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Biomedical subjects

M Koide

Publications and source records attributed to M Koide.

At least 145 records · Page 8Linked to original sources
PubMed1992

Alterations in cellular calcium handling as a result of systemic calcium deficiency in the developing chick embryo: I. Erythrocytes.

Chick embryos rendered calcium (Ca) deficient by shell-less (SL) culture develop hypertension and tachycardia. Since hypocalcemia is accompanied by hypernatremia systemically but not by lower cellular Ca (Koide and Tuan, 1989), we speculate that cellular Ca handling may be altered in the SL embryo, perhaps involving Na transport. Using erythrocytes (RBC) from day-14 SL and normal (NL) embryos as the experimental cell, cellular Ca handling was studied under varying extracellular osmotic and ionic conditions by analyzing 45Ca uptake and cell volume regulation. Two agents, p-chloromercuriphenylsulfonate (PCM), and inosine/iodoacetamide (INI) were used to treat the RBCs to modify plasma membrane ion permeability and to deplete cellular ATP, respectively. Other cellular functions and activities related to Ca homeostasis, including ATP content and Ca(2+)-ATPase activity, were also analyzed. These analyses showed: (1) in NaCl, Ca uptake was similar in NL and SL cells, except after INI treatment, which resulted in slower Ca uptake by the SL cells, (2) in choline and sucrose, Ca uptake by SL RBCs was higher, (3) Ca uptake by RBCs of both embryos changed depending on the osmotic agent (Na < K < or = choline < sucrose), (4) Ca(2+)-ATPase activity was higher in SL RBC, although there was no change in the size or charge of the enzyme, and (5) in any osmotic agent, cellular Na was significantly lower, whereas cellular K was higher, in SL RBC. Based on these results, three features of RBC Ca handling were apparent: (1) Na-Ca exchange was functional and was more active in SL RBCs, (2) Ca uptake was dependent on the total ionic electrochemical gradient but not on bulk H2O movement, and (3) Ca pumping out capacity was directly correlated with Ca(2+)-ATPase activity. Elevated Ca uptake in sucrose-treated SL RBC is therefore indicative of its greater ion permeability. Taken together, these findings indicate that cellular Ca handling of the RBCs of SL chick embryos is characterized by a more active Na-Ca exchange system, greater ion permeability, and higher Ca pumping out capacity, thereby suggesting an up-regulated Ca handling function in the SL RBCs. The abnormal cellular Ca handling may be a direct result of the systemic Ca deficiency of the SL chick embryo and may be functionally related to its hypertension and tachycardia.

Adenosine Triphosphate↗
PubMed1992

Stimulation of protein-tyrosine phosphorylation by endothelin-1 in cultured vascular smooth muscle cells.

In cultured rat aortic smooth cells, endothelin-1 induced tyrosine phosphorylation of at least five proteins with molecular masses of about 79, 77, 73, 45 and 40 kDa in dose- and time-dependent manners. Platelet-derived growth factor also induced tyrosine phosphorylation of the same set of proteins in addition to other proteins including platelet-derived growth factor receptors. This growth factor markedly stimulated DNA synthesis and an increase in cell number in this cell type, but endothelin-1 failed to stimulate these responses under the same conditions. These results demonstrate for the first time that endothelin-1 induces tyrosine phosphorylation of some proteins but suggest that these reactions are not enough to stimulate proliferation of vascular smooth muscle cells.

Animals↗
PubMed1992

Endothelin-1 stimulates tyrosine phosphorylation and the activities of two mitogen-activated protein kinases in cultured vascular smooth muscle cells.

OBJECTIVE: Endothelin-1 (ET-1) has been reported to stimulate the expression of the proto-oncogenes c-fos and c-myc, and to cause DNA synthesis in vascular smooth muscle cells (VSMC). The purpose of this study was to clarify the signalling pathway from ET receptors to the nucleus. DESIGN: Mitogen-activated protein (MAP) kinase, which is activated by various growth factors via phosphorylation of tyrosine and threonine residues, plays important roles as an intermediate in the signalling pathways from growth factor receptors to the ribosomes and nucleus. We examined the effect of ET-1 on the phosphorylation and activation of MAP kinase in cultured VSMC. METHODS: Extracts of ET-1-stimulated VSMC were analysed by one- and two-dimensional gel electrophoresis and anion-exchange column chromatography. Tyrosine-phosphorylated proteins and MAP kinases were detected by immunoblot analyses with anti-phosphotyrosine and anti-MAP kinase antibodies, respectively. The MAP kinase activity was measured using myelin basic protein as a substrate. The MAP kinases were isolated from 32P-labelled VSMC and subjected to phosphoamino acid analysis. RESULTS: ET-1 induced tyrosine phosphorylation of at least five proteins of about 79, 77, 73, 45 and 40 kDa in VSMC. The mobilities of the tyrosine-phosphorylated 45- and 40-kDa proteins were identical with those of the two proteins that were recognized by anti-MAP kinase antibody upon one- and two-dimensional gel electrophoresis. ET-1 stimulated MAP kinase activity in a time-course similar to that of the tyrosine phosphorylation of the 45- and 40-kDa proteins. The ET-1-stimulated MAP kinase activity was resolved almost equally into two peaks upon Mono Q column chromatography (kinase 1 and kinase 2). Kinase 1 and kinase 2 were co-eluted with the tyrosine-phosphorylated 40- and 45-kDa proteins, respectively. The apparent molecular masses of kinase 1 and kinase 2 estimated by MAP kinase assay in polyacrylamide gel were identical with those of tyrosine-phosphorylated 40- and 45-kDa proteins, respectively. Upon phosphoamino acid analysis, ET-1 stimulated phosphorylation of MAP kinases not only on tyrosine but also on threonine residues. CONCLUSIONS: ET-1 induces tyrosine and threonine phosphorylation and the activation of two species of MAP kinases of 40 and 45 kDa in VSMC.

Animals↗
PubMed1992

[Detection of Legionella pneumophila using a nested polymerase chain reaction].

We evaluated the usefulness of a Nested PCR method for detecting Legionella pneumophila. This method resulted in L. pneumophila specific detection as far as we evaluated. The first and second step PCR achieved the sensitivity as small as 10 pg and 10 fg of the target DNA, respectively. In the detection from Legionella seeded sputa, the method could detect 0.1 cfu/ml of the bacteria, and it took about 12 hours to detect the target DNA. We demonstrated that the Nested PCR method was superior in sensitivity and rapidity for isolation of the bacteria to the conventional using low pH treatment and selective media for Legionella.

Base Sequence↗
PubMed1992

Angiotensin II stimulates two myelin basic protein/microtubule-associated protein 2 kinases in cultured vascular smooth muscle cells.

In cultured vascular smooth muscle cells, angiotensin II (Ang II) stimulated a cytosolic protein kinase activity toward myelin basic protein (MBP) in a time- and dose-dependent manner. Phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-dibutyrate also increased the MBP kinase activity. Downregulation of protein kinase C by prolonged treatment of the cells with phorbol 12,13-dibutyrate markedly attenuated the Ang II- and PMA-induced MBP kinase activation. The Ang II- and PMA-stimulated MBP kinase activities were resolved almost equally into two distinct fractions on Mono-Q HR5/5 column chromatography (kinase 1 and kinase 2). The kinase assay in polyacrylamide gel revealed that apparent molecular masses of kinase 1 and kinase 2 were 40 and 45 kd, respectively. Microtubule-associated protein 2 also served as a substrate for both the kinases. Immunoblot analysis with an antiphosphotyrosine antibody suggested that both the kinases were tyrosine-phosphorylated during the action of Ang II. Phosphoamino acid analysis revealed that Ang II and PMA induced phosphorylation of both the kinases on serine/threonine as well as tyrosine residues. Phosphopeptide mapping patterns of kinase 1 and kinase 2 isolated from Ang II-stimulated cells were almost identical with those from PMA-stimulated cells. These results indicate that in vascular smooth muscle cells Ang II activates two species of MBP/microtubule-associated protein 2 kinases mainly through the protein kinase C-signaling pathway and suggest that tyrosine and serine/threonine phosphorylation may be involved in this process.

Angiotensin II↗
PubMed1992

[Inhibitory effect of somatostatin analog, SMS 201-995, on exocrine secretion from isolated rat pancreatic acini].

In vitro effect of somatostatin analog, SMS 201-995 (SMS), on pancreatic exocrine secretion was investigated using isolated rat pancreatic acini. SMS had no effect on basal, cholecystokinin octapeptide (CCK-8)- or secretin-stimulated amylase release. SMS inhibited pancreatic amylase release in response to simultaneous stimulation with secretin and CCK-8 in a dose-dependent manner. Significant inhibition was observed with 10 nM SMS and maximal inhibition with 0.1-1 microM SMS. Amylase release in response to the combination of 100 pM CCK-8, 1 nM secretin and 0.1-1 microM SMS was similar to that to 100 pM CCK-8 alone. Secretin significantly increased acinar cell cAMP content. SMS partially inhibited an increase in cAMP content induced by secretin. The present study has demonstrated, therefore, that SMS directly inhibits the potentiating effect of secretin on exocrine secretion in part by inhibiting an increase in secretin-induced cAMP accumulation in rat pancreatic acinar cells.

Amylases↗
PubMed1992

Experimental studies on cardiovascular disorders and cellular Ca handling in shell-less chick embryos.

1. Chick embryos rendered Ca deficient by long-term culture outside the eggshell develop hypertension/tachycardia. 2. In this study, the hypertension of the shell-less (SL) chick embryo has been characterized by comparing their cardiovascular response to adrenergic drugs and cellular Ca handling function with normal (NL) embryos. 3. Before and after the administration of noradrenaline (NA), phentolamine (PA), isoproterenol (IP), and propranolol (PP), blood pressure and pulse rate were measured. 4. Baseline blood pressure was consistently higher in SL embryos. In both embryos, blood pressure was elevated by NA and PP, but lowered by IP and PA. Pharmacological sensitivity to NA and PA was higher in SL embryos. Serum Ca was lower in SL embryos, while myocardial calcium content was not different. At the cellular level, Ca handling was studied using red blood cells (RBC) from SL and NL embryos under varying osmotic and ionic conditions by analysing 45Ca uptake and cell volume regulation. 5. Two features of RBC Ca uptake were apparent: (i) Na-Ca exchange was functional since Ca uptake was dependent on external Na; (ii) Ca uptake was dependent on total ionic electrochemical gradient. The SL RBC had greater ion permeability, more active Na-Ca exchange function and higher Ca pumping out capacity. 6. These systemic and cellular properties of the SL embryo indicate that it is a novel and useful experimental system to study the relationship between Ca homeostasis and the development of hypertension.

Adenosine Triphosphate↗
PubMed1992

Family history study on hypertension in Japan.

1. Familial aggregation of hypertension was determined in 187 Japanese nuclear families. The necessary data on family and case history particularly concerning the hypertensive status were obtained by personal interviews of outpatients at six local hospitals, using a specifically designed questionnaire. 2. From the family history of the 187 living index children (single ascertainment), 134 pairs of parents were informative. Of 819 children, excluding index cases, the number of hypertensives, normotensives and status unknown were 224, 274 and 187, respectively. Individuals under 30 were classified as unknown. 3. For a child given the birth order, empirical risk to be hypertensive was calculated from the proportion of hypertensives to normotensives plus hypertensives. 4. Irrespective of parental mating types, among a simplex family of hypertension the index cases of the first-born child were more than the others. 5. There was no birth order effect in risk among multiplex families of hypertension. 6. There was no increase of hypertensive offsprings observed when the mother was hypertensive.

Family↗
PubMed1992

[Successful Norwood operation for single left ventricle associated with severe subaortic stenosis, coarctation of aorta and hypoplastic aorta].

A 4 month-old boy who had double-inlet left ventricle, severe subaortic stenosis, hypoplastic ascending aorta and coarctation of the aorta revealed severe respiratory failure. An urgent Norwood operation was done. The procedure consisted of enlargement of the ascending aorta and main pulmonary artery and a systemic-pulmonary shunt using 5 mm PTFE tube. Postoperative course was uneventful.

Aorta↗
PubMed1991

Vasoconstrictor-induced protein-tyrosine phosphorylation in cultured vascular smooth muscle cells.

In cultured rat aortic smooth muscle cells, angiotensin II induced tyrosine phosphorylation of at least 9 proteins with molecular masses of 190, 117, 105, 82, 79, 77, 73, 45 and 40 kDa in time- and dose-dependent manners. Other vasoconstrictors such as [Arg]vasopressin, 5-hydroxytryptamine and norepinephrine induced the tyrosine phosphorylation of the same set of proteins as angiotensin II. The tyrosine phosphorylation of these proteins was mimicked by the protein kinase C-activating phorbol ester, phorbol 12 myristate 13-acetate, and the Ca2+ ionophore, ionomycin. These results demonstrate that the vasoconstrictors stimulate the tyrosine phosphorylation of several proteins in vascular smooth muscle cells and suggest that the tyrosine phosphorylation reactions are the events distal to the activation of protein kinase C and Ca2+ mobilization in the intracellular signalling pathways of the vasoconstrictors.

Angiotensin II↗
PubMed1991

[Isolation of Legionella spp. from cooling tower water in Kinki District, Japan].

Distribution of Legionella spp. were surveyed two different times in Kinki District, Japan. The first time, eighty six building cooling tower waters were collected from Osaka, Hyogo and Nara Prefecture between April and December, 1987. The second time, thirty five waters were studied from Nishinomiya City in Hyogo Prefecture on July, 1989. BMPA alpha agar plate was used as the isolation medium for the first eighty six samples and MWY agar plate for the second thirty five samples. Legionella were isolated from forty two samples (48.8%) of the first eighty six samples. Three different species of Legionella were isolated simultaneously from one sample and two species from eight samples. L. pneumophila serogroup 1 was the most predominant species. Twenty three samples (65.7%) were positive in culture from the second thirty five samples. Three different species of Legionella were isolated simultaneously from three samples and two species from eleven samples. Legionella anisa was more predominant than L. pneumophila serogroup 1 in this study.

Air Conditioning↗
PubMed1991

[Effect of XKJ-001, a crude drug preparation, on body water distribution and water excretion in mice].

The effect of XKJ-001, a crude drug preparation based on Seisho-ekki-to, was investigated on the hematocrit, plasma volume, extracellular and interstitial fluid volumes as well as water excretion in mice. Mice were housed in an animal room maintained at 34 degrees C for 3 d with water and food freely available. While the hematocrit, extracellular and interstitial fluid volumes increased, the plasma volume decreased. These results suggest that the distribution of body water in mice housed at high environmental temperature exhibit the state of water metabolism disorders (Suitai) described in Kampo medicine. After the administration of XKJ-001 (3 g/kg, once a day) for 5 d, mice were housed in an animal room maintained at 34 degrees C for 3 d. The administration of XKJ-001 was allowed to continue on the day 0, day 1 and day 2. XKJ-001 inhibited the increase in hematocrit and the changes in body water distribution of mice induced by high environmental temperature. An effect of XKJ-001 on water excretion in mice was investigated in comparison with hydrochlorothiazide (HTZ). Distilled water (D.W., 100 ml/kg) or bicarbonate saline (B.S., 100 ml/kg) was intraperitoneally injected immediately after the oral administration of XKJ-001 (1.5 g/kg) or HTZ (15 mg/kg). The water excretion was enhanced after 3 h for XKJ-001 and after 6 h for HTZ after the intraperitoneal injection of D.W. As for the intraperitoneal injection of BS, HTZ enhanced the water excretion, however, XKJ-001 exhibited no effect. These results suggest that XKJ-001 has activities on water maldistribution and facilitates the water excretion.

Animals↗
PubMed1991

Small GTP-binding proteins in bovine aortic smooth muscle.

In bovine aortic smooth muscle, GTP-binding activity was equally distributed in the membrane and cytosol fractions. The most abundant GTP-binding proteins (G proteins) in each fraction were purified to near homogeneity and characterized. The most abundant G protein in the membrane fraction had a Mr value of about 22,000 (m22K G) as estimated on sodium dodecyl sulfate-polyacryl-amide gel electrophoresis (SDS-PAGE). m22K G and the human platelet smg p21, a ras p21 like G protein having the same effector domain as ras p21s, were eluted at the same retention time on C4 reversed-phase high performance liquid chromatography (HPLC). Moreover, m22K G was specifically recognized by an anti-smg p21 polyclonal antibody. m22K G was phosphorylated by cyclic AMP-dependent protein kinase with a stoichiometry of one phosphate/molecule of protein. The most abundant G protein in the cytosol fraction had a Mr value of about 21,000 (c21K G) as estimated on SDS-PAGE. c21K G was ADP-ribosylated by botulinum ADP-ribosyltransferase and about 0.4 mol of ADP-ribose was maximally incorporated into 1 mol of c21K G. c21K G and the bovine brain rhoA p21, another ras p21 like G protein, were eluted at the same retention time on C4 reversed-phase HPLC and migrated at the same position on two-dimensional gel electrophoresis. These results indicate that the major G proteins in the membrane and cytosol fractions of bovine aortic smooth muscle are smg p21 and rhoA p21, respectively. Possible roles of these G proteins in vascular smooth muscle are discussed.

Animals↗
PubMed1991

The molecular heterogeneity of the smg-21/Krev-1/rap1 proteins, a GTP-binding protein having the same effector domain as ras p21s, in bovine aortic smooth muscle membranes.

smg p21 is a member of the ras p21/ras p21-like small GTP-binding protein (G protein) superfamily, having the same putative effector domain as ras p21s. In the preceding report, we showed that smg p21 was a major G protein in bovine aortic smooth muscle membranes. Recently, two different smg p21 cDNA clones, designated smg-21A and -B, were isolated from a bovine brain cDNA library. In the present studies, we resolved the bovine aortic smg p21 fraction into two distinct G protein fractions on hydroxyapatite column chromatography and purified them separately to near homogeneity (22K G1 and -2). Both 22K G1 and -2 were specifically recognized by an anti-smg p21 polyclonal antibody. 22K G1 and -2 were identified as smg p21B and -A, respectively, by peptide map and amino acid sequence analyses. Purified smg p21A and -B showed GDP/GTP-binding and GTPase activities similar to each other. The GTPase activities of smg p21A and -B were equally stimulated by smg p21 GTPase activating protein 1 and -2. Moreover, both smg p21A and -B were phosphorylated by cyclic AMP-dependent protein kinase with a stoichiometry of one phosphate/molecule of protein. These results indicate that smg p21A and -B coexist in bovine aortic smooth muscle membranes and suggest that smg p21A and -B may serve as intermediates for cyclic AMP actions.

Amino Acid Sequence↗
PubMed1991

[Localization of HTLV-I-associated antigens in adult T cell leukemia cells and HTLV-I-infected cell line cells].

Localization of HTLV-I-associated antigens was studied in adult T cell leukemia (ATL) cells and HTLV-I-infected cell line cells using monoclonal and human polyclonal antibodies against the viral-related antigens. Two monoclonal antibodies that we obtained by hybridoma technique reacted with HTLV-I-virus core antigens, P19 and P24, respectively. Human anti-HTLV-I-antibodies, which were purified from sera from ATL patients reacted with not only HTLV-I virus particles but also their precursors located in the cytoplasm. In tumor cells freshly isolated from ATL patients, no expression of the virus antigens was observed. When the cells were cultured for several days, the virus antigens were defined in about 3-5% of the cultured cells by the monoclonal antibodies, and in 5-10% by the purified human anti-HTLV-I antibodies. Addition of 5-iodo-2'-deoxyuridine to the culture inhibited cell growth, and at the same time, increased the percentage of the virus antigen-positive cells. Established HTLV-I-infected cell lines showed different cytological profiles from the original ATL cells in the viral replication and morphology.

Antibodies, Monoclonal↗
PubMed1990

Identification of a major GTP-binding protein in bovine aortic smooth muscle cytosol as the rhoA gene product.

In bovine aortic smooth muscle, about 50% of total GTP-binding activity was present in the cytosol fraction. A major GTP-binding protein (G protein) with a Mr value of about 21,000 (21K G) in this fraction was purified to near homogeneity and characterized. 21K G bound maximally about 0.8 mol of [35S]guanosine 5'-(3-O-thio)triphosphate/mol of protein with a Kd value of about 20 nM. 21K G showed GTPase activity with a turnover number of about 0.007 min-1. 21K G was ADP-ribosylated by botulinum ADP-ribosyltransferase and about 0.4 mol of ADP-ribose was maximally incorporated into 1 mol of 21K G. 21K G and the bovine brain rhoA gene product (rhoA p21) were eluted at the same retention time on C4 reversed-phase high performance liquid chromatography and migrated at the same positions on two-dimensional gel electrophoresis. These results indicate that the major G protein in bovine aortic smooth muscle cytosol is rhoA p21.

Adenosine Diphosphate Ribose↗
PubMed1990

Prediction of left ventricular function during supine bicycle ergometer exercise in angina-free patients with old myocardial infarction.

We investigated whether or not left ventricular function during dynamic exercise in angina-free patients with old myocardial infarction could be estimated using resting left ventricular function and noninvasive parameters determined during exercise. We studied 70 patients with old myocardial infarction by measuring hemodynamic parameters during supine multistage bicycle ergometer exercise. Coronary arteriography and left ventriculography were performed: then the left ventricular ejection fraction and left ventricular end-diastolic volume were measured. The parametric changes (delta) between rest and peak exercise were determined. Significant positive correlations were observed between cardiac index (CI) at rest and at peak exercise (r = 0.62, p less than 0.001), as well as between pulmonary artery wedge pressure (PAWP) at rest and at peak exercise (r = 0.72, p less than 0.001). Multiple regression analysis indicated that CI and PAWP at peak exercise as dependent variables were best described by the equations: CI at peak exercise = 1.074 [CIrest] +0.031 [delta HR] + 0.004 [ExD] + 0.018 [LVEF] - 1.560 (r = 0.79, p less than 0.001), PAWP at peak exercise = 0.994 [PAWPrest] - 0.181 [LVEF] + 0.203 [delta DBP] -0.076 [delta HR] -21.488 (r = 0.80, p less than 0.001). These data suggested that CI and PAWP during dynamic exercise in angina-free patients with old myocardial infarction could be predicted using noninvasive parameters, such as increments of blood pressure and heart rate as well as exercise duration, together with data on resting left ventricular function, such as resting CI, resting PAWP, and resting left ventricular ejection fraction (LVEF).

Adult↗