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Biomedical subjects

T Mikami

Publications and source records attributed to T Mikami.

At least 235 records · Page 13Linked to original sources

Cytoplasmic diversity in leaf beet cultivars as revealed by mitochondrial DNA analysis.

Mitochondrial (mt) DNA restriction fragment length polymorphisms are convenient markers for identifying cytoplasmic variation among plant cultivars. In an attempt to detect new cytoplasmic genotypes useful for sugarbeet breeding, we have compared the hybridization patterns of mtDNA from three groups of cultivated beets, viz. leaf beet, garden beet, and fodder beet. Utilized as probes were the two sugarbeet mtDNA clones that were capable of distinguishing normal fertile and different sources of male-sterile cytoplasms from one another. The analysis allowed the identification of four chondriome types among 14 leaf beet cultivars examined. Two out of the four chondriome types were found to be different from the previously described fertile or male-sterile chondriome type. Our results thus indicate that leaf beet cultivars and landraces make up the primary cytoplasmic gene pool of the sugarbeet.

Base Sequence↗

A novel beta-N-acetylglucosaminidase from Streptomyces thermoviolaceus OPC-520: gene cloning, expression, and assignment to family 3 of the glycosyl hydrolases.

A beta-N-acetylglucosaminidase gene (nagA) of Streptomyces thermoviolaceus OPC-520 was cloned in Streptomyces lividans 66. The nucleotide sequence of the gene, which encodes NagA, revealed an open reading frame of 1,896 bp, encoding a protein with an Mr of 66, 329. The deduced primary structure of NagA was confirmed by comparison with the N-terminal amino acid sequence of the cloned beta-N-acetylglucosaminidase expressed by S. lividans. The enzyme shares no sequence similarity with the classical beta-N-acetylglucosaminidases belonging to family 20. However, NagA, which showed no detectable beta-glucosidase activity, revealed homology with microbial beta-glucosidases belonging to family 3; in particular, striking homology with the active-site regions of beta-glucosidases was observed. Thus, the above-mentioned results indicate that NagA from S. thermoviolaceus OPC-520 is classified as a family 3 glycosyl hydrolase. The enzyme activity was optimal at 60 degreesC and pH 5.0, and the apparent Km and Vmax values for p-nitrophenyl-beta-N-acetylglucosamine were 425.7 microM and 24.8 micromol min-1 mg of protein-1, respectively.

Acetylglucosaminidase↗

Apoptosis in feline panleukopenia virus-infected lymphocytes.

Feline panleukopenia virus (FPLV) was shown to induce apoptosis to feline lymphoid cells and to reduce the expression of interleukin-2 receptor alpha on the cells. FPLV-induced apoptosis might be a key element in the pathophysiology of atrophy of lymphoid tissues associated with feline panleukopenia caused by FPLV.

Animals↗

CagA seropositivity associated with development of gastric cancer in a Japanese population.

BACKGROUND/AIMS: Infection with Helicobacter pylori strains possessing the cagA gene is associated with increased risk of gastric cancer of the intestinal type. The aims of this study were to investigate whether CagA seropositivity is associated with increasing risk of gastric cancer in a Japanese population that has a much higher incidence of gastric cancer than western populations. METHODS: Eighty one gastric cancer patients and 81 sex and age matched endoscopically evaluated controls were studied. Histologically, 62 cancers were of the intestinal type and 76 were early gastric cancer. Serum CagA IgG antibodies were assayed by enzyme linked immunosorbent assay (ELISA) using purified recombinant CagA protein as antigen. Polymerase chain reaction (PCR) analysis for cagA in H pylori isolates (n = 80) showed that the CagA ELISA had a sensitivity of 83.3% (controls) and 72.5% (cancers). RESULTS: CagA seropositivity was 60% (49 of 81) in cancer patients and 44% (36 of 81) in controls. The odds ratio for the risk of cancer if CagA seropositive was 1.93 (95% confidence interval (CI) 1.01 to 3.68; p < 0.05). In the 57 H pylori positive cancer patients and their matched H pylori positive controls, the odds ratio for the risk of cancer if CagA seropositive was 2.2 (95% CI 1.04 to 4.65; p < 0.05). CONCLUSIONS: These results suggest that CagA seropositivity is associated with increased risk of gastric cancer in Japanese populations.

Adult↗

Evaluation of Helicobacter pylori vacA genotype in Japanese patients with gastric cancer.

AIM: To examine the vacA genotypes of Helicobacter pylori strains in Japan and to define whether any specific genotype was associated with gastric cancer. METHODS: The allelic variation of vacA gene was studied using a recently introduced polymerase chain reaction based vacA genotyping system. RESULTS: 80 H pylori strains were isolated from gastric biopsies of 40 patients with gastric cancer and 40 control subjects in a Japanese population. All strains were s1/m1 subtype and 79 of 80 strains were classified as s1a subtype. CONCLUSIONS: The recently proposed vacA genotyping system is applicable to Japanese H pylori strains and most strains have the s1a genotype, associated with increased virulence. While the high frequency of s1a/m1 vacA genotype might play a role in the increased incidence of atrophic gastritis and gastric cancer in Japanese subjects, it precludes its use as a predictor of clinical outcome of H pylori infection in Japan.

Adult↗

Use of hemoglobin as an iron source by Bacillus cereus.

The hemoglobin binding activity of Bacillus cereus cells was measured with fluoresceinisothiocyanate (FITC)-conjugated hemoglobin using flow cytometry. Growth of B. cereus was markedly inhibited by the addition of apo-transferrin. B. cereus could not use transferrin-bound iron as an iron source in serum. The growth inhibition was reversed by the addition of a FeCl3 solution, erythrocytes or hemoglobin. B. cereus released hemolysin; these findings suggested that the hemoglobin released from erythrocytes by B. cereus hemolysin binds to B. cereus and is thus used as an iron source.

Apoproteins↗

Metaiodobenzylguanidine and heart rate variability in heart failure.

It is assumed that the low-frequency power (LF) of heart rate variability (HRV) increases with progress of congestive heart failure (CHF), therefore positively correlating with cardiac 123I-metaiodobenzylguanidine (MIBG) washout. It is demonstrated here that HRV, including normalized LF, correlated inversely with MIBG washout and positively with the ratio of heart-to-mediastinum MIBG activity in controls and CHF patients, whereas these correlations were not observed within CHF patients. Thus MIBG washout may increase and HRV including normalized LF may decrease with CHF, although the HRV and MIBG measures may not similarly change in proportion to the severity of the cardiac autonomic dysfunction in CHF.

Adult↗

Treatment of hypertension on the basis of home measurements: a case report.

Should antihypertensive therapy be based on home blood-pressure monitoring in hypertensive patients with marked white-coat phenomenon? A 53-year-old hypertensive woman with marked white-coat phenomenon had been treated on the basis of blood pressure at home without measurements of clinic blood pressure for 5 years. The target of her treatment was changed to clinic blood pressure because of marked electrocardiographic abnormalities, which were subsequently normalized by the treatment given for the office hypertension. Thus, the experience of this case suggests that treatment of cases of hypertension with white-coat phenomenon should not be based only on home recordings.

Antihypertensive Agents↗

Molecular identification of a recent type of canine distemper virus in Japan by restriction fragment length polymorphism.

Restriction fragment length polymorphism analysis was used to differentiate recent field viruses of canine distemper virus (CDU) from vaccine strains. Virus genomes were amplified by using reverse transcriptase-polymerase chain reaction in part of the haemagglutinin gene. After digestion with EcoRV, the PCR products of recent field isolates were cut into two fragments that differ from the uncut form of old strains including all of vaccine strains. This method could be applied to fresh or stored brains, spleens and peripheral blood mononuclear cells of infected dogs. This molecular approach is useful for determining the causative agent of postvaccinated CDV infection.

Animals↗

Seroepidemiological survey of feline retrovirus infections in domestic and leopard cats in northern Vietnam in 1997.

The prevalence of infections with three feline retroviruses (feline immunodeficiency virus (FIV), feline leukemia virus (FeLV) and feline syncytial virus (FSV)) was examined in northern Vietnam in 1997. We collected a total of 77 blood samples from 69 domestic and 8 leopard cats, and examined the presence of anti-FIV and FSV antibodies and FeLV p27 antigen in the plasma samples by the indirect immunofluorescence and/or two commercial kits. None of the samples was positive for FIV and FeLV. The overall positive rate of FSV was 31% and the positive rates among the domestic and leopard cats were 29 and 50%, respectively. We isolated FSV from peripheral blood mononuclear cells of 6 domestic and one leopard cats.

Animals↗

Nucleotide sequences of glycoprotein I and E genes of equine herpesvirus type 4.

The nucleotide sequences of the glycoprotein I (gI) and E (gE) genes of equine herpesvirus type 4 (EHV-4) strain TH20 were determined. The predicted region encoding the EHV-4 gI gene is 1,263 nucleotides, corresponding to a polypeptide of 420 amino acids in length. The predicted region encoding the EHV-4 gE gene is 1,647 nucleotides, corresponding to a polypeptide of 548 amino acids in length. The EHV-4 gI and gE genes show 74% and 85% identity at the amino acid level with those of equine herpesvirus type 1 (EHV-1), respectively. Furthermore, we have found an open reading frame homologous to the EHV-1 gene 75, which overlaps in part with the 3' end of EHV-4 gE gene. These sequence data will be useful for development of a modified live vaccine against equine herpesvirus type 1 and 4 infections.

Amino Acid Sequence↗

Eight-year observation and comparative study of specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) subtypes A and B: terminal acquired immunodeficiency syndrome in a cat infected with FIV petaluma strain.

Three specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) strains Petaluma, TM1 and TM2, respectively were observed for over 8 years. Without showing any significant clinical signs of immunodeficiency syndrome (AIDS) for 8 years and 4 months of asymptomatic phase, the Petaluma-infected cat exhibited severe stomatitis/gingivitis, anorexia, emaciation, hematological and immunological disorders such as severe anemia, lymphopenia, thrombocytopenia, and decrease of CD4/CD8 ratio to 0.075, and finally died with hemoperitoneum at 8 years and 8 months post-infection. Histopathological studies revealed that the cat had systemic lymphoid atrophy and bone marrow disorders indicating acute myelocytic leukemia (aleukemic type). Plasma viral titer of the cat at AIDS phase was considerably high and anti-FIV antibody titer was slightly low as compared with the other FIV-infected cats. In addition, immunoblotting analysis using serially collected serum/plasma samples of these cats revealed that antibodies against FIV proteins were induced in all the infected cats, however in the Petaluma-infected cat anti-Gag antibodies disappeared during the asymptomatic period. These results suggested that plasma viral load and anti-FIV Gag antibody response correlated with disease progression, and supported FIV-infected cats as a suitable animal model of human AIDS.

Acquired Immunodeficiency Syndrome↗

The nucleotide and predicted amino acid sequence of the fusion protein of recent isolates of canine distemper virus in Japan.

Analysis of the molecular properties of fusion (F) proteins of field isolates of canine distemper virus (CDV) by immunoprecipitation analysis revealed an identical molecular mass of F protein of 3 field isolates as well as the Onderstepoort laboratory strain. Sequencing showed that the F gene of a field isolate (the Yanaka strain) shared 90.1% and 95.7% identities with the Onderstepoort strain at nucleotide and amino acid levels, respectively. All of the 13 cysteine residues and 4 potential asparagine-linked glycosylation sites were completely conserved amongst these strains. These results indicate that the F proteins is much less heterogeneous than that observed in the hemagglutinin proteins of CDV.

Amino Acid Sequence↗

Further development of a recombinant feline herpesvirus type 1 vector expressing feline calicivirus immunogenic antigen.

We previously reported the attenuation of thymidine kinase (TK) deficient mutant (C7301dlTK) of feline herpesvirus type 1 (FHV-1) in cats and the construction of a recombinant FHV-1 (C7301dlTK-Cap) inserted a precursor capsid gene of feline calcivirus (FCV) into the TK deletion locus of the C7301dlTK. In this study, we constructed a further improved recombinant FHV-1 (dlTK(gCp)-Cap) carrying a putative FHV-1 gC promoter sequence upstream of the FCV precursor capsid gene of the C7301dlTK-Cap. Growth kinetics of the dlTK(gCp)-Cap in cell cultures was similar to those of C7301dlTK and C7301dlTK-Cap. A strong expression of FCV immunogenic antigen by dlTK(gCp)-Cap was confirmed by indirect immunofluorescence and enzyme-linked immunosorbent assays. In addition, one vaccination with dlTK(gCp)-Cap protected cats more effective against subsequent virulent FCV challenge than that with C7301dlTK-Cap.

Animals↗

Properties and functions of feline herpesvirus type 1 glycoproteins.

Feline herpesvirus type 1 (FHV-1) is a causative agent of feline viral rhinotracheitis and belongs to the subfamily Alphaherpesvirinae of the family Herpesviridae. Since first isolated in 1958 by Crandell and Maurer, FHV-1 is distributed worldwide and is the most clinically significant agent for respiratory infections in cats. In this review, we describe the recent findings with properties and functions of FHV-1 glycoproteins, especially hemagglutinins.

Animals↗

New quantitative methods for detection of feline parvovirus (FPV) and virus neutralizing antibody against FPV using a feline T lymphoid cell line.

Previously, we reported that a feline T lymphoid cell line, FL74 cells, was very sensitive to feline parvovirus (FPV) infection. In the present study, we developed new quantitative methods for detection of FPV and virus neutralizing antibody against FPV using FL74 cells. The methods presented here were very simple and applicable to both canine parvovirus and feline panleukopenia virus.

Animals↗

Diversity of genomic electropherotypes of naturally occurring equine herpesvirus 1 isolates in Argentina.

The genomes of 10 equine herpesvirus 1 (EHV-1) strains isolated in Argentina from 1979 to 1991, and a Japanese HH1 reference strain were compared by restriction endonuclease analysis. Two restriction enzymes, BamHI and BglII, were used and analysis of the electropherotypes did not show significant differences among isolates obtained from horses with different clinical signs. This suggests that the EHV-1 isolates studied, which circulated in Argentina for more than 10 years, belong to a single genotype.

Argentina↗

Identification of carbohydrates on Eimeria stiedai sporozoites and their role in invasion of cultured cells in vitro.

Invasion specificity of Eimeria stiedai sporozoites in cultured cells was examined. Intracellular sporozoites were observed in hepatobiliary epithelial cells, as early as 3 hours post inoculation (p.i.) but the infection rate was monitored for 6 hours. No intracellular parasites were found in rabbit parenchymal hepatocytes and rabbit kidney cells, even on prolonged culturing. In the hepatobiliary epithelial cells inoculated with fixed sporozoites, no intracellular parasite were found. Sporozoites attached on the cell surface of the hepatobiliary epithelial cells fixed with paraformaldehyde but did not penetrate. The carbohydrates present on Eimeria stiedai sporozoites and their functional role in the process of invasion of host cells were also examined. Lectin binding sites on the surface of sporozoites were detected by means of peroxidase-conjugated lectins. Sporozoites showed specific binding with UEA-I and PNA lectins, which bind L-fucose and D-galactose, respectively. Exposure of sporozoites to 100 microg/ml UEA-I significantly reduced their ability to invade primary rabbit hepatobiliary epithelial cells, but similar treatment with PNA had no such effect. Preincubation of these cells in Dulbecco's minimum essential medium containing 10% fetal bovine serum and 1% L-fucose suppressed the invasion activity of the sporozoites, but preincubation of the sporozoites in the same medium without L-fucose had no effect on cell penetration. D-galactose added to the medium had no effect on the invasion activity of sporozoites. These results indicate that L-fucose residues on E. stiedai sporozoites and L-fucose binding sites on host cells both are associated with recognition and/or invasion process.

Animals↗