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H Shibata

Publications and source records attributed to H Shibata.

At least 343 records · Page 19Linked to original sources
PubMed1995

[5'-Nucleotidase].

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5'-Nucleotidase↗
PubMed1994

Inhibition of ATP-sensitive K+ channel by a non-sulfonylurea compound KAD-1229 in a pancreatic beta-cell line, MIN 6 cell.

We studied the mechanism of action of KAD-1229, a non-sulfonylurea compound shown to stimulate insulin secretion, in a glucose responsive insulinoma cell line, MIN 6 cells. In microsomal fraction of MIN 6 cells, KAD-1229 displaced binding of [3H]glibenclamide in a concentration-dependent manner. The dissociation constant and the maximum binding capacity were 0.61 nM and 8.70 pmol/mg.protein, respectively. In inside out configuration of patch-clamp technique, KAD-1229 attenuated the opening of ATP-sensitive K+ channels. The effect of KAD-1229 was detected at 10(-8) M, and 10(-5) M KAD-1229 almost completely blocked the activity of ATP-sensitive K+ channel. When membrane potential was monitored by a perforated mode of patch clamp, KAD-1229 induced depolarization of plasma membrane, which was followed by a burst of action potentials. These action potentials were blocked by cobalt. In a fura-2-loaded single MIN 6 cell, KAD evoked an elevation of intracellular free Ca2+ concentration, [Ca2+]i. The KAD-1229-mediated elevation of [Ca2+]i was attenuated by either removal of extracellular Ca2+ or an addition of nifedipine. Finally, KAD-1229 augmented insulin secretion in MIN 6 cells in a concentration-dependent manner. KAD-1229 also enhanced the effect of glucose and nifedipine inhibited the action of KAD-1229 on insulin secretion. These results indicate that KAD-1229 stimulates insulin secretion by stimulating Ca2+ influx and that, despite the lack of sulfonylurea structure, KAD-1229 binds to sulfonylurea receptors and inhibits the activity of ATP-sensitive K+ channel in MIN 6 cells.

Action Potentials↗
PubMed1994

Activation of PKN, a novel 120-kDa protein kinase with leucine zipper-like sequences, by unsaturated fatty acids and by limited proteolysis.

PKN, a novel protein kinase with catalytic domain homologous to PKC family and unique amino terminal leucine zipper-like sequences, was purified partially from COS7 cells transfected with the cDNA construct encoding human PKN for enzymatic characterization of the enzyme. Using serine containing synthetic peptides based on PKC pseudosubstrate sites as the phosphate acceptors, kinase activities estimated from partially purified PKN were not stimulated by Ca2+/phosphatidylserine/diolein but were activated several-fold to several tens-fold by 40 microM unsaturated fatty acids, such as arachidonic acid, linoleic acid, and oleic acid. Autophosphorylation of the immunoprecipitates using anti-PKN antiserum was also stimulated by various unsaturated fatty acids. Limited proteolysis of PKN with trypsin induced an enhancement of the peptide kinase activity that was almost independent of arachidonic acid.

Amino Acid Sequence↗
PubMed1994

Lactate-dependent killing of Escherichia coli by nitrite plus hydrogen peroxide: a possible role of nitrogen dioxide.

The killing of Escherichia coli by nitrite plus hydrogen peroxide was observed in lactate, but not in phosphate or acetate. Although nitrite or hydrogen peroxide alone caused a slight decrease in bacterial survival, nitrite plus hydrogen peroxide killed bacteria synergistically in time-, dose-, and pH-dependent manners. The killing was increased with decreasing pH. The plot of viable cells versus [nitrous acid] was linear. Among the hydroxyl radical scavengers used, only benzoate and formate at concentrations higher than that of lactate inhibited the killing by nitrite plus hydrogen peroxide, whereas dimethyl sulfoxide enhanced it. The generation of peroxynitrous acid during the reaction of nitrite and hydrogen peroxide was confirmed by the formation of malondialdehyde using deoxyribose as a hydroxyl radical-like oxidant detector. The nitration of glycyl-tryosine was observed only in lactate buffer, but not in phosphate and acetate buffers. Benzoate and formate inhibited the nitration, whereas dimethyl sulfoxide and ethanol enhanced it. No evidence for the formation of nitric oxide and superoxide during the reaction of nitrite and hydrogen peroxide was found. These data suggest that nitrogen dioxide from the decomposition of peroxynitrous acid or secondary oxidants formed from the reaction of peroxynitrous acid with lactate is responsible for the lactate-dependent killing of E. coli induced by the reaction of protonated nitrite and hydrogen peroxide.

Animals↗
PubMed1994

Single amino acid substitution (58Pro-->Ser) in HTLV-I tax results in loss of ras cooperative focus formation in rat embryo fibroblasts.

Two tax genes cloned from a healthy HTLV-I carrier in whom the viral genome is clonally integrated into peripheral CD4+CD8+ cells showed a considerable difference in ras cooperative focus forming ability in rat embryo fibroblasts (REF). Sequence analysis revealed differences in two codons of the two genes. SH-1tax and SH-2tax. Studies using recombinants between these two tax genes showed that the deficiency in ability of SH-1tax to cooperate with ras in focus formation in REF was caused by a 58Pro-->Ser substitution. This amino acid substitution did not affect other tax functions such as colony formation in soft agar, focus formation in Rat-1 cells, immortalization of REF, and transcriptional activation through the CREB/ATF and NFkB/rel pathways. These results suggest that the domain of tax required for cooperative focus formation with ras in primary rat fibroblasts may be different from those required for other tax functions.

Animals↗
PubMed1994

Rate of calcium entry determines the rapid changes in protein kinase C activity in angiotensin II-stimulated adrenal glomerulosa cells.

The present study was conducted to monitor precisely the activity of protein kinase C (PKC) in adrenal glomerulosa cells stimulated by angiotensin II (ANG II). PKC activity in cells was monitored by measuring phosphorylation of a synthetic KRTLRR peptide, a specific substrate for PKC, immediately after the permeabilization of the cells with digitonin [Heasley and Johnson J. Biol. Chem. (1989) 264, 8646-8652]. Addition of 1 nM ANG II induced a gradual increase in KRTLRR peptide phosphorylation, which reached a peak at 30 min, and phosphorylation was sustained thereafter. When the action of ANG II was terminated by adding [Sar1,Ala8]ANG II, a competitive antagonist, both Ca2+ entry and KRTLRR phosphorylation ceased rapidly, whereas diacylglyercol (DAG) content was not changed significantly within 10 min. Similarly, when blockade of Ca2+ entry was achieved by decreasing extracellular Ca2+ to 1 microM or by adding 1 microM nitrendipine, KRTLRR peptide phosphorylation was decreased within 5 min. In addition, restoration of Ca2+ entry was accompanied by an immediate increase in KRTLRR peptide phosphorylation. Under the same condition, DAG content did not change significantly. We then examined the role of the PKC pathway in ANG II-induced aldosterone production. Ro 31-8220 inhibited ANG II-induced KRTLRR phosphorylation without affecting the activity of calmodulin-dependent protein kinase II. In the presence of Ro 31-8220, ANG II-mediated aldosterone production was decreased to approx. 50%. Likewise, intracellular administration of PKC19-36, a sequence corresponding to residues 19-36 of the regulatory domain of PKC known to inhibit PKC activity, attenuated ANG II-mediated activation of PKC and aldosterone output. These results indicate a critical role of Ca2+ entry in the regulation of PKC activity by ANG II.

Aldosterone↗
PubMed1994

Quantitative analyses of the structure-hydrophobicity relationship for N-acetyl di- and tripeptide amides.

The partition coefficient (P) of neutral species and the apparent partition ratio (P') at pH 7 of the ionized form were measured with the 1-octanol/water system for a number of N-acetyl di- and tripeptide amides having un-ionizable and ionizable side chains. Their log values were studied in terms of free-energy-related substituent and substructural parameters using regression analysis to give correlation equations of high quality physicochemically as well as statistically. The intrinsic hydrophobicity of side-chain substituents and their steric effect on the relative solvation of the backbone CONH groups were significant in determining the log P values of the un-ionizable acetyl peptide amides. For the log P value of peptides with polar side-chain substituents, respective indicator variable terms were required to account for the sum of specific effects of substituents such as intramolecular hydrogen-bond formation and the "polar proximity factor" for augmentation of the hydrophobicity. For the log P'(pH 7) value of basic and acidic peptides, the ability of counterionic species to form ion-pairs, the change in the apparent hydrophobicity of ionizable groups from the intrinsic value for their nonionized forms, the effect of ion-pairing itself, and other effects were additionally considered. From the regression coefficients of the parameter terms in correlation equations an effective hydrophobicity index was defined for each side chain, and the application and its limitation were suggested.

Acetylation↗
PubMed1994

Terminal distribution of projections from the retrosplenial area to the retrohippocampal region in the rat, as studied by anterograde transport of biotinylated dextran amine.

The terminal distribution of projections from the retrosplenial area to the retrohippocampal region was examined in the rat with anterograde transport of biotinylated dextran amine. Projections from the retrosplenial granular area (RSG) to the retrohippocampal region terminate predominantly ipsilaterally in layers I, III, V and VI of the presubiculum, layers I and IV-VI of the parasubiculum, the molecular and pyramidal cell layers of the subiculum, and layers I, III, V and VI of the entorhinal area. On the other hand, projections from the retrosplenial agranular area (RSA) terminate predominantly ipsilaterally in layers I and III of the presubiculum and layers V and VI of the entorhinal and perirhinal areas, and ipsilaterally in layers IV-VI of the parasubiculum. The results show that projections from the RSG to the retrohippocampal region are as massive as those from the RSA, and that each retrosplenial area has distinct projection fields in the retrohippocampal region. This suggests that each retrosplenial area may play some distinct functional roles in memory and learning processes such as spatial behavioral learning.

Animals↗
PubMed1994

Temperature high sensitivity of manganese uptake in ileal longitudinal smooth muscle of guinea-pig.

1. Mn2+ (5 mM) inhibited completely the K+ (60 mM)-induced ileal tonic tension to the base line, however, the tension increased progressively to the above level of the original K+ tonic response after 3 hr of the Mn2+ application at 37 degrees C. 2. At 30, 32 or 34 degrees C, the tensions which developed after 3 hr of the addition of 5 mM Mn2+ in the high-K+ medium was 0, 21, 48% of their original K+ tonic levels, respectively. 3. The tension development and manganese uptake in the presence of Mn2+ in the high-K+ medium was highly dependent on the temperature in very narrow range of 32-37 degrees C in the suspending medium.

Animals↗
PubMed1994

Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using the RLGS method.

A new imprinted gene has been discovered in mice using the technique of restriction landmark genomic scanning (RLGS) with methylation sensitive enzymes. Eight out of 3,100 strain-specific NotI and BssHII spots were identified as imprinted in reciprocal F1 hybrids. Subsequently, we isolated a genomic clone for one locus on proximal chromosome 11 near the Glns locus, an imprinted region in uniparental disomic mice, and its corresponding cDNA clone. Expression of this transcript from the paternal allele was established using RT-PCR of reciprocal F1-hybrid mice. The amino-acid sequence deduced from the cDNA showed significant homology to the U2 small nuclear ribonucleoprotein auxiliary factor 35 kDa subunit.

Amino Acid Sequence↗
PubMed1994

Rate of falls and the correlates among elderly people living in an urban community in Japan.

Eight hundred and seven elderly persons aged 65-84 years who were randomly selected from the community were interviewed about falls. The rate of falls in the preceding year was 12.8% in men and 21.5% in women. The difference in the rate between the sexes was statistically significant. The rate in the older group was significantly higher than that in the younger group in both sexes, when they were divided into two major age categories. Multivariate logistic regression analysis showed that contact with a doctor within a month preceding the survey in both sexes, age, history of stroke, visual deficit in men, and urinary or bowel incontinence in women were significantly associated with increased risk of falling. The study reveals that falls tend to occur in physically frail elderly people in the community.

Accidental Falls↗
PubMed1994

A genetic linkage map of the mouse using restriction landmark genomic scanning (RLGS).

We have developed a multiplex method of genome analysis, restriction landmark genomic scanning (RLGS) that has been used to construct genetic maps in mice. Restriction landmarks are end-labeled restriction fragments of genomic DNA that are separated by using high resolution, two-dimensional gel electrophoresis identifying as many as two thousand landmark loci in a single gel. Variation for several hundred of these loci has been identified between laboratory strains and between these strains and Mus spretus. The segregation of more than 1100 RLGS loci has been analyzed in recombinant inbred (RI) strains and in two separate interspecific genetic crosses. Genetic maps have been derived that link 1045 RLGS loci to reference loci on all of the autosomes and the X chromosome of the mouse genome. The RLGS method can be applied to genome analysis in many different organisms to identify genomic loci because it uses end-labeling of restriction landmarks rather than probe hybridization. Different combinations of restriction enzymes yield different sets of RLGS loci providing expanded power for genetic mapping.

Animals↗