Yoshihiko Kominato

Gunma University, Maebashi, Gunma Prefecture, Japan

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Publications (83)223.15 Total impact

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    ABSTRACT: Case history A 3-month-old infant was found dead in his bed. Postmortem CT scan suggested fatty attenuation in the liver parenchyma, but no other potentially fatal changes were found. To clarify the cause of death, a medicolegal autopsy was carried out. Autopsy findings Internal examination confirmed the presence of liver steatosis as well as hepatomegaly. There were no other significant findings including encephalitis or brain edema. Mass spectrometry analysis To clarify the mechanism underlying lipid accumulation in the liver, matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) analysis was conducted. This indicated significant accumulation of C14:1-acylcarnitine in the liver of the deceased, suggesting very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency. Genetic analysis To find the cause of the VLCAD deficiency, genetic analysis of the responsible gene, acyl-CoA dehydrogenase, very long chain (ACADVL), was performed. This revealed two novel mutations that may have accounted for the disease. Conclusion Combination of these data revealed that the liver steatosis in this case might have been caused by VLCAD deficiency based on genetic mutations of ACADVL. Thus, the deceased might have been vulnerable to energy crisis and sudden infant death. The present findings show that MALDI-IMS analysis as well as genetic analysis can be useful for elucidating the cause of death.
    Forensic Science International. 09/2014;
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    ABSTRACT: Introduction The human ether-à-go-go-related gene (hERG) encodes the α-subunit of a cardiac potassium channel. Various mutations of hERG, including missense mutations, have been reported to cause long QT syndrome (LQTS) and severe arrhythmic disorders such as sudden cardiac death. We identified a novel hERG frameshift mutation (hERG(ΔAT)) in the S5-pore region from a LQTS patient who died suddenly and analyzed its genetic profile and the molecular and electrophysiological behaviors of the protein product to assess the pathogenicity of hERG(ΔAT). Methods and results We performed direct sequencing of hERG and evaluated its transcript level by using a whole blood sample from the patient. We performed immunoblotting, immunocytochemistry, and patch-clamp recordings of HEK-293 T cells transfected with hERG(ΔAT), wild-type hERG (hERG(WT)), or both. The patient demonstrated an AT deletion (c.1735_1736del) in hERG and a decrease inhERG mRNA transcripts. HEK-293 T cells showed lower production and cell surface expression of hERG(ΔAT) compared with hERG(WT) protein. In addition, the hERG(ΔAT) protein failed to form functional channels, while the activation kinetics of functional channels, presumably consisting of hERG(WT) subunits, were unaffected. Conclusion The ΔAT mutation may decrease the number of functional hERG channels by impairing the posttranscriptional and posttranslational processing of the mutant product. This decrease may partly explain the cardiac symptoms of the patient who was heterozygous for hERG(ΔAT).
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    ABSTRACT: Background and objectivesAn erythroid cell-specific regulatory element, referred to as the +5.8-kb site, has been identified in the first intron of the human ABO blood group gene. Subsequent studies have revealed involvement of deletion or mutation at the site in phenotypes Am, Bm and ABm. We investigated the molecular mechanisms involved in the A3 and B3 phenotypes. Materials and methodsGenomic DNAs were prepared from peripheral blood of seven A3 individuals and twelve B3 or AB3 individuals, and the nucleotide sequences were investigated using PCR and sequencing. Promoter assays were performed with K562 cells. ResultsTwo single point-mutations at +5893 or +5909 in the site on the A-allele were found in A3 individuals, while promoter assays revealed decreased activity at the site as a result of each substitution. In two B3 individuals, a single point-mutation at −77 in the ABO promoter on the B-allele was found, and the substitution was demonstrated to reduce the promoter activity. Conclusion Nucleotide substitutions in the transcriptional regulatory elements such as the +5.8-kb site and the ABO promoter appear to decrease transcription from the A- and B-alleles, resulting in reduction in A- and B-antigen expression in A3 and B3, respectively.
    Vox Sanguinis 03/2014; · 2.85 Impact Factor
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    ABSTRACT: It is essential for medical students to learn and comprehend human anatomy in three dimensions (3D). With this in mind, a new system was designed in order to integrate anatomical dissections with diagnostic computed tomography (CT) radiology. Cadavers were scanned by CT scanners, and students then consulted the postmortem CT images during cadaver dissection to gain a better understanding of 3D human anatomy and diagnostic radiology. Students used handheld digital imaging and communications in medicine viewers at the bench-side (OsiriX on iPod touch or iPad), which enabled "pixel-to-tissue" direct comparisons of CT images and cadavers. Students had lectures and workshops on diagnostic radiology, and they completed study assignments where they discussed findings in the anatomy laboratory compared with CT radiology findings. This teaching method for gross and radiological anatomy was used beginning in 2009, and it yielded strongly positive student perspectives and significant improvements in radiology skills in later clinical courses. Anat Sci Educ. © 2013 American Association of Anatomists.
    Anatomical Sciences Education 01/2014;
  • 01/2014;
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    ABSTRACT: Blood group-related glycans determining ABO and Lewis blood groups are known to function as attachment factors for most of the norovirus (NoV) strains. To identify binding specificity of each NoV, recombinant norovirus-like particles (VLPs) and human saliva samples with different ABO, Lewis phenotypes and secretor status have been commonly applied. When binding specificities of VLPs prepared from 16 different genotypes of NoVs in GI and GII genogroups were characterized in samples of human gastric mucosa compared to human saliva based on blood group phenotypes, considerable differences were observed for several strains. Novel binding specificities determined by an ELISA using preparations from human gastric mucosa were also ascertained by immunohistochemical analyses using human jejunal mucosa, widely believed to be susceptible to NoV infection. Further, A, B and O(H) blood group substances prepared from porcine and squid tissues were found to be effective for preventing ABO blood group-specific binding of VLPs to both saliva and mucosa samples. Therefore, these blood group substances might have potential for the prevention and treatment of NoV infection.
    PLoS ONE 01/2014; 9(2):e89071. · 3.53 Impact Factor
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    ABSTRACT: The objectives of this study were to evaluate all of the non-synonymous single nucleotide polymorphisms (SNPs) in the DNase 1 and 1L3 genes, potentially implicated in autoimmune diseases, as a functional SNP in terms of alteration of the activity levels. We examined the genotype distributions of the 32 and 20 non-synonymous SNPs in DNASE1 and DNASE1L3, respectively, in the 3 ethnic groups, and the effect of these SNPs on the DNase activities. Among a total of 44 and 25 SNPs including those characterized in our previous studies [Yasuda et al., Int. J. Biochem. Cell Biol. 42 (2010) 1216-1225; Ueki et al. Electrophoresis, 32 (2012) 1465-1472], only 4 and 1, respectively, exhibited genetic heterozygosity in one or all of the ethnic groups examined. On the basis of alterations in the activity levels resulting from the corresponding amino acid substitutions, 11 activity-abolishing and 11 activity-reducing SNPs in DNASE1, and 2 activity-abolishing and 5 activity-reducing SNPs in DNASE1L3 were confirmed as a functional SNP. Phylogenetic analysis showed that all of the amino acid residues in activity-abolishing SNPs were completely or well conserved in animal DNase I and 1L3 proteins. Although almost all of non-synonymous SNPs in both genes that affected the catalytic activity showed extremely low genetic heterogeneity, it seems plausible that a minor allele of 13 activity-abolishing SNPs producing a loss-of-function variant in both the DNase genes would be a direct genetic risk factor for autoimmune diseases. These findings may have clinical implications in relation to the prevalence of autoimmune diseases. This article is protected by copyright. All rights reserved.
    FEBS Journal 11/2013; · 4.25 Impact Factor
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    ABSTRACT: An erythroid cell-specific regulatory element, referred to as the +5·8-kb site, had been identified in the first intron of the human ABO blood group gene. Subsequent studies revealed that either a 5·8-kb deletion including the +5·8-kb site or disruption of a GATA factor binding motif at the site was present in all Bm and ABm individuals examined. We investigated the molecular mechanism of the Am phenotype, which is analogous to the Bm phenotype. Genomic DNAs were prepared from peripheral blood of two Am individuals, and the nucleotide sequences were investigated using PCR and direct sequencing. Electrophoretic mobility shift assay (EMSA) and promoter assay with K562 cells were carried out. A novel 23-bp nucleotide deletion was found at the +5·8-kb site in both individuals. EMSAs demonstrated binding of the transcription factor RUNX1 to the nucleotides within the deletion. Promoter assays showed that the deletion reduced the transcriptional activity of the +5·8-kb site. Deletion of the 23-bp nucleotides including the RUNX1 binding site decreases transcription of the A allele, resulting in the reduction in A antigen expression in the Am phenotype.
    Vox Sanguinis 09/2013; · 2.85 Impact Factor
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    ABSTRACT: BACKGROUND: The ABO blood group is important in blood transfusion. Recently, an erythroid cell-specific regulatory element has been identified in the first intron of ABO using luciferase reporter assays with K562 cells. The erythroid cell-specific regulatory activity of the element was dependent upon GATA-1 binding. In addition, partial deletion of Intron 1 including the element was observed in genomic DNAs obtained from 111 Bm and ABm individuals, except for one, whereas the deletion was never found among 1005 individuals with the common phenotypes. STUDY DESIGN AND METHODS: In this study, further investigation was performed to reveal the underlying mechanism responsible for reduction of B antigen expression in the exceptional Bm individual. Peptide nucleic acid-clamping polymerase chain reaction was carried out to amplify the B-related allele, followed by sequence determination. Electrophoretic mobility assays and promoter assays were performed to examine whether a nucleotide substitution reduced the binding of a transcription factor and induced loss of function of the element. RESULTS: Sequence determination revealed one point mutation of the GATA motif in the element. The electrophoretic mobility shift assays showed that the mutation abolished the binding of GATA transcription factors, and the promoter assays demonstrated complete loss of enhancer activity of the element. CONCLUSION: These observations suggest that the mutation in the GATA motif of the erythroid-specific regulatory element may diminish the binding of GATA transcription factors and down regulate transcriptional activity of the element on the B allele, leading to reduction of B antigen expression in erythroid lineage cells of the Bm individual.
    Transfusion 04/2013; · 3.53 Impact Factor
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    ABSTRACT: INTRODUCTION: The human ether-à-go-go-related gene (hERG) encodes the α-subunit of a cardiac potassium channel. Various mutations of hERG, including missense mutations, have been reported to cause long QT syndrome (LQTS) and severe arrhythmic disorders such as sudden cardiac death. We identified a novel hERG frameshift mutation (hERG(ΔAT)) in the S5-pore region from a LQTS patient who died suddenly and analyzed its genetic profile and the molecular and electrophysiological behaviors of the protein product to assess the pathogenicity of hERG(ΔAT). METHODS AND RESULTS: We performed direct sequencing of hERG and evaluated its transcript level by using a whole blood sample from the patient. We performed immunoblotting, immunocytochemistry, and patch-clamp recordings of HEK-293 T cells transfected with hERG(ΔAT), wild-type hERG (hERG(WT)), or both. The patient demonstrated an AT deletion (c.1735_1736del) in hERG and a decrease in hERG mRNA transcripts. HEK-293 T cells showed lower production and cell surface expression of hERG(ΔAT) compared with hERG(WT) protein. In addition, the hERG(∆AT) protein failed to form functional channels, while the activation kinetics of functional channels, presumably consisting of hERG(WT) subunits, were unaffected. CONCLUSION: The ΔAT mutation may decrease the number of functional hERG channels by impairing the posttranscriptional and posttranslational processing of the mutant product. This decrease may partly explain the cardiac symptoms of the patient who was heterozygous for hERG(ΔAT).
    Deutsche Zeitschrift für die gesamte gerichtliche Medizin 04/2013; 128:105-115. · 2.79 Impact Factor
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    ABSTRACT: An 81-year-old man was found dead 1 month after he had disappeared following a visit to a hot spring resort in early autumn. The body showed severe postmortem changes with advanced skeletonization from the head to the abdomen as well as putrefactive and autolytic changes in the remaining tissues. The thoracic and abdominal organs had been lost. Naked eye examination revealed soft tissue injuries accompanied by ragged edges and characteristic punctures with no signs of vitality, suggesting that these injuries had been due to postmortem animal scavenging. However, bruises were prominent on the anterior parts of both lower extremities. Postmortem computed tomography (PMCT) scan demonstrated subdural hematoma over the right cerebral hemisphere, although the brain itself had undergone putrefactive and autolytic changes. Subsequent autopsy confirmed the presence of a 140 g acute subdural hematoma, which would likely have been fatal. This case illustrates that PMCT is able to yield important information about possible cause of death, even in a partially skeletonized body.
    Legal Medicine 01/2013; 15(1):32–34. · 1.08 Impact Factor
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    ABSTRACT: A 53-year-old man who had suffered from hypertension was found dead on the floor of his room by his roommate on return from work in the summer. The body showed severe postmortem changes with advanced putrefaction and autolysis. No evident injury was detectable anywhere on the body. Postmortem computed tomography (PMCT) scan demonstrated cerebral hemorrhage in the left putamen and infiltration of blood into the third and lateral ventricles, although the brain had undergone putrefactive and autolytic changes including intravascular gas accumulation and partial cerebral settling. Detailed slice examination of the brain at autopsy proved impossible because of its fragility when attempting to remove it from the skull. This case illustrates that PMCT can be useful for demonstrating cerebral hemorrhage even in putrefactive brains, acting as a guide for forensic pathologists when conducting careful examination of a fragile brain.
    Journal of Forensic Radiology and Imaging. 01/2013; 1(4):212–214.
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    ABSTRACT: Five SNPs in the human DNase II gene have been reported to be associated with rheumatoid arthritis (RA). Genotype and haplotype analysis of 14 SNPs, nine SNPs of which reported in the NCBI dbSNP database in addition to these five SNPs, was performed in healthy subjects. The enzymatic activities of the amino acid substituted DNase II corresponding to each SNP and serum DNase II in healthy Japanese, and promoter activities derived from each haplotype of the RA-related SNPs were measured. Significant correlations between genotype in each RA-related SNP and enzymatic activity levels were found; alleles associated with RA exhibited a reduction in serum DNase II activity. Furthermore, the promoter activities of each reporter construct corresponding to predominant haplotypes in three SNPs in the promoter region of the gene exhibited significant correlation with levels of serum DNase II activity. These findings indicate these three SNPs could alter the promoter activity of DNASE2, leading to a decline in DNase II activity in the serum through gene expression. Since the three SNPs in the promoter region of the DNase II gene could affect in vivo DNase II activity through reduction of the promoter activity, it is feasible to identify these SNPs susceptible to RA.
    Electrophoresis 09/2012; 33(18):2852-8. · 3.26 Impact Factor
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    ABSTRACT: Adult height is a highly heritable trait involving multiple genes. Recent genome-wide association studies have identified that SNP rs12338076 in the LHX3-QSOX2 locus, and rs1457595 and rs17032362 in the IGF1 locus are associated with human height in the Japanese population (Okada et al. (2010)). We performed a replication study to examine the associations between these three SNPs and adult height in the Japanese population based on autopsy cases. However, it was not possible to confirm that all these SNPs influenced adult height in the study population. We first conducted a wide-ranging survey of these three SNPs in the above genes using nine different populations including Asians, Africans and Caucasians, and demonstrated that the genotypes of rs12338076 and rs17032362 were distributed in an ethnicity-dependent manner; even within Asian populations, the genotype distributions of the SNPs differed widely. Although there are differences in height distribution between different populations, possibly due to genetic factors and/or gene-environmental interactions, the contradictory results of the association study and ethnic differences in genotype distribution allow us to assume that these height-related SNPs in the genes may contribute to adult height to a slight extent, at least in the Japanese population. It is anticipated that the present information will be useful for developing a reliable tool for personal identification through elucidation of the genetic basis of human height.
    Legal Medicine 04/2012; 14(4):205-8. · 1.08 Impact Factor
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    ABSTRACT: The ABO blood group is of great importance in blood transfusion and organ transplantation. However, the mechanisms regulating human ABO gene expression remain obscure. On the basis of DNase I-hypersensitive sites in and upstream of ABO in K562 cells, in the present study, we prepared reporter plasmid constructs including these sites. Subsequent luciferase assays indicated a novel positive regulatory element in intron 1. This element was shown to enhance ABO promoter activity in an erythroid cell-specific manner. Electrophoretic mobility-shift assays demonstrated that it bound to the tissue-restricted transcription factor GATA-1. Mutation of the GATA motifs to abrogate binding of this factor reduced the regulatory activity of the element. Therefore, GATA-1 appears to be involved in the cell-specific activity of the element. Furthermore, we found that a partial deletion in intron 1 involving the element was associated with B(m) phenotypes. Therefore, it is plausible that deletion of the erythroid cell-specific regulatory element could down-regulate transcription in the B(m) allele, leading to reduction of B-antigen expression in cells of erythroid lineage, but not in mucus-secreting cells. These results support the contention that the enhancer-like element in intron 1 of ABO has a significant function in erythroid cells.
    Blood 03/2012; 119(22):5301-10. · 9.78 Impact Factor
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    ABSTRACT: DNA fragmentation factor beta (DFFB) polypeptide, endonuclease G (EndoG), and Flap endonuclease-1 (FEN-1) are responsible for DNA fragmentation, a hallmark of apoptosis. Although the human homologs of these genes show three, four, and six nonsynonymous single-nucleotide polymorphisms (SNPs), respectively, data on their genotype distributions in populations worldwide are limited. In this context, the objectives of this study were to elucidate the genetic heterogeneity of all these SNPs in wide-ranging populations, and thereby to clarify the genetic background of these apoptosis-related endonucleases in human populations. We investigated the genotype distribution of their SNPs in 13 different populations of healthy Asians, Africans, and Caucasians using novel genotyping methods. Among the 13 SNPs in the 3 genes, only 3 were found to be polymorphic: R196K and K277R in the DFFB gene, and S12L in the EndoG gene. All 6 SNPs in the FEN-1 gene were entirely monoallelic. Although it remains unclear whether each SNP would exert any effect on endonuclease functions, these genes appear to exhibit low degree of genetic heterogeneity with regard to nonsynonymous SNPs. These findings allow us to conclude that human apoptosis-related endonucleases, similarly to other human DNase genes, revealed previously, are well conserved at the protein level during the course of human evolution.
    DNA and cell biology 01/2012; 31(1):36-42. · 2.28 Impact Factor
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    ABSTRACT: A 53-year-old man was found dead after a fire at his residence had been extinguished. Although a pistol was recovered beside the body, external examination was unable to indicate any gunshot wound because of severe charring of the body. Postmortem computed tomography (CT) scan performed prior to autopsy suggested an entrance gunshot wound in the posterior pharynx with loss of soft tissue and an internal bullet path through the right anterior and posterior parts of the occipital bone. Autopsy revealed an entrance gunshot wound with hemorrhage in the soft tissue of the posterior pharynx, massive contusion of the right occipital lobe, and subarachnoid hemorrhage in the right temporal lobe, both occipital lobes and the superior surface of the left cerebellar hemisphere, thus being consistent with the findings of postmortem CT. A carboxyhemoglobin concentration of 5% in blood from the cadaver was consistent with the lack of soot deposition from the larynx to the bronchus. These observations confirmed that death had been caused by an intraoral gunshot resulting in severe brain damage, before the body had been burned.
    Legal Medicine 11/2011; 13(6):286-8. · 1.08 Impact Factor
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    ABSTRACT: Gene expression is driven by promoters, enhancers, silencers, and other cis-regulatory elements upstream and downstream of the gene. Previous studies of the regulation of human ABO gene transcription have focused mainly on the 5' region, including the core promoter and the region proximal to it. However, as the involvement of the 3' flanking region in transcriptional regulation has not yet been examined, we focused on this issue. The 3' region approximately 2.2kb downstream of the ABO gene was PCR-amplified and inserted into a cloning vector, followed by sequence determination and preparation of luciferase reporter vectors. Transient transfections into KATOIII and K562 cells were performed using various reporter plasmids containing the 3' region. The 3' region of the ABO gene, which was characterized by a high degree of sequence repetition, was effectively cloned by a single-copy cloning method. Transfections in KATOIII and K562 cells showed that negative elements were demonstrable within the 3' region. These observations suggest that negative regulatory elements seem to be present in the 3' region of ABO in both epithelial and erythroid lineages. As we had observed a negative region just upstream of the ABO promoter, transcription from ABO could be negatively regulated by repressive regions just upstream of the promoter and downstream of the gene. Further studies of the enhancer will be required for elucidating the molecular basis of ABO gene expression.
    Legal Medicine 01/2011; 13(1):22-9. · 1.08 Impact Factor
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    ABSTRACT: The C/EBP family of proteins represents an important group of bZIP transcription factors that are key to the regulation of essential functions such as cell cycle, hematopoiesis, skeletal development, and host immune responses. They are also intimately associated with tumorigenesis and viral disease. These proteins are regulated at multiple levels, including gene induction, alternative translational initiation, post-translational modification, and protein-protein interaction. This review attempts to integrate recent reports with more than 20 years of previous effort focused on this fascinating collection of regulators.
    Cytokine 01/2011; 54(1):6-19. · 2.52 Impact Factor
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    ABSTRACT: A 22-year-old woman was found dead in her bed, and subsequent postmortem examination was performed using ordinary methods such as external examination, Triage®, and computed tomography (CT) scan which demonstrated a high-density content of the duodenum. Autopsy and quantitative analysis of drugs present in the GI tract showed that high amounts of radiopaque psychotic agents such as fluvoxamine maleate, carbamazepine, and zolpidem tartrate had been responsible for the high-density profile of the duodenum. Postmortem quantitative analysis of drugs in the blood suggested that death had been caused by fatal intoxication with fluvoxamine maleate. Thus, postmortem CT could offer an opportunity to suspect drug intoxication due to radiopaque psychotic agents such as chloral hydrate, phenothiazine, bromovaleryl urea, fluvoxamine maleate, and probably zolpidem tartrate, although it is neither a specific nor a quantitative test for drugs. Therefore, postmortem CT happened to provide clues to investigation of drug intoxication in the present case.
    Legal Medicine 01/2011; 13(1):39-40. · 1.08 Impact Factor

Publication Stats

693 Citations
223.15 Total Impact Points

Institutions

  • 2004–2014
    • Gunma University
      • • Department of Legal Medicine
      • • Graduate School of Medicine
      Maebashi, Gunma Prefecture, Japan
  • 2008–2013
    • University of Toyama
      • Department of Legal Medicine
      Тояма, Toyama, Japan
  • 2004–2013
    • University of Fukui
      • Division of Medical Genetics and Biochemistry
      Hukui, Fukui, Japan
  • 2012
    • Shimane University
      • Department of Legal Medicine
      Matsu, Shimane Prefecture, Japan
  • 2002–2008
    • University of Occupational and Environmental Health
      • School of Medicine
      Kitakyūshū, Fukuoka-ken, Japan
  • 2007
    • Toyama University
      Тояма, Toyama, Japan
  • 1990–2004
    • Toyama Medical and Pharmaceutical University
      Тояма, Toyama, Japan
  • 2003
    • Sanford-Burnham Medical Research Institute
      La Jolla, California, United States
  • 1995
    • Brigham and Women's Hospital
      • Center for Brain Mind Medicine
      Boston, MA, United States