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11. 確定拠出年金における継続投資教育の効果 : 実験による検証

Author

Kitamura, Tomoki, Nakashima, Kunio, 世代間問題研究プロジェクト（世代間問題の経済分析） = Economic Analysis of Intergenerational Issues, Project on Intergenerational Equity, and 一橋大学経済研究所世代間問題研究機構 = Center for Intergenerational Studies, Institute of Economic Research, Hitotsubashi University

Abstract

2008/09/30, 退職給付ビッグバン研究会2008年度年次総会報告要旨, 科学研究費補助金（特別推進研究） = Grant-in-Aid for Specially Promoted Research

Published
2008

34. Construction of a new plasmid vector that can express cloned cDNA in all translational reading frames

Author

Imai Mutsuo, Kato Masanari, Itamura Shigeyuki, Hatanaka Masakazu, Nagai Jim, Nakashima Kunio, Shigesada Katsuya, Tanaka Minoru, and Masuda Naoki

Subjects
Genetic Vectors, Reading frame, Biology, Eukaryotic translation, Start codon, Complementary DNA, Genetics, Animals, Cloning, Molecular, Gene, Binding Sites, Expression vector, Base Sequence, cDNA library, DNA, General Medicine, beta-Galactosidase, Prolactin, Open reading frame, Gene Expression Regulation, Pituitary Gland, Protein Biosynthesis, Cattle, Ribosomes, Plasmids
Abstract

Construction of a bacterial expression vector, pO14001, is described. The vector contains the lac promoteroperator and three sets of ribosome-binding sites (RBSs) tandemly arranged in all possible reading frames. cDNA can be directly cloned downstream from these translational start points in the fixed and proper orientation by using the method of Okayama and Berg [Mol. Cell. Biol. 3 (1982) 280–289]. The open reading frame of any cDNA inserted may be automatically aligned in phase with either of the three ATG start codons, thus enabling its expression with a maximum theoretical probability of unity. Fusion with the lacZ gene (coding for β-galactosidase) has shown that at least two of the three translation initiation sites exhibit high expression capacities and the remaining one can also function at a lower but significant rate. We used the vector to construct a bovine pituitary cDNA library, from which clones coding for prolactin were detected by immunological screening with an efficiency as high as two in three clones. The construction with triple RBSs should also provide a unique experimental model to study the regulation of overlapping translations.

Published
1987
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35. Glycolytic and Gluconeogenic Metabolites and Enzymes in the Liver of Obese-Hyperglycemic Mice (KK) and Alloxan Diabetic Mice

Author

NAKASHIMA, KUNIO

Abstract

Hepatic glycolytic and gluconeogenic metabolites and enzymes were assayed in obese·hyperglycemic mice (KK), alloxan diabetic mice, and the control mice. Alloxan diabetic mice showed distinctly decreased and increased concentrations of metabolites in fed and starved conditions respectively. There were no differences between KK and the control in these conditions. But in glucose load tests, KK showed the retention of hepatic metabolites in the metabolic pathway of glycolysis at five positions. They were glucose, fructose-6-phosphate, glyceraldehyde-3-phosphate, 2-phosphoglycerate, and oxaloacetic acid. This phenomenon was also observed in alloxan diabetes. In alloxan diabetes, glucokinase, and pyruvate kinase activities were lowered while glucose-6-phosphatase and fructose-1, 6-diphosphatase activities were higher by 75% and 45%, respectively. On the contrary, KK mice showed higher glucokinase and hexokinase activities than the control by 20% as well as higher glucose-6-phosphatase activity (50% increase). Concentrations of corticosterone in the serum were the same levels in KK and the control while insulin-like activity was higher in KK. NAD+/NADH ratio in hepatic cytoplasm suggested the increased reaction in the direction of a-glycerophosphate in KK. High level of cytoplasm ATPIAMP ratio was supposed to be the cause of depression of glycolysis. And this might elevate the citrate concentration resulting in increased fatty acid synthesis.

Published
1969

36. 2‐Mercaptoethylamine, a competitive inhibitor of spermidine synthase in mammalian cells

Author

Hibasami, Hiroshige, Kawase, Masahiro, Tsukada, Tetsuya, Maekawa, Satoru, Sakurai, Masaki, and Nakashima, Kunio

Abstract

Spermidine synthase from rat ventral prostate was inhibited by 2‐mercaptoethylamine (MEA). Inhibition of spermidine synthase by MEA was competitive with respect to one of the substrates putrescine, but not competitive with respect to the other substrate decarboxylated S‐adenosylmethionine. MEA markedly depressed spermidine and spermine contents in human erythroid leukemia K562 cells, suggesting that these changes resulted from the inhibitory effect of MEA on spermidine synthase in situ.

Published
1988
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37. Proximity of mRNA5′-region and 18S rRNA in eukaryotic initiation complexes

Author

Nakashima, Kunio, Darzynkiewicz, Edward, and Shatkin, Aaron J.

Abstract

Messenger RNA was covalently linked to 18S ribosomal RNA in eukaryotic 40S and 80S initiation complexes by photoreaction with an RNA cross-linking agent, 4′-substituted psoralen. The sites of interaction of the mRNA capped 5′-region included some 3′-ends of 18S rRNA.

Published
1980
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39. Evaluation of immunoglobulin G subclass antibodies against recombinant Fasciola gigantica cathepsin L1 in an enzyme-linked immunosorbent assay for serodiagnosis of human fasciolosis.

Author

Wongkham C, Tantrawatpan C, Intapan PM, Maleewong W, Wongkham S, and Nakashima K

Subjects
Animals, Antigens, Helminth immunology, Case-Control Studies, Cathepsin L, Cathepsins immunology, Cysteine Endopeptidases immunology, Enzyme-Linked Immunosorbent Assay methods, Enzyme-Linked Immunosorbent Assay standards, Fasciola isolation & purification, Humans, Immunoglobulin G immunology, Predictive Value of Tests, Recombinant Proteins, Sensitivity and Specificity, Serologic Tests methods, Antibodies, Helminth, Antigens, Helminth blood, Cathepsins blood, Cysteine Endopeptidases blood, Fasciola enzymology, Fascioliasis diagnosis, Immunoglobulin G classification
Abstract

A cystatin capture enzyme-linked immunosorbent assay (ELISA) using recombinant Fasciola gigantica cathepsin L1 antigen was developed to detect specific immunoglobulin G (IgG) subclass antibodies (IgG1, IgG2, IgG3, and IgG4) and was evaluated for its diagnostic potential for human fasciolosis. In an analysis of the sera of 13 patients infected with F. gigantica, 209 patients with other parasitic infections, 32 cholangiocarcinoma patients, and 42 healthy controls, the IgG4-ELISA gave the highest diagnostic values. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method based on the detection of IgG4 antibody were 100%, 99.3%, 99.3%, 86.7%, and 100%, respectively. The results revealed that restricting the ELISA to the detection of specific IgG4 antibody enhanced the specificity and accuracy for the serodiagnosis of human fasciolosis.

Published
2005
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40. Deletion of dinucleotide repeat (Delta 14 allele) in the methylthioadenosine phosphorylase (MTAP) promoter and the allelotype of MTAP promoter in the Japanese population.

Author

Kadariya Y, Nishioka J, Nakatani K, Nakashima K, and Nobori T

Subjects
Alleles, Base Sequence, Cell Line, Tumor, Cloning, Molecular, DNA metabolism, Female, Gene Deletion, Genes, Reporter, Genetics, Population, Homozygote, Humans, Luciferases metabolism, Male, Molecular Sequence Data, Neoplasms metabolism, Pedigree, Plasmids metabolism, Polymerase Chain Reaction, Sequence Analysis, DNA, Transfection, Dinucleotide Repeats, Promoter Regions, Genetic, Purine-Nucleoside Phosphorylase genetics
Abstract

5'-deoxy-5'-methylthioadenosine phosphorylase (MTAP) is an enzyme involved in purine and polyamine metabolism and is ubiquitously expressed in normal human tissues and cells. However, this enzyme has been found to be deficient in a variety of human cancers. Although the enzyme deficiency is known to be caused by MTAP gene deletion, human diffuse histiocytic lymphoma cell line DHL-9 without any detectable MTAP activity has been found to possess the intact MTAP gene. These lines of evidence suggested that promoter abnormality might cause the MTAP deficiency in DHL-9. Therefore, we analyzed the MTAP promoter region of DHL-9 and found the deletion of 14 bases in its sequence. We designated the allele lacking (GT)(6)GC as dinucleotide repeat deletion (Delta 14 allele) and determined the effect of the Delta 14 allele on the MTAP promoter activity by a luciferase reporter assay. We have also analyzed the distribution of the Delta 14 allele and wild-type (WT) allele in the Japanese population by PCR assay. A reporter plasmid harboring the Delta 14 allele exhibited luciferase activity comparable to that of a plasmid containing the WT allele. Forty-six (22%) out of 210 people were homozygous for WT allele in the MTAP promoter, whereas 43 (20.5%) were homozygous for Delta 14 allele. The remaining 121 people (57.5%) possessed Delta 14/WT alleles in the MTAP promoter region. These results indicated that the Delta 14 allele has nothing to do with MTAP deficiency in DHL-9. The Delta 14 allele is distributed among the general population irrespective of gender.

Published
2002
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