[Antimicrobial activity of bacteriocin S760 produced by Enterococcus faecium strain LWP760].

Antimicrobial activity of bacteriocin S760 (enterocin) produced by Enterococcusfaecium strain LWP760 was studied. Bacteriocin S760 is a cationic, hydrophobic, and heat stable peptide with the molecular weight of 5.5 kDa and pl of 9.8. Enterocin S760 is shown to inhibit in vitro the growth both of sensitive and resistant to antibacterials gramnegative and grampositive bacteria of 25 species. MICs of the bacteriocin S760 vary between 0.05-1.6 mg/l for Escherichia coli 0157:H117, Salmonella typhimurium, Salmonella enteritidis, Campylobacter jejuni, Yersinia enterocolitica, Yersinia pseudotuberculosis, Listeria monocytogenes and Clostridium perfringens, that are main food-borne pathogens, and from 0.4-1.6 mg/l for Streptococcus pyogenes, Streptococcus pneumoniae and Corynebacterium diphteriae. It is also active against antibioticresistant strains of Staphylococcus aureus, Enterobacter cloacae, Acinetobacter baumannii (with MICs of 0.05-3 mg/l), Klebsiella pneumoniae (with MICs of 6 mg/l), Pseudomonas aeruginosa (with MICs of 0.4-25 mg/1), as well against fungi belonging to species of Candida albicans, Candida krusei and Aspergillus niger (with MICs of 0.1-0.2 mg/l). Enterocin S760 is a novel antimicrobial agents useful in medicine, veterinary and food industry.

[An algorithm for identification of CTX-M-type beta-lactamase genes using restriction fragment length polymorphism analysis of PCR-product].

The algorithm of the identification of the bla(CTX-M) genes coding CTX-M-type beta-lactamases providing resistance to cephalosporins III-IV was developed. This algorithm provides identification of 49 genes of 96 genes presented in the GenBank database so far. Remaining 47 genes can be identified as consisting of small sub-groups composed of 2-6 genes with the exception of sub-group of the bla(CTX-M-14)-like genes composed of 13 genes. The identification of the bla(CTX-M) genes is based on two-step restriction fragment length polymorphism analysis of 544 bp PCR-product (PCR-RFLP). In the first step, determination of subtype (cluster) of the bla(CTX-M) gene occurred using the restriction nuclease Alu I: cluster 1, -2, -8, -9 or -25. Moreover, four genes can be identified just at this step: bla(CTX-M)-59, (cluster 2); bla(CTX-M-63) (cluster 8), bla(CTX-M-45) (cluster 9), and bla(CTX-M-78) (hybrid gene between cluster 2 and cluster 25). At the second step gene identification goes on inside of each cluster separately using a set of 26 restriction nucleases. As a result of the PCR-RFLP-analysis, 23 bla(CTX-M) genes can be identified at the cluster 1, 11 genes--at the cluster 2, 4 genes--at the cluster 8, 9 genes--at the cluster 9, 1 gene--at the cluster 25, and 2 hybrid genes: bla(CTX-M-78) (between clusters 2 and 25), and bla(CTX-M-64) (between clusters 1 and 9). The described algorithm was used for identification of the blac(CTX-M) genes (n = 585) detected in Enterobacteriaceae nosocomial isolates (n = 877), collected from Russial hospitals in 2003-2007. It was shown that major genes belonged to cluster 1 (n = 543), namely--bla(CTX-M-15) gene (n = 515), bla(CTX-M-3) (n = 25), bla(CTX-M-22) (n = 1), bla(CTX-M-23) (n = 1), and bla(CTM-34) (n = 1). Moreover, the genes atributed to cluster 2 were identified: bla(CTX-M-2) (n = 1), and bla(CTX-M-5) (n = 4); and genes belonged to cluster 9: bla(CTX-M-9) (n = 2), and bla(CTX-M-14) (n = 35).

[Genetic determinants of antibacterial resistance among nosocomial Escherichia coli, Klebsiella spp., and Enterobacter spp. isolates collected in Russia within 2003-2007].

Nosocomial bacterial isolates collected within 2003-2004 (n=411) and 2005-2007 (n=422) were highly resistant to cephalosporins III-IV and antibacterials of other groups (aminoglycosides, fluoroquinolons, chloramphenicol, and co-trimoxazole). Genes encoding TEM, SHV, CTX-M, OXA-2, and AmpC types of beta-lactamases (BLs) in the E. coli, Klebsiella spp., and Enterobacter spp. isolates were detected using polymerase chain reaction (PCR). Prevalent CTX-M-type BLs were detected in 85% of the E. coli, 87% of the Klebsiella spp., and 38% of the Enterobacter spp. isolates of the first strain collection and in 94% of the E. coli, 91% of the Klebsiella spp., and 38% of the Enterobacter spp. isolates of the second one. Genes belonging to three subtypes of blacTx-M genes were identified: bla(CTX-M-1) (228 bla(CTX-M-15) and six bla(CTX-M-3) of the first strain collection; 275 bla(CTX-M-15), three bla(CTX-M-3), and one bla(CTX-M-22) of the second one), bla(CTX-M-2) (one bla(CTX-M-5) of the first strain collection and one bla(CTX-M-2) of the second one), bla(CTX-M-9) (17 bla(CTX-M-14) and one bla(CTX-M-9) of the first strain collection; seven bla(CTX-M-14) and one bla(CTX-M-9) of the second one). Three isolates of the first strain collection and one isolate of the second one carried two genes belonging to two different subtypes, i.e., bla(CTX-M-15) and bla(CTX-M-14) simultaneously. The bacterial isolates had high levels of associative resistance to ciprofloxacin, co-trimoxazole, gentamicin, amikacin, and chloramphenicol associated with the resistance gene cassettes aadA1, aadA2, aadA5, aadB, aacA4, aac(6')Ib; dfrA1, dfrA5, dfrA12, dfrA17, cmlA1, ereA2, and catB8 in the class 1 integrons and the resistance gene cassettes dfrA1, sat1, and aadA1 in the class 2 integrons.

[Efficacy of using vitaprost in patients with infertility caused by chronic prostatitis].

The article provides the results of Vitaprost efficacy in 40 infertile men with chronic prostatitis. Vitaprost treatment was found effective, safe and well tolerated.

[Development of a quantitative PCR variant, and use of it for assessing gene expression]. / Razrabotka kolichestvennogo varianta PTsR i primenenie ego dlia otsenki ékspressii genov.

The data on the development of a quantitative PCR technique are presented, including the synthesis of modified primers, the choice of conditions for the removal of reaction-mixture components from the amplified DNA, and the colorimetric detection of the PCR product. The application of this technique to the assessment of the level of gene expression in nonculturable bacterial forms characterized by a drastically reduced metabolism is described.

[Use of molecular-biological methods for identifying brucella in a comparative analysis of strains, isolated from sick dogs]. / Ispol'zovanie molekuliarno-biologicheskikh metodov identifikatsii brutsell v sravnitel'nom analize shtammov, vydelennykh ot bol'nykh sobak.

Ten strains isolated from sick dogs in 1998 in St. Petersburg were studied by traditional and molecular biological methods of Brucella identification. PCR study confirmed that the isolated cultures were Brucellae, and comparative study of the traditional phenotypical characteristics and protein and antigenic composition allowed referring all the isolated strains to B. canis. Traditional identification showed similarity of 7 strains with the reference B. canis strain RM6/66, and 3 strains were similar to B. canis Mex 51 strain. These results confirmed the division of B. canis into two biovars. Polyacrylamide gel electrophoresis with sodium dodecyl sulfate demonstrated the identity of protein profiles of 10 strains isolated from dogs to the reference B. canis RM6/66 strain. Immunoblotting analysis with S- and R-specific rabbit antisera also demonstrated the identity of antigens binding IgG antibodies in the strains isolated from dogs to the reference B. canis RM6/66 strain.

[The mechanism of action and the nature of the factors inducing the formation of uncultivatable forms of Salmonella typhimurium]. / Mekhanizm deistviia i priroda faktorov, indutsiruiushchikh obrazovanie nekul'tiviruemykh form u Salmonella typhimurium.

Approaches to the study of gene expression in the process of transition and prolonged stay of bacterial cells in the uncultivated state have been worked out with the use of the methods of reverse transcription and polymerase chain reaction. The differential expression of genes pqi and stn has been shown to occur in the uncultivated forms of S. typhimurium. The factor inducing the process of the transition of Salmonella cells into the uncultivated state has been found out.

[Electrokinetic potential of Yersinia pestis and Escherichia coli cells with an intact or defective ycaA gene (caf1M) of the fra-operon of plague pathogen]. / Elektrokineticheskii potentsial kletok Yersinia pestis i Escherichia coli s intaktnym ili defektnym genom ucaA (caf1M) fra-operona vozbuditelia chumy.

The dependence of the amount of electrokinetic potential in cells of Escherichia coli and Yersinia pestis, which differ in the rate of reduction of lipopolysaccharide (LPS), on the presence or absence of typical and atypical capsules of Y. pestis, encoded by intact and mutant fra operons, respectively, was studied. The ycaA+ycaF+(caf1 M+caf1+) genotype was shown to be expressed in serological stability of a classical capsular antigen, irrespective of the producer strain, and a decrease in the negative charge of microbial cells compared to their noncapsular variants. Blocking of the synthesis of the product of the ycaA gene of the fra operon resulted in formation of encapsulated bacteria, whose surface electricity and serological characters varied in dependence on the LPS structure. Data obtained support the assumption that a product of the ycA gene stabilizes the conformation of the typical capsule of the plague-causing agent, which was formed from YcaF (Caf1) monomers.

[The concept of the pathogenesis of chronic prostatitis]. / K kontseptsii patogeneza khronicheskogo prostatita.

The examination covered 4250 males (mean age 23,8 years) suffering from acute and chronic urethritides of various origin. In the chronic course of nongonococcal urethritis chronic prostatis (CP) occurred in 44.1-59.5% of the patients. In short-term nongonococcal urethritides CP was registered 3-4 times more rarely, in acute gonorrhea 40 time more rarely compared to chronic disease. Out of 128 males aged 17-24 free of urological diseases CP was detected in 20.3% of the examinees. CP 150 sufferers aged 19-61 had chlamydia in the urethra in 41.6% of cases. It is suggested that urethral infection can contribute to the onset of CP or add to prostatitis of another origin. It may also aggravate the run of noninfectious prostatitis.

[The chrysotherapy of patients with Reiter's disease]. / Krizoterapiia bol'nykh bolezn'iu Reitera.

Chrisanol therapy was provided to 172 men suffering from Reiter's disease, having histocompatibility antigen B35 and its combination with B27 as well as B17 and B11 indicating that the articular syndrome may run a persistent and grave course. Chrisanol therapy brought about a decrease of the activity of complement, lysozyme, bactericidal activity of blood serum, a reduction of the content of serum IgG, and autoimmune reactions parameters. Chrisanol therapy was tolerated well. The ten-year follow-up revealed relapses in 8.3% of the patients.

[Ophthalmological aspects of Reiter's disease]. / Oftal'mologicheskie aspekty bolezni Reitera.

Clinical picture and course of eye involvement were studied in 254 patients with Reiter's disease. Altogether 130 patients suffered from conjunctivitis of varying severity, 23 from keratitis, 21 from episcleritis, 20 from uveitis, 2 from detachment of the retina, and 5 from secondary glaucoma. Chlamydia were detected in one third of conjunctivitis patients in scrapings off the conjunctiva. The eyes may be accidentally infected by agents from the urogenital organs if the patient does not observe the hygienic rules. Uveitis developed as a rule after a prolonged course of Reiter's disease. They were resistant to antichlamydial therapy because of autoimmune factors that underlay the condition pathogenesis. Deposits of immune complexes on antigens of ocular vascular coating were detected with the use of monospecific antiglobulin sera in 7 of 10 patients with active uveitis symptoms. The risk of Reiter's disease development in HLA B27 carriers is 27.17 times higher than the mean incidence of this disease in the population. HLA A9 and B40 antigen combinations occurred 7 times more often in uveitis patients than in reference subjects, A1 and B27 combinations were 4-5 times more incident.

[The characteristics of the immune response to Chlamydia antigens in patients with Reiter's disease]. / Osobennosti immunnogo otveta na antigeny khlamidii bol'nykh bolezn'iu Reitera.

Immunologic tests (complement fixation, microprecipitation reaction, leukocytic agglomeration phenomenon, leukocyte migration inhibition tests) with standard chlamydial antigen were used in examinations of 116 male patients with Reiter's disease, 54 of these with the first (early) and 62 with the second (late) stages of this process. The findings evidence a higher intensity of antichlamydial humoral and cellular immune response in the patients with the first stage of the disease and a much lower intensity of those with the second stage. The pathologic process in patients with the second stage is liable to a prolonged self-maintaining course and is torpid to anti-infection therapy, as against that in the patients with the first stage of the condition; this fact prompts a conclusion that such a course of the process is explained not so much by the infectious agent, as by immune inflammation.

[The prognosis of the course of Reiter's disease with the use of histocompatibility antigens]. / Prognozirovanie techeniia bolezni Reitera s pomoshch'iu antigenov gistosovmestimosti.

Histotyping of 158 patients with Reiter's disease and 583 healthy donors was made. Detection of patients with this association of antigens helps timely diagnosis of the disease and appropriate preventive measures.

[A diagnostic criterion of Reiter's disease]. / Kriterii diagnostiki bolezni Reitera.

Clinical and laboratory criteria of the diagnosis of Reiter's disease are presented, based on many-year clinical and immunologic examinations of 254 male patients with this condition; these criteria help early and reliably diagnose this disease.

[The urogenital inflammatory focus and its treatment in Reiter's disease]. / Urogenital'nyi vospalitel'nyi ochag i ego lechenie pri bolezni Reitera.

The complement fixation test, microprecipitation, leukocyte migration inhibition, leukocyte agglomeration phenomenon, direct and indirect immunofluorescence were used to demonstrate that chronic prostatovesiculitis associated with Reiter's disease may be of immune-induced nature. This requires a differentiated approach to the choice of the treatment policy. The methods of the treatment for the urogenital inflammatory focus in men suffering from Reiter's disease are provided. It is indicated that complete cure of Reiter's disease can be attained only after elimination of inflammation in the urogenital organs.