PubMed Health⌕ Search

Biomedical subjects

A I Kalinovskiĭ

Publications and source records attributed to A I Kalinovskiĭ.

At least 19 recordsLinked to original sources

[Use of specific antibodies labelled with colloid silver particles for detection of Brucella antigens with dot immune analysis].

Fitness of dot immuno-analysis for detection of Brucella antigens labeled with colloid silver is evaluated. Soluble lipopolysaccharides and protein-saccharide antigen and corpuscular antigens of 22 Brucella strains (7 species) pathogenic for humans and animals in the S and R forms were used. The specificity of the method was tested on 10 heterologous microorganisms whose antigens were closely related. The suggested test system is simple, economic, highly sensitive (from 62 thousands to 8 million CFU/ml) and specific, requires no expensive equipment, and is an alternative to enzyme immuno-assay and dot immuno-analysis with gold immunosole.

Animals↗

[Development of a method for alkaline extraction of DNA from Brucella for diagnosing brucellosis using the polymerase chain reaction].

Several methods of alkaline extraction of chromosome DNA from Brucella in the presence of 50 microliters model diagnostic material blood serum are developed for the diagnosis of brucellosis by the polymerase chain reaction (PCR). These methods are based on the capacity of NaOH to effectively denature proteins and destroy Brucella cell wall, thus isolating the genome DNA without exposure to proteolytic enzymes, detergents, deproteinization, or pH neutralization. The first method consists in alkaline lysis by 0.2-1.0 M NaOH followed by DNA precipitation with two ethanol volumes in the presence of 0.1 M NaCl, washing of the resultant precipitate in 80% ethanol, drying of the precipitate, and dissolving in distilled water. The second method includes alkaline lysis in the presence of 0.3 M NaCl with NaOH concentrations of 0.5-4.3 M and the stages of DNA sedimentation, washing of precipitate, its drying and dissolving similar to those in alkaline lysis. The third method consists in alkaline lysis-precipitation by 0.2-05 M NaOH in the presence of 0.1 M NaCl and 64% ethanol, followed by DNA preparation stages similar to those in alkaline lysis. The best results were achieved by alkaline lysis in the presence of 0.3 M NaCl at NaOH concentrations of 0.7 and 2.1 M, which meant theoretical levels of sensitivity 140 and 86 Brucella cells, respectively.

Alkalies↗

[Use of protein-polysaccharide Brucella antigen labeled with colloidal gold for detection of specific antibodies by the dot-immunoassay method].

Antibrucella antibodies were detected by dot-immunoassay with colloid gold label of antigen. The specific antigen was protein-polysaccharide complex (PPC) isolated from vaccine strain Brucella abortus 19 BA by acetic acid hydrolysis of bacterial cells. Dot immunoassay with PPC labeled with colloid gold was effective in testing cattle, rabbit, and human sera for antibrucella antibodies. The proposed test system is simple, economic, highly sensitive and specific, and requires no expensive equipment and reagents, which are its advantages over routine serological tests.

Animals↗

[Analysis of PCR-diagnosis of brucellosis in human].

Polymerase chain reaction (PCR) was used for the diagnosis of brucellosis in humans with different forms of this disease. A high incidence (77.6%) of Brucella infection was revealed in the staff of cattle breeding centers with unfavorable situation with regard to brucellosis. Such a conclusion was made after PCR testing of native human sera. In acute brucellosis of humans amplification of the specific site of brucella DNA in PCR is possible only after extraction of DNA by a procedure adapted for DNA extraction from intact brucella cells. In chronic infection weak amplification of brucella genome DNA fragment was observed in investigation of native sera by the PCR. More expressed amplification product was recorded in PCR with a DNA precipitate from this serum obtained by ethanol precipitation. A still higher level of brucella DNA fragment amplification was observed after DNA extraction from sediment obtained by ethanol precipitation from this serum. These data confirmed the incomplete phagocytosis phenomenon at the early stage of infection, known in brucellosis pathogenesis, and allowed some hypotheses on the pathogenesis of chronic phase of brucellosis infection.

Animals↗

[Detection of Brucella antigens by dot immunoassay using specific antibodies labelled with colloid gold particles].

Dot immunoassay was developed to improve the quality of laboratory diagnosis of brucellosis. Particles of colloid gold were used as a marker of specific antibodies. The method was used for detecting Brucella antigens in artificially contaminated environmental objects (soil and water) and in biological material (milk, blood serum, and visceral homogenates of animals). The sensitivity of the test system was 19.5.10(3)-62.5.10(4) CFU/ml. Specificity of the assay was tested with 10 heterologous antigenically closely related bacterial species. The proposed test system is simple, economic, highly sensitive and specific, and requires no expensive equipment and reagents.

Animals↗

[Experimental anthrax infection in laboratory animals with differing species susceptibility to the causative agent].

In guinea pigs and noninbred white mice, infected subcutaneously with anthrax which resulted in their death, characteristic generalized infection with the hematogenic contamination of their organs and the signs of intoxication and shock could be observed. In inbred white rats (Fisher 344) the invasion and dissemination of B. anthracis are relatively slightly pronounced, the phenomena of intoxication and shock being clearly prevalent.

Animals↗

[Specific intoxication in anthrax infection].

The pathomorphological picture of experimental B. anthracis infection in white rats (strain Fisher-344) essentially corresponds to experimental anthracic intoxication with very moderately pronounced morphological manifestation of B. anthracis invasion. This indicates that specific anthracic intoxication is an essential component of the pathological process in B. anthracis infection.

Animals↗

[Br. ovis sensitivity to antibiotics].

Irrespective of their geographical origin the cultures of Br. ovis are highly sensitive to tetracycline and streptomycin. The bactericidal concentrations of tetracycline for 86.6 per cent of the strains and streptomycin for 93.20 per cent of the strains were 0.02--0.04 and 0.1--0.4 gamma/ml respectively. The bactericidal effect of morphocycline, kanamycin, benzyl-penicillin, lincomycin and monomycin was less pronounced. Polymyxin M had no bactericidal effect on the Brucella even in concentrations of 200 gamma/ml. Antibiotic sensitivity of S-subcultures of Br. ovis and the initial R-variants was studied comparatively. It was found that genetically related S-subcultures of Br. ovis were more resistant to the antibiotics than the initial R-strains.

Anti-Bacterial Agents↗

[Steroid compounds from far Eastern starfishes Henricia aspera and H. tumida].

Six new natural compounds were isolated from two Far Eastern starfish species, Henricia aspera and H. tumida, collected in the Sea of Okhotsk. Two new glycosylated steroid polyols were obtained from H. aspera: asperoside A and asperoside B, which were shown to be (20R,24R,25S)-3-O-(2,3-di-O-methyl-beta-D-xylopyranosyl)-24-methyl-5alpha-cholest-4-ene-3beta,6beta,8,15a,16beta,26-hexaol and (20R,24R,25S,22E)-3-O-(2,4-di-O-methyl-beta-D-xylopyranosyl)-24-methyl-5alpha-cholest-22-ene-3beta,4beta,6beta,8,15alpha,26-hexaol, respectively. Two other glycosylated polyols, tumidoside A, with the structure elucidated as (20R,22E)-3-O-(2,4-di-O-methyl-beta-D-xylopyranosyl)-26,27-di-nor-24-methyl-5alpha-cholest-22-ene-3beta,4beta,6beta,8,15alpha,25-hexaol, and tumidoside B, whose structure was elucidated as (20R,24S)-3-O-(2,3-di-O-methyl-beta-D-xylopyranosyl)-5alpha-cholestan-3beta,4beta,6beta,8,15alpha,24-hexaol, were isolated from the two starfish species. (20R,24S)-Salpha-Cholestan-3beta,6beta,15alpha,24-tetraol and (20R,24S)-5alpha-cholestan-3beta,6beta,8,15alpha,24-pentaol were identified only in H. tumida. The known monoglycosides henricioside H1 and laeviuscolosides H and G were also identified in both species.

Animals↗

[Novel triterpene glycosides from the sponge Ulosa sp].

New triterpene glycosides, ulososides C, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-glucopyranosyl)-32-nor-24-methyllanost- 8(9)-ene-30-oic acid, D, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-N-acetyl-glucosaminopyranosyl)-32-nor- 24-methyllanost-8(9)-ene-30-oic acid, and E, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-glucuronopyranosyl-(1-->2)-alpha-D- arabinopyranosyl-32-nor-24-methyllanost-8(9)-ene-30-oic acid, were isolated from an Ulosa sp. sponge. Their structures were determined by spectral methods and chemical transformations. Specific features of their structures are discussed.

Animals↗

[Steroid compounds from the Far Eastern starfish Diplopteraster multipes].

Sodium salt of (20R)-3 alpha,4 beta-dihydroxycholest-5-ene-21-yl sulfate and disodium salts of (20R)-4 beta-hydoxycholest-5-ene-3 alpha,21-diyl disulfate, (20R)-24-methylcholest-5,24(28)-diene-3 alpha,21-diyl disulfate, (20R)-24-methyl-5 alpha-cholest-24(28)-ene-3 alpha,21-diyl disulfate, (20R)-cholest-5-ene-3 alpha, 21-diyl disulfate, (20R)-5 alpha-cholestane-3 alpha,21-diyl disulfate, and (20R)-3 alpha-hydroxycholest-5-ene-2 beta,21-diyl disulfate were isolated from the far eastern starfish Diplopteraster multipes and characterized. These compounds differ structurally from sulfated polyhydroxysteroids in other starfish species. At the same time, they are typical secondary metabolites of Ophiuroidea and have some structural features characteristic of the ophiuroid-isolated steroids, namely the 3 alpha-hydroxy (or 3 alpha-sulfoxy) and 21-sulfoxy groups. These data support the opinion of some taxonomists that starfishes and ophiuroids are phylogeneteically related classes and are closer to each other than to other classes of the Echinodermata phylum.

Animals↗

[Steroidal polyols from Far Eastern starfish Henricia sanguinolenta and H. leviuscula leviuscula].

Two new asterosaponins, (20R)-3-O-beta-D-(2-O-methylxylopyranosyl)-24-propylcholest-4-ene-3 beta,6 beta, 8, 15 alpha, 16 beta, 29-hexaol (sanguinoside A) and (20R,24S)-3-O-beta-D-(2,3,4-tri-O-methylxylopyranosyl)-5 alpha-cholestane-3 beta, 4 beta, 6 beta, 8, 15 alpha, 24-hexaol (sanguinoside B), were isolated from two species of Pacific Far Eastern Starfish Henricia sanguinolenta and H. leviuscula leviuscula, collected in the Sea of Okhotsk. Both glycosides contain aglycones with pentahy-droxysteroid nuclei of similar structures, which are substituted at the 3-hydroxy group with differently methylated beta-D-xylosyl residues. Sanguinoside A has an unusual structure of its aglycone side chain, whereas sanguinoside B has a unique permethylated carbohydrate chain. In addition, laevisculoside G, a known glycoside, was identified in the H. leviuscula starfish. The structures of the isolated glycosides were established by interpreting their spectral data and by comparing their spectral characteristics with those of known compounds. The English version of the paper. Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 2; see also http://www.maik.ru.

Animals↗

[New steroid glycosides from the starfish Fromia milleporella].

Two new steroid glycosides from the starfish Fromia milleporella collected in the Seychelles were isolated and characterized: milleporoside A, (20R, 24R)-29-O-[3-O-methyl-beta-D-xylopyranosyl-(1-->4)-3-O-methyl-beta-D-xylopyranosyl]-24-ethyl-5alpha-cholestane-3beta,4beta,6alpha,8,15beta,16beta,29-heptaol, and milleporoside B, (20R, 24R)-(22E)-28-O-[3-O-methyl-beta-D-xylopyranosyl-(1-->4)-3-O-methyl-beta-D-xylopyranosyl]-24-methyl-5alpha-cholest-22-ene-3beta,4beta,6alpha,8,15beta,16beta,28-heptaol. The structures of the glycosides were determined from their spectra and a comparison with spectral characteristics of known compounds. These compounds exhibit a moderate cytostatic activity toward the embryos of the sea urchin Strongylocentrotus intermedius.

Animals↗

[Detection of brucella antigens with colloid metal particles as markers for specific antigens].

Gold and silver sols were comparatively approved as markers of specific IgG isolated from hyperimmune Brucella antisera for the detection of brucellar antigens. The sensitivity of the test system using gold immunosol proved to be some higher (3.1-9.8 ng/ml of soluble and 2.0 x 10(4)-5.3 x 10(6) CFU/ml of corpuscular brucellar antigens) than that achieved with the use of silver immunosol (5.7-18.4 ng/ml of soluble and 6.1 x 10(4)-8.0 x 10(6) CFU/ml of corpuscular brucellar antigens). At the same time silver sol was a cheaper and more available marker. Both test systems were found to be highly specific. False positive results were observed only with Yersinia enterocolitica O:9 at high concentrations due to the fact that they had common polysaccharide haptens incorporated into lipopolysaccharides of these microorganisms. The proposed test systems with colloid metal particles used as markers of specific antibodies for the detection of Brucella antigens are technologically simple, economic, rapid, highly sensitive and specific. Their use in combination with other serological methods will make the results of analyses more informative, thus improving the quality of laboratory diagnostics of brucellosis.

Animals↗

[Dot immunoassay with colloidal silver particles as a marker of specific antigen for testing human sera for brucellosis].

Whether the dot immunoassay is suitable for the detection of Brucella antibodies in human sera by using a colloidal silver-labeled Brucella specific antigen as a diagnostic tool is assessed. The antigen was the B. abortus 19BA protein polysaccharide complex isolated by Brucella acetic acid hydrolysis. The dot immunoassay is easy-to-use, cost-effective, highly sensitive, and therefore of more informative value in detecting Brucella antigens than routine serological tests (Huddleson test, Wright agglutination test, passive hemagglutination test, long-term complement fixation test, and Coombs test). It requires no use of expensive equipment and reagents.

Antibodies, Bacterial↗

[Brucellosis in Siberia and the Far East].

Epidemiological analysis has indicated that rat and reindeer brucellosis foci are of definite value in Siberia and the Far East in the liquidation of brucellosis ones. Foci of cattle and reindeer have been first established, evidence has been provided for the epidemiological significance of fifth-biological variant B. ovis and B. suis, as well as the ecological confinement of peculiar B. rangiferi cultures to the brucellosis foci in the Arctic. To plan antibrucellosis efforts, it is necessary to take into account the incidence of human infection, as well as the insidious circulation of the bacillus in the stock farms.

Animals↗