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Biomedical subjects

K Shimada

Publications and source records attributed to K Shimada.

At least 487 records · Page 27Linked to original sources
PubMed1997

Serotonin inhibits nitric oxide synthesis in rat vascular smooth muscle cells stimulated with interleukin-1.

We investigated the effects of serotonin (5-hydroxytryptamine; 5-HT) on nitric oxide (NO) synthesis in vascular smooth muscle cells. We measured the production of nitrite, a stable metabolite of NO, and the expression of inducible NO synthase protein in cultured rat vascular smooth muscle cells. Incubation of the cultures with interleukin-1beta (10 ng/ml) caused a significant increase in nitrite production. 5-HT inhibited nitrite production by interleukin-1beta -stimulated vascular smooth muscle cells in a concentration-dependent manner (10(-8)-10(-5) M). 5-HT-induced inhibition of nitrite production was accompanied by decreased inducible NO synthase protein accumulation in vascular smooth muscle cells. Addition of the 5-HT2 receptor antagonist ketanserin, but not the 5-HT1A receptor antagonist spiroxatrine, inhibited the effect of 5-HT. On the other hand, the 5-HT2 receptor agonist alpha-methyl-5-HT, but not the 5-HT1A receptor agonist (+/-)-8-hydroxy-2-(di-n-propylamino) tetralin, decreased interleukin-1beta-induced nitrite production by vascular smooth muscle cells. 5-HT significantly increased protein kinase C activity in vascular smooth muscle cells, and the protein kinase C inhibitor calphostin C dose-dependently abolished the effect of 5-HT on nitrite production. After protein kinase C activity was functionally depleted by treatment of cells with phorbol 12-myristate 13-acetate for 24 h, the effect of 5-HT was abolished. These results indicate that 5-HT acts on 5-HT2 receptors and inhibits NO synthesis in interleukin-1beta-stimulated vascular smooth muscle cells at least partially through a protein kinase C-dependent pathway.

Animals↗
PubMed1997

SOCS-1/JAB/SSI-1 can bind to and suppress Tec protein-tyrosine kinase.

Tec is the prototype of a recently emerging subfamily among nonreceptor type protein-tyrosine kinases and is known to become tyrosine-phosphorylated and activated by a wide range of cytokine stimulations in hematopoietic cells. Although Tec was recently shown to be involved in the cytokine-driven activation mechanism of c-fos transcription, it is yet obscure how Tec relays the signals from cell surface receptors to the nucleus. To identify signaling molecules acting downstream of Tec, we have looked for Tec-interacting proteins (TIPs) by using the yeast two-hybrid system. Here we report the identification and characterization of a novel protein, TIP3, which has been simultaneously identified by other groups as SOCS-1, JAB, or SSI-1. TIP3 carries one Src homology 2 domain with a sequence similarity to that of CIS. In 293 cells, TIP3 associates with Tec and suppresses its kinase activity. Interestingly, TIP3 can also down-regulate the activity of Jak2 but not that of Lyn. We propose that SOCS-1/JAB/SSI-1/TIP3 is a novel type of negative regulator to a subset of protein-tyrosine kinases.

Amino Acid Sequence↗
PubMed1997

GABAergic neurotransmission in rat taste buds: immunocytochemical study for GABA and GABA transporter subtypes.

Gamma-aminobutyric acid (GABA) is known to be a candidate for the neurotransmitter involved in the sense of taste. We hereby studied GABA and its termination system, GABA transporters, in rat taste buds by immunocytochemical approaches. Immunoblot analysis of three GABA transporter subtypes (GAT1, GAT2 and GAT3) revealed that the immunoreactive bands of GAT2 and GAT3, but not GAT1, were detected in the tongue. GAT3-immunoreactive band was recognized only in the circumvallate papilla containing a large number of taste buds while GAT2-immunoreactive bands were seen in all areas of the tongue. GAT2 immunoreactivity appeared to be specifically in the nerve fibers beneath the lingual epithelium. Both GAT3 and GABA immunoreactivities were detected only in taste buds. A few GAT3-immunoreactive cells were found in a cross-section of each taste bud but most GAT3-immunoreactive cells were localized in the margin of the taste bud. GAT3 was predominantly concentrated in the distal portion of the GAT3-immunoreactive cells. In contrast, GABA-immunoreactive cells were seen more frequently within each taste bud and the immunoreactivity was distributed throughout the perikarya of the cells. These results suggest that the GABA-uptake system is present in the taste buds and the GABAergic neurotransmission involved in the sensation of taste is terminated by the uptake of GABA into certain taste cells via GAT3.

Animals↗
PubMed1997

Promoter of the Na,K-ATPase alpha3 subunit gene is composed of cis elements to which NF-Y and Sp1/Sp3 bind in rat cardiocytes.

Na,K-ATPase alpha subunit has three isoforms whose expression is regulated developmentally and hormonally. Na,K-ATPase alpha3 subunit gene (Atpla3) is expressed only in brain and neonatal heart in a rat. The purpose of this study is to analyze cis-acting elements and trans-acting factors regulating the transcription of Atpla3 in cultured neonatal rat cardiocytes. Transient transfection assays with Atpla3-luciferase chimeric construct and a series of 5' sequential deletion mutations revealed the existence of positive regulatory elements from -74 to -59 and from -59 to -39. A factor was identified to bind across -59 by gel retardation assay. Methylation interference and DNase I footprinting analyses revealed the binding region from -74 to -53 (positive regulatory element (PRE) 1). The binding factor was identified to be NF-Y by gel retardation assay using specific antibody. Gel retardation and methylation interference analyses revealed that factors bind to two other elements from -54 to -43 (PRE2) and from -25 to -13 (PRE3). The binding factors were identified to be Sp1/Sp3 using specific antibodies. The functions of above-mentioned three elements were examined by transient transfection assay with various combinations of mutations. They all regulated the transcription positively and a synergistic enhancement of it was observed. Roles of NF-Y in the transcriptional activation and synergy are discussed.

Animals↗
PubMed1997

Tranilast inhibits contraction of rat aortic smooth muscle.

Recently, the anti-allergic drug tranilast has been shown to reduce the rate of coronary restenosis after percutaneous transluminal coronary angioplasty. In this study, we investigated the effect of tranilast on contraction of and Ca2+ movement in vascular smooth muscle. We measured the isometric force and fura-2-estimated intracellular Ca2+ concentrations ([Ca2+]i) of rat aortic strips. Exposure of aortic strips to tranilast (0-500 microM) dose-dependently inhibited endothelin-1-induced increases in tension and [Ca2+]i elevation of the strips. Similar inhibition by tranilast was observed in response to high K+ stimulation. These results suggest that tranilast inhibits the contraction of vascular smooth muscle by inhibiting Ca2+ mobilization, which might be related to its preventive effect on coronary restenosis after percutaneous transluminal coronary angioplasty.

Animals↗
PubMed1997

Biodistribution of liposomes containing synthetic galactose-terminated diacylglyceryl-poly(ethyleneglycol)s.

We describe the synthesis of biodegradable poly(ethyleneglycol)-coupled galactolipids in which the galactose moiety is separated from a diacylglyceride lipid anchor by poly(ethylene glycol) chains of 10, 20 or 40 oxyethylene residues (PEG10/20/40). These Gal-PEG lipids (Gal-PEG-Lip) were incorporated in the bilayer of liposomes. The surface exposure of the galactose was investigated by aggregation experiments with ricinus communis agglutinin 120. Only the liposomes containing the PEG10 galactolipid aggregated with the lectin. Therefore liposomes were prepared containing Gal-PEG10-Lip and a trace amount of [3H]cholesteryl oleyl ether with an average diameter of approximately 100 nm and injected intravenously into rats. The Gal-PEG10-Lip liposomes were cleared from plasma with a T1/2 of 0.3 h. Identically sized and composed control liposomes without the Gal-PEG10-Lip had a T1/2 of approximately 12 h. The rapid plasma elimination of the Gal-PEG10-Lip liposomes could be attributed entirely to increased uptake by the liver amounting to more than 90% of injected dose. Uptake by the spleen was decreased to less than 1% of injected dose. A single injection of N-acetylgalactosamine 1 min prior to Gal-PEG-Lip liposome administration reduced the initial rate of plasma clearance to control levels. The increased liver uptake was almost entirely attributable to increased uptake by the Kupffer cells. Incorporation of PEG-DSPE in the Gal-PEG10-Lip liposomes only partially reversed the effect of the galactolipid with respect to liver and spleen uptake as well as intrahepatic distribution. These experiments demonstrate that liposome surface-exposed galactose residues, even if attached at the distal end of a poly(ethyleneglycol) chain anchored in the liposomal bilayer are effectively recognized by the galactose particle receptor on the Kupffer cells but fail to achieve significant targeting to the asialoglycoprotein receptor on the hepatocytes.

Acetylgalactosamine↗
PubMed1997

Tranilast inhibits the growth of rat mesangial cells.

We investigated the effects of tranilast on the growth of cultured rat mesangial cells. The number of mesangial cells increased fivefold during a 5-day incubation in RPMI 1640 with 20% fetal bovine serum. The number of cells was significantly lower in the presence of tranilast than in its abscence. Tranilast (0 approximately 500 microM) inhibited platelet-derived growth factor (PDGF)-induced DNA synthesis of rat mesangial cells cultured in RPMI 1640 medium containing 0.5% fetal bovine serum in a dose-dependent manner. The inhibition of DNA synthesis by tranilast was not affected by the presence of indomethacin (1 microg/ml) or N(G)-monomethyl-L-arginine (0.5 mM). Tranilast did not stimulate nitrite oxide synthesis in PDGF-stimulated cells. Mitogen-activated protein kinase activity in mesangial cells was significantly increased by exposure to PDGF, while the effect was significantly suppressed in the presence of tranilast. The present study revealed that tranilast inhibits the growth of rat mesangial cells, independently of nitric oxide or prostacycline synthesis.

Animals↗
PubMed1997

Effect of mitral valvuloplasty in mitral stenosis on coagulation activity.

We investigated the plasma levels of molecular markers for the status of thrombin generation and fibrinolysis in patients with mitral stenosis before and after percutaneous mitral valvuloplasty. Our results show that percutaneous mitral valvuloplasty results in decreased coagulation activity, suggesting that percutaneous mitral valvuloplasty is also useful for prevention of systemic embolism in patients with mitral stenosis.

Blood Coagulation↗
PubMed1997

Characterization of monoglucuronides of vitamin D2 and 25-hydroxyvitamin D2 in rat bile using high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

The characterization of vitamin D2 3-glucuronide, 25-hydroxyvitamin D2 3-glucuronide and 25-hydroxyvitamin D2 25-glucuronide, biliary metabolites obtained from rats dosed with vitamin D2 and 25-hydroxyvitamin D2 per os, was carried out using HPLC-atmospheric pressure chemical ionization (APCI)-MS. The glucuronide obtained from bile specimens was identified by comparison of its chromatographic behaviour with an authentic sample using HPLC-APCI-MS operating in the negative-ion mode. Methylation of the respective fraction with diazomethane gave the methyl ester, which was also confirmed by HPLC-APCI-MS operating in the positive-ion mode. The (M-H)- and (M + NH4)+ ions were monitored in the selected-ion monitoring mode.

25-Hydroxyvitamin D 2↗
PubMed1997

Monocytes modulate the fibrinolytic balance of endothelial cells.

Cultured endothelial cells (ECs) produced a constitutive plasminogen activator inhibitor-I (PAI-1), whereas primary culture of monocytes from blood did not produce a detectable amount of PAI-1. Addition of monocytes to ECs caused the accumulation of a large amount of PAI-1 in the supernatant in a dose- and time-dependent manner. Having almost no effect on the production of tissue-type plasminogen activator (t-PA), monocytes decreased the potential fibrinolytic activity of ECs. The 6 hours conditioned medium obtained from the coculture system between monocytes and either ECs or paraformaldehyde-fixed ECs had almost the same effect on the other ECs to produce PAI-1 and t-PA as monocytes that were direct contact with ECs. In addition, this effect was specifically inhibited by using two antibodies against interleukin-1 beta and tumor necrosis factor-alpha. These results indicate that interleukin-1 beta and tumor necrosis factor-alpha induced by the coculture are mostly responsible for decreasing the fibrinolytic activity of ECs.

Cells, Cultured↗
PubMed1997

Determination of 25-hydroxyvitamin D3 in human plasma by reversed-phase high-performance liquid chromatography with ultraviolet detection.

A method for the determination of 25-hydroxyvitamin D3, the major metabolite of vitamin D3 in human plasma, using a non-radioactive internal standard and reversed-phase high-performance liquid chromatography with UV detection (265 nm) has been developed. The method was applied to the determination of the metabolite in plasma from healthy subjects (n = 25) and from patients with chronic renal failure (n = 12). 25-Hydroxyvitamin D3 3-sulfate, a major conjugated metabolite of 25-hydroxyvitamin D3, was also determined and the correlation between the concentrations of these metabolites was examined. The study showed that almost equal amounts of both compounds were detected in the plasma of healthy subjects, however, in two subjects, the amount of sulfate in the free form was found to be about twice as high as normally detected. In contrast, the free form was predominant in the plasma of patients with chronic renal failure and the sulfate was not detected in four patients.

Calcifediol↗
PubMed1997

Cloning of the cDNA encoding rat Presenilin-1.

We isolated rat presenilin-1 (PS-1; also called S182 previously) cDNA from total brain RNA by using a reverse transcription-polymerase chain reaction (RT-PCR) technique with primers homologous to the conserved sequences of human and mouse PS-1. Rat PS-1 cDNA encoded 468 amino acids (aa) and the deduced aa sequence was highly homologous to those of the human (88.4%) and mouse (92.7%). Northern blot analysis of the rat PS-1 cDNA revealed two mRNA species in rat neurotypic pheochromocytoma and glioma cell lines (PC-12 and C6, respectively) that migrated at rates corresponding to approximately 3.0 and 7.5 kb.

Alzheimer Disease↗
PubMed1997

A selective immunoaffinity chromatography for determination of plasma 1 alpha, 25-dihydroxyvitamin D3: application of specific antibodies raised against a 1 alpha, 25-dihydroxyvitamin D3-bovine serum albumin conjugate linked through the 11 alpha-position.

A selective and simple immunoaffinity chromatography of 1 alpha, 25-dihydroxyvitamin D3 has been developed and found to be a useful pretreatment tool for determining the metabolite in human plasma. A reasonably designed, haptenic derivative, 11 alpha-hemiglutaryloxy-1 alpha, 25-dihydroxyvitamin D3, was linked to bovine serum albumin, and rabbits were immunized repeatedly with the conjugate. The resulting polyclonal antibodies were specific to 1 alpha, 25-dihydroxyvitamin D3, recognizing both the A-ring and the side-chain structures. The antibodies were then immobilized on agarose gel to produce an immunosorbent which was stable and repeatedly usable. A plasma extract prepared with a Chem Elut column was applied to an affinity column containing the immunosorbent. After adequate washing, the adsorbed 1 alpha, 25-dihydroxyvitamin D3 was eluted selectively with a satisfactory recovery rate. This immunoaffinity chromatography enabled a simple radioreceptor assay for human plasma 1 alpha, 25-dihydroxyvitamin D3 which does not require any preparative high-performance liquid chromatography. The mean (+/-SD) values for 30 normal subjects and 8 patients with chronic renal failure were 36.0 (10.2) and 13.1 (2.1) pg/ml, respectively. The present method also gave reliable assay values for the plasma specimens from 1 alpha-hydroxyvitamin D3-administered volunteers, which have conventionally been difficult to measure unless complicated pretreatment is used.

Animals↗
PubMed1997

Endothelin-converting enzyme expression in the rat vascular injury model and human coronary atherosclerosis.

BACKGROUND: Endothelin 1 has been implicated in various human diseases, including atherosclerosis. In this study, we examined the expression and localization of endothelin-converting enzyme-1 (ECE-1), the final key enzyme of endothelin 1 processing, in rat carotid arteries after balloon injury and in human coronary atherosclerotic lesions. METHODS AND RESULTS: ECE-1 mRNA levels and ECE activity in rat balloon-injured arteries started to increase between 2 and 5 days after injury. The endothelin 1 content of tissue in injured arteries was concomitantly increased. Immunohistochemical staining located ECE-1 signals in endothelial cells in uninjured arteries, whereas ECE-1 immunoreactivity was detected in neointimal smooth muscle cells in injured arteries 5 to 14 days after balloon denudation. The size of the neointima was effectively reduced by phosphoramidon, an inhibitor of neutral metalloproteases, including ECE-1. In human coronary atherosclerotic lesions, intense ECE-1 immunoreactivity was detected in subsets of cells embedded in atheromatous plaque that correspond to smooth muscle cells and macrophages, as identified by staining for smooth muscle alpha-actin and CD68 surface marker, respectively. CONCLUSIONS: The present study ascertained that ECE-1 is expressed in neointimal smooth muscle cells in rat balloon-injured arteries and in both smooth muscle cells and macrophages in human coronary atherosclerotic lesions. Blockade of ECE-1 was effective in reducing neointimal formation after balloon injury. Thus, ECE-1 may contribute to the process of injury-induced neointimal formation and atherosclerosis through the autocrine/paracrine effects of endothelin 1.

Aged↗
PubMed1997

Morphological and histological analysis of the thoracic duct at the jugulo-subclavian junction in Japanese cadavers.

The macroscopic arrangement of the termination of the thoracic duct (TD) was examined in detail in Japanese cadavers, and the distribution of various types of collagen, tenascin, laminin, and fibronectin in the framework of the wall of the thoracic duct termination was demonstrated. We identified several branching patterns and pathways of the TD (i.e., those terminating 1) at the venous angle (Type A); 2) at the end of the internal jugular vein (Type B); 3) at the the external jugular vein (Type C); or 4) in a complex with various branches (Type D). The TD often divided into two or three trunks before or after passing beneath the left brachiocephalic vein. Compared with the main trunk of the TD, fewer muscle fibers, elastic fibers, specific compounds of the extracellular matrices, and valvular connective tissues were found near the termination of the TD and the valves of the TD and veins. Smooth muscle cells were arranged irregularly in each region of the TD.

Cadaver↗