Search PubMed⌕ Search

Biomedical subjects

Y Yanagawa

Publications and source records attributed to Y Yanagawa.

At least 73 records · Page 4Linked to original sources

[Shigella dysenteriae strains possessing a new serovar isolated from imported diarrheal cases in Japan].

Two Shigella strains (93-119 and 95-619) isolated from stool cultures of imported diarrheal cases in Japan, did not react to any antisera of the established SHigella serovars. These strains had the typical biochemical characteristics of Shigella dysenteriae, and were biochemically identical to each other. Both strains were positive in the Serény test and other tests for invasiveness; these indicate that they can cause shigellosis in humans. The results of antigenic analysis showed that they did not belong to any of the recognized or provisional serovars, and were serologically indistinguishable. Strain 93-119 is designated as the test strain for this new serovar.

Animals↗

[Biochemical and molecular characterization of Salmonella serovar enteritidis phage type 4 isolated from food poisoning outbreaks in Tokyo].

Since 1989, outbreaks of Salmonella ser. Enteritidis (S. Enteritidis) food poisoning have dramatically increased in Tokyo, and a total of 31 outbreaks has been reported in 1989. Twenty-one of these 31 outbreaks were caused by S. Enteritidis PT34, but 8 outbreaks were caused by S. Enteritidis PT4. After 1990 instead of SE PT34, food poisoning due to PT4, which was a very common phage type in the UK, has increased in Tokyo. Between 1989 and 1995, there were 144 food poisoning outbreaks caused by S. Enteritidis, and 64 of these outbreaks were by due to S. Enteritidis PT4, which was one of the main phage types in Tokyo. To characterize these strains of phage type (PT) 4,293 isolates from patients, and vehicle foods, eggs and environment in Tokyo were examined for plasmid DNA profiles, acid productivity from glycols (propylene and ethylene) and antimicrobial resistance patterns. Plasmid DNA was extracted by Kado's method, and analyzed by agarose gel electrophoresis. The acid productivity from propylene glycol or ethylene glycol were tested using Barsicow medium with 1% propylene glycol or ethylene glycol. Antimicrobial susceptibility to AM, CP, TC, SM, KM, NA, ST, FOM and NFLX was tested by the K-B disc method. The strains of PT4 were further subdivided into 9 types by those epidemiologic marker analysis. The prevalent pattern of PT4 strains was type A plasmid profile carrying only one plasmid (60 kb) and there were 2 kinds of antibiograms. One was SM resistant, while the other was susceptible. A total of 56 (87.5%) of 64 outbreaks was found to have been caused by these types of S. Enteritidis. Several kinds of egg-related foods were suspected as the vehicles of transmission among 24 outbreaks. Especially, in 5 outbreaks, S. Enteritidis strains were isolated both from patients and suspected food which were cooked with egg. This strongly suggests that these foods may be the potential source of infection in S. Enteritidis PT4 outbreaks.

Anti-Bacterial Agents↗

Changes in proteasome levels in spinach (Spinacia oleracea) seeds during imbibition and germination.

Proteasomes are the major cytosolic protease complexes responsible for energy-dependent and extra-lysosomal proteolysis. Ubiquitinated proteins are degraded by the 26S proteasome which is composed of a 20S proteasome and two regulatory complexes. Changes in the 20S and 26S proteasome activity levels and protein abundance in spinach seeds during imbibition and germination were examined by using glycerol density gradient centrifugation. The 26S proteasome activity level decreased transiently during imbibition, reached a minimum one day after starting imbibition, and then increased again. During the period of minimal accumulation, the protein being detected with an anti-26S proteasome antibody shifted to a lower sedimentation coefficient fraction. In contrast, the 20S proteasome activity level increased during imbibition, and remained high for two days. The change in the 20S proteasome level corresponded to the change in the 20S proteasome activity level.

Centrifugation, Density Gradient↗

[A case of traumatic hematoma in the basal ganglia: successful drainage with ultrasound-guided aspiration surgery via burr hole].

A case was reported of massive traumatic hematoma of the basal ganglia which was treated successfully by ultrasound-guided aspiration surgery. A 8-year-old girl was transferred to our hospital 20 minutes after a traffic accident. She was in stupor and had right hemiplegia. A skull x-rays showed right temporal linear fracture and a head CT disclosed left massive putaminal hematoma. Ultrasound-guided aspiration surgery via a burr hole was performed under general anesthesia and 30 ml of hematoma was successfully evacuated. Her symptoms and signs improved gradually and she was discharged on foot 3 weeks after the accident. The clinical feature and mechanism of development of the hematoma in this lesion, and usefulness of ultrasound-guided aspiration surgery were also discussed.

Basal Ganglia↗

Differentiation-dependent enhanced expression of protein phosphatase 2Cbeta in germ cells of mouse seminiferous tubules.

The presence of five distinct isoforms of protein phosphatase 2Cbeta (PP2Cbeta-1 approximately -5) is known. In this study, we demonstrate that the mRNA levels of PP2Cbeta-3, -4 and -5 and PP2Cbeta protein level increased during the course of the first wave of spermatogenesis in neonatal mouse testis. Northern blot and in situ hybridization analyses revealed that PP2Cbeta-3, -4 and -5 were expressed predominantly in pachytene spermatocytes and in more highly differentiated germ cells. The substrate specificity of PP2Cbeta-4 determined with artificial substrates differed from those of PP2Cbeta-3 and -5, suggesting that the difference in the structure of PP2Cbeta-3, -4 and -5 reflect their unique physiological functions in testicular germ cells.

Animals↗

Okadaic acid suppresses neural differentiation-dependent expression of the neurofilament-L gene in P19 embryonal carcinoma cells by post-transcriptional modification.

Mouse P19 embryonal carcinoma cells in aggregation culture in the presence of 10(-6) M retinoic acid followed by monolayer culture differentiate into nerve and glial cells. In this study, we demonstrated that the neurofilament-L (NF-L) mRNA and protein levels of these cells were enhanced in accordance with their retinoic acid-induced neural differentiation. Okadaic acid (OA) treatment of the cells markedly suppressed this differentiation-dependent NF-L gene expression increase and neurite outgrowth of the cells. Similar results were obtained when tautomycin was used instead of OA, suggesting that inhibition of protein phosphatase(s) is involved in the suppression of neural differentiation. OA treatment did not affect the NF-L gene transcription level, determined by the nuclear run-on transcription assay, but it did reduce the stability of both the 3.5- and 2.3-kilobase NF-L mRNAs. The expression and activity levels of protein phosphatase 2A (PP2A) and 2B (PP2B) but not protein phosphatase 1 (PP1) in P19 cells increased in accordance with the enhanced NF-L gene expression. The presence of OA in the culture medium during the course of the neural differentiation caused a reduced PP2A activity but not PP1 and PP2B activities of the cell extracts. On the other hand, both PP1 and PP2B activities but not PP2A activity of cell extracts were suppressed by the addition of cyclosporin A or FK506 in the culture medium. However, both cyclosporin A and FK506 treatments affected neither NF-L gene expression nor neurite outgrowth. These results demonstrate that the OA treatment inhibits the differentiation-dependent increase in NF-L gene expression by destabilizing its mRNAs and suggest that PP2A plays key roles in the differentiation-dependent enhanced expression of the NF-L gene and is the point of the action of OA.

Animals↗

Enhanced UV sensitivity of yeast cells induced by overexpression of Mg(2+)-dependent protein phosphatase alpha (type 2C alpha).

The UV sensitivity of wild-type Saccharomyces cerevisiae cells was increased 2-fold when rat Mg(2+)-dependent protein phosphatase alpha (protein phosphatase type 2C alpha) was overexpressed in the cells. The overexpression of this enzyme rendered the rad 18 mutant (defective in postreplication repair) more UV-sensitive than was observed in the wild-type cells. However, this increase in UV sensitivity disappeared when the host cells had a rad 1 mutation (defective in excision repair). These results suggest that the Mg(2+)-dependent protein phosphatase overexpressed in the yeast cells inhibited their excision repair system.

Animals↗

Molecular mechanism of null expression of aldehyde dehydrogenase-1 in rat liver.

In isozyme systems in general, the pattern of tissue-dependent expression of a given type of isozyme is uniform in various mammalian species. In contrast, a major cytosolic aldehyde dehydrogenase isozyme, termed ALDH1, which is strongly expressed in the livers of humans and other mammals, is hardly detectable in rat liver. Thirteen nucleotides existing in the 5'-promoter region of human, marmoset, and mouse ALDH1 genes are absent in the four rat strains examined. When the 13 nucleotides were deleted from a chloramphenicol acetyltransferase expression construct, which contained the 5'-promoter region of the human ALDH1 gene and a low-background promoterless chloramphenicol acetyltransferase expression vector, the expression activity was severely diminished in human hepatic cells. Thus, deletion of the 13 nucleotides in the promoter region of the gene can account for the lack of ALDH1 expression in rat liver.

Aldehyde Dehydrogenase↗

Pontine glioma with osteoblastic skeletal metastases in a child.

BACKGROUND: The development of systemic metastases from primary intracranial gliomas is rare. We report here a rare case of pontine glioma with osteoblastic skeletal metastases. CASE: This 12-year-old boy presented with a 4-month history of hoarseness, dysphagia, and a progressively ataxic gait. Cranial computed tomography (CT) and magnetic resonance imaging (MRI) revealed a brain stem tumor that was diagnosed as a low grade glioma by stereotactic biopsy. Twelve months later following chemotherapy and radiotherapy, neurologic examination and neuroradiologic studies disclosed a recurrence of the pontine glioma. Skeletal roentgenograms revealed widespread osteoblastic metastases in the skull, vertebral bodies, pelvis, and long bones. A specimen from the iliac bone demonstrated cells that were immunoreactive glial fibrillary acidic protein (GFAP). DISCUSSION: The mechanism of how glioma cells determine their biologic behavior at bony metastatic sites is not known. Infratentorial gliomas, which occur frequently in young patients and demonstrate active bony metabolism, may stimulate osteoblastic cells, and induce osteoblastic changes.

Bone Neoplasms↗

Nosocomial diarrhoea in the elderly due to enterotoxigenic Clostridium perfringens.

To diagnose sporadic diarrhoea due to Clostridium perfringens infection, faecal specimens from elderly patients were examined directly for C.perfringens enterotoxin using reverse passive latex agglutination assay, and then cultured for this organism. C.perfringens isolates from those samples were grouped by slide agglutination and by pulsed-field gel electrophoresis (PFGE). Fifty of the 60 isolates agglutinated with newly raised antiserum WX2 and 38 shared the same genomic PFGE pattern. Characteristics of the epidemics and experimental data suggest that the diarrhoea was caused by a nosocomial spread of C.perfringens, and not by a food-borne outbreak.

Aged↗

[Evolution of susceptibilities of Campylobacter jejuni isolated from diarrhoeal cases to fluoroquinolones in Tokyo].

Recently, the increase in the number of resistant strains of Campylobacter jejuni to fluoroquinolone has been reported in European countries. We also studied antimicrobial susceptibilities of 600 clinical isolates of Campylobacter jejuni isolated during a 6 year period from 1989 through 1994 in four Tokyo Metropolitan Hospitals. The susceptibility to 6 antimicrobial agents, norfloxacin (NFLX), ofloxacin (OFLX), ciprofloxacin (CPFX), nalidixic acid (NA), erythromycin (EM) and tetracycline (TC) were examined. The overall resistant rates were as follows: NFLX, 45 strains (7.5%); OFLX, 45 strains (7.5%); CPFX, 44 strains (7.3%); NA, 62 strains (10.3%); EM, 4 strains (0.6%) and TC, 259 strains (43.2%). The number of resistant strains to fluoroquinolones and NA has increased significantly since 1993 in Japan, but the susceptibility to erythromycin has still remained the same level during the past 6 years. The susceptibility to TC was variable, and MICs gave a bimobal distribution, as pointed out previously. The resistance pattern of NFLX, OFLX, CPFX and NA were observed most frequently in those isolates.

Anti-Infective Agents↗

[Characterization of Salmonella ser. enteritidis phage type 34 isolated from food poisoning outbreaks in Tokyo by epidemiological makers].

Since 1989, outbreaks of Salmonella ser. Enteritidis food poisoning has dramatically increased in Tokyo, and a total of 95 outbreaks has been reported between 1989 and 1993. As the results of examination of phage type by the method of Ward, et al., strains isolated from 31 (32.6%) of 95 outbreaks were phage type 34 which is a very rare type in Europe and America. To characterize these unique strains of phage type (PT) 34, 302 isolates obtained from patients and vehicle foods of the outbreaks, eggs and environments in Tokyo were examined for plasmid DNA profiles and antimicrobial resistance patterns. Plasmid DNA was extracted by the Kado's method, and analysed by agalose gel electrophoresis. Antimicrobial susceptibility was tested for CP, TC, SM, KM, ABPC, NA, SXT, FOM and NFLX by the K-B disc method. The strains of PT 34 were further subdivided to 4 types by plasmid profile and antibiogram. The prevalent pattern of PT 34 strains was type plasmid profile which carried 2 different plasmids (> 250 and 60 kb) and was streptomysin-resistant. A total of 28 (90.3%) of 31 outbreaks were found to be caused by this specific type of Salmonella ser. Enteritidis. Several kinds of egg-related foods were suspected for the vehicles of transmission among 12 outbreaks. Especially, in 5 outbreaks, this specific type (type B plasmid profile and streptomysin resistant) of Salmonella ser. Enteritidis strains were isolated both from patients and incriminated food which were cooked with eggs. The phase type of Salmonella ser. Enteritidis strains isolated from raw egg and poultry or environmental specimens from poultry farm showed that the prevailing phage types were almost the same as those of foods poisoning outbreaks. This strongly suggests that these foods may be considered as a source of infection in Salmonella ser. Enteritidis outbreaks.

Anti-Bacterial Agents↗

The transcriptional regulation of human aldehyde dehydrogenase I gene. The structural and functional analysis of the promoter.

Human cytosolic aldehyde dehydrogenase 1 (ALDH1) plays a role in the biosynthesis of retinoic acid that is a modulator for gene expression and cell differentiation. Northern blot analysis showed that liver tissue, pancreas tissue, hepatoma cells, and genital skin fibroblast cells expressed high levels of ALDH1. Sequence analysis showed that the 5'-flanking region contains a number of putative regulatory elements, such as NF-IL6, HNF-5, GATA binding sites, and putative response elements for interleukin-6, phenobarbital and androgen, in addition to a noncanonical TATA box (ATAAA) and a CCAAT box. Functional characterization of the 5'-regulatory region of the human ALDH1 gene was carried out by a fusion to the chloramphenicol acetyltransferase gene. A construct containing 2.6 kilobase pairs of the 5'-flanking region was efficiently expressed in hepatoma Hep3B cells, but not in erythroleukemic K562 cells or in fibroblast LTK- cells, which do not express ALDH1. Within this region, we define a minimal promoter (-91 to +53) that contains positive regulatory elements. The study using site-directed mutagenesis demonstrated that the CCAAT box region is the major cis-acting element involved in basal ALDH1 promoter activity in Hep3B cells. Gel mobility shift assays showed that NF-Y and other octamer factors bound CCAAT box and an octamer motif sequence, but not GATA site existing in the minimal promoter region. Two additional DNA binding activities associated with the minimal promoter were found in the nuclear extract from Hep3B cells, but not from K562 cells. These results offer the possible molecular mechanism of the cell type-specific expression of ALDH1 gene.

Aldehyde Dehydrogenase↗

Simultaneous expression of human CYP3A7 and N-acetyltransferase in Chinese hamster CHL cells results in high cytotoxicity for carcinogenic heterocyclic amines.

To investigate whether several food-derived heterocyclic amines are activated to genotoxic products in human fetal livers, cell lines stably expressing CYP3A7, a human fetus-specific form of cytochrome P450, NADPH-cytochrome P450 reductase, and human monomorphic or polymorphic N-acetyltransferase (NAT1 or NAT2) were established. The expression of CYP3A7 mRNAs and proteins was determined by RNA blot and immunoblot analyses, respectively. The introduction of CYP3A7 cDNA to CR-68 cells which had been transfected with guinea pig NADPH-cytochrome P450 reductase, NAT1, or NAT2 cDNA resulted in increased sensitivity of the cells to aflatoxin B1 compared to parental cells. The cytotoxicity assay for 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) showed that 7P-145 cells, which expressed the reductase, CYP3A7, and NAT2, were approximately 4-, 30-, and 14-fold more sensitive to respective IQ, MeIQ, and MeIQx than parental CR-68 cells. There were no clear differences in sensitivity to these compounds among CHL, CR-68, and the cells which expressed the reductase and CYP3A7 (7R-54), the reductase and NAT1 (CNM-4), the reductase and NAT2 (CNP-40), and the reductase, NAT1, and CYP3A7 (7M-124). From these results, it was suggested that both CYP3A7 and polymorphic NAT2 are required for mutagenic activation of several heterocyclic amines in human fetal livers.

Amines↗

Molecular cloning and expression of mouse mg(2+)-dependent protein phosphatase beta-4 (type 2C beta-4).

A full-length complementary DNA (cDNA) clone (pTK-3) encoding an isoform of Mg(2+)-dependent protein phosphatase beta (MPP beta-4) was isolated for the first time from a mouse melanocyte cDNA library. It was strongly suggested that the mRNA corresponding to the pTK-3 insert was a splicing variant of a single pre-mRNA that also encodes MPP beta-1 and -2 (T. Terasawa, T. Kobayashi, T. Murakami, M. Ohnishi, S. Kato, O. Tanaka, H. Kondo, H. Yamamoto, T. Takeuchi, and S. Tamura, 1993, Arch. Biochem. Biophys. 307, 342-349). The amino acid sequence of MPP beta-4 differed from those of MPP beta-1 and -2 only at the carboxyl terminal region. Analysis by reverse transcriptase polymerase chain reaction (RT-PCR) revealed that MPP beta-4 mRNA was expressed only in testis and intestine and not in other mouse tissues tested. Specific expression of the mRNA signals of two other isoforms of MPP beta, MPP beta-3 and -5 (a novel isoform), in testis and intestine was also demonstrated by the RT-PCR. The carboxyl terminal region of MPP beta-5 was found to have a chimera structure composed of part of MPP beta-1 and part of MPP beta-3. The recombinant MPP beta-3 and -4 and the putative MPP beta-5 expressed in Escherichia coli cells exhibited Mg(2+)-dependent and okadaic acid-insensitive protein phosphatase activities. It was demonstrated that the mRNA expression levels of MPP beta-3, -4, and -5 alter according to the maturation of mouse testis. These results suggest that the complex structure of MPP beta isoforms and their tissue- and developmental stage-specific expression reflect the variety of their physiological functions.

Alternative Splicing↗