Vol 90, No 6 (2013)

Aim. Evaluation of efficiency of non-thermal plasma as bactericidal agent affecting biofilms formed in vitro and on walls of a root channel. Materials and methods. The multiple antibiotic resistant strain Staphylococcus epidermidis isolated from pulpitis was used. Biofilms formed in vitro on the plastic surface and ex vivo at the walls of the root canal were treated with plasma torch formed by argon:air (9:1) mixture eradiated with 100 kHz electrtomagnetic field. Bacterial viability was determined by plating and by differential Live/Dead labeling. Results. The dose-dependent decrease in living bacteria was demonstrated. The three-step kinetics ofbacterial killing was observed. Total elimination ofup 10 9 CFU/sample was reached at exposition of240 s or more. Conclusion. The non-thermal plasma effectively destroyed bacterial biofilms within root channels.

The system of indication ofbiological pathogens that exists in our country allows to implement efficacious monitoring of epidemiologic situation, timely detect causative agents of infectious and parasitic diseases in material from humans and environmental samples, conduct their detailed identification and take appropriate means to ensure biosafety of the Russian Federation.

Im. Genotyping and phylogenetic analysis of parvovirus B19 isolates isolated on the territories of Northwestern Federal District (NWFD) of Russia. Materials and methods. 61 blood sera and 30 oropharyngeal lavages obtained from patients with maculopapular rash from various territories of NWFD were studied for the presence of parvovirus B19 DNA (PVB19). DNA isolation and amplification was carried out by standard techniques. DNA segment including fragment of non-structural gene NS1 and region of structural gene VP1 (NS1-VP1u, 994 nucleotides) was sequenced, original sequences of oligonucleotide primers were selected for this purpose. Phylogenetic analysis was carried out online on the website http://www.phylogeny.fr. Data for tree construction was obtained from GenBank. Results. PVB19 DNA was detected in 45% ofsamples. PVB19 genome segment was sequenced in 8 samples. All the PVB19 isolates belong to a single cluster of 1A genotype. Isolate 57.12 from Komi Republic is similar to ISR-G strain isolated from Israel. Conclusion. Phylogenetic analysis showed a high degree of genetic similarity between PVB19 isolates circulating on the territories of NWFD, their membership in the most widespread genotype in the world. Local and import cases of parvovirus infection (PVI) were identified. The authors make a conclusion on the necessity to include PVI into the system of rubella and measles control.

Aim. Study of the interrelation between the presence of immune deficiency and development of complications during vaccination of newborns with BCG vaccine. Materials and methods. In 24 children with complications of vaccine process in the form of cold abscess and lymphadenitis indicators of lymphocyte subpopulation levels were studied by flow cytofluorimetry on Beckman Coulter cytofluoriemter by using monoclonal antibodies with markers CD45+CD3+ - T-cell, CD45 +СD3 +СD4 + - T-helpers, CD45 +СD3 +CD8 + - T-supressors-cytotoxic killers, CD45+CD3 -CD16+CD56+ - natural killers, СD45 +CD3 -CD19 + - B-lymphocytes. The level of IgG, IgA, IgM in sera was determined by immune diffusion method in agar by Mancini. Results. In 4 children selective deficiency of IgA, in 5 - hyper-IgM syndrome was detected, which is an innate immunodeficiency and is characterized by the lack of sera IgA, reduction of IgG level and increase of IgM. In 9 children a reduction of CD16+ natural killer lymphocytes was detected, in some cases combined with a reduction of CD8+ T-supressors-cytotoxic killers. Conclusion. The reason of development of complications during BCG administration is the presence of immunodeficiency in children. In these children severe course of the vaccine process, presence of axillary lymphadenitis was observed, therapy of these children continued from 4 to 6 months.

Aim. Detection of early non-bacteriological markers ofburn wound mixed microbial infection. Materials and methods. The level of soluble form of trigger receptor expressed on myeloid cells-1 (sTREM-1) and cytokines in blood sera of 60 burn patients on days 3 - 6 and 10 - 17 after the burn was studied by solid phase enzyme immunoassay. Results. At the early periods ofburn disease the level of sTREM-1 in sera may be a non-bacteriological marker ofburn wound mixed infection: at days 3 - 6 after the injury - higher than 298.8 pg/ml, at days 10 - 17 after the burn - higher than 294.2 pg/ml. Conclusion. Level of sTREM-1 in blood sera of patients with severe thermic injury could be proposed as an additional laboratory marker ofburn wound mixed microbial infection.

Aim. Study the effect of synthetic ligands on the dynamics of TLR9 and BD-2 gene expression in vivo. Materials and methods. Synthetic ligands were used whose sequence was homologous to DNA and RNA human viruses. At days 1, 7 and 28 blood sampling from BALB/c line mice was carried out. Determination of level of TLR9 and BD-2 gene expression was evaluated by using RT-PCR in real-time mode. Results. The following ligands had the maximum effectiveness: tgg-ccc-ccc-ttg-tgg-acc-gg, ccg-gtc-cac-aag-ggg-ggc-ca and tcg-tcg-ttt-tgt-cgt-tgt-cg. At day 1 after the administration of ligands a significant increase of TLR9 gene expression was observed (62, 18 and 13 times, respectively), but at the same time defensin gene expression changed insignificantly. Conclusion. The approach developed may be used to evaluate the effect of these preparations on the system of innate immunity, namely TLR activation.

Aim. Detection of features of functioning of innate and adaptive immunity pathways in patients with Darier erythema annulare centrifugum (EAC). Materials and methods. 14 EAC patients aged 14 - 52 years were examined. The patients were ranked based on therapy variant. The first group consisted of 6 patients who had received Immunovac-VP-4 (Immunovac) against the background of basic therapy; the second group (4 patients) received cagocel against the background of basic therapy; the third group (4 patients) received only basic therapy; the group of healthy individuals consisted of15 individuals. All the patients had the level of cytokines in blood sera determined by solid-phase EIA by using Biosource (Austria) test-systems. Evaluation of TLR expression in peripheral blood mononuclear lymphocytes and keratinocytes was carried out by flow cytometry method by using monoclonal antibodies (Catlag Laboratories, USA) against the corresponding antigens; evaluation of content of lymphocyte subpopulations in blood was carried out by using monoclonal antibodies by flow cytometry method on FacsCalibur flow cytometer (Becton Dickinson, USA). Results. Immunotherapy by Immunovac and cagocel facilitated the increase of CD3+, CD4+, CD8+. Immunovac facilitated a significant increase ofinitially low values ofCD25+, CD95+ and normalization of CD72+; normalization of IgM level. Immunovac increased the level of serum IL-2, induced the increase of IFN-y synthesis in contrast to cagocel and basic therapy, the administration of those caused its decrease. TGF-0 increased during the course of Immunovac immunotherapy and decreased during basic therapy. In the course of basic therapy a significant increase of initially high level of cytokine IL-1 в was observed. Conclusion. Immunovac therapy resulted in correction of content of lymphocyte populations, sera cytokines, facilitating the normalization of immunocompetent cell proliferation processes, activation of NK-cells, macrophages and at the same time suppression of DTH reactions. Immunovac facilitated the enhancement of TLR3, 9 expression in the skin that indicates the inclusion of intracellular receptor mechanisms of innate immunity.

Aim. Detection of properties of the dominant microsymbiont emerging in the course of interaction of associative symbiosis components and ensuring protection of vaginal mucosa from seeding by pathogens. Materials and methods. H 2O 2-producing Lactobacillus spp., Corynebacterium spp., Staphylococcus aureus, Escherichia coli and primary epitheliocytes from lower reproductive tract of women were used. The ability of Lactobacillus spp. to influence changes of synthesis of catalase inhibitors by corynebacteria, bactericidal activity of lysozyme, lactoferrin and thrombocyte antimicrobial protein, antimicrobial activity of primary vaginal epitheliocytes was evaluated. Changes of antagonistic and growth properties of lactobacilli during interaction with secretory products of Corynebacterium spp., S. aureus, E. coli and vaginal epitheliocytes were also studied. Results. Exometabolites of both epithe-liocytes and corynebacteria were revealed to stimulate in most cases lactobacilli biomass growth and increased their antagonistic activity against S. aureus и E. coli. Metabolites of H 2O 2-producing lacto- bacilli increased synthesis of antimicrobial substances by epitheliocytes and potentiated bactericidity of natural resistance factors. Under the influence of lactobacilli metabolites an increase of production of catalase inhibitors by corynebacteria was revealed. Conclusion. Symbiotic interrelations of dominant microorganisms with host organism and associants under the condition of associative symbiosis leading to stimulation of production and potentiating of the effect of antibacterial protection factors are the basis for colonization resistance of vaginal biotope.

A review of recent publications on epidemiology and seroepidemiology of enterovirus type 71 in various regions of the world and authors’ own results of study of seroepidemiology and molecular epidemiology of EV71 in Russia are presented.