Curriculum Vitaes
Profile Information
- Affiliation
- Department of Physiology, Fujita Health University School of Medicine
- Degree
- 博士(医学)(藤田保健衛生大学)
- Other name(s) (e.g. nickname)
- ナカシマアキラ
- J-GLOBAL ID
- 200901085737243854
- researchmap Member ID
- 1000102599
Research Interests
3Research Areas
2Research History
10-
Apr, 2024 - Present
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Apr, 2014 - Mar, 2024
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2012 - Mar, 2024
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2007 - 2014
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2001 - 2011
Committee Memberships
5-
2020 - 2022
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2011 - 2022
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2016 - 2019
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2015 - 2019
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2013 - 2019
Papers
56-
Biochemical and Biophysical Research Communications, 714 149940-149940, Jun, 2024 Peer-reviewed
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Biochemical and biophysical research communications, 703 149698-149698, Apr 9, 2024 Peer-reviewedLast authorThe gene encoding 5'-nucleotidase domain-containing protein 2 (NT5DC2) has been associated with neuropsychiatric disorders related to the abnormality of dopamine activity in the brain. However, its physiological functions remain unclear. In this study, we analyzed the features of NT5DC2 that influence its binding with tyrosine hydroxylase (TH) and its effects on dihydroxyphenylalanine (DOPA) synthesis, using NT5DC2 overexpressed in PC12D cells by the pCMV vector. Western blot analysis revealed that the purified NT5DC2-DYKDDDDK-tag (NT5DC2-tag) protein can bind with the phosphorylated form of recombinant human TH type 1 (rhTH1), apart from the endogenous TH in PC12D cells. Proteomic analysis by mass spectrometry revealed that the purified NT5DC2-tag protein has the potential to bind to 41 proteins with multiple phosphorylation sites in PC12D cells (NT5DC2 binding proteins: positive, 391 sites/41 proteins; and negative, 85 sites/27 proteins). Overexpression of NT5DC2 in PC12D cells decreased DOPA levels in the medium. When the lysate of PC12D cells overexpressing NT5DC2 was incubated at 37 °C, the phosphorylated form of endogenous TH in PC12D cells decreased. This decrease was also detected when phosphorylated rhTH1 was incubated with purified NT5DC2-tag. Overall, our results suggest that NT5DC2 regulates DOPA synthesis by promoting the dephosphorylation of TH, similar to a phosphatase. Therefore, our study provides useful information for understanding various disorders associated with abnormalities in dopamine levels in the brain.
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Journal of Neural Transmission, Mar 20, 2023 Peer-reviewedAbstract The dark pigment neuromelanin (NM) is abundant in cell bodies of dopamine (DA) neurons in the substantia nigra (SN) and norepinephrine (NE) neurons in the locus coeruleus (LC) in the human brain. During the progression of Parkinson’s disease (PD), together with the degeneration of the respective catecholamine (CA) neurons, the NM levels in the SN and LC markedly decrease. However, questions remain among others on how NM is associated with PD and how it is synthesized. The biosynthesis pathway of NM in the human brain has been controversial because the presence of tyrosinase in CA neurons in the SN and LC has been elusive. We propose the following NM synthesis pathway in these CA neurons: (1) Tyrosine is converted by tyrosine hydroxylase (TH) to L-3,4-dihydroxyphenylalanine (L-DOPA), which is converted by aromatic L-amino acid decarboxylase to DA, which in LC neurons is converted by dopamine β-hydroxylase to NE; (2) DA or NE is autoxidized to dopamine quinone (DAQ) or norepinephrine quinone (NEQ); and (3) DAQ or NEQ is converted to eumelanic NM (euNM) and pheomelanic NM (pheoNM) in the absence and presence of cysteine, respectively. This process involves proteins as cysteine source and iron. We also discuss whether the NM amounts per neuromelanin-positive (NM+) CA neuron are higher in PD brain, whether NM quantitatively correlates with neurodegeneration, and whether an active lifestyle may reduce NM formation.
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Frontiers in Endocrinology, 13 941166-941166, Jul 12, 2022 Peer-reviewedHuman stem cell-derived organoid culture enables the in vitro analysis of the cellular function in three-dimensional aggregates mimicking native organs, and also provides a valuable source of specific cell types in the human body. We previously established organoid models of the hypothalamic-pituitary (HP) complex using human pluripotent stem cells. Although the models are suitable for investigating developmental and functional HP interactions, we consider that isolated pituitary cells are also useful for basic and translational research on the pituitary gland, such as stem cell biology and regenerative medicine. To develop a method for the purification of pituitary cells in HP organoids, we performed surface marker profiling of organoid cells derived from human induced pluripotent stem cells (iPSCs). Screening of 332 human cell surface markers and a subsequent immunohistochemical analysis identified epithelial cell adhesion molecule (EpCAM) as a surface marker of anterior pituitary cells, as well as their ectodermal precursors. EpCAM was not expressed on hypothalamic lineages; thus, anterior pituitary cells were successfully enriched by magnetic separation of EpCAM+ cells from iPSC-derived HP organoids. The enriched pituitary population contained functional corticotrophs and their progenitors; the former responded normally to a corticotropin-releasing hormone stimulus. Our findings would extend the applicability of organoid culture as a novel source of human anterior pituitary cells, including stem/progenitor cells and their endocrine descendants.
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eNeuro, Apr 18, 2022 Peer-reviewedHypothalamic melanin-concentrating hormone (MCH) neurons are important regulators of multiple physiological processes, such as sleep, feeding, and memory. Despite the increasing interest in their neuronal functions, the molecular mechanism underlying MCH neuron development remains poorly understood. We report that a three-dimensional culture of mouse embryonic stem cells (mESCs) can generate hypothalamic-like tissues containing MCH-positive neurons, which reproduce morphologic maturation, neuronal connectivity, and neuropeptide/neurotransmitter phenotype of native MCH neurons. Using this in vitro system, we demonstrate that Hedgehog (Hh) signaling serves to produce major neurochemical subtypes of MCH neurons characterized by the presence or absence of cocaine- and amphetamine-regulated transcript (CART). Without exogenous Hh signals, mESCs initially differentiated into dorsal hypothalamic/prethalamic progenitors and finally into MCH+CART+ neurons through a specific intermediate progenitor state. Conversely, activation of the Hh pathway specified ventral hypothalamic progenitors that generate both MCH+CART- and MCH+CART+ neurons. These results suggest that in vivo MCH neurons may originate from multiple cell lineages that arise through early dorsoventral patterning of the hypothalamus. Additionally, we found that Hh signaling supports the differentiation of mESCs into orexin/hypocretin neurons, a well-defined cell group intermingled with MCH neurons in the lateral hypothalamic area (LHA). The present study highlights and improves the utility of mESC culture in the analysis of the developmental programs of specific hypothalamic cell types.Significance StatementA growing body of literature has revealed the importance of hypothalamic melanin-concentrating hormone (MCH) neurons in energy homeostasis and the cognitive function, but their developmental biology remains relatively unknown. To establish a new approach for addressing this issue, we tested the ability of an in vitro differentiation system of mouse embryonic stem cells (mESCs) to recapitulate the development of MCH neurons. The mESC culture robustly generated MCH-positive neurons resembling native neurons in several aspects and provided evidence that Hedgehog (Hh) signaling is a key factor to produce neurochemical subtypes of MCH neurons. Our results demonstrate the suitability of mESC culture as a platform to study the molecular mechanisms underlying the development of MCH neurons and possibly of other hypothalamic cell types.
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International Journal of Molecular Sciences, 23(8) 4176-4176, Apr 10, 2022
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PloS one, 17(11) e0276694, 2022 Peer-reviewedThe hypothalamus is comprised of heterogenous cell populations and includes highly complex neural circuits that regulate the autonomic nerve system. Its dysfunction therefore results in severe endocrine disorders. Although recent experiments have been conducted for in vitro organogenesis of hypothalamic neurons from embryonic stem (ES) or induced pluripotent stem (iPS) cells, whether these stem cell-derived hypothalamic neurons can be useful for regenerative medicine remains unclear. We therefore performed orthotopic transplantation of mouse ES cell (mESC)-derived hypothalamic neurons into adult mouse brains. We generated electrophysiologically functional hypothalamic neurons from mESCs and transplanted them into the supraoptic nucleus of mice. Grafts extended their axons along hypothalamic nerve bundles in host brain, and some of them even projected into the posterior pituitary (PPit), which consists of distal axons of the magnocellular neurons located in hypothalamic supraoptic and paraventricular nuclei. The axonal projections to the PPit were not observed when the mESC-derived hypothalamic neurons were ectopically transplanted into the substantia nigra reticular part. These findings suggest that our stem cell-based orthotopic transplantation approach might contribute to the establishment of regenerative medicine for hypothalamic and pituitary disorders.
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Journal of neural transmission (Vienna, Austria : 1996), Aug 10, 2020 Peer-reviewed5'-Nucleotidase domain-containing protein 2 (NT5DC2) has been revealed by genome-wide association studies (GWAS) as a gene implicated in neuropsychiatric disorders related to the abnormality of dopamine (DA) activity in the brain. Based on its amino acid sequence, NT5DC2 is assumed to be a member of the family of haloacid dehalogenase-type phosphatases; although there is no information about its function and structural conformation. We recently reported that NT5DC2 binds to tyrosine hydroxylase (TH) and that the down-regulation of NT5DC2 tended to increase DA synthesis. In this study, we investigated whether NT5DC2 could regulate the catalytic activity of TH, which converts tyrosine to DOPA, because the phosphorylation level of TH, controlled by protein kinases and phosphatases, is well known to regulate its catalytic activity. The down-regulation of NT5DC2 by siRNA increased mainly DOPA synthesis by TH in PC12D cells, although this down-regulation tended to increase the conversion of DOPA to DA by aromatic L-amino acid decarboxylase. The increased DOPA synthesis should be attributed to the catalytic activity of TH controlled by its phosphorylation, because Western blot analysis revealed that the down-regulation of NT5DC2 tended to increase the level of TH phosphorylated at its Ser residues, but not that of the TH protein. Moreover, the induction of kinase activity by forskolin markedly potentiated the phosphorylation of TH at its Ser40 in PC12D cells having down-regulated NT5DC2. Immunocytochemical analysis of PC12D cells demonstrated that NT5DC2, TH protein, and TH phosphorylated at its Ser40 were predominantly localized in the cytoplasm and that the localization of NT5DC2 and TH proteins partially overlapped. Collectively, our results indicate that NT5DC2 could work to inhibit the DOPA synthesis by decreasing the phosphorylation of TH at its Ser40. We propose that NT5DC2 might decrease this phosphorylation of TH by promoting dephosphorylation or by inhibiting kinase activity.
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Biochemical and biophysical research communications, 516(4) 1060-1065, Sep, 2019 Peer-reviewedTyrosine hydroxylase (TH), which catalyzes the conversion of l-tyrosine to l-DOPA, is the rate-limiting enzyme in the biosynthesis of catecholamines. It is well known that both α-synuclein and 14-3-3 protein family members bind to the TH molecule and regulate phosphorylation of its N-terminus by kinases to control the catalytic activity. In this present study we investigated whether other proteins aside from these 2 proteins might also bind to TH molecules. Nano-LC-MS/MS analysis revealed that 5'-nucleotidase domain-containing protein 2 (NT5DC2), belonging to a family of haloacid dehalogenase-type (HAD) phosphatases, was detected in the immunoprecipitate of PC12D cell lysates that had been reacted with Dynabeads protein G-anti-TH antibody conjugate. Surprisingly, NT5DC2 had already been revealed by Genome-Wide Association Studies (GWAS) as a gene implicated in neuropsychiatric disorders such as schizophrenia, bipolar disorder, which are diseases related to the abnormality of dopamine activity in the brain, although the role that NT5DC2 plays in these diseases remains unknown. Therefore, we investigated the effect of NT5DC2 on the TH molecule. The down-regulation of NT5DC2 by siRNA increased the synthesis of catecholamines (dopamine, noradrenaline, and adrenaline) in PC12D cells. These increases might be attributed to the catalytic activity of TH and not to the intracellular stability of TH, because the intracellular content of TH assessed by Western blotting was not changed by the down-regulation of NT5DC2. Collectively, our results indicate that NT5DC2 inhibited the synthesis of dopamine by decreasing the enzymatic activity of TH.
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Journal of neural transmission (Vienna, Austria : 1996), 126(4) 397-409, Apr, 2019 Peer-reviewed
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Journal of Neural Transmission, 125(1) 9-15, Jan 1, 2018 Peer-reviewed
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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 472(4) 598-602, Apr, 2016 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 122(6) 757-772, Jun, 2015 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 122(2) 187-199, Feb, 2015 Peer-reviewed
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Fujita Medical Journal, 1(1) 1-5, 2015 Peer-reviewedObjectives: According to our previous work, aripiprazole exerted a protective effect on hydrogen peroxide (H2O2)-treated PC12 cells; haloperidol did not. Because aripiprazole has distinct affinities to a set of neurotransmitter receptor subtypes, this study aimed to clarify which subtype is responsible for rescuing cells from 0.25 mM H2O2 exposure.Methods: A set of compounds, which are more specific to each subset of G-protein coupled receptors, wereexamined for their ability to mimic the pharmacological effects of aripiprazole or haloperidol, including their Ki values.The viability of PC12 cells cultured with test compounds with or without H2O2 was assessed using WST-8 reagent.Results: Results from in vitro studies using PC12 cells showed that agonism at serotonin 5-HT2C-receptors based on the antagonism against 5-HT2B-receptors played a significant role in resistingH2O2-induced cell death. However, the use of a specific 5-HT2B-receptor agonist instead of a 5-HT2B-receptor antagonist completely negated the effect of a specific 5-HT2C-receptor agonist. Furthermore, unlike the dopamine D1-receptor specific antagonist, none of the agonists of dopamine D2-, D3-, and D4-receptors ameliorated the cytopathic effects of H2O2.Conclusion: Antagonism at 5-HT2B-receptors is fundamental for the protection of PC12 cells against the cytopathiceffects caused by 0.25 mM H2O2. However, the role of negatively regulated cyclic adenosine monophosphate in this phenomenon requires further investigation.
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日本内分泌学会雑誌, 90(1) 305-305, Apr, 2014 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 121(1) 91-103, Jan, 2014 Peer-reviewed
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日本内分泌学会雑誌, 89(1) 296-296, Apr, 2013 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 120(1) 49-54, Jan, 2013 Peer-reviewed
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Advances in Pharmacology, 68 3-11, 2013 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 119(11) 1327-1342, Nov, 2012 Peer-reviewed
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CELLULAR AND MOLECULAR NEUROBIOLOGY, 32(5) 777-785, Jul, 2012 Peer-reviewed
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CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, 39(7) 599-607, Jul, 2012 Peer-reviewed
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JOURNAL OF NEUROCHEMISTRY, 120(2) 199-201, Jan, 2012 Peer-reviewedInvited
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Construction of Yanegawara-style skills training in our clinical skills laboratory for new residentsIgaku Kyoiku / Medical Education (Japan), 43(3) 211-214, 2012 Peer-reviewed1)To learn the techniques required immediately after the start of clinical practice, new residents were introduced to the skills laboratory during their orientation period.<br>2)We attempted to establish the Yanegawara style, which is an overlapping teaching style in which the second–year residents plan the entire training schedule and simultaneously teach the first–year residents while being supported in their teaching by more senior physicians.<br>3)Training with the new system resulted in greater rapport among all residents as well as a greater feeling of security among first–year residents.
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NEURODEGENERATIVE DISEASES, 10(1-4) 100-103, 2012 Peer-reviewed
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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 407(2) 343-347, Apr, 2011 Peer-reviewed
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42(3) 135-140, 2011 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 117(10) 1139-1153, Oct, 2010 Peer-reviewed
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NEUROSCIENCE LETTERS, 481(2) 126-130, Sep, 2010 Peer-reviewed
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JOURNAL OF NEURAL TRANSMISSION, 116(11) 1355-1362, Nov, 2009 Peer-reviewed
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BRAIN RESEARCH, 1279 9-20, Jul, 2009 Peer-reviewed
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Stress, Neurotransmitters, and Hormones: Neuroendocrine and Genetic Mechanisms, 1148 127-135, 2008 Peer-reviewed
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Igaku Kyoiku / Medical Education (Japan), 39(6) 397-406, 2008 Peer-reviewedAt the Fujita Health University School of Medicine, about 30% of medical students are admitted on the basis of recommendations.To evaluate the performance of these students after admission, a placement test was given to all new students just after the entrance ceremony to examine basic academic abilities.The scores were compared with the number of absences from lectures and with examination results for the first and second years.<BR>1) The 398 students admitted from 2002 through 2005 were classified into three populations: 126 recommended students, 137 students who scored in the top half on the entrance examination, and 135 students who scored in the bottom half.<BR>2) Scores on the placement test were highest for the top-half students, intermediate for the bottom-half students, and lowest for the recommended students.Scores on examinations in the first and second years were highest for the top-half students, intermediate for the recommended students, and lowest for the bottom-half students.<BR>3) The average number of absences from lectures in the first and second years tended to be lower for recommended students than for the top-half or bottom-half students.<BR>4) The examination scores in the second year were correlated with scores in the first year, and the average number of absences in the second year correlated with those in the first year.<BR>5) These results indicate that the motivation of students in each classification to study in the 1st year is, in addition to their basic academic abilities obtained in high school, an important factor affecting their performance in the second year and beyond.
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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 363(3) 817-821, Nov, 2007 Peer-reviewed
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JOURNAL OF NEUROSCIENCE, 26(39) 10068-10078, Sep, 2006 Peer-reviewed
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JOURNAL OF NEUROSCIENCE RESEARCH, 81(1) 110-120, Jul, 2005 Peer-reviewed
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JOURNAL OF NEUROCHEMISTRY, 94(2) 393-404, Jul, 2005 Peer-reviewed
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Clinical and experimental pharmacology & physiology, 32(4) 279-287, Apr, 2005 Peer-reviewed
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BRAIN RESEARCH, 1039(1-2) 116-129, Mar, 2005 Peer-reviewed
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Biochimica et biophysica acta, 1670(3) 181-198, Feb, 2004 Peer-reviewed
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J. Neural Transm., 110 31-50, 2003 Peer-reviewed
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Neuroscience, 116 7-12, 2003 Peer-reviewed
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Clinical and experimental pharmacology & physiology, 29(11) 980-989, Nov, 2002 Peer-reviewed
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JOURNAL OF NEUROCHEMISTRY, 82(1) 202-206, Jul, 2002 Peer-reviewed
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J. Neurochem., 82 202-206, 2002 Peer-reviewed
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Sequence Analysis of Human Tyrosine Hydroxylase Gene Exon 3 of Japanese Patients with Schizophrenia.Molecular Psychiatry, 6 315-319, 2001 Peer-reviewed
Misc.
40Books and Other Publications
9Presentations
28-
Society for Neuroscience 47th Annual Meeting, Washington D.C., USA, Nov, 2017
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he 11th International Conference on Alzheimer's and Parkinson’s Diseases, 2013
Teaching Experience
8-
2023 - Present生理学実習 (愛知学院大学 健康科学部 健康栄養学科)
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1989 - 2024生理学 (藤田医科大学 医学部)
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Physiology (Aichi Medical University)
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病態生理学 (藤田医科大学 医療科学部大学院)
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Human Biology (藤田医科大学 医学部)
Professional Memberships
6Research Projects
7-
科学研究費助成事業, 日本学術振興会, Apr, 2021 - Mar, 2024
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日本学術振興会:科研費, Apr, 2018 - Mar, 2021
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科研費, 日本学術振興会, Apr, 2013 - Mar, 2016
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KAKEN, 日本学術振興会, Apr, 2010 - Mar, 2013
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科研費, 日本学術振興会, Apr, 2006 - Mar, 2008
教育内容・方法の工夫(授業評価等を含む)
1-
件名(英語)生理学授業へのe-learningによる自己学習システムの導入開始年月日(英語)2011/01終了年月日(英語)2011/03概要(英語)e-learning教材「一歩一歩学ぶ生命科学」をmoodleシステムに組み込み、生理学講義受講前の学生に自己学習をさせることにより生理学講義の理解を助けるシステムを構築した。
作成した教科書、教材、参考書
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件名(英語)新訂・生理学実習書、日本生理学会教育委員会編/ 編集委員/ 南江堂終了年月日(英語)2013/10概要(英語)生理学実習を行うための教員・学生のための実習書を作成した(分担編集)
教育方法・教育実践に関する発表、講演等
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件名(英語)生理学授業へのe-learningによる自己学習システムの導入事例の報告終了年月日(英語)2011/10概要(英語)第43回藤田医学会で報告した。
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件名(英語)学生指導で注意していること(藤田保健衛生大学・医学部・指導教員懇談会・講演)終了年月日(英語)2012/07概要(英語)指導教員に対する学生の指導方法について
その他教育活動上特記すべき事項
2-
件名(英語)学内医学教育ワークショップの開催(第27回〜57回、計30回)開始年月日(英語)2008/04終了年月日(英語)2015/09概要(英語)医学教育企画室による学内教員向けワークショップの開催と運営(試験問題の作り方、ポートフォリオ導入、PBLテュータトレーニングなど)
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件名(英語)基礎統合実習の開催(2005年・第1回〜2015年・第11回、学外活動)開始年月日(英語)2005/08終了年月日(英語)2015/08概要(英語)生理学・病理学・生化学など基礎医学系全ての知識を基にして、学生が自ら考えた仮設を実験で検証する「真実を探求」することを目的とした実習を全国の医学生を集めて開催