A In Vitro and In Vivo Study of the Ability of NOD1 Ligands to Activate the Transcriptional Factor NF-kB.

Pattern-recognition receptors (PRR) play a crucial role in the induction of the defense reactions of the immune system against pathogenic bacterial and viral infections. The activation of PRR by specific, highly conserved pathogen-associated molecular patterns (PAMPs) induces numerous immune reactions related both to innate and adaptive immunity. In addition to the well-studied Toll-like receptors, pathogens can be recognized by the receptors belonging to the other PRR families; including NOD-like receptors (NLR). Stimulation of members of NOD-like receptors (NOD1, 2) and Toll-like receptors results in the activation of the transcriptional factor NF-kB regulating gene expression in numerous molecules implicated in the development of proinflammatory reactions. As opposed to Toll-like receptors, the NF-kB-activating ability of NLRs has not been fully studied. In this work, we examine the ability of one member of the NLR family - NOD1 - to activate the main proinflammatory transcriptional factor NF-kB. We also compare the NF-kB-activating ability of NOD1 ligands of a different structure with TLR4,5 ligandsin vitroandin vivo.

[Microbiocenosis of oral cavity in patients from different age groups].

Worsening of state of oral cavity is observed not only in elderly persons, but also in adults aswell as in adolescents and children. The leading role in development of dental caries and periodontal diseases belongs to microorganisms. Oral cavity bacterial landscape was determined in adolescents, adults and elderly people with normal periodont or with gingivitis or periodontitis. The study showed that species and qualitative composition of oral cavity microflora were different in different age groups. In patients with normal periodont, non pathogenic (common commensals) microbial composition was revealed. In abnormal state of oral cavity, its microflora was characterized by the presence of high titers of potentiality pathogenic microorganisms.

[Association of Candida albicans fungi with some opportunistic microorganisms in intestinal dysbiosis in patients of different age groups].

The occurrence of C. albicans in association with opportunistic microorganisms (Staphylococcus aureus and Klebsiella) in intestinal dysbiosis in patents of different age groups (from 6 days to 31 years) was analyzed. The data on the comparative evaluation of the results of the bacteriological examination of patients with intestinal dysbiosis, carried out during the periods of 2000-2001 and 2001-2002 (388 and 467 patients respectively), are presented. During the period of 2000-2001 the detection rate of C. albicans in monoculture was 6-33% of cases (in the examined group). A higher detection rate was registered with respect to the association of C. albicans with staphylococci (42-65% of cases). The associations of C. albicans with staphylococci and Klebsiella were observed in 13-46% of cases. Similar results were registered during the period of 2001-2002, but during this period a reliable decrease in the detection rate of the association of C. albicans with staphylococci (practically by 20% in the group of infants) and the 1 1/2-fold increase of the detection rate of C. albicans in monooulture (in all age groups) were observed.

[Antistaphylococcal activity of the complex of natural cytokines]. / Protivostafilokokkovoe deistvie kompleksa prirodnykh tsitokinov.

The in vitro study of the action of the complex of natural cytokines (CNC), or preparation Superlymph, on different microbial test strains (Staphylococcus aureus, Escherichia coli, yeast-like fungi of the genus Candida) was carried out. CNC suppressed the growth of S. aureus test culture, depending on the concentration of the preparation. The inhibiting effect was observed in the presence of Ca and Mg bivalent cations in the medium. Under the chosen conditions of the experiment CNC did not inhibit the growth of E. coli test cultures, as well as test cultures of yeast-like fungi of the genus Candida. In addition to its antibacterial effect, Superlymph also produced some effect on the release of cathepsin G, the lysosomal enzyme of human leukocyte granules.

[A nutrient medium for the isolation and cultivation of Campylobacter]. / Pitatel'naia sreda dlia vydeleniia i kul'tivirovaniia kampilobakterii.

Campylobacter agar, nutrient medium intended for the isolation of bacteria of the genus Campylobacter from clinical material, has been developed. The composition of the medium includes sprat hydrolysate, aerotolerant additive (ferric sulfate--oxide, sodium pyruvate, sodium pyrosulfite), sodium glutaminate, agar. The selective properties of the medium are ensured by introducing the mixture of antibiotics consisting of polymyxin B, rifampicin, amphotericin B, ristomycin. The balanced composition of Campylobacter agar ensures the aerotolerance of Campylobacter organisms and gives the optimal conditions for their growth when the inoculated material is cultivated in the atmosphere made up of the mixture of three gases (5% of oxygen, 10% of carbon dioxide, 85% of nitrogen), as well as under the conditions of a "candle vessel". The medium suppresses the development of the associative microflora diluted 10(-1). As shown in the trial of the quality of Campylobacter agar by the inoculation of material taken from patients with acute enteric infections, agricultural animals and monkeys, the medium has pronounced selective, properties with regard to extraneous microflora, while ensuring the isolation of Campylobacter on the level of the control medium.

[The epidemiological markers of Pseudomonas aeruginosa strains circulating in different types of hospitals]. / Epidemiologicheskie markery shtammov Pseudomonas aeruginosa, tsirkuliruiushchikh v statsionarakh raznogo profilia.

Pyocinovar and serovar characteristics of 91 P.aeruginosa strains isolated from patients and the environment in a hospital for premature children, a child survey hospital and a neonatological hospital. The leading epidemiological markers of P.aeruginosa strains, among them pyocinovar 883722, serovar 6 (a hospital for premature children), pyocinovar 888888, serovar 4 (a child surgery hospital), pyocinovars 888888 and 861322 (a neonatological hospital).

[Treatment of endophthalmitis with a single intravitreous administration of cefotaxime and gentamicin in an experiment]. / Lechenie éndoftal'mitov odnokratnym intravitreal'nym vvedeniem tsefotaksima i gentamitsina v éksperimente.

Experimental endophthalmitis was treated with single intravitreous administrations of cefotaxime (claforan) and gentamicin. It was found that a single administration of cefotaxime to the vitreous body prevented development of endophthalmitis in rabbits previously infected with Staphylococcus aureus, E. coli and Pseudomonas aeruginosa. Vitrectomy in the treatment of endophthalmitis was shown to be promising and provide satisfactory anatomical and functional results.

[Serotypes of Pseudomonas aeruginosa strains isolated in obstetric institutions in Azerbaijan]. / Serotypy shtammov Pseudomonas aeruginosa, vydelennykh v rodovspomogatel'nykh uchrezhdeniiakh Azerbaidzhanskoi SSR.

Examinations of Pseudomonas aeruginosa strains isolated from clinical material from newborns and puerperae and from the environmental objects (washings off, water) have shown that 45.45% of these strains belonged to serotype 04, 24.55% to serotype 02, and 16.38% to other serotypes. Serotype 04 was the most incident in all the tested samples, it was detectable in newborns' biotopes and on environmental objects. Serotype 02 strains ranked second in incidence; they were detected in clinical material from the newborns but not on environmental objects.

[The therapeutic use of diucifon-activated immunocytes in adoptive transfer in mice with burn trauma complicated by Pseudomonas aeruginosa infection]. / Lechebnoe primenenie aktivirovannykh diutsifonom immunotsitov v usloviiakh adoptivnogo perenosa u myshei s ozhogovoi travmoi, oslozhnennoi sinegnoinoi infektsiei.

The effectiveness of the therapeutic use of diucyphone-stimulated syngeneic spleen cells was studied in mice with a burn trauma complicated by P. aeruginosa infection. For this purpose CBA mice were subjected to a burn of degree IIIa, covering 30% of their body surface, the wound was infected with P. aeruginosa strain, and 24 hours later spleen cells were taken from some of these animals. The spleen cells, incubated for 3 hours with or without diucyphone at a concentration of 10-100 micrograms/ml, were washed and reinfused intravenously to the remaining animals. The injection of diucyphone-treated spleen cells was shown to greatly increase the survival rate of the animals, to accelerate healing processes and to enhance the proliferative response of spleen cells to mitogens (phytohemagglutinin, concanavalin A) and interleukin-2. A conclusion was made on the efficacy of the therapeutic use of diucyphone-activated immunocompetent cells in cases of immunodeficiency induced by burn infection.

[The diagnostic effectiveness of erythrocyte immunoreagents made from different Pseudomonas aeruginosa antigens]. / Diagnosticheskaia éffektivnost' éritrotsitarnykh immunoreagentov iz razlichnykh antigenov Pseudomonas aeruginosa.

The diagnostic effectiveness of the passive hemagglutination (PHA) tests with the use of erythrocyte diagnostica (ED based on exotoxin A (ETA) and poly- and monovalent ED based on lipopolysaccharides (LPS) isolated from 5 most widespread P. aeruginosa serogroups was compared. 97 patients with different purulent septic diseases and 100 practically healthy adult donors were examined. The treatment of serum samples with 2-mercaptoethanol decreased the diagnostic effectiveness of serological examination. The simultaneous determination of the activity of antibodies to ETA and LPS essentially increased the number of positive results of the PHA tests in patients with bacteriologically confirmed P. aeruginosa infection, but not in patients from whom other bacteria were isolated. Considering the sensitivity and specificity of serological examination, as well as the decrease of the volume of necessary work and the consumption of immunoreagents, the optimum test conditions proved to be ensured by the use of two ED: LPS-based polyvalent ED and ETA-based ED.

[The assessment of the toxigenicity of clinical strains of Pseudomonas aeruginosa by an immunoenzyme method using nitrocellulose filters]. / Otsenka toksigennosti klinicheskikh shtammov Pseudomonas aeruginosa immunofermentnym metodom s ispol'zovaniem nitrotselliuloznykh fil'trov.

The enzyme immunoassay (EIA) on nitrocellulose filters was adapted for the detection of exotoxin A in 104 P. aeruginosa strains isolated from patients and the environment in surgical wards in hospitals of Moscow and Alma-Ata. The method was shown to be highly sensitive: it permitted the detection of 5.ng of P. aeruginosa exotoxin A. The screening of 104 P. aeruginosa clinical strains by means of EIA on nitrocellulose filters revealed that these strains exotoxin A in 88.5% of cases.

[Experimental development of aseptic adhesive films]. / Eksperimental'naia razrabotka asepticheskikh lipkikh plenok.

Antibacterial activity of surgical adhesive films containing various antiseptics in the adhesive layer was studied experimentally. It was shown that the adhesive films containing 0.1 to 1% (in terms of active iodine) of iodopyrone had a stable bactericidal action and were effective against to the main causative agents of surgical infections, i. e. Staphylococcus spp., E. coli, Klebsiella spp., Pseudomonas aeruginosa and others.

[Cloning and regulation of gene expression of EcoRV restriction- modification system]. / Klonirovanie i reguliatsiia ékspressii genov sistemy restriktsii i modifikatsii EcoRV.

A number of recombinant plasmids, containing EcoRV restriction-modification genes have been constructed. Individual genes of this system were introduced into plasmids of various incompatibility groups. Promoter regions of genes encoding methylase and restrictase have been cloned and studied. With the use of specialized vector pVE8 it was shown that the efficiency of the endonuclease gene promoter is comparable with early lambda phage promoters and produced about 70% of PL efficiency. The efficiency of the methylase gene promoter region was twice less than the efficiency of the restriction endonuclease gene promoter. Plasmid with restriction endonuclease gene promoter located downstream in relation to the additional regulatable phage lambda promoter PL has been obtained. It enabled us to construct strains 30-40 fold overproducing this enzyme under conditions of inactivation of the temperature sensitive phage repressor c1857. This construction directs the production of a high level (10%) of the total cellular soluble proteins) of the EcoRV restriction enzyme. The factors that influenced the level of enzyme synthesis under induction are discussed.

[Protective properties of anatoxin obtained from a homogeneous preparation of Pseudomonas aeruginosa exotoxin A]. / Protektivnye svoistva anatoksina, poluchennogo iz gomogennogo preparata ékzotoksina A Pseudomonas aeruginosa.

The protective properties of formulated toxoid obtained from the highly purified preparation of P. aeruginosa exotoxin A have been studied in the test of the active immunization of mice. The study has revealed that the preparation when introduced in 1 or 2 injections in a dose of 15 micrograms, shows faint protective potency with respect to P. aeruginosa strains differing in virulence. Immunization with this toxoid in 3 and 4 injections has been found to ensure 60-100% and 50-60% protection of mice infected with P. aeruginosa toxigenic and proteolytic strains respectively. Immunization with toxoid has been found to induce the appearance of short-term antibacterial immunity which loses its capacity to protect the immunized animals, challenged with both toxigenic and proteolytic P. aeruginosa strains, as early as on day 28. The immunization of mice with toxoid in 4 injections has been shown to induce the development of antitoxic immunity capable of neutralizing up to 150 LD50 of purified exotoxin A.

[Erythrocyte diagnostic kits for detection of Pseudomonas aeruginosa exotoxin A]. / Eritrotsitarnye diagnostikumy dlia vyiavleniia ékzotoksina A Pseudomonas aeruginosa.

The use of formulated chick red blood cells loaded with IgG preparations and affinity-purified antibodies, in comparison with initial immune serum to P. aeruginosa exotoxin A (ETA), has been shown to increase the sensitivity of antibody erythrocyte diagnosticum (AbED) 17-fold and to ensure the detection of ETA at a concentration of 1.2 mg of protein per ml. The passive hemagglutination (PHA) test with AbED has proved to be a more sensitive method for the detection of ETA than the antibody neutralization test with the use of antigenic erythrocyte diagnosticum, the latex agglutination test, the coagglutination test and the enzyme immunoassay. The PHA test has permitted the detection of ETA in the culture fluid of 80% of P. aeruginosa cultures under study.

[Use of a polyvalent erythrocyte diagnostic agent for determining antibodies to Pseudomonas aeruginosa in clinical practice]. / Primeneneie polivalentnogo éritrotsitarnogo diagnostikuma dlia opredeleniia antitel k Pseudomonas aeruginosa v klinicheskoi praktike.

The use of polyvalent erythrocyte diagnosticum prepared on the basis of 5 polysaccharide antigens of P. aeruginosa slime, isolated from strains belonging to the most widespread serovars, makes it possible to check up the humoral response of donors after their immunization with P. aeruginosa polyvalent corpuscular vaccine with the aim of obtaining anti-P. aeruginosa donor plasma. Antibody titers, determined in the passive hemagglutination test with the use of the proposed diagnosticum and corresponding to a serum dilution of 1:320 and greater, age tentatively diagnostic, which may be indicative of P. aeruginosa in the development of purulent septic complications in patients. The use of the passive hemagglutination test with the newly developed polyvalent erythrocyte diagnosticum makes it possible to check up the specific response of patients having P. aeruginosa infection in the process of their treatment with anti-P. aeruginosa hyperimmune plasma used as a part of complex therapy.

[Production of an diagnostic erythrocyte agent from Pseudomonas aeruginosa exotoxin A]. / Poluchenie éritrotsitarnogo diagnostikuma iz ékzotoksina A Pseudomonas aeruginosa.

The sensitization of formolized sheep red blood cells with exotoxin A by means of chromium chloride or glutaraldehyde is more effective with respect to their sensitivity in the passive hemagglutination test than loading by means of amidol, tannin and rivanol. The use of chromium chloride decreases the consumption of exotoxin A 2, 8, 16 and 16 times in comparison with the use of amidol, tannin, rivanol or glutaraldehyde respectively. The high specificity of erythrocyte diagnosticum obtained from exotoxin A by means of chromium chloride is indicated in the study of hyperimmune sera to 22 different antigens of enteric bacteria and staphylococci in the passive hemagglutination test and to 10 different enterobacterial and staphylococcal antigens in the antibody neutralization test.

[Protective artificial polysaccharide-protein antigens made from the O-specific polysaccharides of Pseudomonas aeruginosa bacteria of the immunotypes 1, 2 and 7]. / Protektivnye iskusstvennye polisakharid-belkovye antigeny iz O-spetsificheskikh polisakharidov bakterii Pseudomonas aeruginosa immunotipov 1, 2 i 7.

O-specific polysaccharide (PS) obtained from P. aeruginosa lipopolysaccharide (LPS), immunotypes 1, 2 and 7, were condensed in the presence of aminated bovine serum albumin (amBSA). As a result, artificial polysaccharide-protein antigens were synthesized and their protective properties were studied by the subcutaneous immunization of mice with their subsequent challenge with the cells of the homologous strain injected intraperitoneally in a dose of 3-5 LD50. The most effective immunization was achieved by using homologous LPS, taken in a subimmunogenic dose, as adjuvant. Heterologous LPS, taken in the same doses, showed no stimulating effect. The protective properties of the conjugates were also found to depend on the length of the PS chain. The conjugate of amBSA and PS of immunotype 2 with a molecular weight of about 10 KD was found to possess the highest protective potency. The decrease of the molecular weight of PS of immunotype 7 from 50 KD to 10 KD by selective acid hydrolysis led to obtaining the conjugate with high protective potency. The possibility of using the structurally linked pair antigen-adjuvant for the development of artificial vaccines is discussed.

[Isolation and primary characteristics of elastase from a clinical strain of Pseudomonas aeruginosa]. / Poluchenie i pervichnaia kharakteristika élastazy iz klinicheskogo shtamma Pseudomonas aeruginosa.

The homogeneous preparation of elastase has been obtained from P. aeruginosa clinical strain. The molecular weight of the isolated enzyme is 33,000 daltons, and its isoelectric point is 6.8. Two media manufactured in this country (dialyzed bovine heart hydrolysate and a dried semisynthetic medium) ensuring good production of the enzyme have been proposed. The optimum time for the cultivation of the producer strain (30-40 hours) has been established. Elastase has been shown to be widely spread among P. aeruginosa clinical strains.