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Biomedical subjects

H Ueda

Publications and source records attributed to H Ueda.

At least 451 records · Page 25Linked to original sources

A subtype of opioid kappa-receptor is coupled to inhibition of Gi1-mediated phospholipase C activity in the guinea pig cerebellum.

PLC activity was stimulated either by 1-100 microM of GTP or by 100-3,000 microM Ca2+ in lysed synaptosomal membranes of the guinea pig cerebellum. The kappa-opioid receptor agonist selectively inhibited the PLC activity stimulated by 100 microM GTP, but not by 100-3,000 microM Ca2+. Pretreatment of membranes with PTX abolished such a kappa-agonist-induced inhibition of PLC activity. The reconstitution of Gi1, but not of Go purified from porcine brains with PTX-treated membranes showed a complete recovery of the kappa-agonist-inhibition of PLC activity. These findings suggest that a novel subtype kappa-receptor mediates inhibition of PLC through inhibiting the intrinsic activity of PTX-substrate G-proteins.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Sequences of two cDNAs encoding silkworm homologues of Drosophila melanogaster squid gene.

The squid (sqd) gene of Drosophila melanogaster encodes a few isoforms of a heterogeneous nuclear (hn) RNA-binding protein. We isolated two types of cDNAs coding for homologues of the Sqd protein from the silkworm Bombyx mori. The two predicted amino acid (aa) sequences are identical up to aa 280 and then diverge. The silkworm and fruit fly proteins share 80% homology in the RNA-binding motif region. These cDNAs detect 2.0-, 1.8- and 1-kb mRNAs in the middle and posterior silk glands.

Alternative Splicing↗

Effects of ethyleneglycol chain length of dodecyl polyethyleneglycol monoether on the crystallization of bovine heart cytochrome c oxidase.

Tetragonal crystals that diffracted X-rays up to 5 A resolution were obtained from bovine heart cytochrome c oxidase isolated and solubilized with dodecyl octaethyleneglycol monoether, CH3(CH2)11O(CH2CH2O)8H. Comparison of observed structure factors between data sets each obtained from a different native crystal gave correlation coefficients of 0.92, 0.84 and 0.57 at 10 A, 7 A and 6 A resolution, respectively. The space group and the cell dimensions of the crystal are I4(1) or I4(3) and a = b = 253 A, c = 507 A, respectively. The perfection and stability of the tetragonal crystals are significantly higher than those of the hexagonal crystals of the protein stabilized with Brij-35, CH3(CH2)11O(CH2CH2O)23H (whose details are reported elsewhere). Examination of the effect of ethyleneglycol chain length on the crystallization revealed that only dodecyl polyethyleneglycol monoethers with eight and seven units were appropriate for producing this type of crystal, indicating an optimum size of the detergent for crystallization of the membrane protein.

Animals↗

Lymph flow and lymph node metastasis in esophageal cancer.

This paper delineates which lymph nodes should be dissected due to the high frequency of metastasis associated with different types of primarily lesions of the thoracic esophagus. In cancer involving the upper third of the esophagus (Iu), lymph flow was found to be primary from the superior mediastinal area to the cervical area; in that involving the middle third (Im), it was broadly distributed from the superior, middle, and inferior mediastinal region to the cervical and abdominal regions; and in that involving the lower third (Ei), it tended to extend from the inferior mediastinal region to the abdominal region, with single primary metastatic nodes also being noted in this area. The significance of the "top" nodes, namely, the nodes located along the right recurrent laryngeal nerve in the upper portion of the thorax, was also investigated, and it was confirmed that the prognosis for patients with metastases to both the top nodes and other nodes was unfavorable. An immunohistochemical study on mediastinal lymph flow using the anti-Su-Ps antibody demonstrated interactions between top nodes and cervical and/or thoracic nodes.

Adenocarcinoma↗

A mutation in the glucagon receptor gene (Gly40Ser): heterogeneity in the association with diabetes mellitus.

A possible pathogenic mutation in the glucagon receptor gene causing a Gly to Ser change at codon 40 (Gly40Ser) was reported to be associated and linked with non-insulin-dependent diabetes mellitus (NIDDM), in France and Sardinia. Since the frequency of the mutation (Gly40Ser), about 5% in the French population of familial NIDDM and 8% in randomly chosen diabetic patients in Sardinia, was much higher than that of any of the previously reported mutations in candidate genes, it is important to clarify whether the contribution of this mutation to NIDDM is universal. In this study, we investigated the association of this mutation with diabetes mellitus in a large number of Japanese diabetic patients (383 NIDDM and 53 insulin-dependent diabetic patients) by polymerase chain reaction-restriction fragment length polymorphism analysis. None of the Japanese diabetic patients showed Gly40Ser mutation and the association of this mutation with NIDDM was significantly different (p < 4.10(-5) vs French, p < 3.10(-6) vs Sardinian by Fisher's exact test). The results not only indicate that the mutation plays little, if any, role in susceptibility to diabetes in Japan, but also indicate the genetic heterogeneity in NIDDM and further emphasize the importance of studies on genetic susceptibility to NIDDM and other complex traits in different ethnic groups.

Adolescent↗

The NSY mouse: a new animal model of spontaneous NIDDM with moderate obesity.

The NSY (Nagoya-Shibata-Yasuda) mouse was established as an inbred strain of mouse with spontaneous development of diabetes mellitus, by selective breeding for glucose intolerance from outbred Jcl:ICR mice. NSY mice spontaneously develop diabetes mellitus in an age-dependent manner. The cumulative incidence of diabetes is 98% in males and 31% in females at 48 weeks of age. Neither severe obesity nor extreme hyperinsulinaemia is observed at any age in these mice. Glucose-stimulated insulin secretion was markedly impaired in NSY mice after 24 weeks of age. In contrast, fasting plasma insulin level was higher in male NSY mice than that in male C3H/He mice (545 +/- 73 vs 350 +/- 40 pmol/l, p < 0.05, at 36 weeks of age). Pancreatic insulin content was higher in male NSY mice than that in male C3H/He mice (76 +/- 8 vs 52 +/- 5 ng/mg wet weight, p < 0.05, at 36 weeks of age). Morphologically, no abnormal findings, such as hypertrophy or inflammatory changes in the pancreatic islets, were observed in NSY mice at any age. These data suggest that functional changes of insulin secretion in response to glucose from pancreatic beta cells may contribute to the development of non-insulin-dependent diabetes mellitus (NIDDM) in the NSY mouse. Although insulin sensitivity was not measured, fasting hyperinsulinaemia in NSY mice suggests that insulin resistance may also contribute to the pathogenesis of NIDDM. Since these findings are similar to the pathophysiologic features of human NIDDM patients, the NSY mouse is considered to be useful for investigating the pathogenesis and genetic predisposition to NIDDM.

Aging↗

Seasonal variation in sweating responses of older and younger men.

Eight older (60-65 years) and six younger (20-25 years) men were exposed to a standard heat stress for 60 min in summer, autumn, winter, and spring. The test consisted of placing the lower legs and feet in a 42 degrees C water bath while sitting in constant environmental conditions (30 degrees C and 45% relative humidity). The increase of rectal temperature (delta Tre) was significantly greater (P < 0.05) in autumn, winter, and spring than in summer for the older group, but significantly greater only in winter than in summer for the younger group (P < 0.05). The delta Tre was greater for the older group in all seasons, but of significance only in autumn and spring (P < 0.01). There were no significant season-related differences for metabolic heat production (M) and mean skin temperature (Tsk) during the heat test in the respective groups, although the M and Tsk were lower for the older group in all seasons (P < 0.01). In the older group total body sweating rate (msw) divided by delta Tre (total msw/delta Tre) decreased from summer to winter (P < 0.02) and did not differ between winter and spring, whereas total msw/delta Tre in the younger group increased in spring after decreasing from autumn to winter (P < 0.03). The variations of the value, local sweating rate on the back and thigh divided by delta Tre (back msw/delta Tre and thigh msw/delta Tre), were similar to those of the total msw/delta Tre in each group, except for back msw/delta Tre in the younger group, which did not increase from winter to spring.(ABSTRACT TRUNCATED AT 250 WORDS)

Acclimatization↗

Seasonal variation in physiological responses to mild cold air in young and older men.

Eight men aged 60-65 years and six men aged 20-25 years, wearing only swimming trunks, were exposed to an air temperature of 17 degrees C and 45% R.H. in each of the four seasons. The increase in the rate of metabolic heat production (% delta M) for the older group in the cold test was significantly higher in summer and autumn than in winter and spring (P < 0.05), but did not differ in the young group between seasons. Compared to the young group the % delta M was significantly greater for the older group (due to a marked increase in four individuals) in summer and autumn (P < 0.04). At the end of the period of cold exposure, the decrements of rectal temperature (delta Tre), mean skin temperature (Tsk; due to a marked decrease in four individuals) and foot skin temperature (Tfoot) were significantly greater for the older group compared to the young group at all times of the year (P < 0.003). Seasonal variations in the two groups were similar, e.g., the delta Tre gradually became smaller from summer to winter (P < 0.05) and then increased slightly in the spring (P = 0.07). Tfoot for both groups decreased from summer to autumn (P < 0.01) and remained unchanged subsequently. No seasonal variations were observed for Tsk in either group. The increase in diastolic blood pressure (BPd) during the test was significantly smaller in winter in both groups (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Acclimatization↗

Detection of MspI RFLP in human THY1 gene by the polymerase chain reaction.

THY1 gene encodes a cell surface glycoprotein predominantly expressed in brain and peripheral nerves. Human THY1 gene region on chromosome 11q23 has been implicated in susceptibility to type 1 diabetes (Wong et al., 1991). Two primers derived from the sequences flanking the polymorphic MspI site in intron 2 of the human THY1 gene (Gatti et al., 1988) were selected for RCP to amplify a 566 bp fragment that spans the MspI polymorphism. Polymorphism was detected by MspI digestion of the PCR product.

Base Sequence↗

Risk factors for restenosis after percutaneous transluminal coronary angioplasty: role of lipoprotein (a).

To evaluate serum levels of lipoprotein (a) (Lp[a]) as a predictor of restenosis after percutaneous transluminal coronary angioplasty (PTCA), we evaluated 71 patients who underwent elective single-vessel angioplasty. Patients were divided into two groups according to the presence (n = 24 [34%]; group R) or absence (n = 47 [66%]; group N) of restenosis. Serum insulin levels were similar before and after the glucose challenge test in both groups. The median level of serum Lp(a) was 34.9 mg/dl in group R compared with 19.4 mg/dl in group N (p < 0.01). The frequency of the apo E4 allele was 4 (17%) in group R and 4 (9%) in group N (p = NS). The incidence of restenosis was significantly higher in patients with Lp(a) levels > or = 30 mg/dl than in those with Lp(a) levels < 30 mg/dl (65% vs 26%; p < 0.01). Our results indicate that a serum Lp(a) level > or = 30 mg/dl is a risk factor for restenosis.

Aged↗

Opioid mu- and kappa-receptor mediate phospholipase C activation through Gi1 in Xenopus oocytes.

In the Xenopus oocytes expressing mu- or kappa-opioid receptors, agonist-induced currents were observed only when the oocyte was coinjected with Gi1 alpha RNA and pretreated with K-252a, a potent inhibitor of protein kinases. The evoked currents were abolished by intracellular injection of EGTA or inositol 1,4,5-trisphosphate and the current-voltage relationship revealed that they are mediated through typical calcium-dependent chloride channels. These findings suggest that the mu- and kappa-receptors mediate phospholipase C activation through Gi1 alpha, and that these receptor mechanisms including downstream signalings might be inhibited by phosphorylation in vivo in the Xenopus oocyte.

Animals↗

Protein kinase C inhibitor potentiates the agonist-induced GTPase activity in COS cell membranes expressing delta-opioid receptor.

In COS-7 cell membranes expressing cloned delta-opioid receptor, [D-Ser2, Leu5]enkephalin-Thr6, an opioid delta-agonist, showed no significant stimulation of high-affinity GTPase, while this agonist binding showed a guanine nucleotide sensitivity. Significant stimulation of GTPase activity by this agonist was observed only when the cells were pretreated with 0.1 microM calphostin C, a protein kinase C inhibitor, and when this inhibitor was further added to the reaction mixture at 1 microM. These findings suggest that protein kinase C is involved in the heterologous desensitization of delta-opioid receptor in the cells.

Analgesics↗

Down-regulation of AMPA-type glutamate receptor gene expression during goldfish optic nerve regeneration.

An AMPA-type glutamate receptor cDNA, GFGR52, was cloned from a goldfish retinal cDNA library. GFGR52 is highly homologous to the AMPA D-FLOP-type glutamate receptor. In situ hybridization revealed GFGR52 gene expression in both retinal ganglion cells and optic tectum. This expression was reduced during optic nerve regeneration and this decreased level of gene expression lasted until the reinitiation of synaptogenesis. These findings suggest that interactions between optic nerve and its target, the optic tectum are necessary for maintaining AMPA D-type glutamate receptor gene expression in both retinal and tectal neurons.

Amino Acid Sequence↗

Altered tonic L-3,4-dihydroxyphenylalanine systems in the nucleus tractus solitarii and the rostral ventrolateral medulla of spontaneously hypertensive rats.

We have proposed that L-3,4-dihydroxyphenylalanine (L-DOPA) is a neurotransmitter in the central nervous system [Y. Misu et al. (1995) Adv. Pharmac. 32, 427-459]. L-DOPA as a probable neurotransmitter for the primary baroreceptor afferents tonically functions to mediate cardiodepressor control in the nucleus tractus solitarii and also tonically functions to mediate cardiopressor control in the rostral ventrolateral medulla of rats. We further attempted to clarify whether a transmitter-like L-DOPA system is altered in these areas of adult spontaneously hypertensive rats. By microdialysis in the left nucleus tractus solitarii area, the basal L-DOPA release was lower in spontaneously hypertensive rats than that in Wistar-Kyoto rats. This release was partially reduced by tetrodotoxin (1 microM) to the same absolute levels in the two strains. Tonic neuronal L-DOPA release is impaired in this nucleus of spontaneously hypertensive rats. This impairment is not secondarily due to decrease in formation or increase in decarboxylation of L-DOPA, since tyrosine hydroxylase activity was increased in spontaneously hypertensive rats, compared to Wistar-Kyoto rats, while no difference of L-aromatic amino acid decarboxylase activity was seen in the caudal dorsomedial medulla including the nucleus. L-DOPA (10-300 ng) microinjected into the nucleus produced dose-dependent hypotension and bradycardia. A maximum depressor response of spontaneously hypertensive rats to L-DOPA at higher doses was slightly greater than that of Wistar-Kyoto rats. On the other hand, in the left rostral ventrolateral medulla, the basal L-DOPA release was higher in spontaneously hypertensive rats than that in Wistar-Kyoto rats. This release was also partially reduced by tetrodotoxin to the same absolute levels in the two strains. Tonic neuronal L-DOPA release is enhanced in spontaneously hypertensive rats. This enhancement seems to include partially a decrease in decarboxylation of L-DOPA, since L-aromatic amino acid decarboxylase activity was decreased in spontaneously hypertensive rats compared to Wistar-Kyoto rats, while no difference in tyrosine hydroxylase activity was seen. L-DOPA (10-600 ng) produced dose-dependent hypertension and tachycardia. Importantly, a pressor response of spontaneously hypertensive rats to L-DOPA at lower doses was slightly greater than that of Wistar-Kyoto rats. L-DOPA seems to play a transmitter-like role in blood pressure regulation at levels of the nucleus tractus solitarii and rostral ventrolateral medulla in rats.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Supersensitization of neurochemical responses by L-DOPA and dopamine receptor agonists in the striatum of experimental Parkinson's disease model rats.

We studied the mechanisms of dopamine receptor agonist- and L-DOPA-mediated supersensitization in experimental Parkinson's disease model rats, by measuring in vivo acetylcholine (ACh) release, GTPase activities, and mRNA expression in the striatum of 6-hydroxydopamine-treated rats. D1 agonist (SKF38393) and D2/D3 agonists (bromocriptine and quinpirole) showed more potent stimulation or inhibitions on ACh release in the model rat than in the control. However, quinpirole-evoked stimulation of GTPase activity was enhance in the model rats, compared to the control, while there was no significant enhancement of the bromocriptine-evoked stimulation. On the other hand, L-DOPA at 0.3-10 pM showed a biphasic action including significant inhibition on the GTPase activity in the lesioned striatal membranes, but not in the control. In the RNAase protection assay, neither D1, D2, Gi1 alpha, GoA alpha nor Gs alpha mRNA expression in the model was significantly different from the control. These findings suggest that there is supersensitization of D1 and D2/D3 receptors in the experimental Parkinson's disease model, while the upregulation of their receptors or GTP-binding proteins (G-proteins) to be coupled to their receptors is unlikely involved in major parts of such mechanisms. In addition, the present report provides the first evidence that L-DOPA mediates neurochemical responses in the plasma membranes, possibly through its receptor.

Acetylcholine↗