98 research outputs found

    A One-Step Miniprep for the Isolation of Plasmid DNA and Lambda Phage Particles

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    Plasmid DNA minipreps are fundamental techniques in molecular biology. Current plasmid DNA minipreps use alkali and the anionic detergent SDS in a three-solution format. In addition, alkali minipreps usually require additional column-based purification steps and cannot isolate other extra-chromosomal elements, such as bacteriophages. Non-ionic detergents (NIDs) have been used occasionally as components of multiple-solution plasmid DNA minipreps, but a one-step approach has not been developed. Here, we have established a one-tube, one-solution NID plasmid DNA miniprep, and we show that this approach also isolates bacteriophage lambda particles. NID minipreps are more time-efficient than alkali minipreps, and NID plasmid DNA performs better than alkali DNA in many downstream applications. In fact, NID crude lysate DNA is sufficiently pure to be used in digestion and sequencing reactions. Microscopic analysis showed that the NID procedure fragments E.coli cells into small protoplast-like components, which may, at least in part, explain the effectiveness of this approach. This work demonstrates that one-step NID minipreps are a robust method to generate high quality plasmid DNA, and NID approaches can also isolate bacteriophage lambda particles, outperforming current standard alkali-based minipreps

    Shape selectivity in toluene disproportionation into para-xylene generated by chemical vapor deposition of tetramethoxysilane on MFI zeolite catalyst

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    Dependence of shape selectivity for para-xylene production by toluene disproportionation on conditions of chemical vapor deposition (CVD) of tetramethoxysilane on MFI (ZSM-5) zeolite were investigated in detail. The CVD after pelletization was necessary to obtain 0.7–1 mm particles with high selectivity. The influences of preparation conditions on the selectivity were investigated in detail to find the optimum conditions. The parent zeolite with small number of Brønsted acid sites on the external surface brought the high selectivity after the CVD. The catalyst prepared in the optimized conditions showed the selectivity 99.7% at ca. 10% of the toluene conversion

    COVID-19: challenges faced by Nepalese migrants living in Japan

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    Worldwide, COVID-19 has exacerbated the vulnerability of migrants, impacting many facets of their lives. Nepalese make up one of the largest groups of migrants residing in Japan. Crises, such as the ongoing COVID-19 pandemic could disproportionately affect migrants from low- and middle-income countries like Nepal, widening health and economic inequalities. An in-depth, comprehensive assessment is needed to appraise the diverse problems they encounter. Drawing upon qualitative interviews, this study aimed to identify challenges faced by Nepalese migrants in Japan as a consequence of the COVID-19 pandemic and to discuss their needs to counter these challenges. This qualitative study employed an interpretivist approach to appraise the first-hand experience of Nepalese migrants living in Japan. Fourteen participants (8 males and 6 females, aged 21 to 47 years old) were recruited to participate in semi-structured in-depth telephone interviews (45–60 min) regarding: (a) their perceived current physical and mental health, (b) problems faced as a result of the COVID-19 pandemic, and (c) perception of available and necessary support structures. Purposive and snowball sampling techniques were used to recruit the participants. Interviews were recorded, transcribed, and thematically analyzed. Six themes were identified: 1) experiencing psychosomatic symptoms, 2) adoption of new healthy behaviors, 3) financial hardship, 4) family concerns, 5) reflections on discrimination and 6) reflections of existing support and expectations of support systems. The findings of our study illustrate the specific impact of COVID-19 among Nepalese migrants regarding their unstable employment conditions, perceived lack of social support, possible obligation to send money home, difficulty in accessing services due to the language barrier, and a lack of effective governmental support from Nepal. Pandemic-related adversity has negatively impacted migrants’ mental well-being, exacerbating their vulnerability. Comprehensive and timely support should be provided to the vulnerable migrant population. Effective coordination among relevant parties in both countries, including the governments concerned, should be facilitated.Institute of Medical Care and Societal Health, Japan

    Effect of horizontal strong static magnetic field on swimming behaviour of Paramecium caudatum

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    Effect of horizontal strong static magnetic field on swimming behavior of Paramecium caudatum was studied by using a superconducting magnet. Around a center of a round vessel, random swimming at 0 T and aligned swimming parallel to the magnetic field (MF) of 8 T were observed. Near a wall of the vessel, however, swimming round and round along the wall at 0 T and aligned swimming of turning at right angles upon collision with the wall, which was remarkable around 1~4 T, were detected. It was experimentally revealed that the former MF-induced parallel swimming at the vessel center was caused physicochemically by the parallel magnetic orientation of the cell itself. From magnetic field dependence of the extent of the orientation, the magnetic susceptibility anisotropy (χ‖-χ⊥) was first obtained to be 3.4×10-23 emu cell-1 at 298 K for Paramecium caudatum. The orientation of the cell was considered to result from the magnetic orientation of the cell membrane. On the other hand, although mechanisms of the latter swimming near the vessel wall regardless of the absence and presence of the magnetic field are unclear at present, these experimental results indicate that whether the cell exists near the wall alters magnetic field effect on the swimming in the horizontal magnetic field

    The Catalytic Conversion of 1,2-Propandiol to Propanal on FSM-16 Molded by Wet-Treatment and Pressurization

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    The catalytic conversion of 1,2-propandiol to propanal is examined using FSM-16 particles (0.85-1.70 mm) molded by wet-treatment and pressurization. When FSM-16 was molded with 0.6 g of pressurization and supplied to the catalytic conversion of 1,2-propandiol at 673 K, this system resulted in a 94.8% conversion of 1,2-propandiol and 90.5% selectivity to propanal at 0.25 h on-stream, which was the maximum amount of activity. However, at 4.50 h on-stream, the activity decreased extremely to deactivation 19.9% conversion of 1,2-propandiol and 84.7% selectivity to propanal. In contrast, when FSM-16 molded with wet-treatment (0.15 g) was used for the conversion at 573 K, activity was greatly increased and stable 98.6% conversion of 1,2-propandiol and 56.2% selectivity to propanal at 0.25 h on-stream followed by 91.9% and 52.5%, respectively, at 4.50 h on-stream. The hexagonal structure of FSM-16 was suggested to have contributed to the suitable conversion of 1,2-propandiol to propanal

    Videoconferencing for Home Care Delivery in Japan: Observational Study

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    Telemedicine has been increasingly used in many health care fields, including home care, where patients receive medical care at home. Owing to the current COVID-19 crisis, the value of telemedicine via videoconferencing is more recognized, particularly in allowing immobile patients to continue receiving care. However, the efficacy of telemedicine in home care settings in Japan remains to be fully appraised. This study aims to identify the use and impact of telemedicine in a singular home care delivery setting in Japan. A retrospective observational study was conducted using patient and other administrative records from a home care clinic. We considered patients who were involved in videoconferencing with home care physicians and telepresenters serving patients during 2018 and 2019. We extracted sociodemographic data of the patients and details of the videoconferencing and descriptively illustrated some specific cases. In a home care clinic in Japan, videoconferencing was conducted in 17 cases (involving 14 patients) over a 2-year period. Of all the cases, 12% (2/17) required emergency transfers and were hospitalized. A total of 88% (15/17) of cases remained; 71% (12/17) of cases were found to need extra medication or to go to a medical facility for consultation, whereas 18% (3/17) of cases were found not to be in need of urgent attention and were asked to rest. Problematic symptoms subsequently improved in 82% (14/17) of cases, and only 6% (1/17) of cases were later hospitalized. Telemedicine was deemed effective for assessing patients’ conditions in the home care setting in situations where home visits by a physician cannot be carried out. Our findings indicate that consultations via videoconferencing are safe and effective, suggesting more active use of videoconferencing in other clinical contexts.N/

    Tumor-Derived Microvesicles Induce Proangiogenic Phenotype in Endothelial Cells via Endocytosis

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    Background: Increasing evidence indicates that tumor endothelial cells (TEC) differ from normal endothelial cells (NEC). Our previous reports also showed that TEC were different from NEC. For example, TEC have chromosomal abnormality and proangiogenic properties such as high motility and proliferative activity. However, the mechanism by which TEC acquire a specific character remains unclear. To investigate this mechanism, we focused on tumor-derived microvesicles (TMV). Recent studies have shown that TMV contain numerous types of bioactive molecules and affect normal stromal cells in the tumor microenvironment. However, most of the functional mechanisms of TMV remain unclear. Methodology/Principal Findings: Here we showed that TMV isolated from tumor cells were taken up by NEC through endocytosis. In addition, we found that TMV promoted random motility and tube formation through the activation of the phosphoinositide 3-kinase/Akt pathway in NEC. Moreover, the effects induced by TMV were inhibited by the endocytosis inhibitor dynasore. Our results indicate that TMV could confer proangiogenic properties to NEC partly via endocytosis. Conclusion: We for the first time showed that endocytosis of TMV contributes to tumor angiogenesis. These findings offer new insights into cancer therapies and the crosstalk between tumor and endothelial cells mediated by TMV in the tumor microenvironment

    SORL1 Is Genetically Associated with Late-Onset Alzheimer’s Disease in Japanese, Koreans and Caucasians

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    To discover susceptibility genes of late-onset Alzheimer’s disease (LOAD), we conducted a 3-stage genome-wide association study (GWAS) using three populations: Japanese from the Japanese Genetic Consortium for Alzheimer Disease (JGSCAD), Koreans, and Caucasians from the Alzheimer Disease Genetic Consortium (ADGC). In Stage 1, we evaluated data for 5,877,918 genotyped and imputed SNPs in Japanese cases (n = 1,008) and controls (n = 1,016). Genome-wide significance was observed with 12 SNPs in the APOE region. Seven SNPs from other distinct regions with p-values ,261025 were genotyped in a second Japanese sample (885 cases, 985 controls), and evidence of association was confirmed for one SORL1 SNP (rs3781834, P=7.3361027 in the combined sample). Subsequent analysis combining results for several SORL1 SNPs in the Japanese, Korean (339 cases, 1,129 controls) and Caucasians (11,840 AD cases, 10,931 controls) revealed genome wide significance with rs11218343 (P=1.7761029) and rs3781834 (P=1.0461028). SNPs in previously established AD loci in Caucasians showed strong evidence of association in Japanese including rs3851179 near PICALM (P=1.7161025) and rs744373 near BIN1 (P = 1.3961024). The associated allele for each of these SNPs was the same as in Caucasians. These data demonstrate for the first time genome-wide significance of LOAD with SORL1 and confirm the role of other known loci for LOAD in Japanese. Our study highlights the importance of examining associations in multiple ethnic populations
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