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Biomedical subjects

C Abe

Publications and source records attributed to C Abe.

At least 37 records · Page 2Linked to original sources

Anti-inflammatory effects of mao-bushi-saishin-to in mice and rats.

Effects of Mao-Bushi-Saishin-to (MBS) on anti-inflammatory activities were examined in mice and rats. MBS significantly inhibited the increase in vascular permeability induced by acetic acid, the ear edema induced by arachidonic acid and phorbol ester, and the cutaneous extravasation induced by bradykinin and histamine. MBS, however, was not effective against the serotonin-induced cutaneous permeability increase in mice. MBS significantly inhibited carrageenin-induced hind foot edema and cotton pellet-induced granulation tissue growth in rats. These results show that MBS may exert anti-inflammatory effects through the underlying mechanism(s) of preventing mediator release from mast cells and macrophages.

Acetic Acid↗

[Reinfection tuberculosis at a day-laborers' facility equipped with a sauna].

Restriction-fragment-length polymorphism (RFLP) analysis was used to study one outbreak of reinfection pulmonary tuberculosis at a day-laborers' facility equipped with a sauna. The results were interpreted in conjunction with clinical and epidemiologic data. The DNA prove was derived from the insertion sequence IS 6110. A 63-year-old man who stayed at a day-laborers' facility for eight months was found to have advanced cavitary lung disease with sputum smears strongly positive for acid-fast bacilli. One year later a 46-year-old man staying at the same facility for 24 months was also found to have sputum-positive tuberculosis. The 46-year-old man had undergone treatment for tuberculosis 16 years previously and showed radiographic evidence of previous lung damage from tuberculosis. The second man had no tubercule bacilli in sputum when the 63-year-old man was found to have sputum-positive tuberculosis. The organisms isolated from these two men were both drug sensitive and had a similar RFLP pattern which suggests that 63-year-old man was the source of this outbreak and that pulmonary tuberculosis in the 46-year-old man was caused by exogenous reinfection. We conclude that exogenous reinfection may have been one pattern of tuberculosis transmission in this high-risk environment.

DNA, Bacterial↗

[Multidrug-resistant tuberculosis. 1. Drug susceptibility testing and its standardization].

Drug susceptibility testing is one of the most difficult procedures to standardize in mycobacteriology laboratories. International standardization of the test is needed for comparative evaluation of controlled chemotherapeutic trials, for epidemiological surveys on the prevalence of drug resistance, and for guidance in the treatment of tuberculosis patients. In 1996, a new procedure for drug susceptibility testing was proposed by an ad hoc committee of the Japanese Society for Tuberculosis. The proposal is for a test with the proportion method using an Ogawa egg medium similar to those recommended by the WHO. Resistance is expressed as the percentage of colonies that grow on critical concentrations of the drugs, i.e. 0.2 microgram/ml for isoniazid, 40 micrograms/ml for rifampicin, 10 micrograms/ml for streptomycin and 2.5 micrograms/ml for ethambutol. Strains of tubercle bacilli which are grown on drug-containing media represents more than 1% of the number of colonies that develop on drug-free media are considered to be clinically resistant to that agent. The results are recorded as susceptible (S) or resistant (R) on the laboratory forms. The proportion method using agar- and liquid-based media is used worldwide. In the near future a test with agar- or liquid-based media will be discussed for standardization of the test in Japan, as described by the National Committee for Clinical Laboratory Standards of the United States.

Humans↗

[Recent advance in the isolation of mycobacteria].

Two systems, the newly developed Mycobacteria Growth Indicator Tube (MGIT) and biphasic Septi-Chek AFB based on liquid media, proved to be significantly better than the egg-based solid media for the isolation of mycobacteria from clinical specimens. The isolation of Mycobacterium tuberculosis by MGIT occurred 8 days previous to the isolation by the conventional Ogawa method. These results indicate that the MGIT system is efficient for the recovery of mycobacteria.

Bacteriological Techniques↗

[Drug consultation for patients who present with symptoms like dementia].

It is a lowering of compliance in taking drugs of patients who present with symptoms like dementia that being impairment in a therapy for disease. We started drug consultation to that one at home care, because of improvement of non-compliance in taking drugs. Improvement of consciousness in taking drugs for patients and consciousness in necessity of taking drugs for their family that be brought about by our drug consultation could be made progress of compliance in taking drugs.

Aged↗

A dtsR gene-disrupted mutant of Brevibacterium lactofermentum requires fatty acids for growth and efficiently produces L-glutamate in the presence of an excess of biotin.

A dtsR gene encoding a homolog of the beta subunit of some biotin-containing enzymes suppresses a detergent-sensitive mutation of Brevibacterium lactofermentum (E. Kimura et al., 1996, Biosci. Biotech. Biochem. 60, 1565-1570), which has been used for the fermentative production of L-glutamate. When the dtsR gene was disrupted, the organism exhibited strict fatty acid auxotrophy; oleate or oleate ester, but not palmitate ester or stearate ester, supported the growth of the delta dtsR mutant. Immunoblotting with an anti-DtsR antibody revealed that no intact DtsR was present in the cytosol of the delta dtsR mutant. In the presence of an excess of biotin, the wild type strain did not produce L-glutamate whereas the delta dtsR mutant efficiently produced it. The mechanism underlying the efficient production of L-glutamate by the delta dtsR mutant is discussed as to the possible role of dtsR in fatty acid metabolism.

Bacterial Proteins↗

Mutation in pncA is a major mechanism of pyrazinamide resistance in Mycobacterium tuberculosis.

OBJECTIVE: To characterize the correlation of the mutations in the pncA gene encoding pyrazinamidase (PZase) of Mycobacterium tuberculosis to a loss of PZase activity and development of pyrazinamide (PZA) resistance. DESIGN: The association of PZase activity, minimum inhibitory concentrations (MICs), and mutations in the pncA gene of M. tuberculosis isolated in mostly Asian countries was investigated. RESULTS: One hundred thirty-five out of 168 isolates were PZase positive, and 33 were negative. The MICs of PZA at pH 6.0 were over 400 micrograms/ml for all 33 PZase-negative isolates, while those of PZase-positive isolates were equal to or less than 200 micrograms/ml. Among 33 PZase-negative isolates sequenced, 32 (97%) had mutations within the pncA gene. A mutation was seen in various regions throughout the pncA gene. It was surprising that all three strains of in vitro selected PZA resistant mutants were PZase-positive and showed no change in the pncA gene. These results indicate that additional mechanisms may be involved in PZA resistance. No mutations were observed in all of 135 PZase-positive M. tuberculosis isolates tested, indicating that mutations in the pncA gene could be involved in the loss of PZase activity. CONCLUSIONS: Sequencing analysis of the pncA gene should provide rapid diagnosis of PZA resistant clinical isolates of M. tuberculosis.

Amidohydrolases↗

Characterization of the rpsL and rrs genes of streptomycin-resistant clinical isolates of Mycobacterium tuberculosis in Japan.

Mutations in the rpsL and rrs genes associated with streptomycin resistance in Mycobacterium tuberculosis clinically isolated in Japan were characterized. The rpsL genes of 172 clinical isolates were amplified by PCR and classified into two groups on the basis of MboII restriction digestion. Thirty-three out of 54 (61.1%) streptomycin-highly resistant isolates (MIC > 200 micrograms ml-1) were not digested by MboII. By contrast, the remaining 21 of 54 (38.9%) streptomycin-highly resistant isolates, all of 41 isolates with streptomycin resistance at a lower level (20 micrograms ml-1 < MIC < or = 200 micrograms ml-1), and all of 77 streptomycin-sensitive isolates, were restricted. Thus, all isolates resistant for MboII digestion showed a high level of resistance to streptomycin. Subsequently, the sequence for the rpsL and rrs genes from the 46 isolates were analysed. Eighteen out of 19 (94.7%) streptomycin-highly resistant isolates carried a mutation in any rpsL gene at position 43 or 88, or the rrs gene; 10 out of 17 (58.8%) streptomycin-resistant isolates at a lower level were confirmed to exhibit the mutation of either the mutated rpsL gene at position 88, or the rrs gene. In the total 36 streptomycin-resistant isolates, the mutation of the rpsL or rrs gene was observed in 28 streptomycin-resistant isolates, corresponding to 77.8%, whereas none of the streptomycin-sensitive isolates had mutations in either the rpsL or rrs gene.

Antibiotics, Antitubercular↗

Studies of polymorphic DNA fingerprinting and lipid pattern of Mycobacterium tuberculosis patient isolates in Japan.

Strain differentiation by DNA restriction fragment length polymorphism (RFLP) has been used mainly for the epidemiological purpose of Mycobacterium tuberculosis infection. In this study, we tried to connect the molecular and phenotypic characteristics of M. tuberculosis patient isolates by comparing the DNA fingerprints obtained by RFLP using IS6110 and lipid patterns using two-dimensional thin-layer chromatography (2-D TLC) with silica gel, since M. tuberculosis has a lipid-rich cell envelope which contributes to the virulence and immunomodulatory properties. We found that 66 isolates of M. tuberculosis from tuberculosis patients showed that the occurrence of IS6110 varied from 1 to 24 copies. The IS6110 patterns were highly variable among isolates. Fifty different RFLP patterns were observed, and 12 RFLP patterns were shared by two or more strains. By computerized analysis of the RFLP patterns of M. tuberculosis patient isolates, we found that 95% of the isolates fell into seven clusters, from A to G, with at least two isolates in each (> 30% similarity). Among the cellular lipids, the phospholipid composition did not differ by strain, whereas the glycolipid pattern differed markedly. Especially, the relative concentration of cord factor and sulfolipid, both of which were known as virulent factors, varied by strain. The fingerprints of some strains showed an association between the DNA and glycolipid patterns, even though some of the same DNA fingerprint strains showed differences in lipid patterns. Among the patient isolates, M. tuberculosis strain 249 possessed a specific glycolipid with 2-O-methyl-L-rhamnose and L-rhamnose, which is rarely found in other strains. This glycolipid showed serological activity against the sera of tuberculosis patients, even if the reactivity was not as strong as trehalose dimycolate. It also showed the inhibition of phagosome-lysosome fusion in macrophages, suggesting involvement with virulence. These results suggest that RFLP analysis using IS6110 is useful for clustering the human isolates of M. tuberculosis, however, for further strain differentiation on virulence, a lipid analysis provides more information.

Antibodies, Bacterial↗

Molecular analysis of kanamycin and viomycin resistance in Mycobacterium smegmatis by use of the conjugation system.

We examined the molecular mechanisms of resistance to kanamycin and viomycin in Mycobacterium smegmatis. All of the M. smegmatis strains with high-level kanamycin resistance had a nucleotide substitution from A to G at position 1389 of the 16S rRNA gene (rrs). This position is equivalent to position 1408 of Escherichia coli, and mutation at this position is known to cause aminoglycoside resistance. Mutations from G to A or G to T at position 1473 of the M. smegmatis rrs gene were found in viomycin-resistant mutants which had been designated vicB mutants in our earlier studies. Using the M. smegmatis conjugation system, we confirmed that these mutations indeed contributed to kanamycin and viomycin resistance, and kanamycin susceptibility was dominant over resistance in a heterogenomic strain. Additional experiments showed that three of four Mycobacterium tuberculosis strains with high-level kanamycin resistance had a mutation from A to G at position 1400, which was equivalent to position 1389 of M. smegmatis.

Base Sequence↗

[Serodiagnosis with trehalose-6,6'-dimycolate of pulmonary tuberculosis].

Enzyme-linked immunosorbent assay (ELISA) was used to examine serum antibody levels of mycobacterial antigen in 92 patients with active tuberculosis, 36 with cured tuberculosis, 45 with nontuberculous mycobacteriosis, and as 31 with other diseases. Glycolipid fraction containing mainly cord factor (trehalose-6,6'-mycolate) from Mycobacterium tuberculosis H37Rv were used as ELISA antigen. Overall positive rates of the ELISA tests in the patients with active pulmonary tuberculosis, those with nontuberculous mycobacteriosis, and those with other diseases were 67.4%, 75.6%, and 6.5%, respectively. Patients with tuberculosis and those with nontuberculous mycobacteriosis differed from the control group (p < 0.0001). Higher positive rates were correlated with bacterial loads (smear-positive vs smear-negative, p < 0.01) and with chest roenrgenographic findings (far advanced or other cavitary vs noncavitary, p < 0.01). Because 18 of 36 smear-negative patients (50%) had positive results, we believe that the ELISA test with this antigen can be useful for diagnosis of M. tuberculosis, especially in patients with smear-negative.

Antibodies, Bacterial↗

[Reliability of Amplicor Mycobacteria test for detection of Mycobacterium tuberculosis complex. M. avium and M. intracellulare: a cooperative study among 9 laboratories].

The Amplicor Mycobacteria, a PCR-based assay, is a rapid test for the detection of Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare in clinical samples. To estimate the reliability and reproducibility of the method, a cooperative blind study was conducted among 9 laboratories. Materials used for testing consisted of 105 sputum and 30 water samples containing known numbers of M. bovis BCG, M. avium, M. intracellulare, and samples without bacteria. Only 2 out of the 9 laboratories correctly identified the presence or absence of mycobacterial DNA in all 135 samples. In sputum samples, 6 out of the 9 laboratories detected mycobacterial DNA in all positive samples, and 4 out of the 9 laboratories correctly reported the absence of DNA in the negative samples, indicating the need for good laboratory practice and development of reference reagents to monitor the performance of the whole study, including pretreatment of clinical samples. The main problem was lack of specificity rather than lack of sensitivity. From about half of the laboratories, false-positive results were reported, however, the ratio was below 6%; 1% (1/106 sputum samples) in 3 laboratories, 1.9% (2/105) in 2 laboratories, and 5.7% (6/105) in one laboratory, respectively. These results indicate that the Amplicor Mycobacteria is quite useful for a rapid diagnosis of tuberculosis.

DNA, Bacterial↗

[Drug consultation and at home care--the present situation and the point at issue in our hospital].

We began drug consultation in our hospital and at home care about one year ago, because of progress in the quality of medical care. As the method for drug consultation, we tried to educate patients using our own pamphlets. It was assumed that the education was important not only for the patients but also for their family. Again, it was assumed that the usefulness of a conference with doctors, nurses, MSW and us made for better compliance in taking drugs, assured early detection of side effects and better management of the disease. With home care, it is supposed that the drug consultation will be carried out by clinical pharmacists will make for progress in the quality of medical care.

Home Care Services, Hospital-Based↗

[Rapid diagnosis of tuberculosis].

Two systems, the newly developed Mycobacteria Growth Indicator Tube (MGIT) and biphasic Septi-Chek AFB based on liquid media, proved to be significantly better than the egg-based solid media for the isolation of mycobacteria from clinical specimens. The difference in the rates of isolation of bacteria between the two groups of media was more remarkable with smear-negative specimens. The isolation of the Mycobacterium tuberculosis complex by MGIT occurred 8 days previous to the isolation by the conventional Ogawa method. The mean time for detecting M. tuberculosis complex by Septi-Chek AFB was similar to those of the Ogawa method. A greater difference in isolation time was observed for mycobacteria other than M. tuberculosis (MOTT) isolates. These results indicate that the MGIT and Septi-Chek AFB systems based on liquid media are efficient for the recovery of mycobacteria. PCR and other nucleic acid amplification methods are widely used for the detection of M. tuberculosis in clinical specimens. Although the sensitivities of the Gen-Probe Amplified Mycobacteria Direct Test (MTD) and Amplicor Mycobacteria for the detection of the M. tuberculosis complex appear to be similar to the sensitivity of the culture method using the Septi-Chek AFB, the two methods should be quite useful for rapid detection of M. tuberculosis infections. On the other hand, two cooperative blind studies conducted between 6 to 9 laboratories to estimate the reliability and reproducibility of these two commercially available kits revealed the necessity of good laboratory practice and development of reference reagents to monitor the performance of the whole assay, including pretreatment of clinical specimens. Considerable progress has been made in recent years toward understanding the molecular basis of the resistance to antituberculosis drugs, isoniazid (katG, inhA, ahpC), rifampin (rpoB), pyrazinamide (pncA), streptomycin (rpsL, rrs), ethambutol (embB), and fluoroquinolones (gyrA). Most cases of resistance are related usually to simple nucleotide substitutions rather than to acquisition of new genetic elements. Multidrug-resistant isolates of M. tuberculosis arise as a consequence of sequential accumulation of mutations conferring resistance to single therapeutic agents. The basis of resistance is not able to be explained yet in a substantial percentage of strains (> 90%) for other antituberculosis drugs than rifampin. Further studies are required to fully understand the molecular mechanisms of resistance.

DNA, Bacterial↗

Resistance to antituberculosis drugs in Japan.

SETTING: Five years after the last survey of drug-resistant tuberculosis in Japan, a serious new phenomenon has gradually begun to appear. A nationwide survey was conducted by the Tuberculosis Research Committee. OBJECTIVE: To determine resistance patterns to five anti-tuberculosis drugs and risk factors. DESIGN: Cultures were obtained from patients hospitalized at 38 hospitals in various districts of Japan throughout 6 months, from 1 June through 30 November in 1992. Drug susceptibility testing was carried out in the national reference laboratory. RESULTS AND CONCLUSIONS: Resistance to one or more drugs was found in 5.6% of new cases and 27.8% of recurrent cases (P < 0.001). About 88% of drug resistant isolates from the new cases were resistant to one drug, while 50.8% of the drug resistant isolates from the recurrent cases had resistance to two or more drugs (P < 0.001). Resistance rates to both isoniazid and rifampin in new cases was very low (only 0.14%). Primary drug resistance rates were higher in age groups less than 60 years old, compared to those of 60 years and over (P = 0.05). Compared with the rate in Japanese patients, foreign-born individuals had a higher resistance rate in the recurrent cases (P = 0.034). This survey indicated a similar trend in resistance rates to five antituberculosis drugs to those of the last survey in 1987.

Adolescent↗

Molecular cloning of the Corynebacterium glutamicum ('Brevibacterium lactofermentum' AJ12036) odhA gene encoding a novel type of 2-oxoglutarate dehydrogenase.

The Corynebacterium glutamicum ('Brevibacterium lactofermentum' AJ12036) odhA gene, encoding 2-oxoglutarate dehydrogenase (E1o subunit of the 2-oxoglutarate dehydrogenase complex), has been isolated and identified as an homologous counterpart of the Escherichia coll sucA and Bacillus subtilis odhA genes. The nucleotide sequence of a 4394 bp chromosomal fragment containing the C. glutamicum odhA gene was determined. The odhA gene comprised 3771 bp (1257 codons, including the initiation codon) and a molecular mass of 138656 Da was predicted for the OdhA polypeptide. Northern blot analysis revealed a 3.9 kb transcript. The size of the transcript, together with the presence of a rho-independent terminator-like structure, suggests that C. glutamicum odhA is monocistronic. Cells harbouring plasmids carrying C. glutamicum odhA showed a threefold increase in specific 2-oxoglutarate dehydrogenase complex activity and expression of a protein with an apparent molecular mass of 136 kDa, in good agreement with the predicted size of the OdhA polypeptide. The C-terminal region of the C. glutamicum OdhA protein shows strong sequence similarity to E1os from other organisms. C. glutamicum OdhA has an N-terminal extension not found in previously reported E1os. The amino acid sequence of this extension shows similarity to that of the C-terminal region of dihydrolipoamide S-succinyltransferase (E2o) subunits of 2-oxoglutarate dehydrogenase complexes and dihydrolipoamide S-acetyltransferase (E2p) subunits of pyruvate dehydrogenase complexes. It suggests that the C. glutamicum odhA gene might encode a novel bifunctional protein with E1o and E2o activities.

Amino Acid Sequence↗

[Evaluation of a new rapid detection system for mycobacteria using an oxygen sensitive fluorescent sensor].

A comparison of the rate of recovery and time for detection of mycobacteria from clinical specimens of the newly developed Mycobacteria Growth Indicator Tube (MGIT), the biphasic Septi-Chek, and the egg-based Ogawa medium was made. From the 305 sputum specimens processed, a total of 83 mycobacterial isolates were detected. From these 80 (84.3%) isolates were detected with both the MGIT and Septi-Chek systems, respectively, and 50 (60.2%) were detected by the Ogawa egg method. The difference in the percentages of positive cultures between the two systems based on liquid media and the Ogawa egg method was significant (p < 0.001). The mean time for the detection of the Mycobacterium tuberculosis complex were 16.3 days with the MGIT system, 25.6 days with the Septi-Chek, and 21.5 days with Ogawa egg method. These results indicate that the MGIT system is efficient for the recovery of mycobacteria.

Bacteriological Techniques↗