Results of Structural and Bacteriological Analysis of the Mouse Adrenal Glands in a Sepsis Model.

The article presents the results of histological and bacteriological analysis of the adrenal glands in two models of sepsis caused by intraperitoneal administration of Pseudomonas aeruginosa strains 1623 and 5266 to sexually mature male C57BL/6 mice. In both models, histological changes in the adrenal glands in the dynamics of sepsis consist in cell destruction and decrease in the absolute areas of the zona glomerulosa, columnar part of the zona fasciculata, and medulla, the development of venous congestion in the absence of pronounced signs of leukocytic infiltration. Most adrenocortical cells of the zona glomerulosa and chromaffin cells of the medulla show signs of destruction. The columnar part of the zona fasciculata loses normal architectonics, the cells undergo degeneration and apoptosis, a significant part of cells in the deep layers of the zona fasciculata remain intact, but do not show tinctorial and ultrastructural signs of steroidogenesis. The active growth of P. aeruginosa colonies from adrenal homogenates in both models already in the first hours after infection of animals makes it possible to associate the revealed structural changes in the adrenal glands with the direct negative effect of P. aeruginosa, and high levels of mRNA of proinflammatory cytokines in the adrenal tissues raise the question of the possible synthesis of these modulators of inflammation in the adrenal parenchyma of septic animals.

Antibacterial Activity of Lactobacillus plantarum Supernatant on Non-Fermenting Gram-Negative Bacteria.

We studied the effect of the L. plantarum strain supernatant on the growth of culture and biofilm of non-fermenting bacteria of the genera Pseudomonas, Achromobacter, and Burkholderia. To obtain a supernatant, the culture of L. plantarum was grown for 48 h at 37°C on a Lactic broth nutrient medium with casein peptone, then centrifuged and filtered through a 0.22-µm Millipore filter. Antimicrobial activity was determined by broth microdilution assay. The inhibitory effect of the supernatant on the growth of bacteria of all three genera was demonstrated. The maximum inhibition was observed for P. aeruginosa (by 13 times compared to the control). For bacteria of the Achromobacter and Burkholderia genera, the inhibition was less pronounced: by 7 and 6 times, respectively. The supernatant also inhibited biofilm formation by P. aeruginosa and A. ruhlandii, but did not affect formed biofilm. Thus, the L. plantarum supernatant obtained by us exhibited pronounced antimicrobial activity against non-fermenting bacteria, the causative agents of nosocomial infections, especially in immunocompromised individuals, very often in cystic fibrosis patients.

Comparative Characteristics of the Spleen White Pulp in Experimental Models of Sepsis Caused by Different Strains of Pseudomonas aeruginosa.

We describe reactive changes in the spleen white pulp in male C57BL/6 mice with experimental sepsis induced by intraperitoneal administration of Pseudomonas aeruginosa 1840 (Pa1840) with exotoxin U gene or Pseudomonas aeruginosa 1623 (Pa1623) with exotoxin S gene. Histological analysis and morphometry revealed hypoplasia of the spleen white pulp in mice with sepsis induced by Pa1840, while sepsis caused by Pa1623 was associated with its hyperplasia; with apoptosis of white pulp cells was observed in both cases. The results attest to ambiguous nature of the reactive changes in the white pulp of the spleen in experimental sepsis models initiated by Pa1840 and Pa1623 stains.

Inhibitory Effect of Copper and Zinc Ions on the Growth of Streptococcus pyogenes and Escherichia coli Biofilms.

Analysis of the effect of copper and zinc ions on Streptococcus pyogenes and Escherichia coli biofilms revealed significant differences in the effect of these metals in the form of sulfates or chlorides on biofilm formation. Zinc ions in low doses (salt concentration 0.005 M) inhibited the growth of S. pyogenes biofilms by 1.5 times. After increasing salt concentration to 0.05-0.5 M, the growth of biofilm was reduced by 2.5 times in comparison with the positive control. In case of E. coli biofilms, the inhibition was more pronounced: zinc sulfate in a concentration of 0.005 M reduced its growth by 4.6 times in comparison with the positive control. After increasing salt concentration, the growth of E. coli biofilm decreased by 6.8 times. In case of zinc chloride, zinc ions produced weaker effect and reduced biofilm growth by 2.2 and 5 times, respectively. Copper salts in a concentration of 0.005 M had practically no effect on the growth of S. pyogenes biofilm; with increasing salt concentration, the degree of inhibition was close to the effect of zinc. In case of E. coli biofilm, we observed a slight inhibition of the growth by low doses of copper ions (by 1.4-1.3 times); with increasing salt concentration the effect increased by 5.6 and 2.2 times for copper sulfate and chloride, respectively. Copper and zinc cations had no effect on mature biofilm.

Effects of Miramistin and Phosprenil on Microbial Biofilms.

Effects of Miramistin and Phosprenil on biofilms of S. pyogenes, S. aureus, E. coli, L. acidophilus, and L. plantarum were studied. Significant differences in the effects of these substances on mature biofilms of microorganisms and the process of their formation were observed. Miramistin had significant inhibiting effects on the forming of biofilms and on the formed biofilms of all studied microorganisms. Treatment with Miramistin inhibited biofilm formation by 2-3 times compared to the control. This effect was found already after using of Miramistin in the low doses (3.12 µg/ml). Inhibition of the growth of a formed biofilm was observed only after treatment with Miramistin in the high doses (25-50 µg/ml). Phosprenil in the high doses (15-30 mg/ml) inhibited the forming of biofilms, especially the biofilms of S. pyogenes and L. plantarum (by 3-4.5 times). Treatment of formed biofilms with the agent in doses of 6.0 and 0.6 mg/ml was associated with pronounced stimulation of its growth in S. pyogenes, S. aureus, and L. acidophilus.

Interleukin-6 in Sera of Mice Inoculated with Group A Streptococcus and Cultural Supernatants.

Dynamics of IL-6 level was studied in blood serum of CBA mice receiving intraperitoneal injections of killed and live culture of group A Streptococcus and its supernatants. It was found that administration of killed culture was followed by a significant rise in IL-6 level (by 4.7 times in 1 h and by 9.3 times in 5 h in comparison with the control). By 24 h, cytokine content was below the control. The highest levels of IL-6 were found after treatment with supernatants of Streptococcus cultures (by 10.5 times in 1 h and by 14.9 times in 5 h, in comparison with the control). Administration of live culture was accompanied by an increase in IL-6 concentration by 3.2 times in 3 h. In this experimental series, the maximum level of IL-6 was found in 48 h (by 5.2 times), and then it gradually decreased below the control. Different dynamics of changes in IL-6 level after administration of killed and live cultures of group A Streptococcus may suggests that they activate different signal pathways.

[Antagonistic activity of lactobacilli of the colon].

AIM: Study of antagonistic activity of lactobacilli of the colon against members of its autochthonous bacterial flora and agents of some acute infectious and chronic diseases of the gastrointestinal tract. MATERIALS AND METHODS: Antagonistic activity of 19 lactobacilli cultures against 28 cultures of bacteria belonging to various groups and fungi was evaluated within the framework of specially developed two-stage cultivation technique in the conditions of a combined system. The results of the study were evaluated according to a semi-quantitative scale that allows to put one or the other value of the zone of growth delay of the studied strain culture in compliance with the one or the other (low, moderate, high) level of antagonistic activity of the lactobacillus culture. RESULTS: Lactobacilli of the colon showed selective antagonistic activity against pathogenic enterobacteriae: pronounced against Salmonella enterica Serovar Enteritidis, Shigella flexneri 2b, Yersinia spp., and trace against Salmonella enterica Serovar Typhimurium. The level of antagonistic activity of lactobacilli against a wide range of members of autochthonous bacterial flora varied in a wide range, without revealing connection neither to its belonging to species, nor to its population level, nor to the belonging to group of the antagonistic effect objects. On the other hand a connection was traced with beloriging to a certain microbiota: being quite active against members of its own microbiota, lactobacilli often showed significantly lower level of antagonistic activity against cultures with the same species name isolated from other microbiota. CONCLUSION: In light of the results obtained, level of lactobacilli population may hardly be viewed as the only criteria of their full participation in the process of stabilizing microecological welfare of the colon, that allows to make a complete representation of the level of dysbiotic disorder in the mentioned biotope. With in the framework of rational bacteriological diagnostics of the level of dysbiotic disorders in the colon, evaluation of population level of lactobacilli should be evaluated along with the degree of their antagonistic activity against other components of the same microbiota.

[The role of microflora in etiology of chronic tonsillitis].

Analysis of the results of a bacteriological study involving 218 patients with various forms of chronic tonsillitis (CT) has demonstrated that they were significantly different in terms of the seeding rate of beta-hemolytic group A Streptococci (BHAS). Specifically, these microorganisms were detected in patients with paratonsillar abscess 2.5 times as frequently as in those with uncomplicated forms of chronic tonsilitis. In other words, the former group is at risk of developing such concomitant diseases as post-streptococcal rheumocarditis, glomerulonephritis, and polyarthritis. It was shown that BHAS are capable of forming a biological film at the surface of abiogenic carriers in vitro which may be a principal cause of chronic and recurring tonsillitis.

[Adequacy analysis of bacteriological studies of pyo-inflammatory diseases of ENT organs with fatal outcome].

Results of analysis of the case history patients that died from complications of pyo-inflammatory diseases of ear, nose, and throat in multifield hospitals of Moscow during the last 10 years are presented. Data of microbiological studies for the entire period of each patient's stay in the hospitals are extracted They indicate that bacteriological studies are not prescribed as frequently as needed. As a rule, seeding is conducted after a patient is transferred to the intensive therapy department in 16 or more days following admission to the hospital. The materials seeded are largely biological fluids rather than pathological excreta from primary inflammatory foci.

[The role of bacteriological examination in diagnosis of chronic tonsillitis].

A bacteriological examination was made in 105 patients with various forms of chronic tonsillitis. Eighteen cultures of beta-hemolytic streptococcus (BHSA) were isolated from 36 patients with paratonsillar abscesses. BHSA seeding in paratonsillar abscess was 50.0%. Eight BHSA cultures were isolated from 69 patients with chronic tonsillitis. BHSA seeding in chronic tonsillitis was 11.6%. Adequate set of culture media for BHSA growth was optimized. The technique of the sample collection from tonsillar lacunes was improved. The significance of bacteriological diagnosis of chronic tonsillitis increases considerably due to optimization of out- and intralaboratory tests at the preanalytic stage of the study.

New data on the effect of helicobacter pylori on oxygen metabolism in human neutrophils.

A new phenomenon is discovered: Helicobacter pylori suppressed production of superoxide radical by neutrophils, while intracellular production of oxygen radicals is considerably activated. This phenomenon seems to play an important role in Helicobacter modulation of the inflammatory process in the stomach and persistence of the bacterium in the inflammatory focus.

[The evaluation of the diagnostic value of PCR test systems according to the results of state trials].

The method for the diagnostic value evaluation of preparations, based on the calculation of the accuracy of test results and taking into account the spread of the diagnosed disease (the comparison of the PCR test systems for the diagnostics of hepatitis B and ureaplasmosis) is proposed. As shown in this work, evaluations obtained with the use of this method coincide with those obtained on the basis of prognostic value, but are more convenient in use and provide additional information.

[Production of recombinant fragment of VacA protein and development of non-invasive method for the diagnosis of Helicobacter pylori infection].

Vacuolizing toxin (VacA) Helicobacter pylori is an important factor of pathogenicity of Helicobacter pylori and a basic marker in the diagnosis of helicobacteriosis and related diseases. A coagulation-based diagnostic test-system was elaborated for the detection of VacA in clinical samples. A fragment of vacA was cloned, for the purpose, in Escherichia coli and expressed in preparative quantities; the coded protein was purified and used in raising the diagnostic serum. The thus designed coagulating test-system was successfully tested under the modeling conditions with clinical samples. Therefore, the designed express method can be used for the invasion-free determination of VacA in patients with gastric and duodenal pathologies.

[Pyoinflammatory ENT and upper airway diseases: the importance of the problem and approaches to its management]. / Gnoino-vospalitel'nye zabolevaniia ukha, gorla, nosa i verkhnikh dykhatel'nykh putei: aktual'nost' problemy i puti resheniia.

The problem of pyoinflammatory ENT and upper airway diseases is reviewed. Some new approaches are proposed.

[Effective antibacterial therapy in otorhinolaryngology: bacteriological rationale]. / Bakteriologicheskoe obosnovanie ratsional'noi antibakterial'noi terapii v otorinolaringologii.

The article presents current views on etiopathogenesis of the diseases associated with contamination with microorganisms; reviews inflammatory ENT diseases in terms of current literature on clinical microbiology of these diseases; characterizes microorganisms more or less specific for each relevant nosological entity; analyses basic groups of antibacterial drugs with formulation of bacteriologically grounded principles of initial antibacterial therapy of ENT inflammation.

[Microbiological aspects of effective antibiotic therapy early after operation in chronic maxillary ethmoiditis and tonsillitis]. / Mikrobiologicheskie aspekty ratsional'noi antibiotikoterapii v rannem posleoperatsionnom periode pri khronicheskom gaimoroétmoidite i tonzillite.

Microbiological and chromato-mass-spectrometric examinations were made in 53 patients with chronic maxillary ethmoiditis and 52 patients with chronic tonsillitis. The microflora showed strong resistance to unprotected penicillines and cephalosporins, 100% sensitivity to amoxicilline clavulanate and mupirocine. The role of opportunistic microflora as pathogens of ENT inflammation is growing as well as the percentage of anaerobic microorganisms detected chromato-mass-spectrometrically in chronic inflammation. Administration of mupirocine on polyethylenglicol base early after operation in patients with chronic maxillary ethmoiditis accelerated recovery of the study group patients by 2-3 days vs control patients.