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Biomedical subjects

T Deguchi

Publications and source records attributed to T Deguchi.

At least 109 records · Page 6Linked to original sources

Mycoplasma genitalium in the cervices of Japanese women.

BACKGROUND: Mycoplasma genitalium is considered a cause of nongonococcal urethritis in men. This organism also is a cause of genital infections in women, and has been detected in women attending sexually transmitted disease clinics in the United Kingdom and Denmark, although its prevalence is unknown in Japanese women. GOALS: To determine the prevalence of M. genitalium in the cervices of women with cervicitis or adnexitis as well as in asymptomatic pregnant women in Japan. STUDY DESIGN: Two hundred women who attended obstetric and gynecologic clinics were recruited. Sixty-four women had cervicitis, 53 had adnexitis, and 3 had both. Eighty pregnant women were asymptomatic for infection. Cervical swab specimens were examined for M. genitalium using a polymerase chain reaction-based assay. RESULTS: Five (7.8%) of 64 women with cervicitis and 3 (5.7%) of 53 women with adnexitis were positive for M. genitalium. After exclusion of Chlamydia-positive women, 5 (8.8%) of 57 women with cervicitis, and 2 (4.1%) of 49 women with adnexitis were positive for M. genitalium. In none of 80 asymptomatic pregnant women, including a Chlamydia-positive woman, was M. genitalium detected. Overall, 7 (6.6%) of 106 women with Chlamydia-negative genital infections were positive for the M. genitalium. This prevalence was significantly greater than that in asymptomatic pregnant women (P < 0.05). CONCLUSIONS: A significantly greater prevalence of M. genitalium was demonstrated in Japanese women with Chlamydia-negative cervicitis or adnexitis, compared with that in asymptomatic pregnant women. This study suggests that M. genitalium may play a pathogenic role in a portion of cases with Chlamydia-negative genital infections.

Adult↗

Mutations in the gyrA and parC genes in fluoroquinolone-resistant clinical isolates of Pseudomonas aeruginosa.

We determined partial sequences of the gyrA and parC genes of the fluoroquinolone-susceptible strain ATCC 27853 and 22 clinical isolates of Pseudomonas aeruginosa. While a single amino acid change in GyrA with or without a change in ParC was found in 14 isolates with decreased susceptibility to fluoroquinolones, 3 higher-level fluoroquinolone-resistant isolates had a double amino acid change in GyrA and a single amino acid change in ParC.

Amino Acid Sequence↗

Improved antimicrobial activity of DU-6859a, a new fluoroquinolone, against quinolone-resistant Klebsiella pneumoniae and Enterobacter cloacae isolates with alterations in GyrA and ParC proteins.

MICs of DU-6859a, a novel fluoroquinolone, for 18 Klebsiella pneumoniae isolates and 21 Enterobacter cloacae isolates with altered GyrA or altered GyrA and ParC ranged from < or =0.025 to 6.25 microg/ml and from 0.1 to 3.13 microg/ml, respectively. Based on the MICs at which 90% of the isolates were inhibited for these strains of K. pneumoniae and E. cloacae, DU-6859a exhibited 16- to 256-fold-greater activity than currently available fluoroquinolones.

Anti-Infective Agents↗

Alterations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV in quinolone-resistant clinical isolates of Klebsiella pneumoniae.

We determined a partial sequence of the Klebsiella pneumoniae parC gene, including the region analogous to the quinolone resistance-determining region of the Escherichia coli gyrA gene, and examined 26 clinical strains of K. pneumoniae for an association of alterations in GyrA and ParC with susceptibilities to quinolones. The study suggests that in K. pneumoniae DNA gyrase is a primary target of quinolones and that ParC alterations play a complementary role in the development of higher-level fluoroquinolone resistance.

Amino Acid Sequence↗

In vivo selection of Klebsiella pneumoniae strains with enhanced quinolone resistance during fluoroquinolone treatment of urinary tract infections.

We report two cases of failure of fluoroquinolone treatment of urinary tract infections with Klebsiella pneumoniae strains harboring quinolone resistance-associated alterations in GyrA and ParC and in vivo selection of posttreatment isolates with enhanced fluoroquinolone resistance. Active efflux leading to decreased accumulation of a drug enhanced fluoroquinolone resistance in one posttreatment isolate, and an additional mutation in parC resulting in an additional amino acid change in ParC was associated with increased resistance in the other.

Aged↗

Nylon biodegradation by lignin-degrading fungi.

The biodegradation of nylon by lignin-degrading fungi was investigated. The fungus IZU-154 significantly degraded nylon-66 membrane under ligninolytic conditions. Nuclear magnetic resonance analysis showed that four end groups, CHO, NHCHO, CH3, and CONH2, were formed in the biodegraded nylon-66 membranes, suggesting that nylon-66 was degraded oxidatively.

Basidiomycota↗

Rapid screening of point mutations of the Neisseria gonorrhoeae parC gene associated with resistance to quinolones.

To detect quinolone resistance-associated mutations within the Asp-86, Ser-87, Ser-88, and Glu-91 codons of the Neisseria gonorrhoeae parC gene, we developed a rapid and simple assay based on amplification of the regions of the parC gene containing the mutations sites by PCR and digestion of the PCR products with restriction enzymes. By using the method of primer-specified restriction site modification, artificial SalI, PstI, EcoRI, and HinfI restriction sites were created in the regions containing the Asp-86, Ser-87, Ser-88, and Glu-91 codons, respectively. The mutations generating alterations at Asp-86, Ser-87, Ser-88, and Glu-91 were detected as failures of SalI, PstI, EcoRI, and HinfI to digest the respective PCR products. Fifty-five clinical strains of N. gonorrhoeae were examined for mutations in the parC gene by this assay. Appropriate mutations at either the Asp-86, Ser-87, Ser-88, or Glu-91 codon were detected in each of 11 strains in which a mutation had previously been observed by DNA sequencing. This rapid and simple assay could be a useful device for screening genetic alterations in the parC gene associated with resistance to quinolones in N. gonorrhoeae.

DNA Topoisomerase IV↗

Development of a rapid assay for detecting gyrA mutations in Escherichia coli and determination of incidence of gyrA mutations in clinical strains isolated from patients with complicated urinary tract infections.

The MICs of ofloxacin for 743 strains of Escherichia coli isolated from 1988 to 1994 were determined by testing. The strains were from patients with urinary tract infections complicated by functional or anatomical disorders of the urinary tract. Those determined to be ofloxacin resistant (MIC, > or =12.5 microg/ml) comprised 3 of 395 strains (1.3%) from the 1988 to 1990 group, 2 of 166 strains (1.2%) from the 1991 to 1992 group, and 7 of 182 strains (3.8%) from the 1993 to 1994 group. The incidence of resistant strains increased significantly during this period. The percentage of isolates with moderately decreased susceptibilities to ofloxacin (MIC, 0.39 to 3.13 microg/ml) also rose during the same period. To determine the incidence of gyrA mutations in urinary-tract-derived strains of E. coli, we developed a simple and rapid assay based on PCR amplification of the region of the gyrA gene containing the mutation sites followed by digestion of the PCR product with a restriction enzyme. Using this assay, we examined all 182 strains isolated in 1993 and 1994 for the presence of mutations at Ser-83 and Asp-87 in the gyrA gene. Of these strains, 33 (18.1%) had mutations in the gyrA gene. The incidences of mutations at Ser-83, at Asp-87, and at both codons were 10.4 (19 strains), 4.4 (8 strains), and 3.3% (6 strains), respectively. To determine the correlation of the mutations in the gyrA gene with susceptibilities to quinolones (nalidixic acid, ofloxacin, norfloxacin, and ciprofloxacin), we further examined 116 strains for which the MICs of ofloxacin were > or =0.2 microg/ml that were chosen from the isolates in the 1988 to 1992 group. The MICs of nalidixic acid for the strains without mutations at either Ser-83 or Asp-87 were < or =25 microg/ml, whereas those for the strains with single mutations or double mutations were from 50 to >800 microg/ml. For the fluoroquinolones, significant differences in the distributions of the MICs were observed among the strains without mutations, with single mutations, and with double mutations. The accumulation of mutations in the gyrA gene was associated with an increase in fluoroquinolone resistance. Ofloxacin MICs for the majority of the strains with single and double mutations were 0.39 to 3.13 and 6.25 to 100 microg/ml, respectively. This study demonstrates a chronological increase in the percentage of not only highly fluoroquinolone-resistant strains, corresponding to those with double mutations in the gyrA gene, but also strains with moderately decreased susceptibilities to fluoroquinolones, corresponding to those with single mutations. This increase in the incidence of strains with a single mutation in the gyrA gene portends a further increase in the incidence of strains with clinically significant resistance to fluoroquinolones.

Anti-Infective Agents↗

Antinociceptive and antidepressant-like profiles of BL-2401, a novel enkephalinase inhibitor, in mice and rats.

To clarify the properties of BL-2401 ((+/-)-3-[2-benzyl-3-(propionylthio) propionyl]amino-5-methylbenzoic acid), a novel enkephalinase inhibitor, we examined its antinociceptive and antidepressant-like activities after oral administration, along with their association with endogenous opioid systems. BL-2401 produced an antinociceptive effect after oral administration in the mouse phenylbenzoquinone writhing test (ED50: 12.4 mg/kg) and the rat acetic acid writhing test (ED50: 55.8 mg/kg), the antinociceptive effect being antagonized by naloxone hydrochloride. BL-2401 also relieved arthritis-induced hyperalgesia in rats. In the mouse hot-plate and tail pressure tests, BL-2401 showed significant but modest antinociception at higher doses (200 and 400 mg/kg). In addition, BL-2401 (100 mg/kg) produced a naloxone-reversible antidepressant-like effect in the mouse forced swimming test. As for the mechanism of the action, the active metabolite of BL-2401, BL-2240 ((+/-)-3-(2-benzyl-3-mercaptopropionyl) amino-5-methylbenzoic acid), selectively inhibited enkephalinase in vitro (IC50: 5.2 nM). Oral administration of BL-2401 to mice significantly inhibited the enkephalinase activity in the striatum and also potentiated the antinociceptive effect of (D-Ala2,Met5)-enkephalin given intracisternally. These findings indicate that BL-2401 is an orally active enkephalinase inhibitor and may produce antinociceptive and antidepressant-like effects in association with endogenous opioid systems.

Administration, Oral↗

Prevalence of Mycoplasma genitalium in asymptomatic men in Japan.

We aim to clarify the prevalence of Mycoplasma genitalium in asymptomatic men in Japan. First-catch urine specimens were obtained from 187 asymptomatic Japanese men and examined for the presence of M. genitalium using a polymerase chain reaction (PCR)-based assay. Two (1.1%) of 187 first-catch urine specimens were positive for M. genitalium. The prevalence of M. genitalium in urine specimens of asymptomatic men in Japan is lower than that in asymptomatic men in the UK (6%) and Denmark (9%).

Adult↗

[Changes in tissue and blood polyamines during N-butyl-N-(4-hydroxybutyl) nitrosamine-induced bladder carcinogenesis in rats].

BACKGROUND: Polyamine are recognized as cell growth factors. We studied in order to determine whether alterations in the levels of tissue and blood polyamines were useful biochemical markers for bladder tumor. METHODS: The concentrations of three polyamines, diamine, spermidine and spermine, in urinary bladder and blood were determined during N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced bladder carcinogenesis in male F344 rats. At 5 weeks of age, rats were given 0.05% BBN in the drinking water for 20 weeks. RESULTS: BBN induced bladder hyperplasia in 4 of 5 rats at 8 weeks, papillomas in 2 of 5 rats at 12 weeks, and transitional cell carcinoma in all the rats by 20 weeks. The levels of total polyamine in both bladder and blood of the rats during 12-20 weeks were significantly higher than those of the control animals given water alone. The elevation of total polyamine was mainly due to the increase of spermidine of the three polyamines, which was coincident with the incidence of bladder tumors. CONCLUSION: The results indicated that the polyamines are excellent biochemical markers for bladder tumors.

Animals↗

[Changes in tissue and blood polyamine levels following chemotherapy in rats with urinary bladder carcinoma induced by N-butyl-N-(4-hydroxybutyl) nitosamine in rats].

BACKGROUND: Polyamines are recognized as cell growth factors. We attempted to determine whether alterations in the levels of tissue and blood polyamines were useful biochemical makers for monitoring the efficacy of the chemotherapy for bladder tumors. METHODS: The concentrations of three polyamines of diamine, spermidine and spermine in urinary bladder and blood were determined in male F344 rats with urinary bladder carcinoma induced by N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN), following chemotherapy with cisplatin, methotrexate and pirarubicin. RESULTS: Bladder carcinoma was observed in 5 of 20 rats of the chemotherapeutic group, and 16 of 20 rats of the control group given saline alone. The levels of spermidine, spermine and total polyamine in both bladder and blood of the treated rats were significantly lower than those of the control rats. CONCLUSION: The study suggested that the levels of tissue and blood polyamines could be used as biochemical markers for monitoring the efficacy of the chemotherapy for bladder tumors.

Animals↗

[Analysis of induction of MDR1 gene expression by anticancer chemotherapy in bladder cancer].

Using a reverse transcriptase-polymerase chain reaction (RT-PCR)-based quantitative analysis method, we investigated MDR1 mRNA expression levels in 58 bladder cancer specimens to determine whether MDR1 gene expression was induced or enhanced in bladder cancers during chemotherapy. In bladder cancer specimens which were obtained from patients treated with anticancer drugs, significantly higher expression levels of MDR1 mRNA were observed than in those from patients not treated with any anticancer drugs (p = 0.0134, Mann-Whitney U test). From 14 patients who had bladder cancer, clinical specimens were obtained before and after neoadjuvant intra-arterial chemotherapy. The expression levels of MDR1 mRNA were significantly higher in the post-treatment specimens than in the pre-treatment specimens (p = 0.0298, Wilcoxon signed-rank test). Of these 14 patients, 7 patients exhibited increased levels of MDR1 mRNA expression after chemotherapy. In 6 patients, there were no changes in the MDR1 mRNA expression levels before and after chemotherapy. Only one patient exhibited decreased levels of MDR1 mRNA expression after chemotherapy. No significant correlations were observed, between MDR1 mRNA expression levels and effect of the chemotherapy determined microscopically, dosage of anticancer drugs, or patient outcome. In conclusion, this study indicates that MDR1 gene expression in bladder cancers is induced and enhanced during chemotherapy. This overexpression of the MDR1 gene may contribute to resistance to anticancer drugs after repeated chemotherapy.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

[Usefulness of DNA ploidy, AgNORs, PCNA and c-erbB-2 as predictors of prognosis in patients with renal cell carcinoma].

Seventy one patients with renal cell carcinomas were examined for a variety of markers associated with tumor malignancy: nuclear DNA ploidy, AgNORs, PCNA and c-erbB-2. Usefulness of the markers in predicting the prognosis was studied by analyzing the relationship between each of these markers and the prognosis of the patients with renal cell carcinomas. DNA ploidy was analyzed by flow cytometry. AgNORs were stained by the silver colloid method. PCNA and c-erbB-2 were detected by immunohistochemistry. In all the patients examined, DNA ploidy, AgNORs, PCNA and c-erbB-2 were significant predictors of the prognosis. Of the patients with grade 2 carcinomas, the survival rate was significantly higher in the patients with the PCNA-positive cells of lower than 35.0% than in those with the positive cells of more than 35.0%. The patients without the expression of c-erbB-2 exhibited a significantly higher survival rate than those with the expression. In the patients with grade 2 carcinomas, however, neither DNA ploidy nor AgNoRs was a significant predictor of the prognosis. These findings suggest that PCNA and c-erbB-2 provide more accurate information than the others to understand the biological characteristics of the grade 2 carcinomas and are useful in predicting the prognosis of the patients with grade 2 renal cell carcinomas.

Adult↗

Calbindin-D28k-immunoreactivity in the trigeminal ganglion neurons and molar tooth pulp of the rat.

The cell body size and coexpression of carbonic anhydrase (CA), calretinin (CR) and calcitonin gene-related peptide (CGRP) of primary neurons with calbindin-D28k (CB) was examined in the trigeminal ganglion (TG) of the rat. CB-immunoreactive (-ir) cells were mostly large and preferentially distributed in the maxillary and mandibular divisions of the TG. 48% of CB-ir TG cells exhibited enzyme CA activity. 10% of CB-ir TG cells contained CR-ir. Most TG cells coexpressing CB- and CR-irs were localized to the maxillary and mandibular divisions and exhibited CA activity. 6.5% of CB-ir TG cells coexisted with CGRP-ir. 46% of TG cells coexpressing CB and CGRP exhibited CA activity. The innervation of the molar tooth pulp by CB-ir TG primary neurons was also examined. CB-ir thick and smooth nerve fibers projected from the root pulp to the pulp horn and the roof of the pulp chamber, where they became thinner and rarely entered the subodontoblastic layer. However, they could not be traced to the odontoblastic layer, predentin or dentine. The distribution pattern of CB-ir pulpal fibers was different from that of CR-ir ones. The trigeminal neurons cells retrogradely labeled with fast blue (FB) from the maxillary molar tooth pulp contained CB- and CR-irs. 23% and 1% of the labeled cells were immunoreactive for CB and CR, respectively. The coexpression of CB- and CR-immunoreactivities (-irs) in FB-labeled cells was negligible. An immunoelectron microscopic method revealed that 21% of pulpal nerve fibers were immunoreactive for CB, and that all CB-ir nerve fibers in the root pulp were myelinated. The present study indicated that the tooth pulp primary neurons contained CB-ir but did not coexpress CB- and CR-irs and that these neurons projected their myelinated axons to the pulp.

Animals↗

Cytogenetic analyses of hepatocellular carcinoma by in situ hybridization with a chromosome-specific DNA probe.

BACKGROUND: Numerical chromosome analysis has been established in solid tumors by using in situ hybridization (ISH) with a chromosome-specific probe. We analyzed human hepatocellular carcinoma (HCC) by ISH for chromosome 17 and investigated the correlation of its copy number with histologic malignancy, proliferative activity, p53 mutation, and DNA ploidy. METHODS: Chromosome 17 was hybridized with a pericentromere-specific DNA probe directly on the tumor cells isolated from paraffin blocks of 25 surgically resected HCCs. Proliferative activity was measured by Ki-67 immunohistochemistry, p53 mutation was analyzed by p53 immunohistochemistry, and DNA ploidy was estimated by cytofluorometry. RESULTS: Forty-four percent of the 25 HCCs showed numerical abnormality of chromosome 17. Many disomic cases had a less malignant histology, whereas many polysomic cases had a more malignant histology. The Ki-67 positive index of polysomic cases was higher than that of disomic cases. In 22 cases (88.0%), the copy number of chromosome 17 was well matched with DNA ploidy. However, the numerical abnormality of chromosome 17 did not show a significant correlation with p53 mutation. Two of four HCCs that showed histologic heterogeneity were also heterogenous on ploidy pattern and the copy number of chromosome 17. Conversely, there was one case in which only ISH could demonstrate heterogeneity, although the other features exhibited homogeneity. CONCLUSIONS: Numerical chromosome abnormalities correlated with the increase of histologic malignancy proliferative activity, and DNA ploidy. Moreover, ISH analysis was useful in assessing the intratumoral heterogeneity in HCC, especially when current methods failed to detect it. Thus, ISH provides information on important biologic features, such as malignant potential and intratumoral heterogeneity, in HCC.

Adult↗

Cytodifferentiation of the odontoclast prior to the shedding of human deciduous teeth: an ultrastructural and cytochemical study.

BACKGROUND: In human deciduous teeth, odontoclastic resorption takes place at the pulpal surface of the coronal dentine prior to shedding, and this resorption shows clear time-related histological changes (Sahara et al., 1992). METHODS: Using this phenomenon as an observation system, we examined the cytodifferentiation of human odontoclasts by light and electron microscopy. For a histochemical marker of odontoclast differentiation and function, tartrate-resistant acid phosphatase (TRAP) activity was determined by light and electron microscopic enzyme histochemistry. RESULTS: As root resorption neared completion, TRAP-positive mononuclear cells were initially detected in the pulp chamber. They had abundant mitochondria, small lysosomes, and moderately developed rough endoplasmic reticulum throughout their cytoplasm. In these mononuclear cells, TRAP activity was localized in compartments of the biosynthetic pathway, i.e., in cisternae of the endoplasmic reticulum and Golgi lamellae, as well as small lysosomes. The TRAP-positive mononuclear cells first made contact with the predentine surface by their elongated cellular processes. After attachment, they spread out along the predentine surface and developed specialized membrane structures, clear zones, and ruffled borders. Next, they fused with each other on the predentine surface and formed typical multinucleate odontoclasts. After termination of their resorption function, the odontoclasts lost their ruffled borders and became detached from the resorbed surface. Most of the detached odontoclasts had numerous large pale vacuoles and secondary lysosomes and appeared to be in the process of degeneration. CONCLUSIONS: The present study demonstrates that: (1) odontoclasts differentiated from TRAP-positive mononuclear cells, which presumably originate from circulating progenitor cells, (2) membrane specialization of odontoclasts, i.e., development of a clear zone and ruffled border, is induced following their contact with the resorption surface, (3) multinucleation of odontoclasts takes place only after their attachment to the resorption surface, (4) mature multinucleate odontoclasts can resorb predentine as well as dentine in the same way as osteoclasts resorb bone, and (5) at the end of the resorption, odontoclasts gradually lose their ruffled borders and become detached from the resorbed surface.

Acid Phosphatase↗