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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 SCs.It 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 bodies.Regarding lifestyle diseases,we have already several therapeutic options,and thus,development of human pluripotent stem cell-based therapeutics tends to be avoided.Nevertheless,human pluripotent stem cells can contribute to the development of new therapeutics in this field.As 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 rejected.In 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 effects.Another utility of human pluripotent stem cells is the provision of an innovative tool for drug discovery that are otherwise unavailable.For 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 problems.However,BA can easily be produced from human pluripotent stem cells.In 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

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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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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) VECs.METHODS: 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 the regulation of VEC-VSMC interactions. We foun

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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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 arteriostenosis.Type-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?Matrix.The 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 arteries.Histologies of the femoral arteries were examined over time.The presence of human iPSdECs was checked by immunostaining studies using an antibody that specifically recognizes human VECs.Degrees of stenosis of the femoral arteries were calculated after three weeks.To 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 immunosuppressants.RESULTS:Because immunocompromised mice showed unexpected resistance to WI-induced arteriostenosis,we performed xenotransplantation experiments using immunocompetent mice along with immunosuppressant administrations.After one week,luminal surfaces of the WI-operated arteries were completely covered by human iPSdECs,showing the efficacy of our novel transplantation technique.After three weeks,type-IiPSdECs-transplanted arteries underwent total stenosis,while type-II-iPSdECs-transplanted arteries remained intact.However,untransplanted arteries of immunosuppressant-treated mice also remained intact by unknown reasons.We found that transplanted human VECs had already been replaced by murine endothelial cells by this time,indicating that a transie

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 but they were up-regulated in pathologica arterie

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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Stochastic model predictive braking control for heavy-duty commercial vehicles during uncertain brake pressure and road profile conditions

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Control Theory and Technology 2022年第2期20卷 248-262页

作者： Ryota nakahara Kazuma Sekiguchi Kenichiro Nonaka Masahiro Takasugi Hiroki Hasebe Kenichi Matsubara Graduate School of Integrative Science and Engineering Tokyo City UniversityTamazutsumiSetagaya-kuTokyo 1588557Japan Nabtesco Automotive Co. Ltd.HirakawachoChiyoda-kuTokyo 1020093Japan

When heavy-duty commercial vehicles(HDCVs)must engage in emergency braking,uncertain conditions such as the brake pressure and road profile variations will inevitably affect braking control.To minimize these uncertainties,we propose a combined longitudinal and lateral controller method based on stochastic model predictive control(SMPC)that is achieved via Chebyshev–Cantelli inequality.In our method,SMPC calculates braking control inputs based on a finite time prediction that is achieved by solving stochastic programming elements,including chance constraints.To accomplish this,SMPC explicitly describes the probabilistic uncertainties to be used when designing a robust control strategy.The main contribution of this paper is the proposal of a braking control formulation that is robust against probabilistic friction circle uncertainty effects.More specifically,the use of Chebyshev–Cantelli inequality suppresses road profile influences,which have characteristics that are different from the Gaussian distribution,thereby improving both braking robustness and control performance against statistical disturbances.Additionally,since the Kalman filtering(KF)algorithm is used to obtain the expectation and covariance used for calculating deterministic transformed chance constraints,the SMPC is reformulated as a KF embedded deterministic MPC.Herein,the effectiveness of our proposed method is verified via a MATLAB/Simulink and TruckSim co-simulation.

关键词： Heavy-duty commercial vehicle Brake system Stochastic model predictive control Road profile

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Fragmented QRS complex with an additional R-wave attenuated by short RR interval in a patient with acute pulmonary embolism and cardiogenic shock

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Journal of Geriatric Cardiology 2022年第12期19卷 995-1000页

作者： Koji Takahashi Hiromasa nakahara Eiji Arimitsu Satoshi Imamine Yoshiyasu Obata Kimio Nakanishi Yoshiyasu Taniguchi Maiko Amano Chika Omori Takafumi Okura Department of Community Emergency Medicine Ehime University Graduate School of MedicineEhimeJapan Department of Cardiology Yawatahama City General HospitalEhimeJapan Department of Internal Medicine Oozu City HospitalEhimeJapan Department of Clinical Laboratory Oozu City HospitalEhimeJapan

Registries and hospital discharge datasets of unselected patients with pulmonary embolism show 30-day all-cause mortality rates of approximately 10%.[1]Clinical findings from histories and physical examinations,such as age,systemic blood pressure,and respiratory rate at presentation,classify patients into risk classes of increasing risk of death and other adverse medical outcomes.

关键词： mortality patients Clinical

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Study on the optical property and biocompatibility of a tissue engineering cornea

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International Journal of Ophthalmology(English edition) 2012年第1期5卷 45-49页

作者： Yukiko nakahara Dwight Xuan Department of Medical Engineering Faculty of Health ScienceJunshin Gakuen UniversityChikushioka1-1-1Fukuoka 815-8510Japan Brooke Army Medical Center 3851 Roger Brooke Drive Fort Sam Houston TX 78234 USA

AIM: To study the optical property and biocompatibility of a tissue engineering cornea. METHODS: The cross-linker of N- (3-Dimethylaminoropyl)-N'ethylcarbodiimide hydrochloride (EDC)/ N-Hydroxysuccinimide (NHS) was mixed with Type I collagen at 10% (weight/volume). The final solution was molded to the shape of a corneal contact lens. The collagen concentrations of 10%, 12.5%, 15%, 17.5% and 20% artificial corneas were tested by UV/vis-spectroscopy for their transparency compared with normal rat cornea. 10-0 sutures were knotted on the edges of substitute to measure the corneal buttons's mechanical properties. Normal rat corneal tissue primary culture on the collagen scaffold was observed in 4 weeks. Histopathologic examinations were performed after 4 weeks of in vitro culturing. RESULTS: The collagen scaffold appearance was similar to that of soft contact lens. With the increase of collagen concentration, the transparency of artificial corneal buttons was diminished, but the toughness of the scaffold was enhanced. The scaffold transparency in the 10% concentration collagen group resembled normal rat cornea. To knot and embed the scaffold under the microscope, 20% concentration collagen group was more effective during implantation than lower concentrations of collagen group. In the first 3 weeks, corneal cell proliferation was highly active. The shapes of cells that grew on the substitute had no significant difference when compared with the cells before they were moved to the scaffold. However, on the fortieth day, most cells detached from the scaffold and died. Histopathologic examination of the primary culture scaffold revealed well grown corneal cells tightly attached to the scaffold in the former culturing. CONCLUSION: Collagen scaffold can be molded to the shape of soft contact corneal lens with NHS/EDC. The biological stability and biocompatibility of collagen from animal species may be used as material in preparing to engineer artificial corneal scaffold.

关键词： tissue engineering collagen cross-linking scaffold primary culture in vitro optical property biocompatibility

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Benefits of international trauma rotation in South Africa:comparison with a trauma center in Tokyo

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Emergency and Critical Care Medicine 2022年第4期2卷 179-180页

作者： Tsuyoshi Nagao Shinji nakahara Takashi Fujita Tetsuya Sakamoto Elmin Steyn Department of Emergency Medicine Teikyo University School of MedicineTokyoJapan Graduate School of Health Innovation Kanagawa University of Human ServicesKawasakiJapan Department of surgery Stellenbosch University and Tygerberg HospitalCape TownSouth Africa.

International rotations have been an option for young surgeons who face difficulties in obtaining sufficient trauma training in their own countries.One of the authors attended training in trauma surgery in South Africa for 6 months,where there are many trauma caseloads,and could experience the number of emergency operations equivalent to approximately 5 years in Japan.The rotation allows trainees to expose clinical practices intensively and to learn principles and theories of care under experienced academic surgeons.On the other hand,they could contribute to the host hospital as part of the workforce and by assisting young surgical residents.the development of a more systematic and formalized framework for staff exchange between developed and developing countries will provide mutual benefits to both sides.

关键词： Low-and middle-income countries On-the-job training Trauma surgery

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Improved image denoising via RAISR with fewer filters

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Computational Visual Media 2021年第4期7卷 499-511页

作者： Theingi Zin Yusuke nakahara Takuro Yamaguchi Masaaki Ikehara Department of Electronics and Electrical Engineering Keio UniversityYokohama-shi223-8522Japan

In recent years,accurate Gaussian noise removal has attracted considerable attention for mobile applications,as in smart phones.Accurate conventional denoising methods have the potential ability to improve denoising performance with no additional time.Therefore,we propose a rapid post-processing method for Gaussian noise removal in this paper.Block matching and 3D filtering and weighted nuclear norm minimization are utilized to suppress noise.Although these nonlocal image denoising methods have quantitatively high performance,some fine image details are lacking due to the loss of high frequency information.To tackle this problem,an improvement to the pioneering RAISR approach(rapid and accurate image super-resolution),is applied to rapidly post-process the denoised image.It gives performance comparable to state-of-the-art super-resolution techniques at low computational cost,preserving important image structures well.Our modification is to reduce the hash classes for the patches extracted from the denoised image and the pixels from the ground truth to 18 filters by two improvements:geometric conversion and reduction of the strength classes.In addition,following RAISR,the census transform is exploited by blending the image processed by noise removal methods with the filtered one to achieve artifact-free results.Experimental results demonstrate that higher quality and more pleasant visual results can be achieved than by other methods,efficiently and with low memory requirements.

关键词： block matching and 3D filtering weighted nuclear norm minimization super-resolution geometric conversion census transform

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