[Molecular genetic analysis of Pseudomonas aeruginosa strains isolated from environment and patients in health care facilities].

AIM: Analysis of genetic heterogeneity of Pseudomonas aeruginosa strains isolated in and out of hospitals. MATERIALS AND METHODS: To study the genetic diversity of 36 strains of P. aeruginosa plasmid analysis, random amplified polymorphic DNA (RAPD) technique as well as polymerase chain reaction for detection of virulence genes algD, lasB, toxA, plcH, plcN, exoS, nan1, and nan2. RESULTS: Epidemically important strains were found in different ecological niches. It was shown that these virulence factors could play important roles in pathogenesis of infection. CONCLUSION: RAPD technique was effective for analysis of P. aeruginosa isolates. Number of studied typing bands differed between related isolates for each random primer.

[Comparative assessment of DNA extraction methods for identification of glanders and melioidosis etiological agents by PCR].

Pathogenic Burkholderia are considered as a cause of dangerous infections and potential agents of bioterrorism. Comparative assessment of different methods of extraction and purification of DNA for PCR analysis of pure cultures and samples contaminated by etiological agents of glanders and melioidosis was performed. Samples of soil and food artificially contaminated by pathogenic Burkholderia as well as organs of infected animals were tested. DNA was extracted by methods of boiling, nucleosorption with presence of guanidine thiocyanate, guanidine thiocyanatephenol extraction, guanidine thiocyanate-phenol extraction with additional purification of DNA by nucleosorption. Amplification was performed by "Flash" technique and detector of fluorescence was used for analysis of PCR products. Utilization of the recommended methods of preparation depending on the nature of sample let to detect by the "Flash" technique the etiological agents of glanders and melioidosis in concentration =10(3) microbial cells per ml. Choice of DNA extraction and purification methods is determined by type of a sample and presence in it of admixtures inhibiting PCR.

[The use of multilocus sequence typing (MLST) and randomly amplified polymorphic DNA (RAPD) for the differentiation between strains of Burkholderia mallei].

Burkholderia mallei is highly pathogenic microorganism for both humans and animals. In this work, the possibility of the use of the genotyping method for differentiation between strains of B. mallei was studied. A collection of 14 isolates of B. mallei was characterized using randomly amplified polymorphic DNA (RAPD) and multilocus sequence typing (MLST). RAPD was the best method used for detecting strain differences of B. mallei. It was suggested that this method would be an increasingly useful molecular epidemiological tool.

[The use of polymerase chain reaction for detection of the agents of glanders and melioidosis using experimental infection].

Glanders and melioidosis are severe infectious diseases of people and animals. The causative agents of these infections refer to the potential agents of bioterrorism of group B. In this work the possibility of use of flagellin-based primers for the identification of B. mallei and B. pseudomallei and for diagnosis of experimental glanders and melioidosis was studied. The obtained results permit to make a conclusion that PCR using the developed primers may be recommended for the incorporation in the scheme of laboratory diagnosis of glanders and melioidosis both for the identification of clean cultures and in experimental clinical material.

[Use of PCR for identification of Burkholderia mallei]. / Ispol'zovanie PTsR dlia identifikatsii Burkholderia mallei.

Stimuli of glanders belong to the potential agents of biological terror. The possibility to use various primers in the identification of B. mallei was investigated and the significance of polymerase chain reaction (PCR) was defined within the scheme of laboratory glanders diagnosis in the offered paper. The constructed amplifying test-systems can be used to detect the glanders both in the environmental objects contaminated with B. mallei and in experimental clinical material.

[Aortic endothelium and serum lipoproteins in rats during the restorative period after 30-day hypokinesia]. / Endotelii aorty i lipoproteidy syvorotki krovi krys v vosstanovitel'nom periode posle 30-sutochnoi gipokinezii.

Experiments in 49 albino rats indicated that serum beta-lipoprotein levels were increased on days 1 to 8 and 24 to 29 of readaptation following 30-day restriction of motor activity of the animals kept in the individual cramped cages. The alpha-lipoprotein concentrations were decreased in initial periods and elevated on days 18 to 23 of restoration. In the course of readaptation, the aortic endothelial layer showed greater lymphocyte and monocyte counts and larger number of endotheliocytes with karyopyknosis, karyolysis, and larger number of mitotically divided and binuclear cells. The extent of endotheliocytes and their cytoplasm and nucleus was persistently lower on day 3 to 55 of readaptation.

[Morphometric analysis of aortic endothelium and serum lipoproteins in rats during readaptation after 15-day hypokinesia]. / Morfometricheskii analiz éndoteliia aorty i lipoproteidy krovi u krys v period readaptatsii posle 15 sut gipokinezii.

On hypokinesia day 15, the serum concentration of beta-lipoproteins increased and that of alpha-lipoproteins decreased, and the aortic endothelium was damaged. During the recovery period, the amount of pre-beta-lipoproteins was normal. After hypokinesia the concentration of alpha-lipoproteins increased and returned to normal by the 10th day while that of beta-lipoproteins remained lower for a long time. During the recovery period the number of endothelial cells showing karyopyknosis and karyolysis was increased. At later recovery-periods, the surface area of endothelial cells and their cytoplasm increased and the nucleus/plasma ratio decreased. However, lesions in the structure of the endothelial layer were less expressed than during hypokinesia. In the recovery period, the number of binuclear cells and the number of mitoses grew suggesting greater activity of regeneratory processes.