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Iu V Vertiev

Publications and source records attributed to Iu V Vertiev.

At least 19 recordsLinked to original sources
PubMed2000

[Detection of toxin-producing pathogenic bacterial strains by polymerase chain reaction].

Polymerase chain reaction (PCR) was used for detection of pathogenic Clostridium botulinum, Clostridium perfringens, Clostridium difficile, and Escherichia coli. With this aim in view, primers to botulinic toxins types A, B, C1, D, E, F, and G, perfringens enterotoxin, difficile toxin, and types 1 and 2 Shigella-like toxins were chosen and synthesized. Optimal amplification conditions were selected for each pair of primers, with DNA and the respective agent as the reaction mixture matrices. PCR was highly specific and sensitive in all cases. Its sensitivity was 10-100 cells/sample. Among the tested C. botulinum, C. perfringens, C. difficile, and E. coli strains, specific amplification products of expected size were observed only in the strains containing the respective toxin genes. These findings recommend the use of these methods in clinical microbiology. Strains containing type 2 Shigella-like toxin gene were detected among E. coli strains isolated from patients with the hemolytic uremic syndrome, which for the first time indicates that the problem with E. coli epidemic strain O157 is valid for Russia. As a result of our studies, test systems for detection of types A, B, C, D, E, F, and G C. botulinum strains, C. perfringens and C. difficile, and E. coli O157 strains are now available.

Bacterial Toxins↗
PubMed2000

[Effective expression of fragments of a botulinum neurotoxin type A gene, coding for the L-chain and H-chain in E. coli, with formation of products causing protective immunity to administration of the toxin].

Native Clostridium botulinum gene coding for type A neurotoxin has been used to construct recombinant derivatives coding separately for L and H polypeptide chains of the toxin. The gene derivatives have been cloned into an expression vector pET28b in E. coli BL21 (DE3) cells. The recombinant L and H proteins seem to be the major individual proteins after IPTG induction of the recombinant cells. Each of the proteins has been accumulated only in inclusion bodies. The recombinant L chain (but not H chain) has been successfully resolubilized. Each of the proteins contains six His residues on the N terminus which allows purification on Ni-agarose columns with high yield. No toxic effect has been observed for both L and H chains after injection of 10 micrograms of recombinant preparations purified from inclusion bodies. Moreover, the injection resulted in an increase in the titer of specific antibodies which protected mice from 1 DLM of type A native botulinum neurotoxin. Hence, the recombinant neurotoxin protein derivatives which are present in E. coli inclusion bodies can be a source of material for producing diagnostic and therapeutic sera against type A botulinum neurotoxin.

Base Sequence↗
PubMed1991

[The splitting of the acceptor proteins of the protein kinase system in eukaryotic cells by Legionella cytolysin].

In the process of protein kinase reaction carried out in the mixture consisting of tris-HCl buffer, EDTA, MgCl2, gamma-32P-ATP and the cytoplasmic fraction of rabbit pulmonary cells the phosphorylation of proteins with molecular weights of 150 and 55 kD took place. The addition of L. pneumophila culture fluid to the reaction mixture resulted in the splitting of phosphorylated proteins with the formation of the component having a molecular weight of 45 kD. These disturbances in protein kinase reaction were found to occur due to the involvement of Legionella cytoplasm, a previously characterized protein with a molecular weight of 37 kD, into the process. In this connection, the participation of cytolysin in the pathogenesis of Legionella infection may also be considered from the viewpoint of the effect produced by cytolysin on the regulatory processes affecting the metabolism of target cells.

Animals↗
PubMed1990

[Localization of the structural gene for botulinum neurotoxin type A using synthetic DNA probe for neurotoxin light chain].

To obtain the information on the genetic control of toxin production in the botulism causative agents, the oligonucleotides were synthesized as the molecular probes by translation of the amino acid sequence of the botulinic type A neurotoxin. The optimal conditions for hybridization of botulinic DNA with the synthetic DNA probes were determined and the probes specificity was demonstrated. The DNA fragments homologous to the probes used were shown to belong to bacterial genome, but not to bacteriophage one.

Amino Acid Sequence↗
PubMed1990

[The toxin-producing capacity of populations of Vibrio cholerae of different origins].

As the result of the study of toxin formation in 165 V. eltor strains having different virulence, most of the virulent cultures have shown stable toxin production, though 5-10% of their colonies have proved to be nontoxigenic. In faintly virulent strains toxin production was found to be unstable, which is indicative of the heterogeneity of their population composition as regards their capacity for toxin formation. The population of avirulent strains consists mainly of nontoxigenic clones (95.7%).

Animals↗
PubMed1988

[Cytolysin as an immunoserological marker of Legionella pneumophila].

Coagglutination test (CoaT), radioimmunoassay (RIA) and ELISA were used for detection of cytolysin produced by L. pneumophila. The sensitivity of RIA was 100 ng/ml, CoaT--10-20 ng/ml, ELISA--1 ng/ml. To determine cytolysin production among various strains and species of Legionella, the authors studied bacterial ultrasonic lysates. All 11 strains of L. pneumophila tested were cytolysin-positive. L. longbeachae, L. bozemanii and L. dumoffii strains failed to produce cytolysin. The authors believe that an antigen of cytolysin can be used for identification of L. pneumophila.

Agglutination Tests↗
PubMed1988

[Enterotoxigenic Escherichia in patients with acute intestinal infection].

Out of 4,082 Escherichia strains isolated from 989 hospitalized patients with acute enteric infections 2,979 strains belonged to 139 O-groups and the remaining strains could not by typed with O-sera (O1-O164) or were in the serological R-form. 19.3% of the strains isolated from 354 patients showed a positive reaction for thermolabile enterotoxin (TLET) determined in the radioimmunoassay. Escherichia with pronounced reaction for TLET were isolated from 5.5% of the patients. The serological picture of Escherichia with a positive reaction for TLET comprised 78 O-groups, showed practically no variations in children of different age and adult patients and was related to the intensity of reaction for TLET. The number of enterotoxigenic Escherichia greatly varied within individual O-groups: from 3.1 +/- 3.63% (O11) to 100 +/- 9.81% (O114) of the total number of known strains in these O-groups. In most O-groups the number of enterotoxigenic strains varied from 10% to 50%. The most widely spread serovars of etiologically important enterotoxigenic Escherichia belonged to 24 O-groups and were often isolated as monocultures. The capacity for producing TLET (and, probably, some other antigenically related substances) in widely spread among Escherichia; still, the determination of enterotoxicity is a necessary, but not sufficient criterion of their etiological importance in acute enteric infections.

Acute Disease↗
PubMed1988

[Characteristics of the cytolysin of Legionella pneumophila].

A thermolabile cytolysin was purified from liquid culture of L. pneumophila. Its homogenicity was determined by the Ouchterlony double immunodiffusion and SDS-electrophoresis in polyacrylamide gel. The molecular weight of cytolysin was ca. 37 kDa. Analysis of amino acid composition revealed a high proportion of aromatic, dicarbonic amino acids, and methionine. The minimal cytolytic concentration for CHO cells and erythrocytes was ca. 1 microgram/ml: Purified cytolysin in doses of 10-60 micrograms caused haemorrhage and necrosis when injected i. c. into guinea pigs.

Amino Acids↗
PubMed1987

[Immunochemical characteristics of the subunits of cholera enterotoxin and of thermolabile Escherichia coli enterotoxins of various origins].

Antigenic determinants of subunits A and B of cholera enterotoxin (CT), heat-labile enterotoxin from the human E. coli strain (hLT) and heat-labile enterotoxin from the porcine E. coli strain (pLT) were analysed by Ouchterlony double gel-diffusion test against antisera to B subunits of three toxins and antisera to three holotoxins. The results have shown the existence of the following antigenic determinants: in subunits B-1. antigenic determinants, common for B subunits of all three enterotoxins-B(chp); 2. group antigenic determinants, common for B subunits of two toxins in the pair-B(ch), B(hp); 3. antigenic determinants, unique for B subunits of each CT, hLT, pLT-(B(c), B(h), B(p); in subunits A.-1. antigenic determinants, common for A subunits of all three enterotoxins-A.(chp); 2. group antigenic determinants, common for A subunits of two enterotoxins (hLT and pLT/-A(hp); 3. antigenic determinants, unique for A subunit of CT-A(c). On the basis of these results antigenic formulas for subunits of CT, hLT, pLT were proposed.

Animals↗
PubMed1987

[Construction of recombinant plasmids encoding the biosynthesis of the beta-subunit of cholera toxin].

The structure of the cholera toxin operon and the location of A and B toxin subunits have been studied by the Southern blot hybridization on filters. The gene coding for the synthesis of the cholera toxin B-subunit has been cloned in the vector plasmid pBR322. The structural gene of A-subunit has been partially deleted by the restriction endonuclease Bal31 digestion. The size of the 250 b. p. deletion has been defined by electron microscopy. The production of the cholera toxin B-subunit in Escherichia coli K12 cells has been studied.

Cholera Toxin↗
PubMed1987

[Characteristics of the B subunit of a thermolabile Escherichia coli enterotoxin produced by the A-B+ E. coli strain].

The preparation and identification of B subunit of thermolabile enterotoxin produced by A-B+ gene-containing strain are described. The E. coli strain studied is shown to produce protein identical in its molecular properties and antigenic specificity to B subunit obtained from the whole thermolabile enterotoxin. Partial antigenic affinity between B subunits of thermolabile enterotoxin obtained from different sources and B subunit from cholera enterotoxin has been established in immunochemical studies. Electrophoretic and immunochemical analysis has confirmed the absence of A-subunit admixtures in B-subunit preparation obtained from /A-B+/E. coli strain.

Animals↗
PubMed1986

[Conditions for Legionella pneumophila cultivation that affect cytotoxin production].

Experiments in CHO cell cultures have demonstrated the cytotoxic action of culture filtrates obtained after growing L. pneumophilia strain Philadelphia I, serogroup 1, in a liquid culture medium. This cytotoxic activity has been found to differ in its character from that described in earlier works. The dynamics of the accumulation of cytotoxin in the culture medium has been studied, and the work aimed at the determination of the limits of some cultivation parameters, within which the maximum toxin formation is observed, has been carried out.

Animals↗
PubMed1986

[Ability of Citrobacter freundii strains isolated in acute intestinal infections to produce LT-enterotoxin].

C. freundii enteropathogenic strains were found to be capable of producing choleroform thermolabile enterotoxin. Thus, in the study of 96 C. freundii strains 38 enterotoxin-producing cultures (39.5%) were revealed by means of the molecular-biological techniques and 29 such cultures (30.0%), by means of the radioimmunoassay (RIA). 100% coincidence was noted in the results of tests for enterotoxigenicity, made by means of RIA or hybridization techniques with the use of the LT-probe containing a cloned fragment with the gene coding the synthesis of LT-enterotoxin in enterotoxigenic Escherichia coli. At the same time only 29 out of 38 Citrobacter strains found to be positive in the hybridization tests, yielded the positive result when tested in RIA for the presence of LT-enterotoxin. This fact should be taken into consideration in the determination of enterotoxin-producing cultures isolated in acute enteric infections, as the method of genetic probing is capable of bringing out the genetic information in bacteria even in the absence of its phenotypical expression.

Acute Disease↗
PubMed1985

[Trial of different nutrient media for the production of an intracellular thermolabile enterotoxin by Escherichia coli strains H74-114 and 86].

The conditions of cultivation, ensuring the maximum accumulation of intracellular thermolabile enterotoxin in the cultures of two E. coli strains of different origin, have been studied. Culture media manufactured in the USSR have been selected and the conditions of cultivation, necessary for obtaining intracellular thermolabile enterotoxin in preparative amounts, have been established. Under these conditions the yield of thermolabile enterotoxin in 1.4 mg per 1 liter of culture medium for strain H74-114 and 1.0 mg per 1 liter of culture medium for strain 86.

Culture Media↗
PubMed1985

[Isolation and characteristics of the thermolabile enterotoxin from Escherichia coli strain H74-114].

The modified method for isolation and purification of LT-enterotoxin from Escherichia coli strain H74-114 has been proposed. Modification is based on exclusion of concentration of bacterial cell lysates by 3-5 fold dilution and changing PH during chromatography. The proposed modification permitted to increase the yield of purified and active product 8-10 fold. The obtained preparations are homogenic due to PAAG-SDS data and immunochemical analysis. B-component of TL-enterotoxin is shown to consist of 4 subunits B in contrast to B-component of choleragen, consisting of 5 subunits. The mol mass of A component of both toxins is identical but in contrast to choleragen the component of LT-enterotoxin is not subject to proteolysis.

Bacterial Toxins↗
PubMed1985

[Determination of toxigenicity of Escherichia coli and Vibrio cholerae strains by radioimmunologic method].

A solid phase variant of radioimmunoassay has been elaborated for screening toxin-producing strains of E. coli and V. cholerae grown on agar plates. The method is based on the ability of cholera-like toxins to be absorbed on nitrocellulose filters and their further identification with the use of homologous sera and [125I]-A protein from staphylococci. Sensitivity of the method reaches 20 pg. The proposed technique permits identification of intracellular enterotoxin and is aimed at a massive screening of E. coli strains, NAG-vibrios and V. cholerae strains for toxin production.

Enterotoxins↗
PubMed1984

[Exotoxin A production during Pseudomonas aeruginosa PA-7 cultivation in Martin's broth].

The activity of exotoxin A in culture filtrates prepared from cultures obtained by growing P. aeruginosa strains PA-7 and PA-103 in Martin's broth containing iron at a concentration of 0.08 microgram/ml, 0,05 M sodium glutamate and 1% of glycerin has been shown to be 1.5 times higher than that in filtrates prepared from cultures obtained by growing the above strains in a medium containing soybean tryptic digestion (USA). The optimun conditions for the production of exotoxin A by these strains are achieved during their cultivation in a fermenter at a temperature of 32 degrees C for 18 hours with simultaneous stirring (800 r. p. m.) and oxygenation (450 m3/h). Under these conditions the biological activity of the filtrates is 200 LD50/ml, their ADP-ribosyltransferase activity is 9500 c. p. m. and a sharply defined precipitation line appears in the double diffusion test in gel with monospecific antiserum to purified toxin, used in a dilution of 1:8.

ADP Ribose Transferases↗