患者由来人工多能性幹細胞を用いた遺伝子治療の至適化戦略

学位の種別: 課程博士審査委員会委員 : （主査）東京大学教授 中内 啓光, 東京大学准教授 佐藤 均, 東京大学教授 武川 睦寛, 東京大学教授 吉田 進昭, 東京大学准教授 高橋 聡University of Tokyo(東京大学

Paxillin facilitates timely neurite initiation on soft-substrate environments by interacting with the endocytic machinery.

Neurite initiation is the first step in neuronal development and occurs spontaneously in soft tissue environments. Although the mechanisms regulating the morphology of migratory cells on rigid substrates in cell culture are widely known, how soft environments modulate neurite initiation remains elusive. Using hydrogel cultures, pharmacologic inhibition, and genetic approaches, we reveal that paxillin-linked endocytosis and adhesion are components of a bistable switch controlling neurite initiation in a substrate modulus-dependent manner. On soft substrates, most paxillin binds to endocytic factors and facilitates vesicle invagination, elevating neuritogenic Rac1 activity and expression of genes encoding the endocytic machinery. By contrast, on rigid substrates, cells develop extensive adhesions, increase RhoA activity and sequester paxillin from the endocytic machinery, thereby delaying neurite initiation. Our results highlight paxillin as a core molecule in substrate modulus-controlled morphogenesis and define a mechanism whereby neuronal cells respond to environments exhibiting varying mechanical properties

Construction and prokaryotic expression of the fusion gene PRRSV GP5 and Mycobacterium bovis Hsp70

Porcine reproductive and respiratory syndrome (PRRS) is an economically important swine disease that has devastated the swine industry worldwide. Vaccination with live attenuated vaccine or inactivated vaccine is the main treatment to control PRRS. However, the disadvantages such as virulence resumption of the attenuated vaccine and low immunogenicity of the inactivated vaccine call for a more efficient and safer genetically engineered vaccine. In this study, the structural protein GP5 of the PRRS virus (PRRSV), one of the major protective antigens which stimulates a protective immune response was selected to develop a genetically engineered subunit vaccine. In order to promote the immune reaction of the host to GP5, heat shock protein 70 (Hsp70) was selected as immuno-adjuvant to enhance PRRSV GP5 immunogenicity. The Hsp70 gene was amplified by PCR from attenuated Mycobacterium bovis, and the PRRSV GP5 gene was amplified by RT-PCR from the total RNA of PRRSV SCQ strain which was isolated, identified and maintained by the Animal Biotechnological Center, Sichuan Agricultural University, China. The fusion expressing plasmid pET32-GP5-Hsp70 was constructed and expressed in Escherichia coli BL21. Ni2+-chelating resin was used to purify the His-tagged fusion protein expressed under optimized expressing conditions. The rabbit anti-GP5-Hsp70 fusion protein antibody was made, and Western blot assay verified the successful expression of the fusion protein, making it possible for further investigation whether Hsp70 could improve the immunogenicity of the PRRSV GP5 subunit vaccine, or evaluating the immunogenicity of the GP5-Hsp70 subunit vaccine.Keywords: Porcine reproductive and respiratory syndrome virus (PRRSV) GP5 gene, Mycobacterium bovis Hsp70 gene, cloning, prokaryotic expression, identification.African Journal of Biotechnology Vol. 12(30), pp. 4754-476

High expression FUT1 and B3GALT5 is an independent predictor of postoperative recurrence and survival in hepatocellular carcinoma.

Cancer may arise from dedifferentiation of mature cells or maturation-arrested stem cells. Previously we reported that definitive endoderm from which liver was derived, expressed Globo H, SSEA-3 and SSEA-4. In this study, we examined the expression of their biosynthetic enzymes, FUT1, FUT2, B3GALT5 and ST3GAL2, in 135 hepatocellular carcinoma (HCC) tissues by qRT-PCR. High expression of either FUT1 or B3GALT5 was significantly associated with advanced stages and poor outcome. Kaplan Meier survival analysis showed significantly shorter relapse-free survival (RFS) for those with high expression of either FUT1 or B3GALT5 (P = 0.024 and 0.001, respectively) and shorter overall survival (OS) for those with high expression of B3GALT5 (P = 0.017). Combination of FUT1 and B3GALT5 revealed that high expression of both genes had poorer RFS and OS than the others (P < 0.001). Moreover, multivariable Cox regression analysis identified the combination of B3GALT5 and FUT1 as an independent predictor for RFS (HR: 2.370, 95% CI: 1.505-3.731, P < 0.001) and OS (HR: 2.153, 95% CI: 1.188-3.902, P = 0.012) in HCC. In addition, the presence of Globo H, SSEA-3 and SSEA-4 in some HCC tissues and their absence in normal liver was established by immunohistochemistry staining and mass spectrometric analysis

利用日、月及年降雨量估算年降雨沖蝕指數

The Universal Soil Loss Equation (USLE) is presently one of the most widely used models for evaluating soil erosion. The rainfall amount is often used to estimate the R factor for Universal Soil Loss Equation (USLE) modeling. Daily, monthly, and annual rainfall data are commonly available. The purpose of the present study is to establish three models for estimating rainfall erosivity based on daily, monthly, and annual precipitation, respectively, and to identify the most suitable model in the Ailiao River watershed located in southern Taiwan. The data set consists of 2266 storm events, monitored by 6 rainfall stations there. The results show that the 6 stations indicate strong positive relationship (r2 > 0.75) between annual rainfall erosivity and annual rainfall with a 99% confidence level, which means the simplified estimation methods based on annual precipitation are useful for predicting long-term annual rainfall erosivity in most of locations in the Ailiao River watershed. The results also show that annual and monthly precipitation correlate with erosivity better than does daily precipitation based on the analysis results of root mean square error (RMSE) and mean absolute percentage error (MAPE) in the Ailiao River watershed.降雨沖蝕指數是通用土壤流失公式 (universal soil loss equation, USLE) 重要的參數之一，由於計算降雨沖蝕指數需要30 分鐘或更短量測間距的雨量資料，但大部分地區只有日、月或年雨量資料。因此利用何種降雨資料推估年平均降雨沖蝕指數較為接近30 分鐘雨量資料計算的年平均降雨沖蝕指數，是本研究的重點。本研究以隘寮溪集水區為研究區域，蒐集區域內6 個雨量站10 年 (2002-2011 年)10 分鐘等間隔的降雨資料，計算年平均降雨沖蝕指數 (Ryc)，另以日、月及年降雨資料，建立三種降雨量與降雨沖蝕指數關係式，利用RMSE、MAPE 及Bias 等誤差分析方法，評估三種降雨沖蝕指數推估關係式之適用性。研究結果顯示，利用年降雨資料推估年降雨沖蝕指數的適用性較好。本研究結果可提供缺乏短量測間距雨量資料地區年降雨沖蝕指數推估之參考依據

Regular exercise and the trajectory of health-related quality of life among Taiwanese adults: a cohort study analysis 2006–2014

Abstract Background Physical activity is related to health-related quality of life, but little evidence from multiple waves of panel data in Asian countries area available. This study aims to explore the impacts of different degree of regular exercise on the trajectories of physical and mental dimensions of health-related quality of life (HRQOL) for community-dwelling Taiwanese adults during 2006–2014. Methods Data were derived from the “Landseed Integrated Outreaching Neighborhood Screening (LIONS)” study, with 6182 adults enrolled at the baseline and subsequently followed up to three times till 2014. Linear mixed-effects modeling approach was employed to evaluate the growth curve models of HRQOL (with 16,281 observations) by linear & quadratic time effects, regular exercise (5-level moderate-intensity physical activity), and major influential factors of HRQOL. Results Regular exercise showed significantly positive dose-response effects on physical HRQOL (β =1.27~2.54), and regular exercise of 150 min or more showed positive effects on mental HRQOL (β = 1.55~2.03). Besides, irregular exercise could also improve both physical and mental HRQOL (β = 1.27 & β = 0.87). However, such effects were not significant over time (at time slope) on HRQOL. In addition, physical and mental HRQOL improved across time (β = 1.01 and 1.49, respectively), but the time quadratic effect would significantly offset a little bit on physical dimension (β = − 0.22). Moreover, being female, increasing age, living alone, or poorer health status were related to lower physical HRQOL; and being younger, living alone, or poorer health status were associated with lower mental HRQOL. Conclusions The positive dose-response relationship between regular exercise and HRQOL or its domains was demonstrated for community-dwelling Taiwanese adults. Thus, a regular exercise habit (better ≧150 min per week) is advised for community-based healthcare professionals and the government to incorporate into health promotion strategies and plans.https://deepblue.lib.umich.edu/bitstream/2027.42/152207/1/12889_2019_Article_7662.pd

Microwave amplification via interfering multi-photon processes in a half-waveguide quantum electrodynamics system

We investigate the amplification of a microwave probe signal by a superconducting artificial atom, a transmon, strongly coupled to the end of a one-dimensional semi-infinite transmission line. The end of the transmission line acts as a mirror for microwave fields. Due to the weak anharmonicity of the artificial atom, a strong pump field creates multi-photon excitations among the dressed states. Transitions between these dressed states, Rabi sidebands, give rise to either amplification or attenuation of the weak probe. We obtain a maximum amplitude amplification of about 18 %, higher than in any previous experiment with a single artificial atom, due to constructive interference between Rabi sidebands. We also characterize the noise properties of the system by measuring the spectrum of spontaneous emission

Biobutanol production in a Clostridium acetobutylicum biofilm reactor integrated with simultaneous product recovery by adsorption

BACKGROUND: Clostridium acetobutylicum can propagate on fibrous matrices and form biofilms that have improved butanol tolerance and a high fermentation rate and can be repeatedly used. Previously, a novel macroporous resin, KA-I, was synthesized in our laboratory and was demonstrated to be a good adsorbent with high selectivity and capacity for butanol recovery from a model solution. Based on these results, we aimed to develop a process integrating a biofilm reactor with simultaneous product recovery using the KA-I resin to maximize the production efficiency of biobutanol. RESULTS: KA-I showed great affinity for butanol and butyrate and could selectively enhance acetoin production at the expense of acetone during the fermentation. The biofilm reactor exhibited high productivity with considerably low broth turbidity during repeated batch fermentations. By maintaining the butanol level above 6.5 g/L in the biofilm reactor, butyrate adsorption by the KA-I resin was effectively reduced. Co-adsorption of acetone by the resin improved the fermentation performance. By redox modulation with methyl viologen (MV), the butanol-acetone ratio and the total product yield increased. An equivalent solvent titer of 96.5 to 130.7 g/L was achieved with a productivity of 1.0 to 1.5 g · L(-1) · h(-1). The solvent concentration and productivity increased by 4 to 6-fold and 3 to 5-fold, respectively, compared to traditional batch fermentation using planktonic culture. CONCLUSIONS: Compared to the conventional process, the integrated process dramatically improved the productivity and reduced the energy consumption as well as water usage in biobutanol production. While genetic engineering focuses on strain improvement to enhance butanol production, process development can fully exploit the productivity of a strain and maximize the production efficiency