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K Takeda

Publications and source records attributed to K Takeda.

At least 811 records · Page 45Linked to original sources

Secretion of peptidylglycine alpha-amidating monooxygenase (PAM) from rat salivary glands.

Peptidylglycine alpha-amidating monooxygenase (PAM) is a regulating enzyme to synthesize the biologically active hormones having carboxy-terminal amide. In the present study we investigated secretion of the enzyme from rat saliva. Property of PAM in the saliva was similar to that in the submandibular gland. Both enzymes showed similar pH optimum at 5.0 and optimal ascorbic acid concentration at 2.5 mM. But molecular size of PAM in the saliva was 75 kDa in the gel permeation chromatography on Superose 12 column, while the size in the submandibular gland was 25 kDa. After the treatment with trypsin, PAM in the saliva was converted to a small size molecule, which is similar to the size in rat submandibular gland. These and other data indicate that a native molecular size of PAM is secreted into saliva and plays some physiological roles.

Aging↗

Effects of phenol compounds, glutathione analogues and a diuretic drug on glutathione S-transferase, glutathione reductase and glutathione peroxidase from canine erythrocytes.

1. Phenol compounds (ellagic acid, quercetin and purpurogallin), glutathione analogues (S-hexylglutathione and S-octylglutathione) and a diuretic drug (ethacrynic acid) were compared for their inhibitory effects on glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GSH-Px) in the canine erythrocytes. 2. All these compounds inhibited GST activity; quercetin was found to be the most potent inhibitor. 3. Ellagic acid, purpurogallin, quercetin and ethacrynic acid inhibited GR activity; S-hexylglutathione and S-octylglutathione had no effect on GR and GSH-Px activities. 4. Quercetin and purpurogallin inhibited GST non-competitively toward glutathione, whereas ellagic acid showed a competitive inhibition. Ellagic acid and purpurogallin inhibited GR non-competitively toward oxidized glutathione.

Animals↗

Kanji word reading process analysed by positron emission tomography.

Positron emission tomography, involving the H2(15)O injection technique, was used to study changes in regional cerebral blood flow during Japanese kanji word reading. Three-times intrasubject averaging analysis revealed that the main cortical responses occurred in the bilateral (left-side dominant) posterior inferior temporal area for visual input. We believe the left posterior inferior temporal area processes highly complex morphological features of kanji. For language output, left-side dominant activation of the posterior inferior frontal gyrus and the basal ganglia was found.

Adult↗

Blood vessels and immunoreactive substance P-containing nerve fibers in rat skin treated topically with clobetasol propionate, a corticosteroid.

After applying topically a cream (0.1 ml) containing corticosteroid (clobetasol propionate), on rat back skin, we examined the morphological alterations of blood vessels, substance P-containing nerve fibers, and cutaneous mast cells. After 3, 6, 10, 15, 30, and 60 min and 4 h, the skin treated was cut out with a sharp knife after killing the animals. The skin pieces were processed into conventional histological sections cut vertically and examined by staining immunohistochemically with anti-substance P serum, by staining with toluidine blue for mast cell granules, and by estimating morphometrically the average areas of vascular cavity and the number of substance P fibers in the dermis. In the dermis and subcutaneous tissue of untreated skin, we found many immunoreactive SP-containing nerve fibers and mast cells in close association. Three to ten min after the treatment, the average area of the vascular cavities steadily increased, and SP-positive fibers became less frequent in the dermis. In concomitant with those events, cutaneous mast cells discharged their granules. Thereafter, the average area of vascular cavities gradually decreased to a minimum at 4 h after the treatment. In contrast, both SP-containing fibers and mast cells reestablished their initial states after the same duration.

Administration, Topical↗

Maintenance of androgen-, glucocorticoid- or estrogen-responsive growth in shionogi carcinoma 115 subline sustained in castrated mice with high dose of estrogen for 30 generations (3 years).

Shionogi carcinoma 115 (SC115), an androgen-dependent mouse mammary tumor, rapidly loses its androgen responsiveness after androgen withdrawal. The growth of this tumor can also be stimulated by high doses of estrogen or glucocorticoid. In the present study, the maintenance of hormone-responsive growth of SC115 tumors with a high dose of estrogen was examined in castrated male mice using an SC115 subline obtained by serial transplantations of SC115 tumors in estrogen-treated castrated mice for 3 years (30 generations) (subline E2). Seed tumors from both SC115 and subline E2 could rapidly grow in castrated mice given daily injections of testosterone propionate (TP), 17 beta-estradiol (E2), or dexamethasone (Dex) (100 micrograms/mouse/day) but not in those given vehicle alone. Although SC115 and subline-E2 tumors grown with TP or Dex showed temporary regression after steroid withdrawal, the tumors grown with E2 did not show such temporary regression. The TP-, E2-, or Dex-induced growth of subline-E2 tumors was almost the same as that of the original SC115 tumors. However, responsiveness to androgen, estrogen or glucocorticoid of both tumors disappeared within one passage in steroid-depleted castrated mice. The present findings demonstrate that the loss of responsiveness to androgen as well as to high doses of estrogen or glucocorticoid of SC115 tumors can be prevented in castrated mice not only with androgen but also with high doses of estrogen.

Animals↗

Analysis of collagen gene expression by cultured fibroblasts in morphoea.

Collagen gene expression was studied in dermal fibroblasts derived from the inflammatory and sclerotic skin lesions of patients with localized or generalized morphoea. The levels of mRNA for type I collagen in early-passage fibroblasts derived from inflammatory lesions were higher than those obtained from the uninvolved skin, whereas those fibroblasts obtained from sclerotic lesions were unaltered. No alteration in type I collagen mRNA levels was observed in late-passage fibroblasts derived from the inflammatory lesions. The relative rate of collagen to total protein synthesis in early-passage fibroblasts derived from inflammatory lesions was higher than that of fibroblasts from uninvolved skin, while no alteration or a slight decrease was observed in fibroblasts from the sclerotic lesions. The data suggest that the inflammatory reactions induce increased collagen synthesis by fibroblasts in the skin in scleroderma.

Actins↗

Regulatory role of the GTP-binding protein, G(o), in the mechanism of exocytosis in adrenal chromaffin cells.

To elucidate the possible involvement of GTP-binding proteins (G proteins) in the mechanism of exocytosis, we studied effects of pertussis toxin (PTX), guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S), and antibodies against the G proteins (Gi and G(o)) on the secretory function of bovine adrenal chromaffin cells. Pretreatment of chromaffin cells with PTX resulted in an increase in acetylcholine-evoked catecholamine release. High K(+)-, histamine-, or gamma-aminobutyric acid-evoked catecholamine release was also potentiated by PTX pretreatment. The concentration of extracellular Ca2+ required for maximal release by 10(-4) M acetylcholine was decreased significantly in PTX-treated cells. In digitonin-permeabilized cells, PTX pretreatment resulted in a decrease of the half-maximal concentration (Km) of Ca2+ required for exocytosis with no significant change in the maximal stimulation (Vmax). Exposure of permeabilized cells to GTP-gamma-S (a nonhydrolyzable GTP analogue) inhibited Ca(2+)-dependent exocytosis by reducing the affinity for Ca2+. The effects of PTX pretreatment were mimicked by treatment of permeabilized cells with polyclonal antibodies selective for the alpha subunit of the PTX-sensitive G protein, G(o). Treatment with similar antibodies against the alpha subunit of Gi had no effect. These findings suggest that G(o) directly controls the Ca(2+)-triggered process in the machinery of exocytosis by lowering the affinity of the unknown target for Ca2+.

Animals↗

Examination of bacterial characteristics of anaerobic membrane bioreactors in three pilot-scale plants for treating low-strength wastewater by application of the colony-forming-curve analysis method.

Characteristic sludge ecosystems arising in anaerobic membrane bioreactors of three pilot-scale plants treating low-strength (less than 1 g of biological oxygen demand per liter) sewage or soybean-processing wastewater were examined by analysis of the colony-forming-curves (CFC) obtained by counting colonies at suitable intervals. The wastewaters, containing high amounts of suspended solids (SS) (SS/chemical oxygen demand ratio, 0.51 to 0.80), were treated by using two types of bioreactors: (i) a hydrolyzation reactor for solubilization and acidification of SS in wastewater and (ii) a methane fermentation reactor for producing methane. The colony counts for the two sewage treatment plants continued to increase even after 3 weeks of incubation, whereas those for soybean-processing wastewater reached an approximately constant level within 3 weeks of incubation. The CFCs were analyzed by correlating the rate of colony appearance on roll tubes with the physiological types of bacteria present in the bioreactors. It was found that there were large numbers of slow-colony-forming anaerobic bacteria within the bioreactors and that the viable populations consisted of a few groups with different growth rates. It is considered that the slow-growing colonies appearing after 10 days of incubation were the dominant microflora in the sewage treated by hydrolyzation reactors. In particular, highly concentrated sludge (30.0 g of mixed-liquor volatile SS per liter) retained by the membrane separation module contained a large number of such bacteria. Slow-growing colonies of these bacteria could be counted by using a sludge extract medium prepared from only the supernatant of autoclaved sludge. In addition, the highest colony counts were almost always obtained with the sludge extract medium, meaning that most of the anaerobic bacteria in these sludges have complex nutrient requirements for growth. This report also indicates the usefulness of application of the CFC analysis method to the study of bacterial populations of anaerobic treatment systems.

Journal Article↗

Recessive inheritance of thyroid hormone resistance caused by complete deletion of the protein-coding region of the thyroid hormone receptor-beta gene.

Generalized resistance to thyroid hormone is a syndrome of reduced responsiveness of target tissues to thyroid hormone. The determination of amino acid sequences of the human thyroid receptor-beta (hTR beta), deduced from cDNA sequencing, has enabled evaluation of the genetic basis for this syndrome. Distinct point mutations in the ligand-binding domain of hTR beta have been identified in affected members of unrelated families, producing single amino acid substitutions that result in products with decreased or no hormone-binding activity. Inheritance in these families was autosomal dominant. We now report the molecular basis of generalized resistance to thyroid hormone in a consanguineous family unique for its autosomal recessive mode of inheritance. Deletion of the entire coding region of both hTR beta alleles in homozygous affected members of the family was demonstrated by the failure to amplify the coding exons 3-8 by the polymerase chain reaction using primers specific for flanking intronic sequences and by the demonstration of the presence of only two noncoding exons in Southern blots hybridized with exon-specific probes. As expected, obligate heterozygotes were phenotypically normal, since, in contrast to alleles with point mutations, the deleted allele could not act in a dominant negative fashion. Survival and maintenance of a euthyroid state are presumably mediated through expression of the hTR alpha gene, present in affected subjects, and the maintenance of high thyroid hormone levels. Furthermore, the clinical manifestations were relatively more mild that those observed in a homozygous patient with a single amino acid deletion in the hTR beta gene.

Amino Acid Sequence↗

Rapid localization of mutations in the thyroid hormone receptor-beta gene by denaturing gradient gel electrophoresis in 18 families with thyroid hormone resistance.

Generalized resistance to thyroid hormone (GRTH) is an inherited syndrome of reduced tissue responsiveness to thyroid hormone. Point mutations in the human thyroid hormone receptor-beta (hTR beta) gene of these patients, causing single amino acid substitutions, appear be different in unrelated individuals affected by the same syndrome. To localize mutations in the hTR beta gene, GC-clamped DNA fragments from affected individuals belonging to 21 families with GRTH were generated by the polymerase chain reaction and analyzed by denaturing gradient gel electrophoresis (DGGE). Putative mutations in the hTR beta gene of 18 unrelated individuals with GRTH were identified, and their nature has been confirmed in 9 by sequencing. All were in the hormone-binding domain of the receptor, and 13 of 18 mutations were in its center (exon 7). In 3 families we were unable to identify mutations in hTR beta, suggesting the existence of mutations at other loci, possibly the hTR alpha gene or other proteins involved in the thyroid hormone-dependent transactivation system. Sequencing of DNA fragments negative for the presence of putative mutations by DEEG confirmed the absence of sequence differences. DGGE of amplified DNA fragments can rapidly and reliably localize the sites of mutations in the hTR beta gene of patients with GRTH. The procedure enabled mapping the regions in the hTR beta harboring mutations associated with GRTH.

Base Sequence↗

Epithelial hyperplasia and decreased colloid content of the thyroid gland in triiodothyronine-predominant Graves' disease.

Patients with T3-predominant Graves' disease have an increased serum T3 level despite a normal or even lower level of serum T4 caused by antithyroid drug treatment. This study investigated the morphological characteristics of the thyroid gland in this type of hyperthyroidism. Ultrasound showed a similar thyroid echogenicity in both T3-predominant and ordinary Graves' disease, but thyroid weight was significantly greater in T3-predominant Graves' disease than in the controls [91 +/- 37 g vs. 48 +/- 17 g (mean +/- SD); P < 0.01]. The height of thyroid follicular epithelial cells were significantly greater in T3-predominant Graves' disease than in the controls (9.6 +/- 1.0 vs. 6.4 +/- 1.1 microns), but the epithelial height did not correlate significantly with the serum TSH receptor antibody titers. Epithelial cells occupied 26.4 +/- 10.9% of the total thyroid tissue in T3-predominant Graves' disease, while they occupied only 14.5 +/- 6.8% in the controls. Intracolloidal vacuoles also occupied a significantly greater area in the T3-predominant group (6.6 +/- 3.9% vs. 1.9 +/- 1.0% for controls). These findings suggest that the thyroid gland is more active in T3-predominant Graves' disease and has a more rapid turnover of intrathyroidal iodine than in the ordinary type of Graves' disease.

Adolescent↗

Japanese crossed Wernicke's aphasia.

A right-handed Japanese man with no personal or family history of left-handedness developed severe Wernicke's aphasia, a mild constructional disorder, and slight left hemiparesis. MRI revealed infarction in the territory of the righ middle cerebral artery, including areas homologous to Broca's and Wernicke's areas. The cerebral blood flow in these areas remained diminished even after language activation. The most likely explanation is that language production occurred in the left Broca's area, while language comprehension occurred in the right Wernicke's area (a dissociated aphasia).

Aphasia, Wernicke↗

Characterization of a neutral polysaccharide having activity on the reticuloendothelial system from the rhizome of Curcuma longa.

A neutral polysaccharide, named ukonan D, was isolated from the rhizome of Curcuma longa L. It produced a single band on electrophoresis and a single peak on gel chromatography, and its molecular mass was estimated to be 28,000. It showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test. Ukonan D is composed of L-arabinose: D-galactose: D-glucose: D-mannose in the molar ratio of 1:1:12:0.2, in addition to small amounts of peptide moiety. Methylation analysis, carbon-13 nuclear magnetic resonance and enzymic degradation studies indicated that its structural features include mainly both alpha-1,5-linked L-arabino-beta-3,6-branched D-galactan type and alpha-4,6-branched D-glucan type structural units. The influence of degradation with alpha-amylase followed by the elimination of glucan side chains on its immunological activity was discussed.

Animals↗

Stimulation of Na+/H+ exchange induced by angiotensin II in cultured rat vascular smooth muscle cells: role of Ca2+ and C-kinase.

Changes in cytosolic calcium concentration ([Ca2+]i) have been implicated in the regulation of intracellular pH (pHi) in several cell types. In the present study we investigated the regulatory mechanism of Na+/H+ exchange induced by angiotensin II (AII) in cultured rat vascular smooth muscle cells (VSMCs). Serially passaged VSMCs from Sprague-Dawley rat thoracic aorta were grown on coverslips and loaded with the pH-sensitive fluorescent indicator 2',7'-bis-(carboxyethyl)-5,6-carboxyfluorescein (BCECF). In HCO(3-)-free Ringer solution, pH 7.40, the resting pHi was 7.21 +/- 0.02 (n = 21). A biphasic response was seen after exposure of these cells to AII: an initial transient a acidification, followed by sustained alkalization. The magnitude of alkalization was dose-dependent. AII-mediated acidification was completely inhibited by [Sar1-IIe5-Gly8]AII, but amiloride had no effect. In contrast, the alkalization induced by AII was abolished by both amiloride and Na(+)-free medium. In Ca(2+)-free medium, the AII-induced alkalization was partially blocked and verapamil also caused partial inhibition. Since AII activates phospholipase C in VSMCs, we examined whether AII would increase Na+/H+ exchange by activation of protein kinase C. An inhibitor of protein kinase C, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), partially inhibited the alkalization induced by AII. These results indicate that AII stimulates cytoplasmic alkalization via an amiloride-sensitive Na+/H+ exchange system in cultured rat VSMCs, and that this AII-stimulated Na+/H+ exchange is mediated by Ca(2+)-dependent and protein kinase C-dependent mechanisms.

Amiloride↗

Effect of alpha 1-adrenoceptor blockade on the left ventricular force-length relationship in dogs.

We used the left ventricular (LV) end-systolic force-diameter (Fes-Des) relation to evaluate the effect of an alpha1-adrenoceptor antagonist (bunazosin hydrochloride) on the contractility of the beta-blocked left ventricle. Nine adult mongrel dogs were instrumented with ultrasonic crystals to measure LV diameter and a micromanometer to measure LV pressure. Beta-adrenergic and vagal blockade was induced with intravenous propranolol (2 mg/kg) and atropine (0.2 mg/kg), respectively, and preload was decreased by inferior vena caval occlusion. The slope (Ec) and extrapolated diameter intercept (Do) of the LV Fes-Des relation were derived from end-systolic data obtained in the control state (after beta-blockade) and after bunazosin infusion (1 mg/kg). Ec was used as a new index of LV contractility. After bunazosin infusion, the heart rate and Ec were decreased by 7 and 22%, respectively, in comparison with the control state, whereas Do did not change. These results indicate that alpha1-adrenoceptor blockade significantly reduces myocardial contractility in the beta-blocked canine heart, perhaps by decreasing the intracellular calcium concentration and/or myosin ATPase activity.

Adrenergic alpha-Antagonists↗

Role of sympathetic nerve inhibition in the vasodepressor effect of bromocriptine in normotensive and hypertensive rats.

This study aimed to elucidate the mechanism of the hypotensive effect of bromocriptine (BRC), and to investigate whether or not the effects of BRC on the sympathetic nervous system are altered in hypertension. BRC was administered intravenously to normotensive and spontaneously hypertensive rats (SHR). It elicited hypotensive effects dose-dependently in urethane-anaesthetized normotensive rats, an effect which was antagonized with metoclopramide. Pretreatment with intravenous hexamethonium attenuated the hypotensive effect of BRC. BRC decreased plasma norepinephrine (NE) without inhibiting the sympathetic nerve spikes recorded from the postganglionic sympathetic nerve bundle. The hypotensive effect of BRC was significantly greater in SHR than in Wistar Kyoto Rats (WKY). Decrease in NE by BRC was also significantly greater in SHR than in WKY. These results suggest that the hypotensive effect of BRC is induced by suppression of NE release, not by inhibition of sympathetic nerve spikes, and that the dopaminergic presynaptic inhibition is attenuated in SHR.

Animals↗

Giant fistula between the right coronary artery and the left ventricle: diagnostic significance of right posterior oblique chest radiograph.

Congenital coronary artery fistulas are uncommon. The prevalence of coronary artery fistulas in adults has been estimated as 0.13% [1]. Most of these fistulas communicate with the right atrium or ventricle or with the pulmonary artery; those emptying into the left atrium or ventricle are quite rare [2, 3]. Usually, patients with coronary artery fistula do not have clinical symptoms, and their chest radiographs do not show the corresponding shadow because the fistula is hemodynamically insignificant. When the fistula is large, however, chest radiographs show the secondary hemodynamic changes, such as cardiac enlargement or increased pulmonary vasculature, depending on the emptying site. Moreover, the silhouette of the dilated tortuous vessel itself can be seen on the chest radiograph. We have examined four patients, each of whom had a giant fistula in the right coronary artery emptying into the left ventricle, proved by coronary angiography. In this essay, we illustrate the chest radiographs of these patients and describe the radiologic features that permit a specific diagnosis or suggest a coronary artery fistula.

Adult↗