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Human pluripotent stem cells:Towards therapeutic development for the treatment of lifestyle diseases

World Journal of Stem Cells 2016年第2期8卷 56-61页

作者： Miwako Nishio Masako Nakahara Akira Yuo kumiko saeki Research Institute National Center for Global Health and Medicine PRESTO Japan Science and Technology Agency

There are two types of human pluripotent stem cells: Embryonic stem cells(ESCs) and induced pluripotent stem cells(iPSCs),both of which launched themselves on clinical trials after having taken measures to overcome problems: Blocking rejections by immunosuppressants regarding ESCs and minimizing the risk of tumorigenicity by depleting exogenous gene components regarding iP *** is generally assumed that clinical applications of human pluripotent stem cells should be limited to those cases where there are no alternative measures for treatments because of the risk in transplanting those cells to living *** lifestyle diseases,we have already several therapeutic options,and thus,development of human pluripotent stem cell-based therapeutics tends to be ***,human pluripotent stem cells can contribute to the development of new therapeutics in this *** we will show,there is a case where only a short-term presence of human pluripotent stem-derived cells can exert long-term therapeutic effects even after they are *** those cases,immunologically rejections of ESC-or allogenic iP SC-derived cells may produce beneficial outcomes by nullifying the risk of tumorigenesis without deterioration of therapeutic *** utility of human pluripotent stem cells is the provision of an innovative tool for drug discovery that are otherwise *** example,clinical specimens of human classical brown adipocytes(BAs),which has been attracting a great deal of attention as a new target of drug discovery for the treatment of metabolic disorders,are unobtainable from living individuals due to scarcity,fragility and ethical ***,BA can easily be produced from human pluripotent stem *** this review,we will contemplate potential contribution of human pluripotent stem cells to therapeutic development for lifestyle diseases.

关键词： Arteriostenosis Human embryonic stem cells Glucose intolerance Human induced pluripotent stem cells Brown adipose tissue

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p38 mitogen-activated protein kinase regulates type-Ⅰ vs type-Ⅱ phenotyping of human vascular endothelial cells

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World Journal of Translational Medicine 2015年第3期4卷 101-112页

作者： Masako Nakahara Miwako Nishio Koichi saeki Akira Yuo kumiko saeki Department of Disease Control Research Institute National Center for Global Health and Medicine Section of Cell Engineering Department of Basic Research DNAVEC Center ID Pharma Co. Ltd. PRESTO Japan Science and Technology Agency

AIM: To identify kinases involved in phenotype regulation of vascular endothelial cells(VECs): Proproliferative G-protein signaling 5(RGS5)^(high)(typeⅠ) vs anti-proliferative RGS5^(low)(typeⅡ) ***: Proteomic kinase assays were performed to identify the crucial kinase involved in the phenotype regulation of human VECs using typeⅠ VECs, which promotes the proliferation of human vascular smooth muscle cells(VSMCs), and typeⅡ VECs, which suppress the proliferation of human VSMCs. The assays were performed using multiple pairs of typeⅠ and typeⅡ VECs to obtain the least number of candidates. The involvement of the candidate kinases was verified by evaluating the effects of their specific inhibitors on the phenotype regulation of human VECs as well as the expression levels of regulator of RGS5, which is the causative gene for the "typeⅡ to typeⅠ" phenotype conversion of human VECs. RESULTS: p38α mitogen-activated protein kinase(p38α MAPK) was the only kinase that showed distinctive activities between typeⅠ and typeⅡ VECs: p38α MAPK activities were low and high in type-Ⅰand typeⅡ VECs, respectively. We found that an enforced expression of RGS5 indeed lowered p38α MAPK activitiesin typeⅡ VECs. Furthermore, treatments with a p38α MAPK inhibitor nullified the anti-proliferative potential in typeⅡ VECs. Interestingly, MAPK inhibitor treatments enhanced the induction of RGS5 gene. Thus, there is a vicious cycle between "RGS5 induction" and "p38α MAPK inhibition", which can explain the unidirectional process in the stress-induced "typeⅡ to typeⅠ" conversions of human VECs. To understand the upstream signaling of RGS5, which is known as an inhibitory molecule against the G protein-coupled receptor(GPCR)-mediated signaling, we examined the effects of RGS5 overexpression on the signaling events from sphingosine-1-phosphate(S1P) to N-cadherin, because S1 P receptors belong to the GPCR family gene and N-cadherin, one of their downstream effectors, is reportedly involved in

关键词： vascular endothelial cells vascular smooth muscle cells proteomic kinase assay p38αmitogenactivated protein kinase regulator of G-protein signaling 5 sphingosine-1-phosphate N-cadherin

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Additional attention to combination antiretroviral therapy-related lipodystrophy

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World Journal of Virology 2017年第3期6卷 49-52页

作者： Norihiko Kobayashi Masako Nakahara Masako Oka kumiko saeki Department of Disease Control Research Institute National Center for Global Health and Medicine

The occurrence of lipodystrophy in patients taking antihuman immunodeficiency virus(HIV) medications is a serious problem as it is irreversible even after drug withdrawal. Although it was first recognized in patients taking proteinase inhibitors, other types of anti-HIV agents can also cause lipodystrophy. In a recent publication by Jones et al entitled "Highly active antiretroviral therapy dysregulates proliferation and differentiation of human pre-adipocytes" in World Journal of Virology, it was reported that simultaneous treatment of human subcutaneous adipocytes with anti-HIV drugs with different mechanisms of action synergistically exerted antiadipogenesis effects in vitro, warning us to take utmost care in every case receiving combination antiretroviral therapy(cART). For elucidation of the molecular basis for cART-related lipodystrophy, multi-faceted approaches should be taken, based on a deeper understanding of the development and organization of adipose tissues.

关键词： Combination antiretroviral therapy Lipodystrophy Protease inhibitor Reverse transcriptase inhibitor Human immunodeficiency virus

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Pro-vs anti-stenotic capacities of type-Ⅰ vs type-Ⅱ human induced pluripotent-derived endothelial cells

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World Journal of Translational Medicine 2015年第3期4卷 113-122页

作者： Miwako Nishio Masako Nakahara Koichi saeki Katsuhito Fujiu Hiroshi Iwata Ichiro Manabe Akira Yuo kumiko saeki Department of Disease Control Research Institute National Center for Global Health and Medicine Section of Cell Engineering Department of Basic Research DNAVEC Center ID Pharma Co. Ltd. Department of Cardiovascular Medicine Graduate School of Medicine the University of Tokyo Translational Systems Biology and Medicine Initiative the University of Tokyo PRESTO Japan Science and Technology Agency

AIM:To verify in vivo relevance of the categorization of human vascular endothelial cells(VECs)into type-I(proproliferative)and type-II(anti-proliferative).METHODS:Endothelial layers of murine femoral arteries were removed by wire injury(WI)operation,a common technique to induce ***-I and type-II VECs produced from human induced pluripotent stem cells(iPSCs),whose characters were previously determined by their effects on the proliferation of vascular smooth muscle cells in in vitro co-culture experiments,were mixed with Matrigel?*** mixtures were injected into subcutaneous spaces around WI-operated femoral arteries for the transplanted human iPSC-derived VECs(iPSdECs)to take a route to the luminal surface via vasa vasorum,a nutrient microvessel for larger *** of the femoral arteries were examined over *** presence of human iPSdECs was checked by immunostaining studies using an antibody that specifically recognizes human *** of stenosis of the femoral arteries were calculated after three *** determine the optimal experimental condition,xenotransplantation experiments were performed under various conditions using immunocompromised mice as well as immunocompetent mice with or without administration of ***:Because immunocompromised mice showed unexpected resistance to WI-induced arteriostenosis,we performed xenotransplantation experiments using immunocompetent mice along with immunosuppressant *** one week,luminal surfaces of the WI-operated arteries were completely covered by human iPSdECs,showing the efficacy of our novel transplantation *** three weeks,type-IiPSdECs-transplanted arteries underwent total stenosis,while type-II-iPSdECs-transplanted arteries remained ***,untransplanted arteries of immunosuppressant-treated mice also remained intact by unknown *** found that transplanted human VECs had already been replaced by murine endothel

关键词： Vascular endothelial cells Vasa vasorum Arteriostenosis Wire injury Human induced pluripotent stem cells

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New categorization of human vascular endothelial cells by pro-vs anti-proliferative phenotypes

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World Journal of Translational Medicine 2015年第3期4卷 88-100页

作者： Miwako Nishio Masako Nakahara Chikako Sato Koichi saeki Hidenori Akutsu Akihiro Umezawa Kazuyuki Tobe Kazuki Yasuda Akira Yuo kumiko saeki Department of Disease Control Research Institute National Center for Global Health and Medicine Section of Cell Engineering Department of Basic Research DNAVEC Center ID Pharma Co. Ltd. PRESTO Japan Science and Technology Agency Department of Reproductive Biology National Research Institute for Child Health and Development First Department of Internal Medicine School of Medicine Toyama University Department of Metabolic Disorder Diabetes Research Center Research Institute National Center for Global Health and Medicine

AIM: To integrally understand the effects of human vascular endothelial cells(VECs) on the proliferation of vascular smooth muscle cells(VSMCs).METHODS: Various kinds of human VECs of different origins were co-cultured with human aortic smooth muscle cells, a representative of human VSMCs. To exclude the irrelevant effects due to growth competition between VECs and VSMCs, the proliferation of VECs had previously been arrested via a low-dose gamma rayirradiation. To discriminately analyze the proliferation of VSMCs from that of VECs, the former cells were labeled with red fluorescent dye while the latter cells were labeled with green fluorescent dye before performing coculture experiments. After 4 d, total cells were harvested and subjected to flow cytometric analyses. Decrements in red fluorescence intensities due to proliferationmediated dilutions were measured and mathematically processed using a specific software to quantitatively evaluate the proliferation of VSMCs. The findings obtained from the flow cytometry-based analyses were further validated by microscopic observations. RESULTS: Commercially available primary cultured human VECs exclusively promoted VSMC proliferation regardless of their tissue origins and we termed these pro-proliferative VECs as "typeⅠ". By contrast, VECs freshly generated from human bone marrow-derived endothelial progenitors cells or human pluripotent stem cells including embryonic stem cells and induced pluripotent stem cells suppressed VSMC proliferation and we termed these anti-proliferative VECs as "typeⅡ". Repetitive subcultures as well as oxidative stress induced "type Ⅱ VECs to typeⅠ" conversion along with an induction of Regulator of G-protein signaling 5(RGS5)Compatibly, anti-oxidant treatments suppressed both the subculture-dependent "typeⅡ to typeⅠ" conversion and an induction of RGS5 gene. Immunostaining studies of clinical specimens indicated that RGS5 protein expressions in endothelial layers were low in norma arteries b

关键词： Vascular endothelial cells Vascular smooth muscle cells Human induced pluripotent stem cells Human embryonic stem cells Regulator of G-protein signaling 5 Oxidative stress

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Large Mesoporous Tin Dioxide Powders for Gas Sensor Materials-Effects of Various Additives on the Mesostructure and the H_2 Sensing Properties

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稀有金属材料与工程 2006年第A3期35卷 455-458页

作者： Takeo Hyodo kumiko Murayama Yasuhiro Shimizu Makoto Egashira Department of Materials Science and Engineering Faculty of EngineeringNagasaki University1-14 Bunkyo-machiNagasaki 852-8521Japan

Preparation of large mesoporous tin dioxide (lm-SnO_2) under various conditions was attempted by utilizing a self-assembly of a triblock copolymer,P123:(EO)_(20)(PO)_(70)(EO)_(20) or F127:(EO)_(106)(PO)_(70)(EO)_(106)(EO:ethylene oxide and PO: propylene oxide).The sensor fabricated from calcined lm-SnO_2 powder,which had been prepared by using P123 as a template,Na_2SnO_3 as a tin source,and TEOS as an additive ([TEOS]/[Na_2SnO_3]=0.5 in the precursor solution),showed the largest response to 1×10^(-3) hydrogen at 350℃among the sensors *** existence of two kinds of SnO_2 particles with different sizes (ca.100 nm and several nm in diameter) may be important to improve the hydrogen sensing properties drastically.

关键词： mesoporous tin dioxide triblock copolymer triethoxyorthosilicate hydrogen gas sensor

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Simultaneous removal of dissolved organic matter and bromide from drinking water source by anion exchange resins for controlling disinfection by-products

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Journal of Environmental Sciences 2014年第6期26卷 1294-1300页

作者： Athit Phetrak Jenyuk Lohwacharin Hiroshi Sakai Michio Murakami kumiko Oguma Satoshi Takizawa Department of Urban Engineering Graduate School of Engineering The University of Tokyo Institute of Industrial Science The University of Tokyo

Anion exchange resins （AERs） with different properties were evaluated for their ability to remove dissolved organic matter （DOM） and bromide, and to reduce disinfection by-product （DBP） formation potentials of water collected from a eutrophic surface water source in Japan. DOM and bromide were simultaneously removed by all selected AERs in batch adsorption experiments. A polyacrylic magnetic ion exchange resin （MIEX） showed faster dissolved organic carbon （DOC） removal than other AERs because it had the smallest resin bead size. Aromatic DOM fractions with molecular weight larger than 1600 Da and fluorescent organic fractions of fulvic acid- and humic acid-like compounds were efficiently removed by all AERs. Polystyrene AERs were more effective in bromide removal than polyacrylic AERs. This result implied that the properties of AERs, i.e. material and resin size, influenced not only DOM removal but also bromide removal efficiency, MIEX showed significant chlorinated DBP removal because it had the highest DOC removal within 30 rain, whereas polystyrene AERs efficiently removed brominated DBPs, especially brominated trihalomethane species. The results suggested that, depending on source water DOM and bromide concentration, selecting a suitable AER is a key factor in effective control of chlorinated and brominated DBPs in drinking water.

关键词： adsorption dissolved organic carbon drinking water treatment ion exchange

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LC-MS/MS method for the quantitation of serum tocilizumab in rheumatoid arthritis patients using rapid tryptic digestion without IgG purification

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Journal of Pharmaceutical Analysis 2022年第6期12卷 852-859页

作者： Takashi Mochizuki Kaito Shibata Takafumi Naito kumiko Shimoyama Noriyoshi Ogawa Masato Maekawa Junichi Kawakami Department of Hospital Pharmacy Hamamatsu University School of MedicineHamamatsu431-3192Japan Department of Pharmacy Shinshu University HospitalMatsumotoNagano390-8621Japan Third Division Department of Internal MedicineHamamatsu University School of MedicineHamamatsu431-3192Japan Department of Laboratory Medicine Hamamatsu University School of MedicineHamamatsu431-3192Japan

The quantitation of serum tocilizumab using liquid chromatography tandem-mass spectrometry(LC-MS/MS)method has not been widely applied in clinical settings because of its time-consuming and costly sample *** present study aimed to develop a validated LC-MS/MS method for detecting serum tocilizumab by utilizing immobilized trypsin without an immunoglobulin G purification step and evaluate its applicability in the treatment of rheumatoid arthritis(RA)patients administered intravenously or subcutaneously with *** tocilizumab-derived signature peptide was deciphered using a nano-LC system coupled to a hybrid quadrupole-orbitrap mass *** serum tocilizumab was rapidly digested by immobilized trypsin for 30 *** chromatographic peak of the signature peptide and that of the internal standard were separated from the serum digests for a total run time of 15 *** calibration curve of serum tocilizumab concentration was linear with a range of 2-200 μg/*** intra-and inter-day accuracy and relative standard deviation(RSD)were 90.7%-109.4%and<10%,*** serum tocilizumab concentrations in the RA patients receiving intravenous and subcutaneous injections were 5.8-28.9 and 2.4-63.5 μg/mL,*** serum tocilizumab concentrations using the current method positively correlated with those using the enzyme-linked immunosorbent assay,although a systematic error was observed between these *** conclusion,a validated LC-MS/MS method with minimal sample pretreatments for monitoring serum tocilizumab concentrations in RA patients was developed.

关键词： Tocilizumab LC-MS/MS Immobilized trypsin IgG purification Proteomics Rheumatoid arthritis

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Examination of in vivo mutagenicity of sodium arsenite and dimethylarsinic acid in gpt delta rats

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Journal of Environmental Sciences 2016年第11期28卷 125-130页

作者： Masaki Fujioka Min Gi Satoko Kawachi kumiko Tatsumi Naomi Ishii Kenichiro Doi Anna Kakehashi Hideki Wanibuchi Department of Molecular Pathology Osaka City University Graduate School of Medicine

Arsenic is a well-known human bladder and liver carcinogen, but its exact mechanism of carcinogenicity is not fully understood. Dimethylarsinic acid（DMAV） is a major urinary metabolite of sodium arsenite（i As~Ⅲ） and induces urinary bladder cancers in rats. DMAVand i As~Ⅲare negative in in vitro mutagenicity tests. However, their in vivo mutagenicities have not been determined. The purpose of present study is to evaluate the in vivo mutagenicities of DMAVand i As~Ⅲin rat urinary bladder epithelium and liver using gpt delta F344 ***-week old male gpt delta F344 rats were randomized into 3 groups and administered 0,92 mg/L DMAV, or 87 mg/L i As~Ⅲ（each 50 mg/L As） for 13 weeks in the drinking water. In the mutation assay, point mutations are detected in the gpt gene by 6-thioguanine selection（gpt assay） and deletion mutations are identified in the red/gam genes by Spi-selection（Spi-assay）. Results of the gpt and Spi-assays showed that DMAVand i As~Ⅲhad no effects on the mutant frequencies or mutation spectrum in urinary bladder epithelium or liver. These findings indicate that DMAVand i As~Ⅲare not mutagenic in urinary bladder epithelium or liver in rats.

关键词： Dimethylarsinic acid Sodium arsenite In vivo mutagenicity Urinary bladder epithelium gpt delta rat

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A Study on Cyclone Aila Recovery in Koyra, Bangladesh:Evaluating the Inclusiveness of Recovery with Respect to Predisaster Vulnerability Reduction

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International Journal of Disaster Risk Science 2018年第1期9卷 28-43页

作者： Md.Shibly Sadik Hajime Nakagawa Rezaur Rahman Rajib Shaw Kenji Kawaike kumiko Fujita Department of Civil and Earth Resources Engineering Kyoto University Center for Environmental and Geographic Information Services (CEGIS) Disaster Prevention Research Institute (DPRI) Kyoto University Institute of Water and Flood Management Bangladesh University of Engineering and Technology Graduate School of Media and Governance Shonan Fujisawa Campus (SFC) Keio University

The need to consider disaster risk reduction at the time of recovery is well-recognized. Viable disaster risk reduction measures should resolve the root causes of predisaster vulnerabilities. Accordingly, we investigated the recovery from the impact of Cyclone Aila in Koyra Upazila, Bangladesh, which was severely damaged by this2009 cyclone. Our research focused on understanding preAila vulnerabilities to cyclone impact and examined the degree of inclusion of vulnerability reduction measures within the recovery process. A composite methodology that included an institutional survey, key informant interviews,collection of the judgment of experts, focus group discussions, and a score-based quantification technique was adopted. Through a process of understanding pre-Aila vulnerabilities, recognition of the root causes of these inherent weaknesses, and identification of appropriate measures for pre-Aila vulnerability reduction, a set of 23 indicators were selected to represent the most desirable vulnerability reduction measures to implement duringrecovery. A score-based technique was applied to measure the degree of inclusion of vulnerability reduction within the recovery with respect to the indicators. The scoring result shows that the degree of inclusion of vulnerability reduction within the recovery was poor. The result specifies that among the 23 indicators of potential vulnerability reduction measures, 10 are completely missing and the rest are only partially included. The overall findings imply that the Koyra community continues to live with a vulnerability similar to that of the pre-Aila period.

关键词： Bangladesh Cyclone Aila Diagnostic analysis Disaster risk reduction Post-disaster reconstruction

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