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Biomedical subjects

T Izawa

Publications and source records attributed to T Izawa.

At least 73 records · Page 4Linked to original sources
PubMed1994

Cyanoamidines. II. Synthesis and pharmacological activity of N-arylalkyl-N'-cyano-3-pyridinecarboxamidines.

A new series of cyanoamidines, N-arylalkyl-N'-cyano-3-pyridinecarboxamidines was synthesized and evaluated for inhibitory effects on 40 mM KCl-induced contraction and norepinephrine (NE)-induced contraction of rat aorta strips. The N-phenethyl cyanoamidine 4c showed potent vasodilatory action. Further in vitro screening program using 4c as a lead compound resulted in the discovery of highly potent N-[2-(2-chlorophenyl)ethyl]-N'-cyano-3-pyridinecarboxamidine (5j). Compound 5j induced the greatest increase in 86Rb+ efflux among cyanoamidine series. Subsequent modification of the pyridine ring of 5j was performed with evaluation for intravenous and oral antihypertensive activities. Introduction of an amino group at the 5-position of the pyridine ring furnished the new potassium channel opener, 5-amino-N-[2-(2-chlorophenyl)ethyl]-N'-cyano-3-pyridinecarboxamidine+ ++ (9e; KRN4884), which showed highly potent antihypertensive activity and a long duration of antihypertensive action after oral administration. KRN4884 is under further development as an antihypertensive agent.

Administration, Oral↗
PubMed1994

[A case of motor and sensory neuropathy associated with metatropic dysplasia].

We reported a case of a fifteen-year-old boy with metatropic dysplasia. He showed short-limbed dwarfism at the age of 1 year and 2 months. Roentgenograms of the bone revealed generalized platyspondyly, enlargement of the epiphyseal-metaphyseal regions of long bones and halberd-shaped pelvis. Scoliosis progressed and he evolved to short-trunk dwarfism. When he was 10 years old, he developed gait disturbance gradually due to muscle weakness of lower extremities. He was examined at the age of 15 years. Motor nerve conduction velocity was reduced to 21 m/sec in left median nerve and 18.5 m/sec in left peroneus nerve. Needle EMG showed neurogenic pattern. We performed sural nerve biopsy and obtained histopathological findings. Fiber density of myelinated axons decreased. Electron microscopic examination revealed onion bulb formation surrounding a myelinated fiber and a storage in the rough-surfaced endoplasmic reticulum of Schwann cell. These inclusions were similar to those of cartilage of his long bone. We supposed that the neuropathy would have relation to this bone dysplasia.

Adolescent↗
PubMed1993

Vasorelaxant action of Ki1769, a new pyridinecarboximidamide, in isolated porcine coronary artery.

The characteristics of KRN2391 (N-cyano-N'-(2-nitroxyethyl)-3-pyridinecarboximidamide monomethansulfonate) and its phenethyl and 2-hydroxyethyl derivatives (Ki1769 and Ki3315) were studied in isolated porcine coronary arteries. KRN2391, Ki1769 and Ki3315 produced concentration-dependent relaxation of coronary arteries contracted by 25 mM KCl and the order of relaxant potency was KRN2391 > Ki1769 > Ki3315. At the maximum effect, KRN2391 produced nearly complete relaxation but Ki1769 produced about 66% relaxation. The maximum effect of Ki3315 could not be obtained because of its solubility. The relaxation induced by KRN2391 was antagonized by glibenclamide and methylene blue but relaxations caused by Ki1769 and Ki3315 were antagonized by glibenclamide alone. The antagonistic effect of glibenclamide on Ki1769- and Ki3315-induced relaxations was more potent than that on KRN2391-induced relaxation. KRN2391 induced relaxation of coronary arteries contracted by 40 mM KCl in a concentration-dependent manner but the effect of KRN2391 was smaller against 40 mM KCl-induced contractions than against 25 mM KCl-induced contractions. Ki1769 had almost no effect on coronary arteries contracted by 40 mM KCl. These results suggest that pyridinecarboximidamide derivatives which do not possess a nitroxyl group have vasodilating ability based on a K+ channel opening action.

Animals↗
PubMed1993

Plant bZIP protein DNA binding specificity.

Plant bZIP proteins exhibit a relaxed DNA-binding specificity for DNA sequence motifs containing an ACGT core. Gel mobility shift experiments employing ten different recombinant plant bZIP proteins demonstrated that nucleotides flanking the ACGT core affected binding specificity and identified three different types of ACGT elements: G-box, CACGTG; C-box, GACGTC; and A-box, TACGTA, motifs. These ten different bZIP proteins could be categorized into three groups according to their qualitative and quantitative specificity for G-box and C-box elements. Dissociation constant values (Kd values) of these bZIP proteins for high affinity G-box and C-box elements and reciprocal competition gel mobility shift assays confirmed our classification scheme. Group 1 proteins exhibit a stronger binding affinity for G-box elements, group 2 proteins bind both G-box and C-box motifs with comparable binding affinity, whereas the group 3 proteins display a stronger binding affinity for C-box oligonucleotides. Studies using a panel of G-box and C-box oligonucleotides differing in their flanking sequences identified high affinity binding sites. All ten plant bZIP proteins examined, except TGA1a, exhibited type A G-box binding activity preferring class I G-box elements. In contrast to the situation observed for G-box elements, C-box motifs displayed a very much more stringent flanking nucleotide requirement for binding activity. Protein/DNA binding experiments using scanning mutants of a high affinity G-box element and G-box/C-box hybrid elements demonstrated that bZIP protein binding activity depends upon the affinity of protein dimer subunits for ACGT half-sites. Information provided by our systematic analysis of plant bZIP DNA binding specificity can be used to identify high affinity binding sites for the plant bZIP proteins studied here. Assuming that only high affinity bZIP binding sites are likely to function in vivo, identification of these sites will allow us to predict which genes are activated by a particular bZIP protein.

Base Sequence↗
PubMed1993

Trans-activation and stable integration of the maize transposable element Ds cotransfected with the Ac transposase gene in transgenic rice plants.

To develop an efficient gene tagging system in rice, a plasmid was constructed carrying a non-autonomous maize Ds element in the untranslated leader sequence of a hygromycin B resistance gene fused with the 35S promoter of cauliflower mosaic virus. This plasmid was cotransfected by electroporation into rice protoplasts together with a plasmid containing the maize Ac transposase gene transcribed from the 35S promoter. Five lines of evidence obtained from the analyses of hygromycin B-resistant calli, regenerated plants and their progeny showed that the introduced Ds was trans-activated by the Ac transposase gene in rice. (1) Cotransfection of the two plasmids is necessary for generation of hygromycin B resistant transformants. (2) Ds excision sites are detected by Southern blot hybridization. (3) Characteristic sequence alterations are found at Ds excision sites. (4) Newly integrated Ds is detected in the rice genome. (5) Generation of 8 bp target duplications is observed at the Ds integration sites on the rice chromosomes. Our results also show that Ds can be trans-activated by the transiently expressed Ac transposase at early stages of protoplast culture and integrated stably into the rice genome, while the cotransfected Ac transposase gene is not integrated. Segregation data from such a transgenic rice plant carrying no Ac transposase gene showed that four Ds copies were stably integrated into three different chromosomes, one of which also contained the functional hph gene restored by Ds excision. The results indicate that a dispersed distribution of Ds throughout genomes not bearing the active Ac transposase gene can be achieved by simultaneous transfection with Ds and the Ac transposase gene.

Base Sequence↗
PubMed1993

Effect of endurance training on angiogenic activity in skeletal muscles.

The effect of endurance training on angiogenic activity in skeletal muscle was investigated using cell culture. The exercised rats were trained on a rodent treadmill, 5 days a week for 9 weeks. After the training program, a significant increase (7.8%) in mass was found in soleus muscle (MS), although we noted no apparent increase in gastrocnemius muscle and extensor digitorum longus muscle. The extracts from all the muscles (especially from the MS) of trained rats considerably increased the capillary growth in an in vitro angiogenesis model, in which microvascular fragments and myofibroblastic (Mf) cells isolated from lipid tissues were grown in co-culture. In addition, the extract from the MS of trained rats increased the DNA synthesis and growth of bovine capillary endothelial (BCE) cells, although the extract did not stimulate the growth of Mf cells. Moreover, the extract from the MS of trained rats notably enhanced the activity of plasminogen activator in the conditioned medium of BCE cells compared with that of sedentary rats. On the other hand, extracts from the MS of both sedentary and trained rats increased the synthesis of collagen by Mf cells and metalloproteinase activity in the conditioned medium of BCE cells to the same degree. These results suggest that endurance training increases the angiogenic activity in skeletal muscles.

Animals↗
PubMed1993

Effect of running training on uncoupling protein mRNA expression in rat brown adipose tissue.

The effect was investigated of endurance training on the expression of uncoupling protein (UCP) mRNA in brown adipose tissue (BAT) of rats. The exercised rats were trained on a rodent treadmill for 5 days per week and a total of 9 weeks. After the training programme, a marked decrease in BAT mass was found in terms of weight or weight per unit body weight; there was a corresponding decrease in DNA content and a downward trend in RNA and glycogen levels. The UCP mRNA was present at a markedly decreased level in BAT of trained animals. In consideration of the reduced levels of mRNAs for hormone-sensitive lipase and acyl-CoA synthetase, the brown adipose tissue investigated appeared to be in a relatively atrophied and thermogenically quiescent state.

Adipose Tissue, Brown↗
PubMed1993

Recovery of beta-receptors and adenylate cyclase from desensitization induced by short term heat exposure in rat parotid glands.

1. The recovery of rat parotid beta-adrenergic receptors (beta-AR) and adenylate cyclase (AC) from heat (33 degrees C)-induced desensitization was studied. 2. Down-regulated cell surface beta-AR and AC activity in response to isoprenaline (IPR) returned to the control level 120 hr after the termination of heat exposure. 3. However, beta-AR in parotid crude membranes increased over the control level for 48-120 hr. 4. Coupling between beta-AR and G protein(s) was attenuated at 120 hr. 5. These data suggest that beta-AR on the cell surface, but not those internalized, can transduce biological responses.

Adenylyl Cyclase Inhibitors↗
PubMed1993

Pharmacological analysis of the vasodepressor effect of KRN2391 in pithed rats.

1. The antagonism by glibenclamide of the vasodepressor effects of KRN2391, cromakalim and nitroglycerin was compared in pithed rats with blood pressure supported by an infusion of phenylephrine. 2. Cumulative administration of KRN2391 (3-300 micrograms/kg, i.v.), cromakalim (3-300 micrograms/kg, i.v.) and nitroglycerin (1-300 micrograms/kg, i.v.) produced dose-dependent decreases in diastolic blood pressure. 3. In rats given glibenclamide (20 mg/kg, i.v.) the dose-vasodepressor curves for KRN2391 and cromakalim were shifted to the right. However, glibenclamide had no effect on the vasodepressor effect of nitroglycerin. 4. The ED50mmHg values increased about 5.9 fold for KRN2391 and 9.5 fold for cromakalim in glibenclamide-treated rats. 5. These results suggest that the vasodepressor effect of KRN2391 is due to both glibenclamide-sensitive and insensitive mechanisms. This glibenclamide-insensitive effect of KRN2391 is thought to reflect its nitrate action.

Animals↗
PubMed1993

In vivo adaptative regulation of muscarinic receptors and muscarinic stimulation-induced Ca2+ mobilization during short-term heat exposure in rat parotid glands.

1. Adaptation of muscarinic receptors (MR)--muscarinic stimulation--induced intracellular Ca2+ mobilization during short-heat exposure (33 degrees C). 2. Heat-exposure for 48 hr decreased the carbachol (CCh)-stimulated cytosolic Ca2+ concentration increase. 3. The number of MR on cell surface increased transiently at 24 hr with a subsequent decrease at 48 hr. 4. CCh-stimulated inositol triphosphate (IP3) formation decreased at 48 hr. 5. In saponin-permeabilized cells, 1,4,5-IP3-induced 45Ca2+ release decreased at 24 hr. 6. The data suggest that the adaptation for increased muscarinic stimulation occurs at IP3 generating sites as well as at intracellular IP3 receptor sites during heat exposure.

Acetylcholine↗
PubMed1993

Effects of KRN2391, nicorandil and diltiazem on the changes in the electrocardiogram caused by endothelin-1 in anaesthetized rats.

1. The effect of KRN2391, a novel vasodilator, on the changes of electrocardiogram caused by endothelin-1 (ET-1) was studied in anaesthetized rats and compared with the effects of nicorandil and diltiazem. In addition, the effect of KRN2391 on the action potential of guinea-pig papillary muscle was studied. 2. The intracoronary administration (i.c.) of ET-1 (5 micrograms) induced not only ST segment elevation of the electrocardiogram due to contraction of the coronary artery, but also arrhythmias involving atrioventricular block (A-V block), ventricular premature contraction (VPC) and ventricular fibrillation (VF), and resulted in death in most animals. However, the administration of methacholine (3 micrograms, i.c.) produced ST segment elevation alone without developing arrhythmias. 3. Pretreatment with intravenous administration of KRN2391 (30 micrograms kg-1) inhibited the ST segment elevation and the development of arrhythmias induced by ET-1, and decreased the incidence of death. 4. Nicorandil (1000 micrograms kg-1) prevented the ST segment elevation without suppression of the occurrence of VF. Diltiazem (100 micrograms kg-1) suppressed both the ST segment elevation and the occurrence of VF but not other arrhythmias. Nicorandil at 3000 micrograms kg-1 and diltiazem at 300 micrograms kg-1 produced not only a suppression of ST segment elevation and VF incidence but also a decrease in the occurrence of arrhythmias. These doses of nicorandil and diltiazem produced a decrease in death in a dose-dependent manner. 5. KRN2391 (10 and 30 micrograms kg-1), nicorandil (1000 and 3000 micrograms kg-1) and diltiazem (100 and 300 micrograms kg-1) significantly decreased mean blood pressure in a dose-dependent manner. Heart rate was decreased by nicorandil (3000 microg kg-1) and diltiazem (100 and 300 microg kg-1) but was not affected byKRN2391 (10 and 30 microg kg-1).6. KRN2391 (30 microM) significantly shortened the action potential duration of guinea-pig ventricle at 50% and 90% repolarization (APD50 and APD90). The effect of KRN2391 was inhibited by a K+channel blocker, glibenclamide (30 microM).7. These results suggest that the occurrence of ST segment elevation and arrhythmias induced by ET-1 are due to a dual direct action on both coronary vascular smooth muscle and myocardium. Therefore,the protective effects of KRN2391, nicorandil and diltiazem on ET-l-induced heart disorders appear to be due to their direct actions on coronary vascular smooth muscle and the myocardium.

Action Potentials↗
PubMed1993

Intralysosomal pH and release of lysosomal enzymes in the rat liver after exhaustive exercise.

The mechanism underlying exhaustive exercise-induced release of lysosomal enzymes was studied in the rat liver. Exhaustive exercise resulted in the release of beta-glucuronidase and cathepsin D, but not beta-glucosidase and acid phosphatase, into the blood and cytosol, suggesting that the release of lysosomal enzymes is not due to disruption of lysosomal membranes. The intralysosomal pH of the liver, which was approximately 5.5 at the resting level, rose significantly after exhaustive exercise to pH 6.3. In vitro, beta-glucuronidase and cathepsin D were released at an intralysosomal pH exceeding 6.2. In contrast, beta-glucosidase and acid phosphatase were not released. The elevation of intralysosomal pH reduced the aggregation of beta-glucuronidase and cathepsin D. The rate of ammonia accumulation increased markedly in the lysosome-enriched subcellular fraction after exercise. There was a positive relationship between the rate of ammonia accumulation and the elevation of intralysosomal pH in vitro. Lysosomes isolated after exhaustive exercise showed significantly increased osmotic fragility. Our findings suggest that, during exhaustive exercise, the accumulation of ammonia in lysosomes leads to the elevation of intralysosomal pH, resulting in the reduced aggregation of certain lysosomal enzymes. Thus, less aggregated lysosomal enzymes may be released into the cytosol through the lysosomal membrane, the permeability of which has been increased.

Acid Phosphatase↗
PubMed1993

Effect of insulin on adipocyte lipolysis in exercise-trained rats.

The effect of exercise training on the antilipolytic action of insulin was studied in rat adipocytes. Exercise training enhanced lipolysis induced by norepinephrine. Insulin dose dependently inhibited norepinephrine- (1 microM) stimulated lipolysis in both groups. Its inhibition rate was significantly greater in the trained than in the control group. Thus, exercise training enhanced the antilipolytic action of insulin. In the control group, insulin (1,000 microU/ml) reduced the displacement rate of [3H]CGP-12177 binding to adipocytes by low concentrations of (-)-norepinephrine. The slope factor without insulin was 0.76, whereas that with insulin was 0.95. In the trained group, insulin did not affect the competition binding of (-)-norepinephrine for [3H]CGP-12177. The displacement rate of [3H]CGP-12177 binding from adipocytes by low concentrations of (-)-norepinephrine was significantly greater in the trained than in the control group. The number of surface beta-adrenergic receptors per adipocyte was smaller in the trained than in the control group. Cilostamide, which blocks the antilipolytic action of insulin, restored lipolysis in both groups. The recovery rate was significantly greater in the trained than in the control group. These findings suggest that the enhanced antilipolytic action by insulin in the trained group occurs at a site distal to the binding of norepinephrine to beta-adrenergic receptors and that it is due to the increased activity of particulate low-Michaelis constant phosphodiesterase.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1993

Mechanism of isoprenaline-stimulated diacylglycerol formation in rat parotid acinar cells.

The kinetics and mechanism of sn-1,2-diacylglycerol (DAG) formation induced by isoprenaline were studied in rat parotid acinar cells. DAG accumulation induced by 100 microM isoprenaline reached its maximum at 1 min, rapidly decreased (about 50%) at 5 min and then remained constant for 30 min. DAG accumulation 1 min after isoprenaline treatment was dose-dependent. Either propranolol or phentolamine inhibited isoprenaline-stimulated DAG accumulation in a dose-dependent manner. Addition of a vasoactive intestinal polypeptide, forskolin, or dibutyryl cyclic AMP had no effect on DAG accumulation. Isoprenaline did not cause the release of [3H]choline or [3H]ethanolamine metabolites into the medium. Based on the kinetics of DAG formation and [32P]phosphoinositide breakdown, we conclude that isoprenaline-induced DAG formation was mainly related to the hydrolysis of [32P]phosphatidylinositol 4,5-bisphosphate ([32P]PIP2). These results suggest that the effect of isoprenaline on DAG formation is mediated by alpha 1-adrenoceptor activation, that it is not related to the increase in cyclic AMP, and that it is closely related to PIP2 hydrolysis.

Alkaloids↗
PubMed1993

Lack of the stimulatory effect of guanine nucleotide on diacylglycerol generation in permeabilized adipocytes from diabetic rats.

Insulin caused an 8- or a 3-fold increase in lipogenesis in control rats (C) or diabetic rats (DM), respectively. Following insulin treatment for DM, insulin resistance was clearly reversed. Phospholipase C (PLC) caused a 4-fold increase in lipogenesis in C, but not in DM. Insulin treatment partially restored PLC-induced lipogenesis. Insulin or PLC increased protein kinase C (PKC) activity in the membrane fraction in C, but not in DM. Insulin treatment partially restored insulin- or PLC-stimulated PKC activity. 5'-Guanylylimidodiphosphate (Gpp(NH)p) exerted a stimulatory effect on diacylglycerol (DAG) generation in permeabilized adipocytes from C, but not in DM. Insulin treatment partially restored the stimulatory effect of Gpp(NH)p. These findings suggest that a particular G protein(s) is involved in the regulation of DAG generation in adipocytes, and that diabetes leads to a functional or quantitative abnormality in G protein and G protein-PLC. Insulin therapy partially restored G protein-PLC dependent glucose uptake.

Adipocytes↗
PubMed1992

Genetically engineered rice resistant to rice stripe virus, an insect-transmitted virus.

The coat protein (CP) gene of rice stripe virus was introduced into two japonica varieties of rice by electroporation of protoplasts. The resultant transgenic plants expressed the CP at high levels (up to 0.5% of total soluble protein) and exhibited a significant level of resistance to virus infection. Plants derived from selfed progeny of the primary transformants also expressed the CP and showed viral resistance, indicating stable transmission of the CP gene and the trait of resistance to the next generation. Moreover, the virally encoded strip disease-specific protein was not detected in transgenic plants expressing CP 8 weeks after inoculation, indicating protection before viral multiplication. These studies demonstrated that CP-mediated resistance to virus infection can be extended to cereals and to the viruses transmitted by an insect vector (planthopper).

Base Sequence↗
PubMed1992

Mechanism of carbachol-stimulated diacylglycerol formation in rat parotid acinar cells.

We studied the relationship between phosphoinositide hydrolysis, phosphatidylcholine hydrolysis, and sn-1,2-diacylglycerol (DAG) formation in response to carbachol stimulation in rat parotid acinar cells. Previously, we demonstrated that DAG formation stimulated with 1 microM carbachol was biphasic: the first peak occurred at 5 min and the second one at 20 min. It was also demonstrated that the second peak was regulated in part by a calmodulin/protein kinase C-dependent mechanism. Based on the kinetic analysis of DAG formation and [32P]phosphoinositide breakdown, the first peak of carbachol (1 microM)-stimulated DAG accumulation was found to be related to the breakdown of [32P]phosphatidylinositol 4-monophosphate ([32P]PIP) and [32P]phosphatidylinositol 4,5-bisphosphate ([32P]PIP2). The second peak was found to be related to [32P]PIP2 breakdown. Carbachol stimulated the release of [3H]phosphocholine into the medium, indicating that the predominant pathway for phosphatidylcholine hydrolysis was via phospholipase C. Moreover, carbachol stimulated the release of [3H]choline metabolites in a time- and dose-dependent manner. This agonist slightly stimulated the release of [3H]ethanolamine metabolites. A calmodulin/protein kinase C-dependent mechanism was also studied and was found to be involved in carbachol-stimulated phosphatidylcholine hydrolysis; W-7, a calmodulin inhibitor and staurosporine, a protein kinase C inhibitor, inhibited the carbachol (1-microM)-induced release of [3H]choline metabolites at 20 min in a dose-dependent manner, but did not have inhibitory effects at 5 min. These results suggest that the first peak of DAG accumulation induced by carbachol is predominantly associated with the breakdown [32P]PIP and of [32P]PIP2 and that the second peak is predominantly associated with [32P]PIP2 breakdown and phosphatidylcholine hydrolysis.

Alkaloids↗