PubMed Health⌕ Search

Biomedical subjects

M Ehara

Publications and source records attributed to M Ehara.

At least 37 records · Page 2Linked to original sources
PubMed1997

Algae or protozoa: phylogenetic position of euglenophytes and dinoflagellates as inferred from mitochondrial sequences.

The chloroplasts of euglenophytes and dinoflagellates have been suggested to be the vestiges of endosymbiotic algae acquired during the process of evolution. However, the evolutionary positions of these organisms are still inconclusive, and they have been tentatively classified as both algae and protozoa. A representative gene of the mitochondrial genome, cytochrome oxidase subunit I (coxI), was chosen and sequenced to clarify the phylogenetic positions of four dinoflagellates, two euglenophytes and one apicomplexan protist. This is the first report of mitochondrial DNA sequences for dinoflagellates and euglenophytes. Our COXI tree shows clearly that dinoflagellates are closely linked to apicomplexan parasites but not with algae. Euglenophytes and algae appear to be only remotely related, with euglenophytes sharing a possible evolutionary link with kinetoplastids. The COXI tree is in general agreement with the tree based on the nuclear encoded small subunit of ribosomal RNA (SSU rRNA) genes, but conflicts with that based on plastid genes. These results support the interpretation that chloroplasts present in euglenophytes and dinoflagellates were captured from algae through endosymbioses, while their mitochondria were inherited from the host cell. We suggest that dinoflagellates and euglenophytes were originally heterotrophic protists and that their chloroplasts are remnants of endosymbiotic algae.

Animals↗
PubMed1997

A deviant mitochondrial genetic code in prymnesiophytes (yellow-algae): UGA codon for tryptophan.

The sequence of a representative mitochondrial gene COXI, encoding cytochrome c oxidase subunit I, was determined in five species that cover all the orders of the Prymnesiophyta with the exception of the Pavlovales. Through this analysis, we noticed that the 'stop' codon UGA appears frequently and, specifically, at conserved tryptophan (Trp) sites of the gene. We showed these sites were not edited in the corresponding mRNA in one of these species, Isochrysis galbana. Therefore, it is most likely that the UGA codon is used for Trp, and not as a stop codon, in prymnesiophytes. All the analyzed prymnesiophytes made a tight cluster on the COXI phylogenetic tree which includes representative species of green-algae, land plants, yellow-green algae, eustigmatophytes and a red-alga. This suggests a monophyletic origin for the prymnesiophytes. The same deviant genetic code, i.e. UGA for Trp, has also been found in the red-alga, Chondrus crispus. In spite of the fact that this red-alga and the prymnesiophytes, share the same deviant genetic code for Trp, close affinity between the two groups was not statistically supported by the phylogenetic analysis of COXI sequences.

Base Sequence↗
PubMed1997

Morphological features of a filamentous phage of Vibrio cholerae O139 Bengal.

A filamentous phage was isolated from carrier strain AI-1841 of Vibrio cholerae 0139 Bengal and thus was termed fs phage. The phage was measured to be approximately 1 microm in length and 6 nm in width. One end of the phage was slightly tapered and had a fibrous appendage. The plaques developed on strain AI-4450 of V. cholerae 0139 were small and turbid. The phage grew in strain AI-4450 and reached a size of 10(8) to 10(9) pfu/ml at 5 hr after infection without inducing any lysis of the host bacteria. The group of phages attached on rod-shaped materials like fimbriae of this bacteria, with their fibrous appendages at the pointed end, were often found in the phage-infected culture. The anti-fimbrial serum effectively inhibited the infection of fs phage to the host strain AI-4450. We thus concluded that the phage can be adsorbed on fimbriae with a fibrous appendage on the pointed end of the phage filament.

Adsorption↗
PubMed1996

A classical strain of Vibrio cholerae with diminished ability to process the proteolytically sensitive site in the A subunit of cholera toxin.

Vibrio cholerae O1, No. 31, a strain isolated from a patient with mild diarrhea, produced mainly the unnicked cholera toxin. The amount of toxin that had accumulated in the cells was approximately 200 times lower than that secreted into the culture medium. When the unnicked toxin was purified by three successive column chromatographies and then extracted from the polyacrylamide gel, the unnicked toxin showed two bands corresponding to the A and B subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the A1 fragment was detected by trypsinization. Biological and enzymatic activities of the purified toxin with trypsinization were identical to those of cholera toxin from V. cholerae 569B as seen in the rabbit skin permeability test and the NAD:agmatine ADP-ribosyltransferase assay. DNA sequences of the A and B subunits were identical to those of the A- and B-subunit genes from the El Tor 2125 and classical 0395 strains, respectively. These data suggest that the wild V. cholerae strain, No. 31, produces a toxin identical to toxins previously reported in the literature and secretes it without accumulation in the cell, as is the case with other strains. However, strain No. 31's ability to nick the toxin is diminished compared with such abilities of other strains.

Animals↗
PubMed1994

Cloning and sequencing of the gene encoding Vibrio cholerae O1 fimbrial subunit (fimbrillin).

The gene encoding an 18 kDa fimbrial subunit of Vibrio cholerae O1 was identified in a fimbriate strain Bgd17. Mixed oligoprimers were prepared based on the amino acid sequence of the N-terminus and that from a cyanogen bromide-cleaved fragment of the fimbrillin. A PCR-amplified 185 bp DNA fragment was sequenced. This 185 bp fragment was further extended to 540 bp to 3' and 5' termini by RNA-PCR using a primer containing a random hexamer at its 3' end. This fragment did not contain the stop codons. It was further extended by a gene walking method using Eco RI cassette and its primers. Finally a 660 bp fragment was obtained and sequenced. This fragment contained the complete open reading frame of the structural subunit of the fimbriae, composed of 169 amino acids with a molecular mass of 17435.65 and a leader sequence of 6 or 9 amino acids. The deduced amino acid sequence of the polypeptide encoded by the gene, designated fimA, displayed a highly conserved sequence of MKXXXGFTLI EL of type 4 fimbriae.

Amino Acid Sequence↗
PubMed1994

The effect on enterotoxicity of protease purified from Vibrio cholerae O1.

The effect on enterotoxicity of protease purified from Vibrio cholerae O1 was investigated by the inoculation of live vibrio cells into protease-treated loops of the ileal loop model. Fluid accumulation ratios in the protease-treated loops were elevated in a dose-dependent manner by challenge with live vibrio cells but not by that with toxin. An enhancement effect of protease on enterotoxicity was observed in both serotypes of V. cholerae O1 and V. cholerae non-O1. It is suggested, therefore, that the enterotoxicity was enhanced by treatment with protease when live vibrio cells were inoculated into the ileal loops of rabbits.

Animals↗
PubMed1993

Immunogenicity of Vibrio cholerae O1 fimbriae in animal and human cholera.

Parenteral immunization with either formalin-fixed whole cells of the fimbriate Bgd17 strain or purified fimbriae protected against Vibrio cholerae O1 infection in rabbits, independent of biotype and serotype. Parenteral immunization of adult rabbits with purified fimbriae prior to V. cholerae O1 challenge resulted in a reduction of 2 to 3 orders of magnitude in the number of bacteria recovered from the small intestines of immunized rabbits in comparison to non-immunized controls. IgG and IgA antibodies against fimbrillin of V. cholerae O1 were detected in the convalescent sera of patients with cholera; however, little fimbrial antigen was detected in the commercially available cholera vaccines when examined by polyclonal and monoclonal antibodies against fimbriae. These data suggest that fimbrial hemagglutinin is a major adhesin of V. cholerae O1 and that parenteral immunization with fimbriae generates a specific immune response in the gut that may serve as one means of mitigating subsequent V. cholerae O1 gut infection.

Animals↗
PubMed1992

The protease from Vibrio cholerae nicks arginine at position 192 from the N-terminus of the heat-labile enterotoxin a subunit from enterotoxigenic Escherichia coli.

It was examined where a protease purified from Vibrio cholerae might nick the heat-labile enterotoxin (LT) A subunit from enterotoxigenic Escherichia coli. LT was digested by the protease and contained a fragment which had the same mobility on SDS-PAGE as that of the A1 fragment of LT digested by trypsin. The biological activity of LT by this protease was also identical to that of LT by trypsin. The amino acid sequence of the N-terminus of the A2-like fragment was Thr-Ser-Thr-Gly, which corresponded to the sequence from 193 to 196 of the A subunit. These data suggest that this protease, like trypsin, nicks arginine at position 192 from the N-terminus of the A subunit and that the biological activation of LT by this protease is similar to that by trypsin.

Amino Acid Sequence↗
PubMed1992

Studies on novel pili from Shigella flexneri. I. Detection of pili and hemagglutination activity.

Pili were detected using electron microscopy in clinical isolates of Shigella flexneri which had been continuously subcultivated in liquid media. Morphologically, the pili appeared as thin, flexible, cylindrical structures of up to 2-5 microns in length and about 3-5 nm in diameter. Two strains showed mannose-resistant (MR) hemagglutination to fresh fowl erythrocytes (type 4), and one to tannic acid-treated horse erythrocyte (type 3) pili. These pili are novel and different from the mannose-sensitive (MS) type 1 pili described by Duguid and Gillies.

Animals↗
PubMed1992

The fimbriae-like structures on V. cholerae isolated in Kenya.

For Vibrio cholerae 01 to overcome the normal intestinal clearing mechanisms and facilitate colonization in the host gut, fine filamentous structures covering the surface of all the cell must exist. These fimbriae-like structures were investigated in this study. Selection of the K23 V. cholerae 01 biotype el-tor isolated in Kisumu, Kenya, was based on its proteinase activity and ability to agglutinate 0.2M ammonium sulphate. This strain was cultured in modified tryptone broth pH 8.4, at 37 degrees C overnight. Electron microscopy of the strain revealed the presence of fimbriae-like structures which varied in number. Some cells showed more than 200, whereas in other cells only several filamentous structures were detected. It is suggested that these structures could be a possible candidate for a cholera vaccine.

Evaluation Studies as Topic↗
PubMed1990

Diethylcarbamazine, antifilarial drug, inhibits microtubule polymerization and disrupts preformed microtubules.

The effect of diethylcarbamazine (DEC) on microtubules was studied by using microtubule protein prepared from porcine brain. DEC inhibited assembly of microtubules and disassembled preformed microtubules in vitro. When the reassembled or disassembled products were examined in the presence of DEC by electron microscopy, ribbon-microtubules were frequently observed. Subsequently, the effect of DEC on the cytoplasmic microtubules complex was studied. The cells used in our study were LLC-MK2. DEC inhibited proliferation of these cells, and cells grown in the presence of DEC were likely to separate from each other and became round in shape. Immunofluorescence microscopy revealed that the cells exposed to DEC were devoid of the delicate pattern of the cytoplasmic microtubule complex.

Animals↗
PubMed1990

Study of enzyme inhibitory activities by dipyridamole.

Inhibitory activities of Dipyridamole (DPM, 2,6-bis(diethanolamino)-4,8-dipiperidinopyrimido(5,4-d)py rim idine) against xanthine oxidase (XO), carbonic anhydorase (CA) and monoamine oxidase (MAO) were studied, in vitro. DPM did not inhibit XO and CA, but it strongly inhibited MAO. The type of inhibition by DPM against MAO with respect to benzylamine as a substrate was uncompetitive.

Carbonic Anhydrase Inhibitors↗
PubMed1989

Detection of IgA protease from Haemophilus influenzae by immunoblotting.

IgA protease produced by various strains of Haemophilus influenzae can digest serum IgA and yield its fragments which can react with anti-IgA serum. We assayed IgA protease activity by detecting the digests of IgA by SDS-PAGE and immunoblotting. The digests were separated with SDS-PAGE, transferred to nitrocellulose membranes and detected with anti- (alpha chain of human IgA, its Fab and its Fc) immunoglobulin conjugated peroxidases. Using this method, we can determine which type of IgA protease is produced by various of H. influenzae strains. All the 20 strains isolated from respiratory tracts produced IgA protease.

Blotting, Western↗
PubMed1989

Pili of Vibrio cholerae O1 biotype E1 Tor: a comparative study on adhesive and non-adhesive strains.

Pili were found on the cell surface of non-adhesive Vibrio cholerae O1 Biotype E1 Tor as well as the adhesive strain. Purified pili of the adhesive and non-adhesive strains were morphologically, electrophoretically, and immunologically, indistinguishable from each other. The molecular weights of both pilin (subunit protein of the pilus) were about 16,000 daltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These 16 kDa pili are different from the pilus colonization factor, which is a 20.5 kDa protein, reported by Taylor et al. The 16 kDa pili of Vibrio cholerae O1 Biotype E1 Tor have hemagglutinating activity, but may have no role in colonization, because non-adhesive strains also have such pili.

Animals↗
PubMed1987

Purification and partial characterization of fimbriae of Vibrio cholerae O1.

Fimbriae of Vibrio cholerae O1 were purified from a strain of the classical biotype, Inaba serotype (Bgd 17), and a strain of the El Tor biotype, Ogawa serotype (K23), grown on TCG agar medium by the following procedure; homogenization of the cell suspension to detach fimbriae, ultracentrifugation to remove remaining cells and their debris, concentration of the supernatant containing fimbriae with ultrafiltration, and 20 to 40% sucrose linear gradient centrifugation of the concentrated material. The fimbriae in both preparations were flexible, long fibres readily distinguishable under the electron microscope from those of CFA/I, CFA/II seen in ETEC strains. Their structural subunit was a protein of 16 kdaltons. Fimbriae isolated from both serotypes and biotypes shared antigenic determinants.

Antigens, Bacterial↗
PubMed1986

The characterization of Vibrio cholerae isolated in Kenya in 1983.

A total of 245 strains of Vibrio cholerae 01 and two strains of V. cholerae non-01 were isolated and collected from diarrhoeal patients in Homa Bay District Hospital and the other medical facilities in Nyanza Province, Kenya in 1983. The majority of V. cholerae 01 tested were Ogawa type (with the exception of nine Inaba type), biotype E1 Tor (except one untypable strain) and Celebes original type (except one cured type). Haemolytic activity to sheep red blood cells was detected in 75.5% of isolates. Out of 245 strains of V. cholerae 01, 184 were resistant to tetracycline, streptomycin and ampicillin. All were sensitive to chloramphenicol and nalidixic acid. Only one strain of V. cholerae 01 was sensitive to all five antimicrobial agents tested. An environmental cholera survey was done after the cholera outbreak subsided. Twenty strains of V. cholerae non-01 were isolated from water samples in Nyanza Province but none of V. cholerae 01 was isolated.

Anti-Bacterial Agents↗