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Biomedical subjects

T Miwatani

Publications and source records attributed to T Miwatani.

At least 181 records · Page 10Linked to original sources

Comparison of bacterial cardiotoxins: thermostable direct hemolysin from Vibrio parahaemolyticus, streptolysin O and hemolysin from Listeria monocytogenes.

Some properties of the bacterial cardiotoxins, thermostable direct hemolysin from Vibrio parahaemolyticus (vibriolysin), and streptolysin O and hemolysin from Listeria monocytogenes (listeriolysin), were compared. These toxins had rapid lethal effects on mice when injected intravenously. The electrocardiographic changes of rats after intravenous injections of these toxins were very similar, showing bradycardia and inhibition of atrio-ventricular conduction. These toxins also caused cessation of the spontaneous beating and degeneration of cultured foetal mouse heart cells. When equal hemolytic units of these three toxins were administered, vibriolysin had the most potent effects on mice and cultured mouse heart cells. Differences in the kinetics of the hemolysis by each toxin and in the effects of cholesterol of their hemolytic actions suggest that the mode of action of vibriolysin is different from those of streptolysin O and listeriolysin.

Bacterial Toxins↗

Evidence that the multi-drug-resistance of Streptococcus faecalis BIO-4R is not carried by a plasmid.

The multi-drug-resistant strain Streptococcus faecalis BIO-4R was studied to see if it carried a plasmid responsible for antibiotic resistance. From results indicating that the antibiotic resistance of S. faecalis BIO-4R was not transferred to recipient bacteria, that the organism did not produce enzymes which inactivated antibiotics, and that the presence of covalently closed circular DNA was not demonstrated by dye-cesium chloride buoyant density gradient centrifugation, it was concluded that the organism did not carry such a plasmid. Studies on polyphenylalanine synthesis by cell-free extracts of the oganism showed that its ribosomes were resistant to chloramphenicol, tetracycline, streptomycin and kanamycin. These results, although rather indirectly, support the above notion that the multi-drug-resistance of S. faecalis BIO-4R is not mediated by a plasmid.

Anti-Bacterial Agents↗

Effect of zinc ion on the hemolytic activity of thermostable direct hemolysin from Vibrio parahaemolyticus, streptolysin O, and Triton X-100.

The effect of Zn2+ on hemolysis induced by the thermostable direct hemolysin from Vibrio parahaemolyticus (vibriolysin) streptolysin O, and Triton X-100 was studied. We found that in certain buffers, such as tris(hydroxymethyl)-aminomethane-hydrochloride, boric acid-borax, and N-hydroxyethyl piperazine-N'-2-ethanesulfonic acid-sodium hydroxide, hemoglobins released from erythrocytes were easily precipitated by addition of Zn2+, thus resulting in a false inhibition of hemolysin by Zn2+ when hemolysis was assayed by measuring absorbance at 540 nm of released hemoglobins. Under experimental conditions in which hemoglobin was not precipitated, hemolysis induced by streptolysin O was inhibited by Zn2+, whereas that induced by vibriolysin and Triton X-100 was not. Thus, we concluded that the mode of inhibitory action of Zn2+ on hemolysis was not due to a reversible alteration in the state of the lipid bilayer of erythrocytes membranes as proposed by Avigad and Bernheimer (Infect. Immun. 13:1378-1381, 1976).

Buffers↗

Toxin from the culture filtrate of Shigella dysenteriae that causes morphological changes in Chinese hamster ovary cells and is distinct from the neurotoxin.

A toxin that causes morphological changes in Chinese hamster ovary cells was isolated from a culture filtrate of Shigella dysenteriae 1. It was partially purified by successive column chromatography on diethylaminoethyl-cellulose, Bio-Gel A-5m, and hydroxylapatite. The toxin was separated from neurotoxin by diethylaminoethyl-cellulose column chromatography and was not lethal for mice.

Bacterial Toxins↗

Deoxyribonucleic acid strand breaks during freeze-drying and their repair in Escherichia coli.

Freeze-drying of Escherichia coli cells caused strand breaks of deoxyribonucleic acid (DNA) in both radiation-sensitive and -resistant strains. However, in the radiation-resistant strain E. coli B/r the damaged DNA was repaired after rehydration, whereas in the radiation-sensitive strain E. coli Bs-1 the damaged DNA was not repaired and the DNA was degraded. Repeated freeze-drying did not break the damaged DNA into smaller pieces.

DNA Repair↗

Effect of hemolysin produced by Vibrio parahaemolyticus on membrane conductance and mechanical tension of rabbit myocardium.

The mechanisms of the action of hemolysin extracted from Vibrio parahaemolyticus in the S-A node and right atrium cells of rabbit were studied by means of the single sucrose gap and isometric tension recording methods. Hemolysin caused the membrane to depolarize reversibly without affecting the action potential generating mechanism. Lowering of [Na+]o inhibited membrane depolarization in the presence of hemolysin while the readmission of normal Tyrode solution induced depolarization. Tetrodotoxin (TTX) barely antagonized the depolarizing action of hemolysin but slowed the rate of development of depolarization. Therefore, this depolarization is considered to be primarily due to the increase in conductance to Na which TTX may not block. The dose-response relationship was obtained by measuring a change in membrane resistance. The concentration necessary to yield one-half of the maximum reduction of the membrane was determined to by 7.5 micrograms/ml. Accumulation of Na within the cell may be responsible for an increase of twitch tension observed during the action of a low concentration of hemolysin. On the other hand, a higher concentration of hemolysin seemed to promote exchange of intracellar Na with extracellular Ca, especially when the Na concentration of the perfusing solution was reduced, and led to stronger contracture.

Animals↗

Identification of lethal toxin with the thermostable direct hemolysin produced by Vibrio parahaemolyticus, and some physicochemical properties of the purified toxin.

Lethal toxin was purified extensively from the culture filtrate of a Kanagawa phenomenon-positive strain of Vibrio parahaemolyticus. The purified toxin was a protein, and its homogeneity was demonstrated by sodium dodecyl sulfate-polyacrylamide gel disc electrophoresis and analytical ultracentrifugation. It was demonstrated that the thermostable direct hemolysin was identical to the lethal toxin and that it was the main, if not only, lethal toxin in the culture filtrate. The purified toxin had a lethal effect when injected into mice either intravenously or intraperitoneally. Its lethal effect was very rapid, a dose of 5 mug of toxin per mouse killing the animals within 1 min. The lethal activity was inhibited by a ganglioside mixture. Some physicochemical properties of the purified toxin are reported.

Animals↗

Demonstration of the cardiotoxicity of the thermostable direct hemolysin (lethal toxin) produced by Vibrio parahaemolyticus.

Intravenous injection of the thermostable direct hemolysin (lethal toxin) produced by Vibrio parahaemolyticus caused rapid death of rats. Studies by electroencephalography and electrocardiography showed that after intravenous injection of the toxin the electroencephalogram remained normal for quite a long time after the heart of the animals had stopped beating. Depression of intraatrial and intraventricular conduction of electrical activation, including atrioventricular block, was observed in electrocardiograms of animals injected with the toxin. The toxin was also found to be toxic to cultured mouse heart cells. When it was added to the medium, the beating rhythm of the cultured heart cells increased temporarily and then soon stopped abruptly. The effect of the toxin on cultured mouse heart cells was blocked by preincubation of the toxin with a ganglioside mixture. From these results it is concluded that the thermostable direct hemolysin (lethal toxin) had cardiotoxic activity, and thus administration of the toxin causes rapid death of animals.

Animals↗

Cytotoxic effect of the thermostable direct hemolysin produced by Vibrio parahaemolyticus on FL cells.

The thermostable direct hemolysin produced by Vibrio parahaemolyticus showed cytotoxic activity on FL cells derived from human amniotic membrane. Scanning electron micrographs of the whole cells showed that the microvilli on the cell surface decreased in number and changed in shape on treatment with the hemolysin. Most of the microvilli disappeared before death of the cells, as judged from the results of staining the cells with trypan blue and measuring release of alkaline phosphatase from the cells. Electron micrographs of thin sections ofthe cells showed that the cytoplasm of the cells was not significantly affected by treatment with sublethal amounts of hemolysin, even when the microvilli on the cell surface were significantly affected. Lethal amounts of hemolysin affected the cytoplasm and caused disarovilli on the cell surface are affected by treatment with the hemolysin before cytoxic effects develop.

Alkaline Phosphatase↗

Inactivation of the biological activities of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside Gt1.

Biological activities of the thermostable direct hemolysin produced by Vibrio parahaemolyticus, such as its hemolytic activity and lethal activity, were inhibited by neuraminidase-sensitive gangliosides, of which GT1 ganglioside was the most inhibitory. Neuraminidase-resistant gangliosides did not affect the activities of the hemolysin. Results showed that horse erythrocytes, which are resistant to the hemolysin, do not contain the neuraminidase-sensitive gangliosides GT1 and GD1a. Therefore, we propose that neuraminidase-sensitive gangliosides, and especially GT1 ganglioside, may be the receptor sites on the membranes for the thermostable direct hemolysin of V. parahaemolyticus.

Binding Sites, Antibody↗