M Watanabe

Shimane University, Matsu, Shimane Prefecture, Japan

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Publications (315)1080.87 Total impact

  • Clinical and Experimental Immunology - CLIN EXP IMMUNOL. 01/2008; 100(3):446-451.
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    ABSTRACT: Purpose: CYP3A7 gene expression is increased in some adults as those with hepatocellular carcinoma, although this P450 is dominant with fetus or neonate, while CYP3A4 dominant with normal adult. We hypothesized the discrepancy of promoter sequence in up-stream between these P450 genes play important role, and attempted to investigate this in the cell line, especially focusing on p53.Method: (1) HepG2 and Huh7 (human hepatocellular carcinoma), Huh6 (hepatoblastoma), and other cancer cell lines were purchased and cultured to examine the expression of CYP3A7 mRNA and p53 protein. (2) p53 sense (SE) and p53 antisense (AS) were designed by the human p53 cDNA sequence report (GenBank), and of those influence in HepG2 cells on expression of CYP3A7 mRNA, CYP3A4 mRNA with RT-PCR method, and also of p53 protein with Western blot analysis were analyzed, respectively. Results: (1) HepG2 and Huh7 cell line expressed increased CYP3A7 mRNA and p53 protein, while other cell lines did not remarkably. (2) p53 AS decreased CYP3A7 mRNA and p53, while not with p53 SE in HepG2 cells.Discussion: There is p53 recognizing region in up-stream of 5` promoter region of CYP3A7 gene (-328 to -337bp), though not of CYP3A4 gene. This fact and the present results will suggest the importance of p53 in CYP3A7 regulation in liver tissue.
    Clinical Pharmacology &#38 Therapeutics 01/2004; 75(2):P17-P17. · 6.85 Impact Factor
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    ABSTRACT: Precise positioning, which is the base technology for ultra-precise treatments, is highly essential in semiconductor LSI circuits, bio-cells, molecular sensors, and quantum devices. Conflicting performances, high-speed, long-stroke, and high-resolution have been achieved by using die nonresonant ultrasonic motor (NRUSM) controlled under the piezoelectric effect. One of the notable features of the NRUSM is do the frequency of the driving voltage can be varied in a wide range, which means the motion control of the NRUSM can be done by both the amplitude and frequency The maximum stage feed velocity is 140 mm/s with average acceleration of 2200 mm/s<sup>2</sup> in the 100 mm stroke under open-loop control. 100 nm step response with ±0.69 nm positioning accuracy is completed in 0.35 s under closed-loop control.
    Ultrasonics Symposium, 2002. Proceedings. 2002 IEEE; 11/2002
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    ABSTRACT: In the present study, morphological changes of the exocrine pancreas in rats after pancreatic duct ligation were examined with light microscopy (hematoxylin-eosin, TUNEL, and PCNA staining) and scanning electron microscopy in order to elucidate the effects of increased pancreatic duct pressure. On the fifth day after pancreatic duct ligation, ductular proliferation, periductal fibrosis, and disappearance of acini were observed. TUNEL and PCNA staining demonstrated many apoptotic acinar cells and proliferating ductal cells immediately after ligation, which reached a maximal number on the 2nd or 3rd day. Tortuous or helical interlobular pancreatic ducts with inner surfaces containing many crater-like depressions and long cilia were found after ligation. These changes were almost identical to those observed in the pancreatic tissue of model chronic pancreatitis rats, WBN/Kob rats, and stroke-prone spontaneously hypertensive (SHRSP) rats. In summary, the morphological changes observed after pancreatic duct ligation were similar to those of chronic pancreatitis, therefore, the characteristic changes of pancreatic ducts observed in chronic pancreatitis may be caused by increased pancreatic duct pressure.
    Histology and histopathology 11/2002; 17(4):1033-41. · 2.28 Impact Factor
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    ABSTRACT: The three-dimensional structure of the pancreatic ductular system (from the intercalated duct to the intercellular secretory canaliculus) is controversial and unclear. The aim of this study is to reveal the three-dimensional structure of the pancreatic ductular sysytem at the onset of pancreatitis. One day following rat pancreatic duct ligation, dilated lumina from the pancreatic ductular system were reconstructed by light microscopic and scanning electron microscopic examination of pancreatic tissue serial sections. The existence of the intra-acinar duct, which is formed only by centroacinar cells and interconnects the adjacent central lumina in an acinus, was demonstrated. The intercellular secretory canaliculi, which are the terminal parts of the pancreatic ductular system, anastomose and end blindly in the intercellular space located between adjacent lateral surfaces of the acinar cells. The intercalated ducts, the intra-acinar ducts, the central lumina, and the intercellular secretory canaliculi are arranged together in a complex connecting and branching system. However, there were no anastomoses found among the central lumina or acini.
    Histology and histopathology 02/2002; 17(1):107-12. · 2.28 Impact Factor
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    ABSTRACT: Helicobacter pylori infection induces T helper-1 immune responses in inflamed mucosa. However, the role of T cell-mediated cytotoxicity in the induction of epithelial apoptosis is still unclear. The aim of this study was to investigate the involvement of the Fas/Fas ligand (Fas/Fas-L) system in the H. pylori-infected gastric corpus. Gastric fundic biopsy specimens were taken from patients with and without H. pylori infection. The expression of Fas and Fas-L was examined by immunohistochemistry and RT-PCR. Subsets of gastric infiltrating T cells in the biopsy specimens were studied by immunohistochemistry and flow cytometry. In histological sections, apoptosis was detected by the TUNEL method. We studied the in vitro expression of Fas-L in peripheral T cells after stimulation with H. pylori antigen and interferon-gamma (IFN-gamma). The Fas-mediated in vitro cytotoxicity of activated T cells was assessed by flow cytometry. The numbers of CD4+ and CD8+ T cells were greater in H. pylori-infected subjects. Fas expression was abundantly increased on fundic gland epithelium, and Fas-L was detected on lamina propria mononuclear cells in H. pylori-infected mucosa. TUNEL-positive epithelial cells were also increased in H. pylori-infected subjects. H. pylori antigen and IFN-gamma induced Fas-L mRNA expression in both CD4+ and CD8+ T cells. In cytotoxic assay, activated T cells induced apoptosis in AGS cells, which could be significantly inhibited by neutralizing Fas-L antibody. T cell-mediated cytotoxicity via Fas/Fas-L signaling may contribute to the induction of apoptosis in gastric epithelial cells during H. pylori infection.
    Helicobacter 01/2002; 6(4):283-93. · 3.51 Impact Factor
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    ABSTRACT: We previously reported that gallic acid (3,4,5-trihydroxybenzoic acid), a naturally occurring plant phenol, can induce apoptosis in four kinds of human lung cancer cell lines in vitro. The present study further investigated the in vivo anti-tumor effects of orally administered gallic acid. Gallic acid reduced cell viability of LL-2 mouse lung cancer cells in vitro dose dependently, with a 50% inhibitory concentration (IC50) value of around 200 microM. C57Black mice were transplanted with LL-2 cells, and administered gallic acid (1 mg/ml in drinking water, ad libitum) and/or cisplatin (4 mg/kg i.p. injection, once a week). The average weight of the transplanted tumors, obtained at 29 days after transplantation, in the mice of control, gallic acid-treated cisplatin-treated and cisplatin plus gallic acid-treated groups was 4.02, 3.65, 3.19 and 1.72 g, respectively. The average tumor weight of the mice treated with cisplatin combined with gallic acid was significantly smaller than that of the control group (p<0.05). The amount of apoptotic cells in the tumor tissues of mice treated with gallic acid and/or cisplatin was significantly higher than those of the control mice. Combination of gallic acid and cisplatin increased the tumor cell apoptosis compared with the treatment with cisplatin alone. The present findings suggest that the combination of gallic acid with an anti-cancer drug, including cisplatin, may be an effective protocol for lung cancer therapy.
    Anti-Cancer Drugs 12/2001; 12(10):847-52. · 2.23 Impact Factor
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    ABSTRACT: Helicobacter pylori (H. pylori) infection is known to affect the gastric microcirculation, and cetraxate is reported to accelerate gastric ulcer healing, possibly by augmenting gastric mucosal blood flow (MBF). The aim of this study is to clarify the effect of H. pylori infection and cetraxate on MBF during gastric ulcer healing. Forty-two patients who had undergone endoscopic mucosal resection (EMR) were studied. Mucosal blood flow was measured by the use of a laser Doppler flowmeter in the surrounding mucosa and at the ulcer margin, before, 1 day, 1 week and 4 weeks after EMR. Helicobacter pylori infection was confirmed by the use of bacterial culture and histology. After EMR, patients were randomly assigned to receive 30 mg lansoprazole (u.i.d; L-regimen) or 30 mg lansoprazole (u.i.d.) with 200 mg cetraxate (q.i.d; LC-regimen) for 4 weeks. The MBF ratio (MBF at ulcer margin/MBF in surrounding mucosa) 1 week after EMR was significantly lower than that before or 4 weeks after EMR only in H. pylori-positive patients treated with the L-regimen. No such decrease in MBF was observed after 1 week in H. pylori-positive patients treated with the LC-regimen or in H. pylori-negative patients. A transient decrease in MBF was detected at the ulcer margin during healing of EMR-induced ulcers in H. pylori-infected patients. Cetraxate seemed to prevent this decrease in MBF.
    Journal of Gastroenterology and Hepatology 12/2001; 16(11):1211-6. · 3.33 Impact Factor
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    ABSTRACT: Nocturnal gastric acid breakthrough (NAB) is defined as an intragastric pH < 4.0 lasting more than 1 h during the night in patients taking a proton pump inhibitor (PPI). Gastroesophageal reflux disease (GERD) patients with nocturnal gastroesophageal acid reflux accompanied by NAB are thought to be refractory to PPI treatment. The aim of this study was to endoscopically identify the patients with predominant nocturnal gastroesophageal acid reflux. The subjects were 37 patients with erosive reflux esophagitis (Los Angeles classification (LA) grade A, 12; B, 10; C, eight; and D, seven cases) and a control group of 20 patients without esophagitis. The results of ambulatory 24 h gastric and esophageal pH monitoring were compared among different grades of esophagitis. Gastroesophageal reflux during 24 h in patients with high-grade esophagitis was more frequent than for patients with low-grade esophagitis or no esophagitis. Although the length of esophageal acid exposure (percentage time with pH < 4.0) in patients with grade A or without esophagitis was longer in the daytime, that in patients with grades C and D was longer during the night. The reason for the delayed nocturnal acid exposure was the longer nocturnal acid clearance in high-grade reflux esophagitis. Nocturnal exposure of the esophagus to acid occurs frequently in patients with LA grades C and D esophagitis. Thus, the existence of NAB with resulting nocturnal acid reflux should be considered when the patient with high-grade esophagitis shows resistance to PPI treatment.
    Journal of Gastroenterology and Hepatology 11/2001; 16(11):1191-6. · 3.33 Impact Factor
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    ABSTRACT: Pierisin-1 is a potent apoptosis-inducing protein derived from the cabbage butterfly, Pieris rapae. It has been shown that pierisin-1 has an A small middle dotB structure-function organization like cholera or diphtheria toxin, where the "A" domain (N-terminal) exhibits ADP-ribosyltransferase activity. The present studies were designed to identify the target molecule for ADP-ribosylation by pierisin-1 in the presence of beta-[adenylate-(32)P]NAD, and we found DNA as the acceptor, but not protein as is the case with other bacteria-derived ADP-ribosylating toxins. ADP-ribosylation of tRNAs from yeast was also catalyzed by pierisin-1, but the efficiency was around 110 of that for calf thymus DNA. Pierisin-1 efficiently catalyzed the ADP-ribosylation of double-stranded DNA containing dG small middle dotdC, but not dA small middle dotdT pairs. The ADP-ribose moiety of NAD was transferred to the amino group at N(2) of 2'-deoxyguanosine to yield N(2)-(alpha-ADP-ribos-1-yl)-2'-deoxyguanosine and its beta form, which were determined by several spectral analyses including (1)H- and (13)C-NMR and mass spectrometry. The chemical structures were also ascertained by the independent synthesis of N(2)-(D-ribos-1-yl)-2'-deoxyguanosine, which is the characteristic moiety of ADP-ribosylated dG. Using the (32)P-postlabeling method, ADP-ribosylated dG could be detected in DNA from pierisin-1-treated HeLa cells, in which apoptosis was easily induced. Thus, the targets for ADP-ribosylation by pierisin-1 were concluded to be 2'-deoxyguanosine residues in DNA. This finding may open a new field regarding the biological significance of ADP-ribosylation.
    Proceedings of the National Academy of Sciences 11/2001; 98(22):12414-9. · 9.74 Impact Factor
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    ABSTRACT: We investigated the effect of antisense oligodeoxynucleotides (AS ODN) against tyrosine hydroxylase (TH) on hypertension and sympathetic nervous system activity in spontaneously hypertensive rats (SHR). Systolic blood pressure (SBP) in SHR treated with TH AS ODN (50, 200 microg/rat, i.v.) was significantly lower than that of control SHR. Epinephrine and norepinephrine levels, TH activity, and TH protein levels in the adrenal medulla of SHR were reduced concomitant with TH AS ODN treatment-induced changes in SBP. In contrast, TH AS ODN (200 microg/rat) had no effect on SBP in Wistar-Kyoto rats (WKY), despite significantly decreased catecholamine levels, TH activity, and TH protein levels. These findings suggest that peripheral systemic injection of TH AS ODN may be effective as hypotensive therapy in SHR.
    Journal of Hypertension 11/2001; 19(10):1769-73. · 3.81 Impact Factor
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    ABSTRACT: Clinical relevance of nucleotide changes in precore and basal core promoters in the hepatitis B virus genome during hepatitis B e antigen seroconversion may be overstated. The authors investigated the existence and changes in the relative proportion of variants to wild virus that occur with seroconversion. Sera from 30 school-aged long-term hepatitis B virus carriers, including 11 tested before and after seroconversion during 1 to 8 years of follow-up, were evaluated for variations in nucleotide sequences of the basal core promoter (T1762 and A1764), precore region (A1869), and carboxyl-terminus of the X region of the hepatitis B virus genome using an amplification refractory mutation detection system with mutant-specific primers. All variants were found to already exist before seroconversion at various wild-type/mutant ratios. The positive rates of these variants were not changed with loss of hepatitis B e antigen. Although there was a relative increase in the concentration of these mutants in wild-type/mutant mixed populations, most patients with only a wild-type population maintained the same pattern after loss of hepatitis B e antigen. Our results indicate that hepatitis B virus exists as a quasi species, and correlations of nucleotide sequences with clinical and serologic findings must be done with caution.
    Journal of Pediatric Gastroenterology and Nutrition 10/2001; 33(3):301-6. · 2.20 Impact Factor
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    ABSTRACT: A new IgE-reactive glycoprotein with a molecular size of 60 kDa was isolated from wheat flour. The N-terminal amino acid sequence of the protein was LDPDESEXVTRYFRIR. The 8th amino acid residue would have been Asn to which the peroxidase-type glycochain was attached. The IgE-binding activity of the glycoprotein was rendered negligible by the enzymatic treatment applied for hypoallergenic flour production.
    Bioscience Biotechnology and Biochemistry 10/2001; 65(9):2102-5. · 1.27 Impact Factor
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    ABSTRACT: Expression of the KAI1 gene, a metastasis-suppressor for prostate cancer, is reduced in all foci of prostatic metastasis. The altered regulatory mechanism is not strongly related to mutations or allelic losses of the KAI1 gene in prostate tumors. Since transcriptional silencing of genes has been found to be caused by epigenetic mechanisms, we have investigated the involvement of this epigenetic regulation of KAI1 expression in prostate cancers. The methylation status of the KAI1 promoter region was examined by restriction-enzyme digestion and sequencing, after amplifying a 331-bp fragment in the GC-rich promoter region from 4 human prostate cancer cell lines treated with bisulfite. The same 4 cell lines were also exposed to various concentrations of the demethylating agent, 5-aza-2'-deoxycytidine (5-AzaC) and / or the histone deacetylase inhibitor, trichostatin A (TSA). To clarify the influence of epigenetic modification on reduced KAI1 mRNA expression in the tumor cells, RT-PCR and northern-blot analyses were performed. Bisulfite-sequencing data showed a few methylated CpG islands in the promoter. RT-PCR analysis of 5-AzaC and / or TSA-treated cells indicated reversal of suppression of KAI1 transcription in two cell lines (PC-3 and DU-145), although the expression could not be detected by northern blots. From these results, it is suggested that epigenetic change is not the main mechanism of KAI1 down-regulation, though there remains a possibility that methylation in a more upstream region might be associated with this regulation.
    Japanese journal of cancer research: Gann 10/2001; 92(9):947-51.
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    ABSTRACT: Brown Norway rats were immunized with gluten, and then fed a diet containing hypoallergenic flour or an amino acid mixture. The rats were then made to inhale a solubilized gluten to induce gluten-specific bronchial asthma. The antibody levels in the serum of rats were measured by ELISA, and cell counts were done on cytospin preparations of bronchoalveolar lavage fluid. Body weight was decreased after allergen challenge in rats fed the amino acid mixture but not in rats fed the hypoallergenic flour. Antibody levels in the serum were significantly lower in rats fed hypoallergenic flour than in those fed the amino acid mixture. Differential cell counts in the bronchoalveolar lavage fluid showed that the numbers of eosinophils, lymphocytes, and neutrophils were significantly lower in rats fed the hypoallergenic flour than in those fed the amino acid mixture. These results suggest that hypoallergenic flour actively suppresses the allergic reactions, probably by inducing oral tolerance.
    Bioscience Biotechnology and Biochemistry 09/2001; 65(8):1729-35. · 1.27 Impact Factor
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    ABSTRACT: The cytoprotective agent, ecabet sodium, inhibits urease activity and growth of Helicobacter pylori. To evaluate the efficacy and safety of ecabet sodium-based eradication of H. pylori infection, compared with a lansoprazole-based regimen, in a randomized multicentre study. A total of 120 H. pylori-positive patients were assigned to one of two treatment regimens for 2 weeks: ecabet sodium 1 g b.d., amoxicillin 500 mg t.d.s. and clarithromycin 400 mg b.d. (EAC: 60 patients); or lansoprazole 30 mg (o.m.) with the same antimicrobial agents (LAC: 60 patients). Cure of infection was assessed by a 13C-urea breath test 1 month after completion of treatment. One patient in the EAC group and two in the LAC group did not complete therapy because of an adverse event, and three did not undergo the 13C-urea breath test. Cure rates for the intention-to-treat, all-patients-treated and per protocol analysis in the EAC group were 85%, 86% and 88%, respectively, whereas those in the LAC group were 85%, 88% and 91%. There were no significant differences in cure rate or adverse events between the two regimens. Ecabet sodium in combination with amoxicillin and clarithromycin is as effective as lansoprazole-based eradication therapy for H. pylori.
    Alimentary Pharmacology & Therapeutics 09/2001; 15(8):1187-91. · 4.55 Impact Factor
  • M Watanabe, N Kohge, T Kaji
    Annals of internal medicine 08/2001; 135(2):142-3. · 13.98 Impact Factor
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    ABSTRACT: Poly(ADP-ribose) polymerase (Parp) monitors DNA strand breaks and poly(ADP-ribosyl)ates nuclear proteins using NAD as a substrate. The participation of Parp in DNA damage responses has been demonstrated by recent studies using Parp knockout mice. On the other hand, accumulated evidence has shown that Parp is involved in the regulation of gene expression and cell differentiation. In this study, the role of Parp in tumorigenesis and differentiation was studied with Parp-/- embryonic stem (ES) cells. When Parp+/+, Parp+/-, and Parp-/- ES cells were injected subcutaneously into nude mice, teratocarcinoma-like tumors developed from ES cells. However, only tumors derived from Parp-/- ES cells showed trophoblast giant cells (TGCs) containing single or multiple megalo-nuclei. These TGCs are located in a large blood-lake like hemorrhage. This example suggests that Parp is not essential for tumor formation, however, it is involved in trophoblastic cell differentiation and could consequently affect tumor phenotype.
    Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 07/2001; 477(1-2):111-7. · 3.90 Impact Factor
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    ABSTRACT: Steroid hormones, especially testosterone, play important roles in the carcinogenesis of prostate cancer, and several studies have reported changes in risk with polymorphisms of genes involved in steroid metabolism. One example is the CYP17 gene, which has a polymorphic T-to-C substitution in the 5'-untranslated region giving rise to A1 (T) and A2 (C) alleles. Steroid 5alpha-reductase type II (SRD5A2), which converts testosterone to the metabolically more active dihydrotestosterone, exhibits 2 polymorphisms: V89L, which substitutes leucine for valine at codon 89, and A49T, which substitutes threonine for alanine at codon 49. We therefore designed a case-control study of 105 prostate-cancer patients and 210 controls with benign prostatic hyperplasia for the purpose of investigating the association between prostate-cancer risk and polymorphisms in the SRD5A2 and CYP17 genes among the Japanese. The frequency of the CYP17 A2/A2 genotype in cases (18.8%) was higher than in controls (14.5%). Compared with the A1/A1 genotype, the odds ratio for the A2/A2 genotype was 2.39 (95% confidence interval 1.04-5.46, p = 0.04). The frequency of the SRD5A2 LL genotype in cases (29.3%) was also slightly higher than in controls (24.6%), but this was not significant. Regarding the A49T polymorphism of SRD5A2, we could not detect the T allele in any of the examined samples. These data suggest a significant association between the CYP17 polymorphism and prostate-cancer risk among the Japanese.
    International Journal of Cancer 07/2001; 92(5):683-6. · 6.20 Impact Factor
  • The American Journal of Gastroenterology 07/2001; 96(6):1941-3. · 7.55 Impact Factor

Publication Stats

6k Citations
1,080.87 Total Impact Points

Institutions

  • 1987–2002
    • Shimane University
      • Department of Internal Medicine
      Matsu, Shimane Prefecture, Japan
  • 2001
    • Hokkaido University
      • Graduate School of Agriculture
      Sapporo-shi, Hokkaido, Japan
    • Gifu University
      Gihu, Gifu, Japan
    • Drexel University
      • Department of Physics
      Philadelphia, Pennsylvania, United States
  • 1997–2001
    • Harvard Medical School
      • Department of Cell Biology
      Boston, Massachusetts, United States
  • 1996–2001
    • The University of Tokyo
      • • Institute of Molecular and Cellular Biosciences
      • • Department of Internal Medicine
      • • Division of Internal Medicine
      Tokyo, Tokyo-to, Japan
  • 1995–2001
    • National Cancer Center, Japan
      • Endoscopy Division
      Edo, Tōkyō, Japan
    • Tokyo Gakugei University
      Koganei, Tōkyō, Japan
    • YAMASA Corporation
      Tiba, Chiba, Japan
  • 1991–2001
    • St. Marianna University School of Medicine
      • Department of Pharmacology
      Kawasaki, Kanagawa-ken, Japan
  • 1988–2001
    • Tottori University
      • • Faculty of Medicine
      • • Third Department of Internal Medicine
      Tottori, Tottori-ken, Japan
  • 1996–2000
    • Mie University
      • • Department of Pathophysiology
      • • Department of Neurology
      • • School of Medicine
      Tu, Mie, Japan
  • 1994–2000
    • National Defense Medical College
      • • Division of Radiology
      • • Department of Surgery
      Tokorozawa, Saitama-ken, Japan
    • National Institute of Health Sciences, Japan
      • Division of Division of Genetics and Mutagenesis
      Edo, Tōkyō, Japan
    • Institute of Environmental Toxicology
      Ibaragi, Ōsaka, Japan
  • 1998–1999
    • Tohoku University
      • Department of Pharmacology
      Japan
    • Keio University
      • Department of Surgery
      Tokyo, Tokyo-to, Japan
  • 1992
    • Osaka Prefectural Institute of Public Health
      Ōsaka, Ōsaka, Japan
  • 1990–1991
    • Finnish Institute of Occupational Health
      Helsinki, Southern Finland Province, Finland
  • 1982–1985
    • National Astronomical Observatory of Japan
      • Division of Optical and Infrared Astronomy
      Edo, Tōkyō, Japan