Quantitative method for the determination of antibiotic-resistant gut bacterial strains for the early diagnosis of chronic arthritis.

Based on the clinical and microbiological monitoring of two groups of children aged 3 to 17 years with acute (n=78) and chronic (n=46) course of reactive arthritis (ReA), a method for early diagnosis of chronic arthritis was developed by determining the number of antibiotic-resistant coprostrains in patients with ReA, characterized by the absence of the need to isolate a pure culture of the pathogen and its identification; inoculation of faeces at a dilution of 10-5 on solid 1.5% GRM-agar with an antibacterial agents used in the treatment of a particular patient, at a minimum inhibitory concentration in the resistance range, followed by incubation and counting of the colonies of microorganisms grown on the plate. A significant relationship between the number of antibiotic-resistant gut bacterial strains and the course of arthritis (acute, chronic) was revealed, and the borderline value of the number of antibiotic-resistant gut bacterial strains was determined - 5×103 CFU/g, which allows differentiating the acute course from the chronic one: in the acute course< 5×103 CFU/g, with chronic - ≥ 5×103 CFU/g. The method allows, at the stage of completion of anti-inflammatory therapy in the active phase of the disease, to identify a risk group for the development of a chronic course of arthritis among patients with ReA, which can contribute to timely therapeutic measures aimed at preventing recurrence of the disease and making the patient disabled.

[Characterization of the microbiota and cytokine profile of sperm plasma in men with chronic bacterial prostatitis].

BACKGROUND: One of the leading causes of the occurrence of chronic bacterial prostatitis (CBP) in men is infection, microecological disorders of the urogenital tract and cytokine-mediated mechanisms of inflammation of the prostate gland, which actualizes a comprehensive study of the clinical and bacteriological features of CBP from the perspective of a symbiotic approach in the framework of a new scientific field - "infectious symbiology". OBJECTIVE: to study the characteristics of spermogram, microbiota, and the cytokine profile in men with chronic bacterial prostatitis (CBP) and CBP complicated by infertility. MATERIALS AND METHODS: A comprehensive study of patients with CBP and CBP complicated by infertility, in comparison with conditionally healthy individuals, was conducted. Species identification of microorganisms was carried out according to biochemical characteristics and the genetic method (sequencing of strains). The biological properties of the microbiota were evaluated: growth properties, biofilm formation, antipeptide activity against the cytokines IL-10, RAIL-1, TNF-, INF- and IL-17 (8 parameters). Immunological parameters of sperm plasma included 13 parameters: the content of cytokines TNF-, INF-, Rail, interleukins (IL) -1, 2, 4, 6, 8, 10, 17, immunoglobulin (Ig) A, lactoferrin and lysozyme. To evaluate sperm plasma, the following quantities were determined: ejaculate volume, pH, sperm plasma liquefaction, total sperm count, sperm count per 1 ml, motility, number of progressively motile, non-progressive motile and motionless spermatozoa, number of round cells, white blood cells, spermatogenesis cells, erythrocytes, erythrocytes, cells, sperm agglutination and aggregation (16 parameters in total). The results are statistically processed. RESULTS: Data were obtained on changes in biofilm formation, antipeptide activity of microbiota (especially pronounced in corynebacteria), sperm plasma cytokine profile (increased TNF , IL-2, 6, 17), as well as IgA and lactoferrin, which can be used to build a prognostic model of reproductive pathology tract of men and their fertile activity. CONCLUSION: The study of the antipeptide activity of microbiota in combination with the cytokine profile of ejaculate allows us to recommend them as a "biotarget" for diagnostic, preventive and therapeutic measures for chronic prostatitis in men, which contribute to solving the medical and social problem of preventing male infertility and contributes to the development of health-saving technologies with incorporating elements of personalized medicine.

[GUT MICROSYMBIOCENOSIS IN CHILDREN WITH REACTIVE ARTHRITIS].

AIM: To study the state of gut microsymbiocenosis in children with reactive arthritis (RA), with the assessment of biofilm formation (BFF) of microsymbionts and the ability to change cytokine levels (their anticyokine activity) in vitro. MATERIALS AND METHODS: The investigation of gut microsymbiocenosis by means of bacteriological method was conducted in 34 children with RA and 25 relatively healthy 3 - 16 year- old children. Microorganisms were identified with the help of MALDI-TOF mass-spectrometry, anticytokine activity (ACA) of microsymbionts - according to Bukharin O.V et al. (2011), biofilm formation - according to O'Toole G.A., Kolter R. (1998). RESULTS: On the ground of species composition differences of gut microbiota discrimination model was created which allowed to separate the group of children with RA from healthy individuals. Microsymbiocenosis of patients with RA was characterized by increasing number of opportunistic microorganisms (OM) (enterobacteria, clostridia, bacteroides, and Candida), BFF and ACA level. CONCLUSION: The obtained data greatly contribute.to the deciphering of spondylo- arthritis and disclose the role of microbial factor under given pathology. Hypercolonisation of human gut with OM, having pronounced ability to BFF and regulating cytokine level, promotes strengthening of arthritogenic potential and serves as additional marker of arthritis development risk in children.

[IMMUNE REGULATORY PROPERTIES OF BIFIDOBACTERIA METABOLITES DURING EUBIOSIS AND DYSBIOSIS OF THE HUMAN COLON].

AIM: Evaluate immune regulatory properties of bifidobacteria metabolites during eubiosis and dysbiosis of the human colon. MATERIALS AND METHODS: Anti-cytokine activity of metabolites of bifidobacteria clinical strains and their ability to influence the production of pro- and anti-inflammatory cytokines (IL-10) by peripheral blood mononuclear cells of healthy humans was studied, taking into account microecological state of the human intestine. Determination of final concentration of cytokines in experimental and control samples was carried out by EIA. RESULTS: Sensitive parameters, that are suitable for evaluation of stability of human intestine microsymbiocenosis, were detected. The level of microbial seeding, concentration of TNF-α and anti-lysozyme activity turned out to be informative for bifidobacteria in eubiosis conditions. The ability of bifidoflora metabolites to influence the production of pro-inflammatory cytokines (INF-γ, TNF-α, IL-8) by human mononuclears was a significant parameter during formation of 1 - 3 degree dysbiosis. CONCLUSION: The maintenance of physiological state of intestine homeostasis is determined by immune regulatory properties of bifidobacteria metabolites, that is realized via their interaction with both cytokines (anti-cytokine activity) and production of cytokines by host immune cells (peripheral blood mononuclears).

[HEALTH STATUS AND DEVELOPMENT OF CHILDREN BORN USING IVF].

In the work the analysis of incidence, the dynamics of physical development during the first year of life among 28 children born using IVF was conducted. It was found out that during the first year of life these children have certain patterns of growth and development, as well as a wide range of diagnosed pathology. Thereby observing children of this category should be optimized.