PRIMARY STRUCTURE AND PRODUCT CHARACTERIZATION OF THE SACCHAROMYCES-CEREVISIAE CHO1-GENE THAT ENCODES PHOSPHATIDYLSERINE SYNTHASE

An open reading frame of 828 base pairs was found in the CHOI gene region of Saccharomyces cerevisiae by nucleotide sequencing analysis. Its enhanced expression with the aid of the PH05 regulatory sequence resulted in an overproduction of a protein with a molecular weight of approximately 30,000, which in turn was converted by proteolysis to active phosphatidylserine synthase, whose molecular weight was approximately 23,000. The larger protein was concluded to be the primary product of the CHOI gene, since its amino-terminal sequence was identical to that deduced from the nucleotide sequence of the above open reading frame, except for the terminal methionine residue. A partial homology in primary structures was noticed between this yeast enzyme and phosphatidylglycerophosphate synthase of Escherichia coli which also uses CDP-diacylglycerol as a substrate. The overproduced phosphatidylserine synthase in both microsomal and extensively purified fractions displayed two different Km values for L-serine, i.e., 0.14 mM at low L-serine concentrations and 9.5 mM at high L-serine concentrations. This may indicate a negatively cooperative regulation of this enzyme activity or the presence of two active components with different affinities for L-serine. This work was supported in part by a Grant-in-Aid for Special Project Research on Intracellular Processing of Biomolecules for the Expression of their Biological Functions from the Ministry of Education, Science and Culture of Japan, and by Special Coordination Funds for Promoting Science and Technology from, the Science and Technology Agency of Japan.textapplication/pdfjournal articl

Observation of b→dγ and Determination of |Vtd/Vts|

journal articl

太陽光発電システムと直接接続可能な小型風力発電装置の開発に関する研究

application/pdf三重大学大学院工学研究科博士前期課程電気電子工学専攻66thesi

Essay about the Ore deposit of Sunokobashi district in the Ashio Mine（足尾銅山簀子橋鉱床調査意見書）

東京帝国大学工科大学種別:卒業論文thesi

マルクスにおける宗教と文明－ マルクスの宗教理解について（六）

1987-03departmental bulletin pape

Astrocyte-Specific Genes Are Generally Demethylated in Neural Precursor Cells Prior to Astrocytic Differentiation

Epigenetic changes are thought to lead to alterations in the property of cells, such as differentiation potential. Neural precursor cells (NPCs) differentiate only into neurons in the midgestational brain, yet they become able to generate astrocytes in the late stage of development. This differentiation-potential switch could be explained by epigenetic changes, since the promoters of astrocyte-specific marker genes, glial fibrillary acidic protein (Gfap) and S100b, have been shown to become demethylated in late-stage NPCs prior to the onset of astrocyte differentiation; however, whether demethylation occurs generally in other astrocyctic genes remains unknown. Here we analyzed DNA methylation changes in mouse NPCs between the mid-(E11.5) and late (E14.5) stage of development by a genome-wide DNA methylation profiling method using microarrays and found that many astrocytic genes are demethylated in late-stage NPCs, enabling the cell to become competent to express these genes. Although these genes are already demethylated in late-stage NPCs, they are not expressed until cells differentiate into astrocytes. Thus, late-stage NPCs have epigenetic potential which can be realized in their expression after astrocyte differentiation.journal articl

Validity of the Intake of Sugars, Amino Acids, and Fatty Acids Estimated Using a Self-administered Food Frequency Questionnaire in Middle-aged and Elderly Japanese: The Japan Public Health Center-based Prospective Study for the Next Generation (JPHC-NEXT) Protocol Area

Background: The Japanese database of food composition was revised in 2020, during which both the number of food items and the number of food items measured for sugars, amino acids, and fatty acids were increased. We evaluated the validity of estimated intakes of sugars, amino acids and fatty acids using a long food frequency questionnaire (long-FFQ) among middle-aged and elderly Japanese. Methods: From 2012 to 2013, 240 men and women aged 40–74 years from five areas in the JPHC-NEXT protocol were asked to respond to the long-FFQ and provide a 12-day weighed food record (WFR) as reference. The long-FFQ, which included 172 food and beverage items and 11 seasonings, was compared with a 3-day WFR, completed during each distinct season, and validity was assessed using Spearman’s correlation coefficients. Results: Percentage differences based on the long-FFQ with the 12-day WFR in men and women varied from −84.4% to 419.6%, and from −75.8% to 623.1% for sugars, −17.5% to 3.8% and −5.8% to 19.6% for amino acids, and −58.5% to 78.8% and −43.4% to 129.3% for fatty acids, respectively. Median values of correlation coefficients for the long-FFQ in men and women were 0.52 and 0.42 for sugars, 0.38 and 0.37 for amino acids, and 0.42 and 0.42 for fatty acids, respectively. Conclusion: The long-FFQ provided reasonable validity in estimating the intakes of sugars, amino acids, and fatty acids in middle-aged and elderly Japanese. Although caution is warranted for some nutrients, these results may be used in future epidemiological studies

IL-17A-mediated enhanced killing of <i>S. pneumoniae</i>.

<p>A and B. Effect of human IL-17A on surface phagocytic killing of <i>S. pneumoniae</i>. A. Isolated neutrophils from healthy adult volunteers were incubated with recombinant human IL-17A at the indicated concentrations and evaluated in a surface phagocytic killing assay with pneumococcal strain 0603; colonies were counted after overnight incubation at 37°C with 5% CO₂. IL-17A induces a dose-dependent enhancement of neutrophil killing of <i>S. pneumoniae</i> (P = 0.01 for 1 or 10 µg of IL-17A vs. no added IL-17A). B. Supernatant obtained from neutrophils after incubation with IL-17A did not have any demonstrable antipneumococcal effect, whereas washed neutrophils after incubation with IL-17A demonstrated enhanced killing. C. Effect of human IL-17A on opsonophagocytic killing of <i>S. pneumoniae</i>. Neutrophils purified from the peripheral blood of healthy adult volunteers were incubated with pneumococci anticapsular antibodies, complement, and a range of concentrations of IL-17A as indicated for 90 minutes, following which viable counts were obtained by plating on blood agar plates. Each line represents a different volunteer. IL-17A enhanced killing of pneumococci in a dose-dependent fashion in 6/6 subjects. *P = 0.016 by Wilcoxon matched pairs test.</p

Role of T-helper-subset-associated cytokines in protection from nasopharyngeal colonization.

<p>A. Mice defective in IFN-γ, IL-4 or IL-17A receptor were immunized as described, then challenged with pneumococcal strain 0603. Mice with IFN-γ or IL-4 deficiency were significantly protected by WCV (P<0.001 vs. respective CT controls) whereas IL-17A receptor deficient mice were not protected (P>0.5 vs. CT). Dashed line represents the lower limit of detection of bacterial colonization. B. Expression of IL-17A from splenocytes of WCV-immunized mice. Cultured splenocytes from mice immunized with WCV (black columns) or CT alone (white columns) were stimulated for 72 hours with medium alone, Concanavalin A (5 µg/ml), WCA (10 µg dry weight) after which IL-17A production was measured by ELISA. Significantly more IL-17A was expressed following WCA stimulation of WCV-immunized vs. CT-immunized mice, although response to concanavalin A was similar. C. Effect of CD4+ T cell depletion upon IL-17A expression from splenocytes. Splenocytes (without or with CD4+ T cell depletion) from mice immunized with WCV were stimulated for 72 hours with medium alone or WCA after which IL-17A was measured by ELISA. IL-17A expression in splenocytes following WCA stimulation was significantly higher in the presence of CD4+ T cells compared to stimulation with medium alone or when CD4+ T cells were depleted. Repletion of CD4+ T cells restored the response. ** P<0.01 compared to cells stimulated with medium alone. D. IL-17A intracellular staining of splenocytes from WCV immunized mice. Splenocytes from WCV immunized mice were stimulated with WCA, blocked with monensin, harvested and stained for CD4+ and intracellular IL-17A as described. There is a statistically significant increase in CD4+ IL-17A positive cells following stimulation with WCA, which is not observed in the CD4- population. No increase in IL-17A positive cells could be detected in cells from unimmunized mice (data not shown). **P = 0.008 for comparison of frequency of IL-17A-positive cells in absence and presence of WCA stimulation among CD4+ cells. Data shown here are representative of three experiments, including at least 5 mice per experiment. E. Expression of IL-17A from NALT of WCV- vs. CT-immunized mice. Cultured splenocytes from mice immunized with WCV (black columns) or CT alone (white columns) were stimulated for 72 hours with medium alone or with WCA (10 µg dry weight) after which IL-17A production was measured by ELISA. Significantly more IL-17A was expressed following WCA stimulation of WCV-immunized vs. CT-immunized mice. **P<0.01 for comparison of IL-17A in WCV vs. CT-immunized mice following stimulation with WCA.</p

Correlation of IL-17A expression and density of nasopharyngeal colonization in mice.

<p>Three weeks after immunization of mice (n = 90) with CT with doses of WCA ranging from 1 to 100 µg, and one week before pneumococcal challenge, blood samples were obtained and stimulated with WCA (10 µg) for 6 days, after which supernatants were collected and assayed for IL-17A concentration. The correlation between density of colonization (cfu/nasal wash) 7 days after challenge and pre-challenge IL-17A expression was evaluated. IL-17A expression was significantly correlated with density of colonization.</p