[The economic prerequisites of implementation of innovative technologies of mass-spectrometry in diagnostic of bacterial infections].

The article presents the results of clinical economical analysis, based on "cost-effectiveness" technology, of MALDI-TOF MS, innovative medical technology of express identification of microorganisms, calculation of incremental indicator and application of notion "willingness-to-pay-threshold". Due to the extension of sanctions against the Russian Federation, this medical equipment for national laboratories becomes difficult to access and expensive, that conditions necessity to scientifically substantiate economical effectiveness of implementation of expensive innovative MALDI-TOF MS technology as instrument to contain global antibiotic resistance increase. The understanding of importance of express identification of microorganisms as well as other positive effects that can be achieved by using modern medical equipment on the basis of mass spectrometry results in improving medical care quality, increasing reputation level of medical institution, greater commitment of physicians and patients to microbiological analysis with purpose of prescription of rational antibiotic therapy and improving population health.

[Serologic study of experimental cholera polymer antigen diagnosticums].

AIM: Evaluation of quality indicators of constructed cholera antigen polymer diagnosticums by using a complex of specific anti-cholera sera. MATERIALS AND METHODS. Cell lysates of cholera vibrio strains Vibrio cholerae cholerae 1395, V. eltor Ogawa 2044, V. eltor Inaba 13020, V. cholerae O139 16064 were sensitins for experimental preparations. 3 sera from cholera patients, normal human sera, cholera O1 (Ogawa, Inaba) commercial horse, cholera O139 commercial rabbit and heterologic sera against shigella, salmonella, escherichia and yersinia as well as experimental cholera rabbit sera against O1 and O139 were used as control. RESULTS: The study established that diagnosticums based on V. cholerae cholerae 1395 and V. cholerae O139 16064 strain sensitins by quality indicators may be used in the future for construction of these diagnosticums. CONCLUSION: Antibody containing preparations--commercial horse O1 sera, rabbit experimental and commercial sera and MCA O139 demonstrating titers not lower than 1/5120-1/10240 may serve as a control of experimental diagnosticums in the absence of human sera from cholera patients.

[Increasing gingivoplasty efficiency by means of free palatal grafts].

The paper presents short and long-term results of surgical treatment of 80 patients with insufficient attached gingiva. The procedures included vestibuloplasty and gingival plasty with perfluorane and free gingival and subepithelial connective tissue grafts. Proposed surgical treatment method with perfluorane proved to reduce the local inflammatory reaction greatly thus decreasing perioperative morbidity and increasing the long term efficacy of treatment.

[Antibioticograms of Vibrio cholerae non-01/non-0139 strains isolated from humans within 1968-2009].

Analysis of the antibioticograms of the Vibrio cholerae non-01/non-0139 strains showed that in the cultures isolated in the Rostov Region in 1968--1975 there were present markers of resistance to ampicillin (7%), kanamycin (15.8%), rifampicin (3.5%) and trimetoprim/sulfamethoxazole (14%). Among the strains isolated in the Ukraine in 1975 14% was resistant to ampicillin. More than a half of the strains isolated in Uzbekistan in 1990 and 2000-2001, in the Arkhangelsk Region in 1999-2000 and in the Kalmykia in 1999-2005 was antibiotic resistant. In the above regions the strains were resistant to ampicillin (12.5-44.4%), kanamycin (11-55%), rifampicin (1.9-12.5%) and trimetoprim/sulfamethoxazole (25-62.5%). Among the cultures isolated in Uzbekistan in 1990 and 2000-2001 25 and 7.8% were resistant to furazolidone and 31.25% was resistant to streptomycin (1990). All the cultures isolated in the Rostov Region in 2005-2009 were resistant to ampicillin, 50% was resistant to ceftazidim, 57% was resistant to streptomycin and furazolidone, 7.2% was resistant to kanamycin and 14% was resistant to trimetoprim/sulfamethoxazole. The studies revealed an increase of the extent of the V.cholerae non-01/non-0139 resistance spectrum within 1968-2009, simultaneous presence of up to 5 diverse resistance markers and a variety of their combinations, that requires the use of antibacterials for the treatment of the diseases due to the vibrios in strict compliance with the pathogen antibioticogram and their early replace by more efficient drugs.

[In vitro induction of transmissive resistance to tetracycline, chloramphenicol and ampicillin chloramphenicol in Vibrio cholera non-O1/non-O139 serogroups isolated within 1990-2005].

Inducible character of resistance to tetracycline, chloramphenicol and ampicillin was investigated in 20 strains of Vibrio cholera non-O1/non-O139 serogroups isolated from inhabitants of Uzbekistan in 1990 (10 strains, ctx+) and in 2001 (5 strains, ctx-) and from inhabitants of Kalmykiya within 2003-2005 (5 strains, ctx-). Eight of the 20 isolates showed not only capacity for induction of the antibiotic resistance, but also its possible self transfer to Escherichia coli and reverse crosses in El Tor V. cholerae P-5879. It was shown that the effect of the antibacterial on the isolates phenotypic susceptibility could increase the resistance markers expression, when the genomes contained sites responsible for their expression, that required constant bacteriological control of the treatment efficacy and the use of the isolates antibioticograms for early replace of the inefficient drug by the efficient one. The prevalence of V. cholerae O1 and non-O1/non-O13 serogroups with multiple resistance to the antibacterial and the genetic potency for the antibiotic resistance development in the pathogen made difficult the choice of efficient drugs for prophylaxis and treatment of diseases caused by V. cholerae.

[Characterization of the adhesive activity of cholera vibrions in mammalian red blood cells as an additional test for assessment of their epidemic significance].

The in vitro study of the adhesive properties of V. cholerae eltor and V. cholerae O139 on a model of mammalian red blood cells revealed a correlation of their adhesive properties, the presence of the ctx AB, tcpA genes, and their hemolytic activity when blood group A (II) red blood cells were used. In the latter case, the strains having the characteristics of ctx(+) tcp(+) Hly(-) were ascertained to have a mean adhesive value (MAV) of > 1.5, a red blood cell involvement coefficient (RBCIC) of > 50%, while those with the characteristics of ctx(-) tcp(-) Hly(+) had a MAV of < 1.5 and a RBCIC of < 50%. In terms of the adhesive activity, the cultures with the genotype ctx tcp(+) Hly(+) is a heterogeneous group and may be low (MAV < 1.5) and high (MAV > 1.5) adhesive. According to the data of our studies, their adhesiveness is associated with the region of identification and independent on its object.

[Virulence of pre-CTXphi-carrying Vibrio cholerae: genotypic and phenotypic characteristics].

AIM: Complex assessment of virulence of cholera vibrios carrying the truncated CTX element (pre-CTXphi prophage). MATERIALS AND METHODS: Twenty-two strainsof Vibriocholerae O1 and non-O1/non-O139 were studied by PCR and laboratory models. RESULTS: Genomes of all strains, besides pre-CTXphi genes, contained genes hapA (hemagglutinin/proteases), cef (CHO cell elongating factor), rtxA (high-molecular cytotoxin), and rtxC (its activator). Nucleotide sequences of rtxA and vgrG genes from ACD domains, genes VPI and VPI-2 from islands of pathogenicity, mshA (mannose-sensitive pili) gene were presented in different combinations. None strains contained shiga-like toxin (slt1) aswell as thermostable direct (tdh) and thermostable direct-related (trh) hemolysin genes of V. parahaemoliticus. On the model of infant rabbits almost all strains caused a significant enteropathogenic effect sometimes resembling cholera effect and in a number of cases dissemination of bacteria into various organs and tissues took place. Cultural supernatants of the majority of strains stipulated cell rounding in CHO cultures (one of them caused cell destruction) and disconnection of cells in McCoy and L-929 dense monolayers as well as increase of skin permeability in Craig's test. Conclusion. Apparently, diarrhea of different severity observed in patients from whom these strains were isolated as well as signs of virulence revealed in the laboratory models were determined by the expression of genes of accessory pathogenicity factors including those detected in the present study.

[Genotyping of gram-negative bacteria isolated from blood in sepsis in patients with hematological diseases].

AIM: Genotyping of hospital strains of gram-negative bacteria isolated from blood of patients with hematological diseases in the presence of sepsis. MATERIAL AND METHODS: Bacterial DNA were studied with pulse-electrophoresis with commercial kits Genepath Reagent Kits (BioRad, USA). Comparison and interpretation of the results were made with application of the computer analysis program Bionumerics, Applied Maths. RESULTS: DNA of bacteria of 152 hospital strains was studied. In genotyping of 26 strains of P. aeruginosa 21 clones was isolated, 15 strains of Enterobacter spp.--12 clones, 31 strains of K. pneumoniae--27 clones, 80 strains of E.coli--79 clones. The greatest number of clones of identical bacteria (19%) was detected in the strains of P. aeruginosa, 17%--Enterobacter spp., 11%--K. pneumoniae, minimal (1%) in the strains of E. coli (p < 0.001). Incidence of isolation of clonal-relative strains among P. aeruginosa isolates was 35% (n = 9), Enterobacter spp.--33% (n = 5), K. pneumoniae--23% (n = 7), E. coli--2% (n = 2). CONCLUSION: Polyclonality of the microorganisms was detected. Frequency of isolation of clones containing identical bacteria in the test bacteria was variable. In clinic there are endogenic and exogenic mechanisms of infection with prevalence of endogenic.

[Hydrogen peroxide as an inductor of uncultivable state of Vibrio cholerae eltor in an experiment].

The influence of hydrogen peroxide on the dynamics of transition into uncultivable state (UCS) and on the reversion of V. cholerae and their subcultures, resistant to hydrogen peroxide, was studied. The transition of the initial cultures in river and distilled water into UCS took place earlier than that in resistant to hydrogen peroxide variants. The capacity for reversion to hydrogen peroxide resistant subcultures preserved, on the average, 2 - 3 times longer. An increase in the level of hydrogen peroxide in uncultivable populations was found to be 2.7 - 4.4 times. Subcultures, resistant to hydrogen peroxide, in the vegetative form had lower characteristics of peroxide concentrations than in uncultivable form (UCF), but somewhat higher than in initial variants. In revertants the concentration of hydrogen peroxide was lower in UCF, but somewhat higher than in vegetative cultures. The dynamics of the formation of UCF by cholera vibrios, with different degree of stability to the action of hydrogen peroxide, the accumulation of hydrogen peroxide in uncultivable populations, the deceleration of transition into uncultivable forms, an accumulation of hydrogen peroxide and an increase in the time of the reversion of clones, resistant to hydrogen peroxide, made it possible to suggest that the accumulation of hydrogen peroxide was possible to make an essential contribution to the formation of UCF of cholera vibrios in an experiment.

[Dynamics of the reversible transition of Vibrio cholerae into the uncultivable state in the presence of organic and inorganic microcosmic components].

The dynamics of the transition of V. cholerae into the uncultivable state in distilled, river and tap water, containing organic and inorganic components added, was studied. As additives, potassium nitrate, potassium phosphate, magnesium sulfate, ammonium chloride, lysine, alpha-ketoglutarate, succinic acid, catalase were used. The study of the influence of biotic factors on transition into the uncultivable state was carried out in the presence of one-celled green algae Scenedesmus quadricauda or infusoria Paramecium caudatum. The linear dependence of speed of transition into the uncultivable form on the concentration of cells was noted. The composition of the microcosmic medium was also found to have some influence on the speed of transition into the uncultivable form and on the reversibility of this process. The presence of organic substances, such as peptone solution or destroyed cells of phyto- and zooplankton, in the microcosmic medium prolonged the time of transition into the uncultivable form and produced a positive effect on the capacity of the population to reversion. In respect of live biotic components, no such dependence was found. Inorganic additives prolonged the time of transition into the uncultivable state, but did not promote reversion.

[Hemolytic activity of Vibrio cholerae eltor and V. cholerae O139 toxigenic and nontoxigenic strains]. / Gemoliticheskaia aktivnost' toksikogennykh i netoksikogennykh shtammov Vibrio cholerae eltor i V.cholerae O139 serogruppy.

A total of 20 ctx- and 16 ctx+ V. cholerae eltor strains, 20 ctx- and 22 ctx+ V. cholerae O139 strains were under study. Hemolytic activity was tested in modified Greig test with sheep, guinea pig and rabbit red blood cells. The comparative study of the hemolytic properties of V. cholerae O1 and O139 under different conditions of cultivation demonstrated their capacity of lysing sheep red blood cells (SRBC) irrespective of the presence of toxigenic properties. A wider spectrum of lytic activity of ctx- strains in Greig test with respect to red blood cells of different animals and the capacity of lysing SRBC, most resistant to the action of toxin, may be due to a considerably greater content of Hly+ clones in their population.

[Role of iron ions in the production of hemolysin by toxigenic and nontoxigenic Vibrio cholerae of different serogroups]. / Rol' ionov zheleza v produktsii gemolizina toksigennymi i netoksigennymi kholernymi vibrionami razlichnykh serogrupp.

The hemolytic activity of ctx- and ctx+ V. cholerae, serogroups eltor and O39, in a medium free of FeCl3 was studied. During the cultivation in this medium, the strains of both V. cholerae serogroups proved to be capable of lysing sheep red blood cells in the Graig test, irrespective of the presence of ctx genes. The cultivation of V. cholerae ctx+ strains of both serogroups under such conditions facilitated the production of hemolysin with the same spectrum of lytic activity as hemolysin produced by ctx- strains.

[Dependence of the expression of the biological features of Vibrio cholerae on the conditions of their cultivation]. / Zavisimost' ékspressii biologicheskikh svoistv kholernykh vibrionov ot uslovii ikh kul'tivirovaniia.

The biological activity of toxigenic and non-toxigenic V. cholerae supernatants was found to depend on the cultivation medium. The use of iron-free tryptone medium made it possible to obtain supernatants of toxigenic V. cholerae with haemolytic activity and destructive action on passaged cell cultures. In the experimental infection of suckling rabbits the influence of the cultivation conditions of V. cholerae on the character and expression of their pathogenic properties was determined. The dissemination of V. cholerae into the internal organs of rabbits after their infection with both toxigenic and non-toxigenic strains correlated neither with the cultivation conditions of these strains, nor with the character of changes in the intestine of the infected animals.

[Indicator strains of cholera vibrions for detection and primary identification of cholera phages]. / Indikatornye shtammy kholernykh vibrionov dlia obnaruzheniia i pervichnoi identifikatsii kholernykh fagov.

A set of test-strains is suggested; it comprises V. eltor P-13169 and V. cholerae 163-0139 designed for the primary identification of unknown previously choleraic bacteriophages.

[Invasive properties of toxigenic and nontoxigenic Vibrio cholerae of different serogroups]. / Invazivnost' toksigennykh i netoksigennykh kholernykh vibrionov razlichnykh serogrupp.

Experimental data on the comparative study of the invasive properties of vct+ Hly- and vct- Hly+ V. cholerae of serogroups 01 and 0139 are presented. Both vct- Hly+ and vct+ Hly- V. cholerae of serogroups 01 and 0139 have been shown to be capable of dissemination into internal organs. No differences in the dissemination of V. cholerae of different serogroups in both immunologically immature and mature experimental animals have been detected.

[Hemolytic activity and toxigenicity of Vibrio cholerae of different serogroups]. / Gemoliticheskaia aktivnost' i toksigennost' Vibrio cholerae razlichnykh serogrupp.

Experimental data on the comparative evaluation of the hemolytic activity of ctx+ Hly- and ctx- Hly+ V. cholerae, serogroups O1 and O139, in the process of their cultivation in different nutrient media are presented. The capacity of ctx+ V. cholerae of both serogroups cultivated under the conditions of iron deficiency, for the production of hemolysin capable of lyzing sheep red blood cells was shown. Hemolysin produced by ctx- strains of V. cholerae was synthesized under any conditions. The study of hemolysin preparations obtained from ctx- and ctx+ strains of V. cholerae, serogroups O1 and O139, revealed that they were biologically and immunologically similar.

[Evaluation of antibiotic sensitivity of pathogenic vibrios of various species]. / Otsenka chuvstvitel'nosti k antibiotikam patogennykh vibrionov raznykh vidov.

The recent increase of the number of antimicrobials and isolation of antibiotic resistant strains from humans and environmental objects is indicative of the necessity of further investigation of antibiotic susceptibility of the representatives of the genus Vibrio pathogenic for man to provide rational therapy of the diseases due to them. Susceptibility of 160 strains of pathogenic vibrios of 9 species to 11 antibiotics and chemotherapeutic drugs was assayed by the method of serial dilutions in agar media. The isolates were shown to be highly susceptible to chloramphenicol, doxycycline, cefotaxime, nalidixic acid and ciprofloxacin which made it possible to consider them as the drugs of choice in the treatment of the diseases caused by the microorganisms. A tendency to form polyantibiotic resistant strains within every species of tested pathogenic vibrios was observed. It conditioned the prospects of further profound study of the phenomenon with the analysis of the genetic determination of antibiotic resistance markers in pathogenic vibrios.

[The characteristics of Vibrio cholerae isolated in Dagestan in 1994]. / Nekotorye osobennosti Vibrio cholerae, vydelennykh v Dagestane v 1994 g.

A high degree of resistance to cholera diagnostic phages and carriership of prophages characteristic of V. cholerae eltor strains vct+ were shown to be the specific features V. cholerae isolated in Daghestan during the period of June-October 1994. Among the strains under study, isolated respectively in 12 and 18 out of 19 regions of Daghestan, a high proportion was found to have resistance to tetracycline (65%) and chloramphenicol (28.6%). Moreover, some strains were found to be resistant to furagin and erythromycin. Out of 242 strains resistant to antibacterial preparations, 163 strains were found to have multiple resistance. Gentamicin, cipropfloxacin and doxicycline were shown to have high in vitro activity with respect to the strains under study.