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Biomedical subjects

T Tanabe

Publications and source records attributed to T Tanabe.

At least 271 records · Page 15Linked to original sources
PubMed1995

Inhibition by interleukin-10 of inducible cyclooxygenase expression in lipopolysaccharide-stimulated monocytes: its underlying mechanism in comparison with interleukin-4.

Both interleukin-10 (IL-10) and IL-4 inhibited the prostanoid synthesis of lipopolysaccharide (LPS)-stimulated human monocytes, and their inhibition was shown to be based on a common mechanism to suppress the gene expression of inducible cyclooxygenase (COX). COX has been shown to exist in at least two distinct isoforms, designated COX-1 and COX-2, and their gene expressions exhibit different profiles. At both the protein and mRNA levels, the expression of COX-1 was constitutive and was not modulated by treatments with LPS, IL-10, or IL-4. In contrast, the expression of COX-2 was observed only after stimulation with LPS. IL-10 and IL-4 significantly inhibited LPS-induced COX-2 expression. Kinetic studies showed that they inhibited COX-2 mRNA expression within 1 hour after stimulation and that maximal inhibition was consistently observed at 5 hours. Moreover, the addition of cycloheximide (CHX) to LPS-stimulated monocytes resulted in a superinduction of COX-2 mRNA, whereas CHX almost abrogated the abilities of IL-10 and IL-4 to inhibit this gene expression. Experiments with actinomycin D showed that both cytokines accelerated the degradation of COX-2 mRNA. Furthermore, nuclear run-on experiments showed that both cytokines modestly inhibited LPS-induced COX-2 gene transcription. Thus, both cytokines seemed to regulate the COX-related pathway in a similar manner, although their receptor systems did not show any structural similarities. Considering recent findings showing that the drugs that exhibit a selective effect on COX-2 may be more preferable in inflammatory conditions, such biologic activities of IL-10 and IL-4 described above may offer useful tools in controlling inflammatory disorders in the future.

Cells, Cultured↗
PubMed1995

Modulation of cloned neuronal calcium channels through membrane-delimited pathway.

Modulation of voltage-gated Ca2+ channels by the receptor coupled GTP-binding proteins (G-proteins) is essential for controlling secretion and muscle contraction. We have expressed cloned Ca2+ channels in dysgenic myotubes to study G-protein modulation through the membrane-delimited pathway. The results obtained by the expression of alpha 1B channels and mutant channels of alpha 1B suggest that the two effects observed in G-protein modulated N-type channels (depression of current and slowing of activation) are through two independent mechanisms. In addition, neither the region linking repeat II and III nor carboxy-terminal region, which were demonstrated in L-type channels to determine some of their specific functions, are directly involved in G-protein modulation. The results obtained by the expression of the alpha 1A channel suggest that this channel is modulated through a novel membrane-delimited pathway that may not involve G-protein activation.

Animals↗
PubMed1995

Human alpha-tocopherol transfer protein: cDNA cloning, expression and chromosomal localization.

alpha-Tocopherol transfer protein (alpha TTP), which specifically binds this vitamin and enhances its transfer between separate membranes, was previously isolated from rat liver cytosol. In the current study we demonstrated the presence of alpha TTP in human liver by isolating its cDNA from a human liver cDNA library. The cDNA for human alpha TTP predicts 278 amino acids with a calculated molecular mass of 31,749, and the sequence exhibits 94% similarity with rat alpha TTP at the amino acid level. The recombinant human alpha TTP expressed in Escherichia coli exhibits both alpha-tocopherol transfer activity in an in vitro assay and cross-reactivity to the anti-(rat alpha TTP) monoclonal antibody. Northern blot analysis revealed that human alpha TTP is expressed in the liver like rat alpha TTP. The human and rat alpha TTPs show structural similarity with other apparently unrelated lipid-binding/transfer proteins, i.e. retinaldehyde-binding protein present in retina, and yeast SEC14 protein, which possesses phosphatidylinositol/phosphatidylcholine transfer activity. Both Southern-blot hybridization of human-hamster somatic cell hybrid lines and fluorescence in situ hybridization revealed a single alpha TTP gene corresponding to the 8q13.1-13.3 region of chromosome 8, which is identical to the locus of a recently described clinical disorder, ataxia with selective vitamin E deficiency (AVED). The relationship between alpha TTP and AVED will be discussed.

Amino Acid Sequence↗
PubMed1995

Co-expression in CHO cells of two muscle proteins involved in excitation-contraction coupling.

Ryanodine receptors and dihydropyridine receptors are located opposite each other at the junctions between sarcoplasmic reticulum and either the surface membrane or the transverse tubules in skeletal muscle. Ryanodine receptors are the calcium release channels of the sarcoplasmic reticulum and their cytoplasmic domains form the feet, connecting sarcoplasmic reticulum to transverse tubules. Dihydropyridine receptors are L-type calcium channels that act as the voltage sensors of excitation-contraction coupling: they sense surface membrane and transverse tubule depolarization and induce opening of the sarcoplasmic reticulum release channels. In skeletal muscle, ryanodine receptors are arranged in extensive arrays and dihydropyridine receptors are grouped into tetrads, which in turn are associated with the four subunits of ryanodine receptors. The disposition allows for a direct interaction between the two sets of molecules. CHO cells were stably transformed with plasmids for skeletal muscle ryanodine receptors and either the skeletal dihydropyridine receptor, or a skeletal-cardiac dihydropyridine receptor chimera (CSk3) which can functionally substitute for the skeletal dihydropyridine receptor, in addition to plasmids for the alpha 2, beta and gamma subunits. RNA blot hybridization gave positive results for all components. Immunoblots, ryanodine binding, electron microscopy and exposure to caffeine show that the expressed ryanodine receptors forms functional tetrameric channels, which are correctly inserted into the endoplasmic reticulum membrane, and form extensive arrays with the same spacings as in skeletal muscle. Since formation of arrays does not require coexpression of dihydropyridine receptors, we conclude that self-aggregation is an independent property of ryanodine receptors. All dihydropyridine receptor-expressing clones show high affinity binding for dihydropyridine and immunolabelling with antibodies against dihydropyridine receptor. The presence of calcium currents with fast kinetics and immunolabelling for dihydropyridine receptors in the surface membrane of CSk3 clones indicate that CSk3-dihydropyridine receptors are appropriately targeted to the cell's plasmalemma. The expressed skeletal-type dihydropyridine receptors, however, remain mostly located within perinuclear membranes. In cells coexpressing functional dihydropyridine receptors and ryanodine receptors, no junctions between feet-bearing endoplasmic reticulum elements and surface membrane are formed, and dihydropyridine receptors do not assemble into tetrads. A separation between dihydropyridine receptors and ryanodine receptors is not unique to CHO cells, but is found also in cardiac muscle, in muscles of invertebrates and, under certain conditions, in skeletal muscle. We suggest that failure to form junctions in co-transfected CHO cell may be due to lack of an essential protein necessary either for the initial docking of the endoplasmic reticulum to the surface membrane or for maintaining the interaction between dihydropyridine receptors and ryanodine receptors. We also conclude that formation of tetrads requires a close interaction between dihydropyridine receptors and ryanodine receptors.

Animals↗
PubMed1995

Regulation of two isozymes of prostaglandin endoperoxide synthase and thromboxane synthase in human monoblastoid cell line U937.

The mechanism responsible for the rapid increase of thromboxane A2 synthesis by cells of the human monoblastoid cell line U937, which were differentiated with 12-O-tetradecanoyl-phorbol-13-acetate, induced by lipopolysaccharide (LPS) was studied. Both RNA blot and immunoblot analyses showed that LPS increased the levels of prostaglandin endoperoxide synthase-1 (PES-1) and -2 (PES-2) in a time-dependent manner, and the modes of induction of the two isozymes differed. The maximum PES-1 mRNA level was 1.6 times higher 36 h after than before stimulation by LPS, and that of PES-2 mRNA was elevated about 20-fold at its peak at 12 h after stimulation. Consequently, the immunoreactive PES-1 and PES-2 protein levels also increased time-dependently after LPS stimulation. However, the effects of LPS on the thromboxane synthase mRNA and protein levels were much less marked. These results indicate that LPS-induced thromboxane synthesis by the differentiated cells was regulated at the levels of the two PES isozymes, predominantly at the PES-2 level.

Arachidonic Acids↗
PubMed1995

Alterations of sympathovagal balance in patients with hypertrophic and dilated cardiomyopathies assessed by spectral analysis of RR interval variability.

Spectral analysis of RR interval variability was performed in 35 ambulatory patients with early hypertrophic cardiomyopathy (HCMa, NYHA class I), 21 hospitalized patients with advanced hypertrophic cardiomyopathy (HCMh, NYHA class II or III), and 18 hospitalized patients with dilated cardiomyopathy (DCMh, NYHA class I, II or III). Twenty-nine ambulatory subjects (COTa) and 20 hospitalized volunteers (COTh) served as normal controls. The RR interval standard deviation (SD), the high-frequency power (HF: 0.15-0.40 Hz) corrected by the mean RR interval (CCVHF) and the normalized unit of the HF power (NUHF) served as markers of vagal modulation. Low-frequency power (LF: 0.04-0.15 Hz) corrected by the mean RR interval (CCVLF) and the normalized unit of LF power (NULF) were markers of sympathetic modulation. The LF/HF ratio was an index of sympathovagal balance. There was no significant difference in the SD, CCVHF, NUHF, CCVLF, NULF or the LF/HF ratio between the HCMa and COTa groups. At night, the SD was lower in the HCMh group relative to the COTh group (P < 0.01). The HCMh group demonstrated lower CCVHF and NUHF values (P < 0.01), higher NULF values (P < 0.01) and higher LF/HF ratios (P < 0.05) at night relative to the COTh group. Two patients who later died suddenly in the HCMh group had markedly reduced CCVHF values (0.2-0.8%) relative to the survivors in the group (mean +/- SD in the morning, afternoon and night, 1.07 +/- 0.43%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Optimum culture conditions for production of exfoliative toxin by Staphylococcus hyicus.

Optimum culture conditions for the production of exfoliative toxin by Staphylococcus hyicus (shET) were examined. High shET activity was obtained from the culture filtrate of HI and TY broth inoculated with S. hyicus. The pH in these two media ranged from 7 to 8.5 during bacterial culture, while the lowest pH in TS and BHI broth was less than 6. shET activity in the culture filtrate from TY broth inoculated with 10(7) CFU of S. hyicus per ml was higher than that in TY broth inoculated with 10(6) and 10(8) CFU of bacteria per ml. When shET activity in the culture filtrate was measured under various shaking conditions, the culture filtrate shaken at 75 oscillations per min had the highest shET activity of the five shaking conditions. shET activity of the culture filtrate of TY broth to which protease inhibitor had been added was the same as that of TY broth without inhibitor. shET activity in a shaking culture in an Erlenmeyer flask was also the same as that in sac culture and that in shaking culture using a shaking (Sakaguchi) flask. shET activity in TY broth supplemented with 100 mM glucose was significantly lower than that in TY broth without glucose. Based on the above results, the optimum culture conditions for the production of shET were as follows: inoculation of 3 x 10(9) CFU of S. hyicus strain P-1 into 300 ml of TY broth in a 2,000-ml Erlenmeyer flask, and incubation at 37 C with shaking at 75 oscillations per min.(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonium Sulfate↗
PubMed1995

Possible receptor for exfoliative toxins produced by Staphylococcus hyicus and Staphylococcus aureus.

Exfoliative toxin produced by Staphylococcus hyicus bound to the GM4-like glycolipid extracted from the skin of 1-day-old chickens but did not bind to glycolipid from adult chickens or suckling mice. Exfoliative toxin produced by Staphylococcus aureus bound to the GM4-like glycolipid extracted from the skin of suckling mice but not to glycolipid from 1-day-old or adult chickens. S. hyicus and S. aureus exfoliative toxins lost their toxicity by preincubation with GM4-like glycolipid from 1-day-old chickens and suckling mice, respectively.

Animals↗
PubMed1995

New markers of remote ischemia in patients with evolving inferior myocardial infarction.

Multivessel disease and decreased left ventricular ejection fraction (LVEF) are believed to be significant predictors of the outcome in patients with acute inferior myocardial infarction (AIMI). We attempted to determine new electrocardiographic (ECG) markers for detecting concomitant left anterior descending (LAD) disease and/or decreased left ventricular function in patients with AIMI. Eighty patients with AIMI were evaluated within 6 h of the onset of symptoms and grouped according to the presence (Group 1) or absence (Group 2) of concomitant LAD disease. All of the patients underwent coronary angiography and left ventriculography 4-6 weeks from the onset of their infarction. We studied the validity of two new ECG markers: S-T depression deeper in lead V5 than in V4 (S-T decreases V5 > V4) and negative U waves (NUs) > 0.5 mm (50 muV) in leads V4-6. The sensitivity and specificity of S-T decreases V5 > V4, NUs in V4-6, or both, in detecting concomitant LAD disease were 56% and 83%, 59% and 87%, and 35% and 98%, respectively. LAD lesions in patients who showed either of these new markers (74% of those with S-T decreases V5 > V4 and 80% of those with NUs in V4-6) were mostly in the proximal segments (AHA segments #6 or #7). Patients with either S-T decreases V5 > V4 or NUs in V4-6 tended to have asynergy in the anterolateral segment, while there was a strong correlation between the asynergy of the anterolateral and septal segments in patients who showed both ECG markers.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

[Effect of ulinastatin on the free radical during cardiopulmonary bypass].

Cardiopulmonary bypass (CPB) involves many factors for generating free radicals. It is difficult to detect free radical directly in clinical situations. Vitamin E is recognized as an intrinsic radical scavenger. We examined the change of vitamin E level to prove indirectly the evidence of free radical generation during CPB. Moreover, we investigated the effect of ulinastatin on the free radical during CPB by determining the vitamin E level. Twenty patients who received cardiac surgery were divided into the membrane oxygenator group and the bubble oxygenator group. In each group, five patients received 12,000 units.kg-1 of ulinastatin 15 min after starting CPB. Blood samples were obtained 15 min after starting CPB and 15 min after reperfusion of coronary artery. In the bubble oxygenator group without ulinastatin, vitamin E level at reperfusion phase decreased significantly compared with that at the start of CPB. In the bubble oxygenator group with ulinastatin, vitamin E level at reperfusion phase showed no decrease compared with the level at the start of CPB. In the membrane oxygenator group, there was no significant change in vitamin E level. Therefore, we suspect that a decrease of vitamin E during CPB with bubble oxygenator demonstrates the evidence of free radical generation and these results suggest that ulinastatin may scavenge free radicals during CPB.

Adult↗
PubMed1994

Characterization of the human gene (TBXAS1) encoding thromboxane synthase.

The gene encoding human thromboxane synthase (TBXAS1) was isolated from a human EMBL3 genomic library using human platelet thromboxane synthase cDNA as a probe. Nucleotide sequencing revealed that the human thromboxane synthase gene spans more than 75 kb and consists of 13 exons and 12 introns, of which the splice donor and acceptor sites conform to the GT/AG rule. The exon-intron boundaries of the thromboxane synthase gene were similar to those of the human cytochrome P450 nifedipine oxidase gene (CYP3A4) except for introns 9 and 10, although the primary sequences of these enzymes exhibited 35.8% identity each other. The 1.2-kb of the 5'-flanking region sequence contained potential binding sites for several transcription factors (AP-1, AP-2, GATA-1, CCAAT box, xenobiotic-response element, PEA-3, LF-A1, myb, basic transcription element and cAMP-response element). Primer-extension analysis indicated the multiple transcription-start sites, and the major start site was identified as an adenine residue located 142 bases upstream of the translation-initiation site. However, neither a typical TATA box nor a typical CAAT box is found within the 100-b upstream of the translation-initiation site. Southern-blot analysis revealed the presence of one copy of the thromboxane synthase gene per haploid genome. Furthermore, a fluorescence in situ hybridization study revealed that the human gene for thromboxane synthase is localized to band q33-q34 of the long arm of chromosome 7. A tissue-distribution study demonstrated that thromboxane synthase mRNA is widely expressed in human tissues and is particularly abundant in peripheral blood leukocyte, spleen, lung and liver. The low but significant levels of mRNA were observed in kidney, placenta and thymus.

Amino Acid Sequence↗
PubMed1994

The cyclic AMP response element plays an essential role in the expression of the human prostaglandin-endoperoxide synthase 2 gene in differentiated U937 monocytic cells.

The promoter activity of 1432 bp upstream of the human prostaglandin-endoperoxide synthase 2 gene (PTGS2) was examined in differentiated U937 monocytic cells expressing prostaglandin-endoperoxide synthase 2 mRNA. Transient transfection experiments were performed using these cells and reporter vectors containing the upstream region of the gene with deletions or site-specific mutations and the luciferase gene. The deletion or destruction of the cyclic AMP response element (nucleotides -59 to -53) markedly reduced the promoter activity of this gene. Electrophoretic mobility shift assays showed that a nuclear protein(s) binding to the cyclic AMP response element was induced during monocytic differentiation of U937 cells. These results indicate that expression of the human prostaglandin-endoperoxide synthase 2 gene in differentiated U937 monocytic cells is regulated by the cyclic AMP response element.

Base Sequence↗