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M Fukuyama

Publications and source records attributed to M Fukuyama.

At least 37 records · Page 2Linked to original sources
PubMed1998

[A case of bacillary dysentery caused by new quinolone-resistant Shigella flexneri 2a].

A 73-year-old male was admitted to our hospital because of detection of Shigella flexneri 2a from his stool. Antimicrobial treatment with levofloxacin (LVFX) was started, but could not eliminate the organism in the stool. In the examination of drug susceptibility, this strain was highly resistant to all new quinolones. The minimal inhibitory concentration of norfloxacin, ofloxacin and ciprofloxacin to this strain was 12.5 micrograms/ml, 6.25 micrograms/ml and 6.25 micrograms/ml, respectively. The dual mutations were detected in the codon 83 and 87 of the gyrA gene by sequencing the quinolone-resistance determining region (QRDR). There was, however, no significant difference between the intracellular uptake of ciprofloxacin in this strain and in the ciprofloxacin-sensitive strain. The amount of ciprofloxacin in this strain unchanged when carbonyl cyanide m-chlorophenyl hydrazone (CCCP) was added. These results suggest that the advanced resistance in Shigella flexneri against new quinolones could be acquired by only this dual mutations without the change of the active efflux mechanism.

4-Quinolones↗
PubMed1997

end-1 encodes an apparent GATA factor that specifies the endoderm precursor in Caenorhabditis elegans embryos.

The endoderm in the nematode Caenorhabditis elegans is clonally derived from the E founder cell. We identified a single genomic region (the endoderm-determining region, or EDR) that is required for the production of the entire C. elegans endoderm. In embryos lacking the EDR, the E cell gives rise to ectoderm and mesoderm instead of endoderm and appears to adopt the fate of its cousin, the C founder cell. end-1, a gene from the EDR, restores endoderm production in EDR deficiency homozygotes. end-1 transcripts are first detectable specifically in the E cell, consistent with a direct role for end-1 in endoderm development. The END-1 protein is an apparent zinc finger-containing GATA transcription factor. As GATA factors have been implicated in endoderm development in other animals, our findings suggest that endoderm may be specified by molecularly conserved mechanisms in triploblastic animals. We propose that end-1, the first zygotic gene known to be involved in the specification of germ layer and founder cell identity in C. elegans, may link maternal genes that regulate the establishment of the endoderm to downstream genes responsible for endoderm differentiation.

Amino Acid Sequence↗
PubMed1996

Identification from a phage display library of peptides that bind to toxic shock syndrome toxin-1 and that inhibit its binding to major histocompatibility complex (MHC) class II molecules.

Phage display technique is a powerful tool with which to identify novel binding sequences for antibody and receptor targets. Few studies, however, have used this technology to select affinity peptides for ligand molecules. Here, we screened a peptide phage library for binding to toxic shock syndrome toxin 1 (TSST-1) to examine whether peptide ligands for TSST-1 which mimic the structure of major histocompatibility complex (MHC) class II receptors could be identified. After three cycles of biopanning, four potent sequences reactive with TSST-1 were isolated (designated phages 2, 3, 8, and 11). Selected phage were found to react specifically with TSST-1 but not with other staphylococcal exotoxins. A synthetic peptide (pep3) corresponding to the most frequently identified sequence (phage3) was shown to inhibit binding of all four isolated phage to TSST-1, suggesting that they bind to a common site on TSST-1. Furthermore, pep3 was shown to compete with MHC class II molecules for binding to TSST-1 in a concentration-dependent manner. Comparison of their sequences with MHC class II molecules revealed that phage8 shared sequence homology with two regions of the beta chain of MHC class II molecules: amino acids 57-62, containing a residue (Tyr-60) involved in TSST-1 binding as suggested by X-ray crystallographic data of TSST-1-MHC class II complex; and amino acids 188-193, a region not previously known as a contact domain. These results suggest that the selected sequences recognized the MHC class II binding site on TSST-1. Thus, affinity selection for peptides binding to ligand molecules (e.g., TSST-1) rather than their cognate receptors (e.g., MHC class II) from a random phage display library represents a useful approach to understanding receptor-ligand interactions.

Amino Acid Sequence↗
PubMed1996

Detection of feline herpesvirus 1 DNA by the nested polymerase chain reaction.

The thymidine kinase region of feline herpesvirus 1 (FHV 1) genome in ocular/nasal swabs from cats with clinical manifestations of upper respiratory disease was amplified by nested polymerase chain reaction (nested PCR). Two primer pairs were prepared for nested PCR. FHV 1 DNA in ocular/nasal swabs was extracted using instaGene-DNA purification matrix. Nested PCR for the FHV 1 culture supernatants was ten times as sensitive as single PCR. On comparing viral isolation with single PCR and nested PCR for the detection of FHV 1 in ocular/nasal secretions, of 5 samples that yielded infectious virus in cell culture, 3 (60%) were positive in single PCR and 5 (100%) were positive in nested PCR. When 22 ocular/nasal swabs that did not yield FHV 1 were assayed, 3 were negative in both single PCR and nested PCR, 2 were positive in both single and nested PCR and 17 were positive in only nested PCR. Thus, FHV 1 was detected in 19/22 (86.4%) by the nested PCR and in 2/22 (9%) by single PCR. These results show that nested PCR is 4.8 (24 positive samples/5 positive samples) times as sensitive as single PCR.

Animals↗
PubMed1996

[In vitro antibacterial activity of pazufloxacin (PZFX) against clinical isolates from infectious enteritis].

Antibacterial activity of pazufloxacin (PZFX) against 65 strains of Shigella spp., 13 strains of Salmonella spp., 7 strains of Escherichia coli, 4 strains of Vibrio parahaemolyticus, 4 strains of V. cholerae O1, 2 strains of Aeromonas spp., 4 strains of Plesionomas shigelloides and 3 strains of V. cholerae non-O1 isolated from patients of infectious enteritis and carriers was compared with that of ofloxacin (OFLX), ciprofloxacin (CPFX), tosufloxacin (TFLX) and nalidixic acid (NA). The MIC90 of PZFX against Shigella spp., Salmonella spp. and E. coli were 0.025, 0.025 and 0.025 micrograms/ml, respectively. The MIC90 of PZFX against Shigella spp. was comparable to that of CPFX, 2-fold higher than that of TFLX and 4-fold lower than that of OFLX. Against Salmonella spp., MIC90 of PZFX was comparable to TFLX and 2- to 8-fold lower than those of CPFX and OFLX. PZFX showed the highest antibacterial activity between the drugs tested. Against other species, the MIC90 values of PZFX were < or = 0.006-0.1 micrograms/ml. No NA-resistant isolates were observed. The antibacterial activity of PZFX against stocked strains (clinical isolates from Aug 1989 to Feb 1991), containing 51 strains of Shigella spp., 50 strains of Salmonella spp., 19 strains of E. coli, 12 strains of V. cholerae O1 and 13 strains of V. parahaemolyticus, was also investigated. The MIC90 of PZFX of each strains were 0.05 micrograms/ml for Shigella spp., 0.05 micrograms/ml for Salmonella spp., 0.1 micrograms/ml for E. coli, 0.0125 micrograms/ml for V. cholerae O1, 0.2 micrograms/ml for V. parahaemolyticus, respectively. As shown above, PZFX has strong antibacterial activity against isolates from infectious enteritis and this suggests the usefulness of PZFX for the treatment of these patients and carriers.

4-Quinolones↗
PubMed1996

[Clinical study of prulifloxacin on infectious enteritis. Japan Research Committee of Prulifloxacin, Research Group on Infectious Enteritis].

Prulifloxacin (PUFX), a new quinolone antimicrobial agent, was administered to a total of 122 patients and carriers to investigate its clinical efficacy, safety and usefulness in infectious enteritis (bacillary dysentery, enteritis caused by Salmonella spp. and enteropathogenic E. coli, cholera and so on). In addition, the minimum inhibitory concentration (MIC) of UFX (active compound) was determined against each clinical isolate, and compared with that of ciprofloxacin (CPFX), ofloxacin (OFLX), tosufloxacin (TFLX) and nalidixic acid (NA). The correlation between the concentration of UFX in feces and the change of the fecal microflora were also investigated when PUFX was administered to the patients with acute infectious enteritis. A daily dose of 400 mg of PUFX was administered orally in two divided doses (morning and evening) for 5 days, with the exception of 7 days administration against salmonella enteritis and 3 days administration against cholera. 84 cases were adapted for evaluating the usefulness. The clinical efficacy was 100% in all the enteritis except salmonella enteritis, in which it was 88.9% (8/9 cases). On the bacteriological efficacy, the elimination rate was 100% in all isolates except Salmonella spp., in which it was 75.0% (12/16 cases). As for the adverse effect, uriticaria in moderate degree was observed in 1 (0.9%) of 109 cases. Abnormal changes in laboratory findings were seen in 3 (3.0%) of 100 cases, consisting of 1 with eosinophilia and 2 with elevated S-GPT, although they were all slight in degree. The usefulness rate was 65.5% (55/84 cases) for "very useful" and 95.2% (80/84 cases) for "very useful" and "useful". MIC90 of UFX against Shigella spp., Salmonella spp., E. coli and V. cholerae, was 0.025, 0.05, 0.025 and 0.05 microgram/ml, respectively. These values were the same as those of CPFX and TFLX, and superior to OFLX and NA. UFX concentrations in feces followed by administration of PUFX in 3 cases with acute infectious enteritis were higher than that of MIC90 of UFX against Shigella spp., Salmonella spp., E. coli and V. cholerae. The changes of the fecal microflora, which influence the efficacy and safety of PUFX, were not observed.

Adult↗
PubMed1995

The absence of evidence of staphylococcal toxin involvement in the pathogenesis of Kawasaki disease.

To detect a causative superantigen and to clarify a possible role for staphylococci in Kawasaki disease (KD), culture supernatants of individual bacterial isolates from 11 acute-stage patients were studied. Toxic shock syndrome toxin-1 (TSST-1) and antibody to TSST-1 and enterotoxins A (SEA), B (SEB), and C (SEC) in acute (mean, day 7) and late convalescent (mean, month 15) sera from 26 patients (12 with coronary artery aneurysms) and 22 age-matched controls were measured. Only 1 of 60 supernatants was mitogenic for human lymphocytes; it was 1 of the 4 Staphylococcus aureus isolates. Mitogenicity was neutralized by sera obtained after administration of intravenous gamma globulin (mean, week 4) but not by late convalescent sera. TSST-1 was detectable in 2 of 26 acute sera and 1 of 22 control sera. No KD but 1 control serum had IgM to TSST-1. IgG seroconversion rates to TSST-1, SEA, SEB, and SEC were 10%, 15%, 21% and 16%, respectively. These data do not support the involvement of toxin-producing staphylococci in KD.

Antibodies, Bacterial↗
PubMed1995

[Properties of a hemolysin produced by group C streptococci isolated from hemolytic streptococcal infection in formosan squirrels].

The hemolysin produced by group C streptococci (GCH) has an isoelectric point (pI) of 5.6 and a molecular weight of 32,000 and shows hemolytic activity in the absence of 2-mercaptoethanol (2-ME). The hemolytic activity of GCH was compared to that of streptolysin S (SLS). These hemolysins did not differ with respect to the binding and release of hemoglobin (Hb). GCH was bound to phospholipids on the membranes of target erythrocytes, followed by the rapid released of K+ and slow Hb release after a relatively long lag time. GCH showed a lower hemolytic efficiency than SLS, reflecting the fact that these hemolysins destroy erythrocytes by slightly different mechanisms.

Animals↗
PubMed1995

[In vitro antibacterial activity of balofloxacin (BLFX) against isolates from patients with bacterial enteritis].

In vitro antibacterial activity of balofloxacin (BLFX), a newly developed fluoroquinoline, was compared with that of norfloxacin (NFLX), ofloxacin (OFLX) and ciplofloxacin (CPFX). Bacterial strains used in this experiment were freshly isolated from patients with infectious enteritis just before BLFX therapy. The isolates were 43 strains of Vibrio cholerae O1, 1 strain of Campylobacter sp., 4 strains of Aeromonas spp., 3 strains of Plesiomonas shigelloides, 1 strain of Vibrio mimicus and 1 strain of Vibrio cholerae non-O1. MIC90 of BLFX against 43 strains of Shigella spp., 13 strains of Salmonella spp. and 9 strains of E. coli were 0.39, 0.39, 0.2 micrograms/ml, respectively. All strains of Aeromonas spp. and P. Shigelloides were inhibited by the concentrations under 0.39 and 0.05 micrograms/ml. MIC90 of BLFX, NFLX, OFLX and CPFX against a total of 79 strains were 0.39, 0.2, 0.2 and 0.05 micrograms/ml, respectively.

Anti-Infective Agents↗
PubMed1995

Phenotypic and genetic diversity of chlorine-resistant Methylobacterium strains isolated from various environments.

Strains of pink-pigmented facultative methylotrophs which were isolated previously from various environments and assigned tentatively to the genus Methylobacterium were characterized in comparison with authentic strains of previously known species of this genus. Most of the isolates derived from chlorinated water supplies exhibited resistance to chlorine, whereas 29 to 40% of the isolates from air, natural aquatic environments, and clinical materials were chlorine resistant. None of the tested authentic strains of Methylobacterium species obtained from culture collections exhibited chlorine resistance. Numerical analysis of phenotypic profiles showed that the test organisms tested were separated from each other except M. organophilum and M. rhodesianum. The chlorine-resistant isolates were randomly distributed among all clusters. The 16S ribosomal DNA (rDNA) sequence-based phylogenetic analyses showed that representatives of the isolates together with known Methylobacterium species formed a line of descent distinct from that of members of related genera in the alpha-2 subclass of the Proteobacteria and were divided into three subclusters within the Methylobacterium group. These results demonstrate that there is phenotypic and genetic diversity among chlorine-resistant Methylobacterium strains within the genus.

Bacterial Typing Techniques↗
PubMed1995

Detection of genomic variability among isolates of Campylobacter jejuni from chickens by crossed-field gel electrophoresis.

Digestion with Sal I facilitated the subclassification of 41 strains of Campylobacter jejuni into seven types, and digestion with Sma I enabled subclassification into twelve types. Sma I was potentially more useful for the detection of variability among the 41 strains, but both restriction enzymes seemed to be potentially useful for detecting variability among crossed-field gel electrophoresis profiles of the strains. The results clearly demonstrated that C. jejuni strains from different sources and with different routes of transmission had invaded the three farms investigated. At farms Sa and Ai, approximately 70% (23 strains) of isolates of C. jejuni (33 strains) were subclassified into the two major genotypes (I and II) on the basis of cleavage profiles with both Sal I and Sma I. These two major genotypes appeared to have invaded, expanded in and occupied the two chicken farms. All nine strains from farm Ai belonged genotype I.

Animals↗
PubMed1994

[In vitro antibacterial activity of fleroxacin (FLRX) against clinical isolates from bacterial enteritis].

Antibacterial activity of fleroxacin (FLRX), a new quinolone antimicrobial, against 36 strains of Shigella app., 14 strains of Salmonella spp., 11 strains of Escherichia coli, 9 strains of Vibrio spp. (including 2 strains of V. cholerae O1), 14 strains of Campylobacter jejuni/coli, 3 strains of Aeromonas spp. and 1 strain of Plesiomonas shigelloides isolated from infectious enteritis patients in this study was determined. Its activity was compared with that of ciprofloxacin (CPFX), norfloxacin (NFLX) and nalidixic acid (NA). The MIC90 values of FLRX were 0.1 microgram/ml against Shigella spp. and E. coli, 0.2 microgram/ml against Salmonella spp. and Vibrio spp., and 12.5 micrograms/ml against C. jejuni/coli MIC90 of FLRX was comparable to that of CPFX and NFLX against Vibrio spp.. Against other species, MIC90 of FLRX were 2- to 4-fold higher than those of CPFX, whereas equal to or 2-fold lower than NFLX. FLRX demonstrated excellent activity against an NA-resistant (MIC: > 100 micrograms/ml) isolate of E. coli, with MIC 0.78 microgram/ml. FLRX showed 8-fold higher activity than NA against other strains. The antibacterial activity of FLRX was compared with that of NA against stocked strains (clinical isolates from August 1989 to February 1991), consisting of 11 strains of Shigella spp., 10 strains of Salmonella spp., 8 strains of E. coli, 10 strains of V. cholerae O1, 10 strains of V. parahaemolyticus and 14 strains of C. jejuni/coli. MICs of FLRX were 0.78 and 12.5-25 micrograms/ml against Shigella spp. and C. jejuni/coli that showed resistance of NA (MIC: > or = 100 micrograms/ml), respectively. Based on the above, although the absolute MICs are low against E. coli and shigella spp., a value of 0.78 micrograms/ml for FLRX suggested that such strains should be considered to be resistant.

Campylobacter jejuni↗
PubMed1994

[Contamination of healthy livestock with verocytotoxin producing Escherichia coli (VTEC), the first isolation from goats in Japan].

As part of the investigation on the source and route of infection with Verocytotoxin producing Escherichia coli (VTEC) in human beings, isolation of VTEC was attempted using fresh feces collected from healthy livestock (cattle, swine and goat) raised in Sagamihra, Yokohama and Hiratsuka cities from October, 1991 to March, 1992. VTEC could be isolated from 1 (1.0%) of 105 swine, 2 (3.6%) of 55 cattle and 2 (15.4%) of 13 goats. VTEC was isolated for the first time from goats in Japan. The combinations of the serotype and toxin type of the isolated strains were O116:H21 (VT2) and O163:H19 (VT2) for those isolated from the cattle, OUT:H19 (VT2vp) for that from the swine and OUT:H21 (VT1) for those from the goats. Since VTEC isolated from the cattle and goats were found to produce VT of the same serotypes as human VTEC, domestic animals were considered to be involved as a source of VTEC infection in human beings.

Animals↗
PubMed1994

Rapid assay for detection of toxic shock syndrome toxin 1 from human sera.

A noncompetitive enzyme-linked immunosorbent assay that enables the quantitation of toxic shock syndrome toxin 1 (TSST-1) to as little as 30 pg/ml and the detection of TSST-1 to 10 pg/ml in phosphate-buffered saline including 33% human serum or plasma was developed. It takes only 3 h to complete this assay after plate preparation. In this study, 64 human serum samples obtained from 30 patients with toxic shock syndrome or toxic shock syndrome-like symptoms were subjected to testing for the detection of TSST-1. With a cutoff level for TSST-1 of less than 100 pg/ml, 28 samples obtained from 12 patients were positive for TSST-1. The mean and maximum concentrations for these TSST-1-positive samples were 440 and 5,450 pg/ml, respectively. Of these 12 patients, 8 were Staphylococcus aureus culture positive, 3 were negative upon bacterial culturing, and 1 had no cultures done.

Adolescent↗
PubMed1994

Shear-induced platelet aggregation in cerebral ischemia.

BACKGROUND AND PURPOSE: Recent evidence has suggested that shear-induced platelet aggregation is an important mechanism of thrombosis at arterial bifurcations or stenoses. We measured shear-induced platelet aggregation with a new apparatus in patients with cerebral ischemia and also studied correlations with other hemostatic parameters as well as the effect of antiplatelet agents. METHODS: The subjects were 75 patients with cerebral ischemia and 26 control subjects. Platelet aggregation was induced in citrated platelet-rich plasma by a high shear stress (108 dynes/cm2) that was applied by means of a cone-plate streaming chamber based on turbidimetry. We studied the correlation of test results with hemostatic parameters and also the effects of antiplatelet agents. RESULTS: Compared with the control subjects, an increase of shear-induced platelet aggregation was observed in 21 patients with atherothrombotic stroke and 12 with transient ischemic attacks, but not in 11 with cardioembolic stroke or 31 with lacunar stroke. There was no significant correlation of shear-induced platelet aggregation with platelet count, agonist-induced platelet aggregation, fibrinogen level, or beta-thromboglobulin level. The extent of shear-induced aggregation was not correlated with von Willebrand factor antigen levels but was significantly correlated with the amounts of larger von Willebrand factor multimers. Oral aspirin (81 mg/d) did not inhibit shear-induced platelet aggregation, whereas oral ticlopidine (200 mg/d) significantly inhibited it. CONCLUSIONS: These results indicate that shear-induced platelet aggregation is increased in patients with atherothrombotic stroke and transient ischemic attacks, is correlated with the increase of larger von Willebrand factor multimers, and is corrected by ticlopidine but not by low-dose aspirin.

Adult↗
PubMed1994

Macro-fingerprinting analysis at the chromosomal genomic DNA level of isolates of thermophilic Campylobacter coli and C. jejuni, by pulsed-field gel electrophoresis.

Chromosomal genomic DNA from 25 strains of thermophilic Campylobacter coli and C. jejuni were analysed after pulsed-field gel electrophoresis (PFGE) of restriction fragments generated by SalI and SmaI to evaluate the utility of macro-fingerprinting analysis and to assess the possible subtyping of the isolates. Two rare-cutting restriction enzymes, SalI and SmaI, allowed the subclassification of eight strains of C. coli into six types. SalI facilitated the subclassification of seventeen strains of C. jejuni into thirteen types and SmaI the subclassification of seventeen strains into twelve types. The diversity in the PFGE profiles was demonstrated and, therefore, the two enzymes were found to be potentially useful for detecting variability of PFGE profiles among strains of C. coli and C. jejuni. The two enzymes and PFGE may be useful in molecular epidemiology.

Animals↗
PubMed1993

Transmembrane calcium influx associated with von Willebrand factor binding to GP Ib in the initiation of shear-induced platelet aggregation.

We found that the binding of multimeric vWF to GP Ib under a shear force of 108 dynes/cm2 resulted in the transmembrane flux of Ca2+ ions with a two- to three-fold increase in their intracellular concentration ([Ca2+]i). The blockage of this event, obtained by inhibiting the vWF-GP Ib interaction, suppressed aggregation. In contrast, the blockage of vWF binding to GP IIb-IIIa, as well as the prevention of activation caused by increased intracellular cAMP levels, inhibited aggregation but had no significant effect on [Ca2+]i increase. A monomeric recombinant fragment of vWF containing the GP Ib-binding domain of the molecule (residues 445-733) prevented all effects mediated by multimeric vWF but, by itself, failed to support the increase in [Ca2+]i and aggregation. These results suggest that the binding of multimeric vWF to GP Ib initiates platelets aggregation induced by high shear stress by mediating a transmembrane flux of Ca2+ ions, perhaps through a receptor-dependent calcium channel. The increase in [Ca2+]i may act as an intracellular message and cause the activation of GP IIb-IIIa; the latter receptor then binds vWF and mediates irreversible aggregation.

Biological Transport↗