[Effect of Fitofron on the persistence factors of opportunistic pathogens in urine isolated from patients with urinary tract infections].

AIM: To perform an experimental evaluation of the effect of Phytofron, used for the treatment of urinary tract infections, on the ability of opportunistic pathogens to inactivate innate immunity factors (lysozyme, pro- and anti-inflammatory cytokines) and form biofilms. MATERIALS AND METHODS: In vitro experiments were carried out on clinical isolates from urine of patients with pyelonephritis and cystitis: Escherichia coli, Staphylococcus aureus, S. haemolyticus, S. epidermidis, Enterococcus faecalis. The effect of Fitofron NPO FarmVILAR (Russia) on the anticytokine activity of bacteria against regulatory cytokines (IL4, IL6, IL8, TNF and IL17A) was determined by enzyme immunoassay, while anti-lysozyme trait and the ability to form biofilms was evaluated by the photometric method. RESULTS: The inhibitory effect of Fitofron on the ability of opportunistic microorganisms to inactivate innate immunity factors (lysozyme) and form biofilms, as well as the predominant inhibition of the studied cytokines, was experimentally established. CONCLUSION: Inhibition of the persistence factors of opportunistic pathogens by Fitofron, documented in vitro, can be considered as one of the possible mechanisms of its biological activity in vivo.

[Effect of Prostanorm on the persistent potential of microorganisms isolated from patients with chronic bacterial prostatitis].

AIM: To determine the effect of the Prostanorm on the persistent potential of microorganisms isolated from the prostatic secretion of patients with chronic bacterial prostatitis. MATERIALS AND METHODS: In vitro experiments were carried out on uropathogens, isolated from prostate secretions in patients with chronic bacterial prostatitis, including Escherichia coli, Staphylococcus aureus, S. haemolyticus, S. epidermidis, Enterococcus faecalis. The effect of Prostanorm (liquid extract for oral use) of NPO FarmVILAR (Russia) on the anti-cytokine activity of bacteria against regulatory cytokines (IL4, IL6, IL8, TNF and IL17A) was evaluated by enzyme immunoassay, while anti-lysozyme trait and the ability to form biofilms was evaluated by the photometric method. RESULTS: Inhibitory effect of Prostanorm on the anti-lysocyme activity and the ability of microorganisms to form biofilms was found. A decrease in the anticytokine activity against IL17A and TNFa in all studied species of microorganisms was seen, including anticytokine activity against IL4, IL6, IL8 in Escherichia coli and against IL6 and IL8 in coagulase-negative staphylococci. CONCLUSION: The ability of Prostanorm to reduce the persistence factors of opportunistic microorganisms in vitro is one of the possible mechanisms, proving its prostatotropic action, bacteriostatic and anti-inflammatory activity.

Diagnostic cytokine marker of male infertility - interleukin 4.

Analysis of the study is to assess the diagnostic significance of cytokines in the sperm plasma of men of reproductive age (20 - 45 years) of two groups: of patients with chronic bacterial prostatitis, not complicated by infertility and with loss of fertility. The study of sperm plasma - the WHO standard. Determination of the level of cytokines in seminal plasma - by enzyme immunoassay («Cytokine¼, Russia). Two methods of mathematical statistics were used: discriminant analysis and classification trees (decision trees).The similarity of interpretations of discriminant analysis and decision tree was noted, where the main role in both cases belongs to the cytokine IL-4. The level of sperm IL-4 in combination with therapeutic monitoring can be used for the medical management of patients with chronic prostatitis in order to prevent the development of infertility and to develop methods for screening diagnostics of fertility disorders in men.

[Antibiotic resistance of strains of gram-positive cocci isolated from prostate secretion in men with chronic bacterial prostatitis].

OBJECTIVE: to characterize the antibiotic resistance of gram-positive cocci strains isolated from the prostate secretion in men with chronic bacterial prostatitis at the level of phenotype and genotype. MATERIALS AND METHODS: Bacteria were isolated from the prostate secretion of men of reproductive age (20-45 years) with chronic bacterial prostatitis by conventional bacteriological method. The type of microorganisms was determined by MALDI-TOF mass spectrometry. Resistance to 16 antibiotics of 31 strains of E. faecalis and 91 cultures of coagulase-negative staphylococci (CoNS) was determined by the disk diffusion method. Antibiotic resistance genes (mecA; blaZ; aac(6)- aph (2); ant (4)-Ia; aph (3)-IIIa; gyrA, grlA) were detected using polymerase chain reaction (PCR ) using selected primers. RESULTS: A high resistance of enterococci to antibacterial drugs was revealed: fluoroquinolones, carbapenens, cephalosporins (with the exception of cefoperazone), gentamicin and oxacillin. It has been established that CoNS are characterized by variable antibiotic resistance, while: isolates of S. epidermidis and S. haemolyticus are resistant to all studied fluoroquinolones and carbapenems; S. warneri to carbapenems and the vast majority of studied cephalosporins; S. saprophyticus - to aminoglycosides. Amoxiclav and cefoperazone are characterized by the highest activity against clinical isolates. Using PCR, the presence of genetic determinants of resistance to aminoglycosides and -lactams was established in the isolates, with a predominance of the studied genes in CoNS. CONCLUSION: For effective antibiotic therapy in chronic bacterial prostatitis, it is necessary to conduct regional monitoring of the resistance of microorganisms to antibacterial drugs.

[Evaluation of antibiotic sensitivity of microorganisms, which were isolated from patients with acute pyelonephritis in the stage of purulent inflammation and located inside red blood cells (in vitro experiment)].

AIM: To study the antibiotic sensitivity of strains of microorganisms isolated from patients with a purulent stage of acute pyelonephritis and located inside the erythrocytes. MATERIALS AND METHODS: In this study, a total of 15 strains of microorganisms isolated from the blood of patients with a purulent form of acute pyelonephritis were used. The bacteria were detected and identified in the blood of patients using a modern molecular genetic method, fluorescent in situ hybridization (FISH), and the classical bacteriological study was done to isolate blood cultures of microorganisms. When studying the effect of antibiotics on the microorganism strains located inside red blood cells, the method of staged lysis was used. RESULTS: It was found that microorganisms isolated from patients with a purulent stage of acute pyelonephritis and, subsequently, inside red blood cells (in vitro experiment) in 80% of cases were resistant to antibacterial drugs of different groups (fluoroquinolones, aminoglycosides, macrolides, phosphonic acid derivatives). Outside of red blood cells, the same pathogens were sensitive to the antibiotics used in the work. Apparently, it is possible to consider the resistance to antibiotics of uropathogens located inside red blood cells as a result of insufficient intracellular concentration of the drug even when it is delivered by the method of stepwise lysis. CONCLUSION: When treating patients with acute pyelonephritis in the stage of purulent inflammation, it is necessary to consider that microorganisms can penetrate into red blood cells. In vitro experiments have shown that microorganisms inside red blood cells exhibit antibiotic resistance to antibacterial drugs of different groups. It is necessary to further study the intraerythrocytic microorganisms in the purulent stage of acute pyelonephritis, as a new pathogenesis mechanism of its development. Perhaps, these results can result in revision of the principles of antibiotic therapy.

[Characteristics of the pathogenic potential of Escherichia coli isolated from patients with calculous pyelonephritis].

OBJECTIVE: Comparative phenotypic and genetic assessment of the pathogenic potential of E. coli strains isolated from patients with calculous pyelonephritis. MATERIALS AND METHODS: 78 strains of E. coli isolated from urine of patients with calculous pyelonephritis in the acute phase (n=58) and in the remission phase (n=20). Escherichia were investigated for the presence of virulence genes papA, pap EF, papGII; afa, bma E, iutA, fyuA, feoB, kspMTII, usp multiplex PCR using selected primers. Phenotypically determined the ability to biofilm formation, antilysozyme, antihemoglobin, anticytokine, adhesive and sIgA-protease activity E. coli. RESULTS: The virulent potential of Escherichia coli at the pheno- and genotype levels was characterized. In strains of E. coli isolated from the urine of patients in the remission phase, the ability to form biofilms was more often and with high values of the trait; and in strains isolated in relapse - adhesive activity, the ability to inactivate pro- and anti-inflammatory cytokines, antihemoglobin activity, and genes encoding aphimbrial adhesin (afa), responsible for the synthesis of siderophore aerobactin (iutA), transporting bivalent iron (feoB). CONCLUSION: The revealed differences in the pheno- and genotypic profiles between the cultures of Escherichia coli isolated from patients with calculous pyelonephritis in the phases of exacerbation and remission make it possible to differentiate the isolated strain and predict the course of the infectious-inflammatory process.

[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.

[The additional criterion for the diagnosis of purulent-destructive forms of acute pyelonephritis].

AIM: to develop an additional criterion for objective diagnosis of purulent-destructive forms of acute pyelonephritis. MATERIALS AND METHODS: A total of 35 blood samples from patients with acute pyelonephritis aged from 19 to 85 years (mean age was 52 years) were studied. For the analysis, a classical bacteriological method of isolating blood culture and a modern molecular genetic method of fluorescent in situ hybridization (FISH) using DNA probes complementary to species-specific regions of the 16S rRNA gene of microorganisms were used. The obtained results were processed using the Statistica 6.0 program. RESULTS: The bacteriological study showed that only in one patient (2.9%) a blood culture was positive, whereas with FISH, polymicrobial infection consisting of two associates was observed in five patients (14.3%), and 207 specific luminescence from different DNA probes were detected in blood samples. The associations of members of the Enterobacteriaceae with S. aureus were 2.4 times more frequent than associations of the Enterobacteriaceae with S. epidermidis, indicating a development of the polymicrobial infection and, accordingly, the development of purulent stage of acute pyelonephritis. CONCLUSION: For the diagnosis of acute pyelonephritis, we suggest to use an additional criterion, which consists in applying of FISH method for the detection and simultaneous identification of bacteria that were adhered on the surface of erythrocytes and located intracellularly. This method may allow for the differential diagnosis of serous and purulent stages of acute pyelonephritis.

[Biological properties of microorganisms isolated from the urine of patients with urolithiasis].

AIM: To define persistent properties and antibiotic resistance of microorganisms isolated from the urine of adult patient undergoing surgery for urolithiasis. MATERIALS AND METHODS: Urine specimens were obtained from the renal pelvis and urinary bladder during percutaneous nephrolithotripsy. Microorganisms that were isolated from the urine were examined for their persistent properties (anti-lysozyme activity, the ability of biofilm formation) and antibiotic resistance using photometric and bacteriological methods. RESULTS: Strains of microorganisms isolated from the urine of patients with urolithiasis have high anti-lysozyme activity and the ability of biofilm formation, and variable antibiotic resistance. These properties should be taken into account when selecting an empirical antibiotic therapy for preventing infectious-inflammatory complications after percutaneous nephrolithotripsy. CONCLUSION: The high level of resistance of microorganisms isolated from the urine of patients with urolithiasis to the studied antibiotics, their ability to inactivate lysozyme and form biofilms may be the cause of the development of postoperative complications.

[Structure of microorganism species cultured from urine of urolithiasis patients].

AIM: To investigate the incidence of microorganisms of different taxonomic groups and their associations in the pelvic and bladder urine of adult urolithiasis patients. MATERIALS AND METHODS: A bacteriological method and metagenomic sequencing were used to investigate the bacterial spectrum of microflora cultured from pelvic and bladder urine sampled during surgical interventions in urolithiasis patients. RESULTS: The both microbiotas had approximately the same spectrum, but in 26.1% of patients it was inconsistent. Metagenomic analysis detected DNA of microorganisms in urine samples which were found free of microflora by the bacteriological method. CONCLUSION: The study findings showed species diversity of microorganisms cultured from pelvic and bladder urine sampled during surgical interventions in urolithiasis patients.