukocytes exposed to microbe-derived polyamines. Materials and methods. Peripheral venous blood samples were obtained from 20 healthy donors. To assess radical production, a luminol-dependent chemiluminescence reaction was carried out with blood leukocytes pre-incubated with cadaverine (0.01 M) and putrescine (0.01 M), measured on a Luminoskan Ascent Thermo Labsystems luminometer (USA) for 180 min. For statistical analysis, an integral chemiluminescence index was used for the entire measurement period (RLU). Results and discussion. It was shown that cadaverine has a stimulating effect on the leukocyte potential to produce radicals (averaged area under the curve is 6.70.7 r.u., p 0.05). Putrescine had little effect on the radical-producing activity of human cells (2.80.4 r.u., p 0.05). This might be due to the direct influence of polycations on the mechanisms of radical generation, as well as increased activity of leukocyte diamine oxidase catalyzing the conversion of diamines into aminoaldehyde, which is accompanied by the release of hydrogen peroxide reacting with luminol. In addition, the enzyme inactivates other compounds, such as histamine leading to formation of a less pronounced clinical picture. Polyamines, particularly cadaverine and putrescine, can be referred to the factors remodeling the metabolic activity of the host leukocytes, which is aimed at maintaining the viability and increasing the number of microorganisms. Conclusion. Thus, cadaverine and putrescine produced by microorganisms, depending on the conditions of the microenvironment, might be evidently considered as mediators of the mild or asymptomatic course of inflammatory diseases, which contributes to underlying persistent potential of bacteria.

Objective. To assess the phagocytic activity of peripheral blood and exudate leukocytes in patients with facial phlegmon. Currently, relatively little attention is paid to assessing the functional activity of peripheral blood and exudate leukocytes in facial phlegmon, the incidence of which is characterized by an increase in the number of cases and, especially, among patients with comorbid pathology. Materials and methods. To study the phagocytic activity of leukocytes using a method based on the assessment of the absorption of formalinized sheep erythrocytes by neutrophils and monocytes, the samples of peripheral blood and exudate were obtained from 18 patients diagnosed facial phlegmon. Blood leukocytes obtained from 29 healthy donors were used as a comparison group. Results. It was shown that for patients with phlegmon of the face, an increase in the phagocytic activity of peripheral blood leukocytes is characteristic. There was observed a redistribution of leukocytes according to the number of absorbed objects towards an increase in actively phagocytizing cells 4438 721 per 1 l (in the comparison group 297 67 per 1 l; p 0.05). However, after migration to the foci of the pathological process, leukocytes lost their phagocytic activity and the number of phagocytic cells decreased to 35.0 8.3 % (for peripheral blood leukocytes of the same patients 64.3 5.4 %; p 0.05). Changes in the phagocytic activity of exudate leukocytes affected both neutrophils and monocytes. Conclusions. Thus, in patients with phlegmon of the face, changes in the phagocytic activity of blood leukocytes are probably associated with the indirect influence of cytokines, and in the focus of inflammation, leukocytes come under pressure from microorganisms. In such a situation, it is necessary to select an effective immunotropic therapy.

BACKGROUND: For today, the most important and discussed issue in the professional medical community is the problem of prevention and treatment of a new coronaviral infection (COVID-19). The main reason for the non-decreasing increase in morbidity and mortality is the absence of an etiotropic drug. In our study, it is proposed to use a previously available drug for the treatment of tick-borne encephalitis, ribonuclease A, obtained from the pancreas of cattle. AIM: The aim of investigation was to study the antiviral activity of RNаse A against SARS-CoV-2 in in vitro experiments. MATERIALS AND METHODS: The experiment used samples of 50 patients with a confirmed (by PCR) primary diagnosis of a new coronoviral infection COVID-19. The preparation for the study was served by ribonuclease A (neoFroxx GmbH, Germany) at a concentration of 0.5; 1; 5 and 10 mg/ml, incubated at 4 and 37C, the exposure was 20 minutes, 20 hours. A set of reagents OTT-PCR-RV-SARS-CoV-2 (Syntol, Russia) was used as test systems. RESULTS: of the current study is the revealed antiviral activity of ribonuclease A at a minmal concentration of 0.5 mg/ml during incubation from 20 minutes to 20 hours, in the temperature range of 437C. CONCLUSIONS: The data obtained in the in vitro study confirmed the ability of ribonuclease A to destroy viral RNA, which suggests the possible use of the drug both for the treatment of patients and for the treatment of environmental objects.

Currently, in the diagnosis of diseases, a decisive place is given to laboratory methods, which should be informative, relatively simple to perform and rapid. The article describes the approbation of a method for rapid detection of Escherichia coli and bacteria of Escherichia coli group in the oral cavity. Research involved 44 volunteers, who were sampled from the oral cavity, followed by incubation in Koda’s medium. The study used oral (n=11) and gingival fluids (n=11); smears-prints from the oral mucosa (n=11); dental biofilm (n=11). After 24 hours, the change in color and transparency of the medium was assessed. The preservation of the initial green color and transparency by the medium meant the absence of E. coli and bacteria of Escherichia coli group in the sample. A change in the color of the medium to yellow, turbidity and / or the formation of bubbles indicated the presence of E. coli and bacteria of Escherichia coli group. In parallel, the material was inoculated onto Endo agar, followed by identification of strains to species. As a result of the study, a complete coincidence of the results of the classical bacteriological method and the method using Koda medium was shown. In the latter case, a significant advantage is the speed of obtaining the result (18-20 hours), in contrast to the classical method, the interpretation of the results of which is available only after 72 hours or more. All of this is in line with the state of the art in clinical microbiology and rapid diagnosis based on «point-of-care testing / doctor’s office» diagnostic principle. The presented method can be successfully applied in clinical practice for topical diagnosis of microorganisms E. coli and bacteria of Escherichia coli group in the oral cavity.

Objective. The aim of investigation was to evaluate the bacteriolytic activity of the peripheral blood of healthy donors against S. aureus. Currently, much attention is paid to the involvement of opportunistic microorganisms in the development of infectious and inflammatory diseases, among which one of the leading places is occupied by the processes of staphylococcal etiology. Staphylococcus aureus possesses a unique spectrum of pathogenic factors, which, together with intracellular persistence, allow staphylococci to avoid exposure to immune factors and other agents. Materials and methods. The bacteriolytic activity of the peripheral blood of 32 healthy donors, as well as the ability of the whole blood and serum to destroy biofilms, were evaluated. The phagocytic activity of peripheral blood monocytes and neutrophils and the ability to produce hydroxyl radicals were analyzed. For opsonization of S. aureus, a commercial immunoglobulin G or donor serum was used. Results. It was shown that the whole peripheral blood practically does not have a significant effect on the number of viable cells of S. aureus. However, freshly obtained blood serum significantly destroys the biofilm. It has been established that a fifth part of peripheral blood leukocytes is absorbed by S. aureus. After opsonization of microbial cells with immunoglobulin G, the indices of phagocytic activity of monocytes and neutrophils did not change significantly. When using freshly obtained serum for opsonization of objects, a stimulating effect on the production of hydroxyl radicals by leukocytes was revealed (2758.7 725.3 and 870.6 197.4 related light units, respectively; p 0.05). After heating the serum at 56 C, the stimulating effect was leveled (1091.1 234.7 related light units; p 0.05 for samples with non-opsonized objects). In general, the obtained data indicate that the complement components can be recognized as the most effective system for the elimination of S. aureus. Conclusions. Thus, S. aureus uniquely adapted to the human body that allows staphylococci to persist for a long time without clinical manifestations. It can be assumed that among the factors of the immune system, the proteins of the complement system, which destroy both S. aureus cells and the biofilm matrix, probably have the most effective bactericidal action. However, the effectiveness of this system depends on the protein-synthesizing function of the liver, the availability of microorganisms to the action of complement.

Objective. To study the possibility of using LED blue, red, and green radiations of a new apparatus to eliminate prokaryotic cells and maintain human cell viability. Currently, laser technologies are widely used in medical practice. However, there are relatively few studies on the effect of laser radiation with similar parameters on the viability of prokaryotic and eukaryotic cells. Materials and methods. As a radiation generator, the LFDT-02 diode medical laser of a new type from the Perm Scientific and Production Instrument-Making Company was used. Staphylococcus aureus 66G cultures were irradiated with a different wavelengths laser (exposure time 2 min, power 22 mV). After that, the viability of microorganisms was evaluated by direct seeding on nutrient media. To study the effect of laser radiation on macroorganism cells, fasting samples of buccal epithelium were obtained in 17 practically healthy volunteers aged 2025 years with an intact periodontium and dentition, without somatic pathology. To assess cell viability, a vital dye was used 0.1 % trypan blue solution. The exposure time and power do not differ from those for the studied S. aureus strain. To assess the morphological changes of epithelial cells, preparations were prepared for microscopy, which, after fixation, were stained according to the Romanovsky-Giemsa method. Results. When evaluating the effect of laser radiation, especially in a clinical setting, it is necessary to take into account its possible effect not only on cells of conditionally pathogenic microorganisms, but also on human cells. In the present study, it was shown that laser radiation with a power of 22 mV allows one to achieve a bacteriostatic effect against S. aureus but it also significantly reduces the viability of buccal epithelial cells, that can be considered, on the one hand, as a side effect of laser radiation and, on the other hand, the removal of dying cells filled with microorganisms, can enhance the effect of ongoing therapy. Conclusions. Based on the results obtained, it can be concluded that the laser radiation of a diode laser with a wavelength of 405 nm (blue) and 650 nm (red) has a pronounced bacteriostatic effect. However, a significant damaging effect of red radiation on macroorganism cells was found.

A trocar-based implantation technique was suggested and used to study the biocompatibility of biomedical synthetic materials. The advantages of the suggested technique are the simplicity of its implementation and a reduction of injuries and postoperative complications in experimental animals. Another advantage is the optimization of the dynamic observation of the experimental animals’ response in vivo without removing them from the experiment.

Aim. To study and compare the features of formation of biofilms S. aureus and S. epidermidis on synthetic materials from polyvinylchloride and polystyrene. Materials and methods. In the experiments, S. aureus and S. epidermidis strains as well as polymeric synthetic materials were used. Expression of biofilm formation of these strains on polystyrene and polyvinylchloride was studied. Biofilm formation was determined in sterile flat-bottomed tablets. Into a part of tablet holes, pieces of polyvinylchloride measuring 551 mm were placed. They were cultivated at the temperature 37 C for 2448 hours. Results. On polyvinylchloride, biofilm-forming ability of staphylococci was practically absent. S. aureus is more active in formation of biofilms as compared with S. epidermidis. As incubation period is increased, S. aureus elevates biofilm-forming activity but S. epidermidis does not. Conclusions. It was established that one and the same microorganisms are different in manifestation of biofilm-forming activity on polystyrene and polyvinylchloride that can be conditioned by various chemical structure of these materials.

Microbial biofilms are heterogeneous, moving and constantly changing communities of microorganisms, often of various taxons. Approaches to study and assessment anti-biofilm drugs widely available today do not adequately assess their effects, while the results of studying the interaction of drugs with components of the film composition can provide them the right choice. The aim of investigation was to test a new method of morphological evaluation of biofilms. To form biofilms, we used an approach when the slide was placed at an angle of 30o-45o relatively to the Petri dish, and a suspension of test strains S. epidermidis in peptone broth was poured into the space between the Petri dish and the slide. A sterile cotton swab moistened with distilled water was placed next to the glass slide to create optimal humidity. The system was placed in a thermostat for 24 hours. The formed films were examined under a microscope using the DCM 310 video eyepiece and the Scope photo x86,3.1.312 program that allowed to conduct a complete morphometric study of the film: select layers, channels, cavities and make measurements, and then save the results on electronic media in jpg file format. Microscopy of the stained slides revealed that the biofilm has a layered structure. In each image obtained using a video eyepiece, it was possible to differentiate 4 layers. From the border of the two media to the inside: the fragmentation layer, the dense layer, the matrix substance layer, and the last one - the persistence layer. Channels of different diameters (from 10 to 24 microns) are observed across the entire thickness of the biofilm. Thus, used approach allows us to visualize and evaluate the structure of microbial biofilm, measure the thickness of layers and channel diameters. In addition, this method can be used to study the effect of antimicrobial drugs on bacterial films.

Microorganisms are able to form biofilms on surfaces of biotic and abiotic nature. In turn, in human biotopes there are optimal conditions for the implementation of biofilm-forming activity. Moreover, in medical practice, polymeric materials are often used for drainage or prosthetics, which can also be successfully colonized by bacteria. However, in laboratory practice, the formation of biofilms is usually evaluated on glass or polystyrene. The purpose of the study is to evaluate the methodological features of studying the biofilm-forming activity of microorganisms on the surface of synthetic polymeric materials. We used strains of Staphylococcus aureus ATCC 25923, Escherichia coli K-12, Candida albicans ATCC 10231, as well as synthetic polymeric materials - DentLight Flow light-curing composite material (nano-hybrid fluid composite; Russia), glass ionomer chemical curing Fuji 1 (Japan), cement for temporary fixation of orthopedic constructions TempBond NE (USA), acrylic, polyurethane and polyvinyl chloride. The formation of biofilms in flat-bottomed ELISA plates in this study was considered as a control group. If the polymer belonged to cold curing materials, sterile flat-bottomed tablets were used, the bottom of which was filled with a thin layer of plastic. After hardening of the plastic, biofilms were formed in the tablets. In the second series of experiments, hot cured materials cut into equal parts 5×5×1 mm in size were placed in the wells of a plate and again used to determine biofilm formation with subsequent coloring. To extract the dye, the pieces were transferred to a new plate to exclude the amount of film biomass formed on the walls of the plate wells. In both cases, cultivation was carried out at 37° C for 24-48 hours. The biomass of the film was stained with fuchsin. Statistical data processing was performed using t-Student criterion. For the threshold level of significance, the value p

Aim.To assess the growth kinetics and biofilm-forming activity of Candida albicans strains when cultivated in nutrient medium with different concentrations of lysocim. Materials and methods. Growth kinetics of strains was determined by changes in optic density of suspension at 620 nm, and biofilms detected them by their staining with fuchsin. The results were registered on photometer. Results.Lysocim changes the growth kinetics of C. albicans, reducing the duration of exponential and prolonging the stationary phases of growth. It was established that increased lysocim concentration positively influences the proliferative activity of the fungi. As lysocim concentration is increased, C. albicans biofilm-forming activity is elevating. Conclusions. Thus, the increased level of lysocim contributes to transition of fungi into the plankton form of growth that should be taken into consideration when treating patients with candidiasis.

Aim. To study the characteristic features of long-term facial phlegmon morbidity in Perm Krai with assessment of the efficiency of infusion therapy using indices of endogenous intoxication expression. Materials and methods. Manifestations of epidemic process of facial phlegmons were studied on the basis of a detailed retrospective analysis of sickness rate among the inhabitants of Perm Krai for the period of 2011–2016. In group 1, all patients besides standard therapy, underwent infusion therapy, and in group 2 – standard therapy alone. Prior to and after the therapy, peripheral blood cellular composition was assessed, and leukocytic indices of intoxication were calculated. Results. A long-term dynamics of facial phlegmon morbidity was established to show the irregularity of process expression according to years. Growth of the quantity of patients is provided by increase in the number of cases with complicated forms. Besides, males play a significant role in formation of sickness rate: the chances for the development of phlegmon among them are 1.49 times higher than in females. In the age structure of morbidity, prevail persons aged 21–30 and 31–50. A half of patients have endogenous intoxication. The course of infusion therapy essentially reduces the expression of endogenous intoxication. Conclusions. There is observed a tendency to growth of facial phlegmon morbidity that is probably connected with changes in etiological structure of pathogenic agents as well as with formation of pathosymbiosis of some opportunistic pathogenic microorganisms. As a rule, phlegmon is being developed against the background of endogenous intoxication, conditioned by both microbial metabolites and tissue destruction products. The efficiency of correction of endogenous intoxication using infusion therapy is shown.

Aim.To assess the growth properties and formation of E. сoli biofilms with different concentrations of cholesterol available. Materials and methods. E. coli strains were cultivated in the meat-peptone broth with cholesterol added in the concentration of 3, 5, 7 and 9 mmol/l as well as in the nutrient medium without cholesterol. The growth parameters and cholesterol concentration in the samples prior to and after cultivating microorganisms and the effect of cholesterol on biofilm-forming activity of E. coli was determined. Results. Cholesterol was shown to have no bactericidal effect on E. coli, however, it is able to change the duration of growth phases of the strains that is, possibly, caused by adaptation to the presence of cholesterol and switching of metabolic pathways. Cholesterol has a stimulating effect on accumulation of biomass by the test-strain and its biofilm-forming activity. Conclusions. In the study performed, the ability of E. coli to metabolize human cholesterol was shown.

Nowadays, in the presence of wide diagnostic possibilities, laboratory diagnostics of microecological disorders of vaginal biotope are often limited to clinical data, microscopic examination results and the use of a culture method. However, with such a complex it is impossible to get an answer about the functional activity of microorganisms. The aim of investigation was to evaluate the information content of a combined study of growth parameters and the ability to produce lactic acid by clinical strains of Lactobacillus spp. to characterize the state of the microecology of the cervical-vaginal biotope. Materials and methods. Studied the growth kinetics of strains of lactobacilli isolated from the detachable posterior vaginal fornix. The concentration of lactic acid in the medium was determined using the “Lactic Acid - Olvex” kit (Russia). Samples were taken every 12 hours of cultivation. Statistical analysis of the results was performed using the methods of descriptive statistics, Student’s t-test. Results. It has been shown that 44% of Lactobacillus spp. to 72 hours of cultivation did not reach the phase of exponential growth. The remaining strains from 12-24 hours passed into the phase of exponential growth. In the production of lactic acid, the strains that are in the lag-phase did not differ from each other, since they practically did not synthesize this metabolite. Among the remaining strains that are in the phase of exponential growth, some did not produce lactic acid, others increased their lactate production every hour. Part of the strains reached the peak of acid production by 36 hours and by 72 hours some decrease in synthetic activity was observed. Conclusion. It has been shown that for most vaginal strains of Lactobacillus spp. characteristic variability of the duration of the adaptation period and the level of functional activity. In addition, only a small number of clinical strains produced lactic acid 24 hours after the start of cultivation. Therefore, to adequately assess the microecological status of the vaginal biotope, microscopic examination of both native material and cultures isolated on special nutrient media is not enough. It seems that, along with the use of modern methods of genetic analysis, the determination of in vitro growth characteristics, primarily lag-phase duration, and lactate production by lactic acid bacteria strains can clarify many issues related to the formation of dysbiotic states, in particular, in the vaginal biotope, and will also serve to increase the effectiveness of the prescribed treatment.

Microbial biofilms are heterogeneous, moving and constantly changing communities of microorganisms, often of various taxons. Approaches to study and assessment anti-biofilm drugs widely available today do not adequately assess their effects, while the results of studying the interaction of drugs with components of the film composition can provide them the right choice. The aim of investigation was to test a new method of morphological evaluation of biofilms. To form biofilms, we used an approach when the slide was placed at an angle of 30o-45o relatively to the Petri dish, and a suspension of test strains S. epidermidis in peptone broth was poured into the space between the Petri dish and the slide. A sterile cotton swab moistened with distilled water was placed next to the glass slide to create optimal humidity. The system was placed in a thermostat for 24 hours. The formed films were examined under a microscope using the DCM 310 video eyepiece and the Scope photo x86,3.1.312 program that allowed to conduct a complete morphometric study of the film: select layers, channels, cavities and make measurements, and then save the results on electronic media in jpg file format. Microscopy of the stained slides revealed that the biofilm has a layered structure. In each image obtained using a video eyepiece, it was possible to differentiate 4 layers. From the border of the two media to the inside: the fragmentation layer, the dense layer, the matrix substance layer, and the last one - the persistence layer. Channels of different diameters (from 10 to 24 microns) are observed across the entire thickness of the biofilm. Thus, used approach allows us to visualize and evaluate the structure of microbial biofilm, measure the thickness of layers and channel diameters. In addition, this method can be used to study the effect of antimicrobial drugs on bacterial films.

Despite the long history of the study, laboratory diagnosis of gonococcal infection remains a complex task that does not have a clearly regulated effective solution. Aim of investigation was to assess the species diversity of the microbiota of the genital tract of men and women with suspected acute genital gonococcal infection (AGGI) using test systems of Russian manufacturers. A study of samples of the contents of the urethra of 69 men and posterior vaginal fornix fluids of 33 women of reproductive age with characteristic clinical manifestations and a presumptive diagnosis of AGGI was made. Cultivation was carried out using elective culture media with subsequent identification of strains by biochemical properties. Detection of DNA of Neisseria gonorrhoeae, Staphylococcus spp., Streptococcus spp., Enterobacteriaceae, Gardnerella vaginalis, Atopobium vaginae, Lactobacillus spp. performed by PCR using Vektor-Best and InterLab Service kits (Russia). All patients were divided into groups according to the results of the bacteriological method and PCR. A metagenomic study of 16S ribosomal RNA samples was performed on the Illumina MiSeq platform using the MiSeq Reagent Kits v3 kit (600-Cycle Kit). Statistical analysis of the data was performed using criterion x2. As a result of a laboratory study, the presumptive clinical diagnosis of «AGGI» found its bacteriological confirmation in 35.3% of cases only, among which fragments of the N. gonorrhoeae genome were detected in 63.9% of the samples only. Moreover, a wide variety of microorganisms in the genital tract of both men and women was found in metagenomic analysis. However, this technique does not allow us to assess the viability of the detected bacteria, and the microflora spectrum is excessively wide. In addition, the high level of genetic polymorphism of different strains of N. gonorrhoeae complicates the interpretation of the results. Deciphering the composition of microbiota allows the use of InterLab Service kits. The decoding of the etiology of purulent-inflammatory processes in the genital tract, which presents serious difficulties, is greatly facilitated by the use of Russian kits for molecular genetic analysis, which, in our opinion, provide the necessary and sufficient information for practice.

Currently, laser technology is widely used in medicine. However, relatively few studies on the simultaneous effect of laser radiation with similar parameters on the viability and functional activity of prokaryotic and eukaryotic cells. When evaluating the effect of laser radiation, especially in a clinical setting, it is necessary to take into account its possible effect not only on cells of conditionally pathogenic microorganisms, but also on human cells. In this study, it was shown that laser radiation of a diode laser with a wavelength of 405 nm (blue) and 650 nm (red) has a pronounced bacteriostatic effect. Thus, red laser radiation with a power of 22 mV allows you to achieve a bacteriostatic effect against Staphylococcus aureus, but it significantly reduces the viability of buccal epithelial cells. The results can be considered on the one hand as a side effect of laser radiation, and on the other hand, the removal of dying cells filled with microorganisms can enhance the effect of the therapy.

The study was carried out to assess the prospects for the combined use of leukocyte and cytokine indices in the bile duct obstruction of different genesis.We have performed a study of the patients with mechanical jaundice, either due to tumor (14 persons), or non-tumor genesis (16 persons). Concentrations of cytokines (IL-1, IL-4, IL-8, TNFα) were determined in blood serum before surgical intervention. The total activity of endogenous inflammatory mediators in the serum of patients was estimated as an index of inflammatory activity (IVA), which was calculated using the following formula: IVA = (IL-1 + TNFα)/IL-4. In addition, a number of leukocyte indices were calculated. The cellular component in the index estimation makes it possible to clarify the intensity of the inflammatory reaction, and to suggest the direction of the functional changes among the immunocompetent cells.It was shown that IVA in presence of malignancy is significantly higher than in benign conditions. Overactivation of immune response in tumors may be caused by a significant predominance of pro-nflammatory cytokines. Moreover, in cases of malignant tumor growth, both with or without cholangitis, the index scores of humoral and cellular components let us suggest the some problems with regeneration, due to pro-inflammatory domination of immune response, which does not ensure activation of the corresponding reactions. It has been established that the leukocyte intoxication index (LII) is increased,in patients with a combination of malignant disorder and cholangitis, and the immunoreactivity index (MDI) is lowered. In absence of cholangitis, the opposite pattern is observed, when LII is slightly lowered and MDI is elevated. Correlation analysis for the patients with cholangitis revealed a high association of cytokine imbalance and endogenous intoxication. A complex of cytokine and leukocyte indices is promising, when each of them reflects a certain part of the process, and their combination gives an integral picture of the pathology and allows to predict development of the disease. Such an approach, seems to increase the diagnostic significance of these immunological indices, primarily, the cytokine production, and may be used when predicting currence of complications in the surgical treatment of the disease.

It is known that the concentration of alpha-amylase in saliva can determine its catalytic activity, the decrease of which occurs during various pathological processes in the oral cavity. The overwhelming number of methods for determining the catalytic activity of enzymes involve the use of a large volume of reagents and samples, which makes it difficult to study saliva in large groups. The purpose of the study is to evaluate the possibility of using the microvariation of the reaction to determine the activity of saliva alpha-amylase, as well as to analyze the dependence of the enzyme activity on its concentration. Materials and methods. Saliva was obtained from 15 people with intact periodontal disease and the dentition, without somatic pathology. For in vitro studies, alpha-amylase solutions were prepared with an enzyme concentration of 10; five; 2.5; one; 0.5 and 0.25 mg / ml ex tempore. To save samples and reagents, the volume of the reaction participants was proportionally reduced. The further analysis procedure was carried out according to the instructions of the manufacturer of the «AMYLASE-VITAL» reagent kit to determine the activity of alpha-amylase. Statistical analysis of the results was performed using the Student’s t-test in the program Statistica 7.0. Results. The comparability of the results of determining the activity of alpha-amylase using the classical and microplate variants of the reaction is shown. With an increase in alpha-amylase concentration from 0 to 2.5 mg / ml, a directly proportional increase in enzyme activity is observed. In the case of an increase in the concentration of alpha-amylase above 2.5 mg / ml, a decrease in its activity is shown, which may be due to the precipitation of a part of the enzyme. The activity of the enzyme in saliva of practically healthy individuals using the microvariation of the reaction was 528.6 ± 2.4 U / l. In conclusion the use of a microvariant of the reaction for determining the activity of alpha-amylase may be justified for a large number of subjects. A linear dependence of the enzyme activity on its concentration in the range of 0-2.5 mg / ml is shown.

Introduction. It is known that the number of patients with phlegmons does not tend to decrease, which can be due to both disease pathomorphosis and changes in the epidemiological features of this pathology. In addition, a significant contribution to the development of the disease makes the characteristics of the comorbidity background. The aim of the study was to assess the clinical and epidemiological aspects of the morbidity of the Perm city population with the maxillofacial area phlegmons. Materials and methods. A retrospective analysis of 137 case reports ofpatients with facial phlegmon was carried out. Results. It is shown that young and middle-aged patients dominated in the age structure of the disease. Phlegmons often localized in 1-2 spaces. In more than half the cases, the phlegmons was in the submandibular space. Among the etiological factors of the disease, the leading place is occupied by gram-positive cocci and their associations. An essential role of anaerobic microflora in the pathogenesis of phlegmons has been established. Microorganisms of the Enterobacteriaceae family play a significant role in the formation of inter-microbial associations in the development of this pathology. Often phlegmons of the maxillofacial region are found in patients with concomitant somatic pathology. With a comorbid component, more frequent detection of microorganism associations is indicated. In addition, such patients enter the hospital much later. Conclusion. Thus, when assessing clinical and epidemiological data, it is shown that the phlegmons of the maxillofacial region occur, as a rule, in young and middle age, and also in the presence of comorbid pathology. A significant contribution to the development of the disease is made by associations of microorganisms, which may include a change in the severity ofpathogenicity factors.

Current methods of biofilm imaging do not support a differentiated assessment of its composition, since it is not possible to establish a substrate stained with crystal violet, as this dye can form complexes with both intracellular and extracellular structures. This approach does not adequately assess the anti-biofilm effects of drugs, while the results of studying the interaction of drugs with biofilm components can ensure their most correct choice. The aim of investigation was to study the possibility of applying the original modification of the current method to determine the ratio of the cellular part and the matrix of biofilms of gram-positive microorganisms. The biofilm components were analyzed using a two-step approach, when prepared biofilms of gram-positive microorganisms were stained with crystal violet for 5 minutes, followed by fixing the dye in bacterial cells with iodine solution, and then the colored products were dissolved with 95% alcohol: matrix components for 1 minute, total biofilm for 15 minutes, after which the composition of biofilms was estimated by the formula: M=(OP1/OP15)×100, Kb=100-M, where M is the proportion of the matrix,%; Kb - the proportion of the cellular component,%; OP1 - optical density of samples, when alcohol was allowed to dissolve the colored product for no more than 1 minute; OP15 - was the optical density of samples, when alcohol is allowed to dissolve the colored product for 15 minutes. It was shown that in the composition of the biofilm formed by the collection strain, the proportion of the matrix was 13.2%, and the cellular component accounted for 86.8%. When the same strain cultivated in the presence of an antibiotic, an increase in the biofilm matrix was observed, which is probably due to the compensatory response of the microorganism to the action of the antibiotic. The proposed approach to the study of biofilms makes it possible to evaluate its component composition. Obtaining additional information in this way can provide, inter alia, an increase in the effectiveness of antimicrobial therapy while reducing the study time.

The aim of research - a comparative study of the microflora of the vagina and bowel in women with secondary infertility and gastrointestinal distress. Materials and methods. We examined gut and vaginal microbiota in 17 infertile women according to the presence or absence of gastrointestinal diseases. We used a standard procedure for microbiological examination. Results. Certain changes of bacterial load and composition in both reservoirs were established. They allowed us to conclude the same trends in characterizing of the developing dysbiosis. Conclusion. Thus, the simultaneous studying of vaginal and gut microflora seems to be rational during the examination of women with childbearing disorders. It could possibly increase the treatment efficacy and/or prevent the development of some pathology.