Vol 94, No 3 (2017)

Aim. To study the production of cytokins on the model of peripheral blood lymphocytes under the activity of Bifidobacterium bifidum 791 strain induced by Lactobacillus fermentum 90T-C4, Escherichia coli 157 and Staphylococcus aureus 209 metabolites. Materials and methods. Reference strains of «self» and «поп-self» types of bacteria were used in the investigation. «Self/non-self» microbial recognition method (Bukharin O.V., Perunova N.B., 2011). Mononuclear leukocytes were isolated from the blood of healthy donors by gradient centrifugation in ficoll-verographin density gradient (Pharmacia, Sweden). Production of pro-(IFN-y, TNF-a, IL-6, IL-17) and anti-inflammatory (IL-10) cytokins was investigated in mononuclear culture by ELISA method. The results are statistically processed. Results. Similarities in the direction of lymphocyte reaction and «self» and «поп-self» microbial differentiation of bifidobacteria were found. It was determined that in reaction to «поп-self» reference cultures the lymphocytes increased pro-inflammatory potential and increased anti-inflammatory potential in reaction to «self» bacteria. Preliminary co-incubation of bifidobacteria with L.fermentum metabolites 90T-C4 increased anti-inflammatory effect of B. bifldum 791, whereas lymphocyte reaction to E. coli and staphylococcus induced bifidobacteria was changed to pro-inflammatory. Conclusion. Combined unidirectional influence of microbiota and its metabolic activity on cytokine level might enhance defence effect of intestinal immune response. The capacity of bifidoflora to carry out primary selection of microsymbionts on account of intermicrobial «recognition» and differentiated exposure to lymphocyte pro- and anti-inflammatory potential evidences the key role of bifidoflora in the human intestine homeostasis maintenance.

Aim. The work was performed with the purpose to study antibacterial action of millimolar concentrations of copper and zinc ions, used as sulfates or chlorides in the cultures of S.pyogenes and S.agalactiae. Materials and methods. Suspensions of the S.pyogenes and S.agalactiae bacteria which contained 10⁸ CFU/ml were sown by the lawns into the standard Petri dishes coated with the supplemented Nutrient Agar. 30 min later the salt solutions of copper or zinc were added by the 5 pi drops on the surfaces of the lawns with use of 36-channel stamp replicator. The salt solutions contained copper or zinc at the concentrations ranged between 5xl0^-3 M to 5xl0^_1 M of the metals. Then the dishes containing bacterial cultures were incubated for 24 hrs at 37°C followed by measuring diameter of the area of culture growth inhibition. The probes of material obtained from the centers of the stunting areas were passed into the centrifuge tubes with the supplemented Nutrient Broth, incubated for up to five days at 37°C and tested for the Broth clarity. Results. In presence of the metal concentrations ranged between 50 to 500 mM the inhibitory action of zinc sulfate towards S.pyogenes bacteria was 1.5 - 1.7 times higher (p<0.001) than the action of copper sulfate. In presence of 100 and 500 mM of the metals the action of zinc chloride was 1.2 - 1.3 limes higher (p<0.05 - 0.1) than the effects of copper chloride. In presence of 100 and 500 mM of copper used as chloride its antibacterial action towards S.agalactiae bacteria was 8 - 13 per cent higher (p>0.1) than the effects of zinc chloride. Passages of the material obtained from the areas of culture growth inhibition demonstrate clarity of the Broth in 67 - 85 per cent (S.pyogenes) or 48 - 67 per cent (S.agalactiae) probes with samples from the areas treated with copper salts, and in 30 per cent probes with samples from the areas treated with the salts of zinc. Conclusion. Inhibitors’ action of copper and zinc ions used in millimolar concentrations in the cultures of S.pyogenes and S.agalactiae was established. S.pyogenes, bacteria demonstrate higher than S.agalactiae sensitivity to the action of zinc ions measured by the area of the growth inhibition. Also, in a comparison with S.agalactiae, the S.pyogenes bacteria undergo bactericidal action of zinc ions in a higher part of observations.

Aim. Study the intensity of circulation of HCV in Khabarovsk Region and genetic variants of the virus circulating in the region. Materials and methods. 940 sera samples obtained from individuals with unestablished risk factors for HCV infection in polyclinic of the Far-Eastern Regional Centre for Prophylaxis and Control of AIDS (Khabarovsk) were studied. Individuals of the following age groups were included into the study: 1 - 4, 5 - 9,10 - 14,15 - 19,20 - 29, 30 - 39, 40 - 49, 50 - 59, older than 60 years; each group included approximately 100 individuals. Anti-HCV was determined in all the samples, HCV RNA was determined in samples positive for anti-HCV. Phylogenetic analysis with a time scale for the detected genovariants of HCV was carried out for reconstruction of the history of prevalence of HCV in Khabarovsk Region. Trees were constructed using BEAST vl.8.2 program by 2 genome sequences - core (1043 bp) and NS5B (389 bp). Results. The mean prevalence of anti-HCV among the examined individuals was 9.3% (87/940). The proportion of individuals with a current HCV-infection (presence of HCV RNA) was 4.2% (39/940). A sharp increase of HCV prevalence was observed beginning from the age group 20 - 29, where frequency of anti-HCV was 13.1% (13/99) that is 6 times higher compared with the previous age group of 15 - 19 (2,2%, p<0.01). The highest frequency of detection of HCV markers was observed in the age group 40 - 49 (20.8 and 9.4% for anti-HCV and HCV RNA, respectively). The frequency of detection of anti-HCV and HCV RNA in the 30-39 group was 20.2 and 8.9%, respectively, and among individuals older than 60-16.8 and 7.4%, respectively. Genotype lb was determined for 64.2% of HCV isolates, 28.3% of isolates belonged to genotype 3a, and 3 isolates were similar to genotype la, 2a and 2c members. Genotype lb was introduced into Khabarovsk Region around 55 - 65 years ago from Japan and the USA, around 15 years later - genotype 3a. Importation of novel HCV genotype 3a strains from Europe took place many times, these strains are present across the whole Russia. Conclusion. The data obtained have demonstrated high prevalence of HCV in the Khabarovsk Region, that has formed due to a prolonged circulation of the virus in the region.

Aim. Determination of prevalence of markers of hepatitis A and E among migrant workers, who had recently arrived in Russian Federation from countries with various degree of endemicity regarding this infection. Materials and methods. Sera samples from 1333 migrant workers, who had recently arrived in Russian Federation from Uzbekistan (n=464), Tadzhikistan (n=415), Ukraine |n=308) and Moldova (n=146) were studied. Anti-HAV IgG, anti-HEV IgM and IgG were determined using commercial ELISA tests. HEV RNA was determined in samples positive for anti-HEV IgM by RT-PCR with degenerate primers to HEV genome sequence coding capsid protein. Anti-HEV IgG were detected significantly more frequently among migrants from Uzbekistan and Tajikistan compared with migrants from the Ukraine and Moldova (25,4 and 43,1% versus 7,8 and 12,3%, respectively, p<0,05), that reflects the degree of endemicity regarding hepatitis E from regions of origin of the examined individuals. Frequency of detection of anti-HAV IgG also depended on the country of origin - seropositive individuals were present significantly less frequently among migrants from the Ukraine compared with individuals who had arrived from Moldova, Uzbekistan and Tajikistan (70,1% versus91,8; 98,7; 99,8%, respectively, p<0,05). Anti-HEV IgM that give evidence on the current or recent infection were detected among migrants from Uzbekistan, Tajikistan, Ukraine and Moldova with approximately the same frequency - 3,9; 7,8; 5,8 и 6,8%, respectively. Viral RNA was detected in none of the samples positive for anti-HEV IgM that could be related to the short period of viremia during hepatitis E. Conclusion. A significant portion of migrant workers arriving in Russian Federation have anamnestic antibodies against HAV and H EV, and this parameter reflects the degree of endemicity of the country of origin for migrants. High frequency of detection of anti-HEV IgM in migrant workers, that gives evidence on the current and recent infection, allows to assume high possibility of import of H EV into Russian Federation, and this necessitates introduction of acute HEV infection markers into algorithm of examination.

Aim. Expansion of arsenal of means capable of inhibiting production of staphylococci entero-toxins (SE) and having an ability to eliminate them from biological substrates, as well as reducing the growth of staphylococci. Materials and methods. Reference strain of Staphylococcus aureus FRI 722 was used as SE producer type A (SEA), S. aureus S6 715H - as SE type В producer (SEB). Polymethylsiloxane polyhydrate (PMSPH) was used at concentrations of 1.82, 9.09 and 18.2%. Results. By using gel double diffusion method and ELISA we have established that a 18.2% solution of PMSPH (enterosgel; PMSPH - 70 g, purified water - 30 g per 100 g of the product) is an optimal concentration for inhibition of production of staphylococcus enterotoxin type A by 100 and more times, and production of staphylococci enterotoxin type В - by more than 300 times. Conclusion. PMSPH is able to eliminate staphylococci enterotoxins type A and В from biological substrates for more than 50% and significantly reduce growth of staphylococci.

Aim. Experimental evaluation of effect of E. coli LPS fractions on phagocyte and metabolic activity of blood neutrophils of laboratory mice with induced immune deficiency. Materials and methods. Phagocyte activity was evaluated by phagocyte number (PN), phagocyte index (PI) and integral phagocyte index (IPI), intensity of metabolism and energetic processes of enzyme systems - by test of tetrazolium nitro blue (TNB), mean cytochemical coefficient (MCC) in spontaneous and induced tests and stimulation index (SI). Results. LPS-3 substance administration into mice with secondary immune deficiency has resulted in a significantly highest increase of PN (15.8%), IPI (17.7%), TNB-IN (10.3%), cytochemical coefficients MCC-IN (u.) and IS (u.) - 14.8 and 10.9%, respectively, compared with the parameters of immune deficient mice that had received licopid. Conclusion. Some fractions of E. coli M17 polysaccharide have immune stimulating activity.

Aim. Determine the prevalence of antibodies to hepatitis C virus (anti-HCV) among conditionally healthy population of Russian Federation. Materials and methods. Sera samples obtained from conditionally healthy population of the Russian Federation from 5 geographically distant regions with different levels of morbidity for hepatitis C were studied: Sverdlovsk, Rostov, Moscow regions, Tyva and Sakha (Yakutia) Republics. 4764 samples were studied in total obtained from individuals of 10 age groups: younger than 1, 1 - 4, 5 - 9, 10 - 14,15 - 19, 20 - 29, 30 - 39, 40 - 49,50 - 59, olderthan 60 years. Anti-HCV were determined by commercial ELISA. Results. The total prevalence of anti-HCV in 5 regions of Russian Federation was 2.6% (126/4764). The highest frequency of detection of anti-HCV was detected in Tyva and Yakutia Republics and was 3.3%, in other regions this parameter varied from 1,7 (Moscow region) to 3% in Sverdlovsk region. Peak parameters of anti-HCV detection among population in various regions were connected with various age groups: olderthan 60 years in Tyva and Yakutia Republics (11 and 9.9%, respectively), 40 - 49 in Moscow region (6.2%), 1 - 4 and 30 - 39 years in Sverdlovsk region (5.9 and 4.9%, respectively) and 20 - 39 in Rostov region (3.9%). Conclusion. Results of anti-HCV detection in various age cohorts of conditionally healthy population of Russian Federation give evidence regarding high morbidity with HCV of almost all the age groups in the interval from 20 to older than 60 years. Age cohorts were detected in each of the studied regions for which inclusion into regional screening programs is feasible.

Bacterial lipopolysaccharides (LPS) are highly toxic molecules released during the lysis of bacterial cells. They play important role in the pathogenesis of sepsis, and can contaminate pharmaceuticals, so removing them from aqueous solutions and biological fluids is an extremely important task. Structure of LPS and its toxicity for various animals are presented in this review. Various low- and high-molecular ligands, suitable for efficient binding and removal LPS from solutions are studied and demonstrated. The main attention is paid to the relationship between the chemical structure of the ligand and its ability to form strong complexes with LPS and the principles of creating selective ligands for the depyrogenation of pharmaceutical substances and the creation of hemoperfusion columns for the sepsis therapy.