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Biomedical subjects

T Ashida

Publications and source records attributed to T Ashida.

At least 109 records · Page 6Linked to original sources

Differential modulation of norepinephrine-induced contractile response by ryanodine and verapamil in the isolated aortic ring of spontaneously hypertensive and Wistar-Kyoto rats.

Using ryanodine and verapamil, we compared the relative contributions of SR Ca2+ release and gated Ca2+ entry in arterial contractions induced by norepinephrine (NE) between spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Aortic rings of 10 SHR and 10 WKY aged 10-12 weeks were superfused in small water-jacketed tissue chambers with physiological salt solution and isometric tension was measured. The inhibition of the NE (3 x 10(-8) M)-induced contraction of aortic rings by ryanodine (10 microM) was significantly greater in WKY (40.1 +/- 6.9%) than in SHR (2.2 +/- 9.0%) (p less than 0.01). The inhibition of the NE-induced contraction by verapamil (10 microM) in the presence or absence of ryanodine (10 microM) was significantly greater in SHR than in WKY. The residual ryanodine and verapamil-insensitive component of NE-induced contraction was significantly greater in WKY than in SHR. Caffeine (5 mM)-induced contraction in the presence of verapamil and phentolamine was significantly smaller in SHR than in WKY. These results suggest that gated Ca2+ entry plays a more important role in Ca2+ control and that ryanodine-sensitive Ca2+ store is smaller, or the ryanodine receptor is altered in these tissues of SHR compared with those of WKY.

Animals↗

Signal transduction mechanisms of Ia induction in B cells by interleukin 4 and immunoglobulin receptors.

Molecular mechanisms of signal transduction through receptors for interleukin 4 (IL-4) are still largely unknown. To elucidate the second messenger(s) of IL-4 action in mature B cells, we performed blocking experiments with inhibitors of various aspects of cellular responses, using Ia-inducing activity of IL-4 as a readout system. In the event, only agents that are shown to inhibit calcium ion (Ca2+) release from intracellular stores, such as 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester (TMB-8) and ryanodine, could block the IL-4 action in a dose-dependent fashion. These results suggest that the process leading to the final expression of IL-4 action may be mediated, at some point, by the release of Ca2+ from intracellular stores. In the parallel experiments with antiimmunoglobulin (Ig) antibody, we found that amiloride, an inhibitor of Na+/H+ pump, blocks the Ia induction by anti-IgM antibody. Thus the Na+/H+ exchange system activated by anti-Ig antibody may be present in mature B cells.

Animals↗

Sodium sensitive and sodium retaining hypertension.

The differences between sodium sensitive and sodium retaining hypertension were theoretically considered using a water tank model of body fluid volume-blood pressure regulation. If an outlet valve is attached to a tank with a base area corresponding to the reciprocal of total peripheral resistance (TPR) and water is poured into this tank at a rate corresponding to the amount of Na+ intake, then equilibrium should be achieved at a certain water level, volume and output from the outlet, which represent mean arterial pressure (MAP), cardiac output (CO) and urinary Na+ excretion. The height of the outlet from the tank bottom and the size cross-sectional area, of the outlet correspond to the x-intercept and slope of the renal function (pressure-natriuresis) curve, respectively. In both nonsodium sensitive hypertension, due to the shift of the curve toward a higher blood pressure level (elevated height of the outlet) without change in the slope (size of the outlet), and sodium sensitive hypertension, due to the depressed slope of the curve (reduced outlet size), not only MAP (water level) but also CO (water volume) are increased, resulting in sodium retaining hypertension, if TPR (reciprocal of base area) remained unchanged, while CO is relatively unchanged, resulting in nonsodium retaining hypertension, if TPR is elevated. Thus, the MAP and its sensitivity to sodium intake is determined by the renal function curve. Since body fluid volume is determined by both the renal function curve and TPR, however, changes in TPR during the development of hypertension is a major factor in determining whether or not the body fluid volume has to change only a small amount or a large amount. Therefore, the sodium sensitivity of blood pressure and sodium retention must be considered separately.

Blood Pressure↗

Preliminary crystallographic study of phospholipase A2 from the venom of Trimeresurus flavoviridis (habu snake).

Crystallization and a preliminary crystallographic study of Trimeresurus flavoviridis (habu snake) phospholipase A2 (PLA2) were carried out. Although crystals were obtained from various solutions, crystals suitable for X-ray analysis could be obtained from polyethylene glycol solutions only when a repeated seeding technique was applied starting from twinned crystals. The crystal is monoclinic with space group P21, with a = 44.1, b = 55.7, c = 48.8 A, and beta = 92.4 degrees. An asymmetric unit contains a dimer consisting of two identical subunits made of 122 amino acids. The crystal reflects X-rays beyond 2.5 A. A Pt derivative gave a good isomorphous crystal.

Animals↗

Crystallization and preliminary crystallographic study of bacterial alpha-amylases.

Crystallization of bacterial alpha-amylases has been achieved by the hanging-drop vapor diffusion method. The crystals of Bacillus licheniformis and B. licheniformis 584 amylases are isomorphous to each other. The crystals of B. licheniformis amylase belong to the tetragonal system, space group P4(2)2(1)2 with cell dimensions of a = 119.3 and c = 85.4 A. The asymmetric unit contains one molecule of amylase, with a volume per molecular mass, Vm, of 2.75 A3/Da. The crystals of B. licheniformis and B. licheniformis 584 amylases diffract beyond 2.5 A resolution and are suitable for X-ray diffraction analysis. The crystals of B. amyloliquefaciens amylase are orthorhombic, and have space group C222(1), with cell dimensions of a = 154, b = 298, and c = 90 A. The asymmetric unit contains three to five molecules. In the crystallization of B. licheniformis and B. licheniformis 584 amylases, the addition of EDTA was indispensable to obtain large single crystals, while it had an adverse effect on the crystallization of B. amyloliquefaciens amylase, producing a large amount of small crystals.

Bacillus↗

[Analysis of subsets of colonic mucosal lymphocyte in ulcerative colitis by two color flow cytometry].

To assess how the immunological events occur in the colonic mucosa in patients with ulcerative colitis, it is thought to be important to evaluate the subpopulations and/or subsets of mucosal lymphocytes. In this point of view, we assayed those by lymphocyte isolation techniques and two color flow cytometry. Although our results showed no disease-specific abnormalities of the percentages of CD3, CD4, CD8, CD20, and HLA-DR (+) cells in PBL (peripheral blood lymphocyte) nor CML (colonic mucosal lymphocyte), these subsets of CML appeared to be altered according to the grade of severity of inflammation. In our cases, the HLA-DR (+) cell and CD4 population were larger in severely inflamed mucosa. Furthermore, fluorescence intensities of HLA-DR antigen of CD20 population in CML were greater than those in PBL. These results suggest that the B cell-mediated mechanisms may play an important role in maintaining the severe inflammation, and the clinical significance of these studies are discussed.

Adolescent↗

[Detection of proliferative cells in colorectal carcinomas and adenomas by monoclonal antibody to DNA polymerase alpha].

DNA polymerase alpha is an endogenous DNA replication enzyme expressed in all cells in a proliferation cycle. An immunoperoxidase method and the monoclonal antibody to DNA polymerase alpha were used to identify proliferating cells in colorectal carcinomas (n = 35) and adenomas (n = 43). The labeling index (L.I.) in colorectal carcinomas was 51.6%, being significantly higher than 28.6% in adenomas. The L.I. in colorectal carcinomas correlated with clinical staging (stage I: 33.1%, stage II and III: 49.5%, stage IV and V: 66.9%). Furthermore, the L.I. had a tendency to elevate as carcinoma deeply invaded (pm: 25.8%, ss-s or a1-a2: 52.2%, si or ai: 67.5%). The L.I. in adenoma was related to the degree of atypia. The L.I. in adenomas with mild atypia, with moderate atypia, and cancer in adenoma were 18.3%, 31.5%, and 47.0%, respectively. And the L.I. of cancer in adenoma had no significant difference in advanced carcinomas (47.0% vs 51.6%). These results suggest that the L.I. is useful as a marker for evaluating the degree of biological malignancy of human colorectal carcinomas and the degree of histopathological atypia of adenomas.

Adenocarcinoma↗

[A case of alder (Alnus sieboldiana) pollinosis and its prevalence among the residents of a densely forested area].

The first patient with alder (Alnus sieboldiana Matsumura) pollinosis in Japan visited our clinic in April 1987. The patient was a 54 year old housewife who had lived in Kobe at the foot of Mt. Rokko since 1972, and had been suffering from the pollinosis for three years. In 1988, prevalence studies of A. sieboldiana in densely planted area close to the patient's house were performed using pollen count, immunological examinations and a population survey by questionnaire. A remarkably high prevalence rate of the pollinosis was demonstrated.

Adolescent↗

Alleviation of autoimmunity in BXSB mice by monoclonal alloantibody to Ly-5 (CD45).

One of the hallmarks of immunological abnormalities in mice that are prone to syndromes resembling systemic lupus erythematosus is polyclonal activation of B cells and a switch of serum immunoglobulins or autoantibodies from IgM to IgG that coincides with onset of autoimmune disease. Therefore, we investigated whether the development of autoimmunity can be alleviated in such mice by administration of monoclonal alloantibody directed against Ly-5, because this was known to reduce levels of polyclonal IgG antibody without affecting other immunoglobulin isotypes. Our results indicate that at least some forms of autoimmune disease may be effectively treated by thus reducing the concentration of IgG autoantibody in this manner.

Animals↗

Clinical application of sodium-23 nuclear magnetic resonance for measurement of red cell sodium concentrations.

Red cell sodium (RBC-Na+) concentrations were measured using 23Na nuclear magnetic resonance (NMR), without the destruction of erythrocyte membranes. Subjects were categorized into four groups: 20 normotensive subjects (NT group), 20 age-matched essential hypertensive patients (EHT group), 10 patients with primary aldosteronism (PA group), and 18 patients treated with digoxin (DIG group). Although RBC-Na+ concentrations were similar between the NT group (6.14 +/- 0.80 (Mean +/- SD) mmol/l) and the EHT group (5.92 +/- 0.99), they were significantly higher in both the PA group (7.55 +/- 0.88, p less than 0.001) and the DIG group (8.43 +/- 3.81, p less than 0.02). In the PA group, RBC-Na+ concentrations decreased significantly after resection of the adenoma, and there was an inverse relationship between serum potassium and RBC-Na+ concentrations (r = -0.65, p less than 0.01). In the DIG group, RBC-Na+ concentrations tended to increase in proportion to serum digoxin levels (r = 0.53, p less than 0.05). These results support the view that RBC-Na+ concentrations are determined primarily by Na+/K+-pump activity of red cell membranes. This study showed also that Na+ NMR is an useful method determining intracellular Na+ concentrations.

Digoxin↗

Hypotensive mechanisms of antihypertensives based on the water tank model.

Mechanisms of hypotensive action of antihypertensive drugs were theoretically considered using the water tank model on body fluid volume-blood pressure regulation. If a cock is attached to a tank with a base area corresponding to the reciprocal of total peripheral resistance (TPR) and water is poured into this tank at a rate corresponding to the amount of Na+ intake, then equilibrium should be achieved at a certain water level, volume, and output from the cock, which represent mean arterial pressure (MAP), cardiac output (CO), and urinary Na+ excretion rate. The height of the cock from the tank bottom and the size of the cock correspond to the x-intercept and slope of the renal function curve (pressure-natriuresis relationship). Vasodilators lowered the cock height and enlarged the base area (reduced TPR), resulting in an increase in the volume (CO). beta-Blockers not only lowered the height, but also decreased the size of the cock. Because they do not alter TPR (base area), CO (volume) must be reduced. Converting-enzyme inhibitors had the same effects on the height and size of the cock as beta-blockers, but enlarged the base area (reduced TPR), resulting in an increase in the volume (CO). Diuretics increased the cock size without affecting the cock height or the base area, resulting in a decrease in CO. The effects of antihypertensive drugs on the cock (kidney) seemed essential in their hypotensive action, similarly as abnormality of the renal function curve is essential in the genesis of hypertension. These analyses can illustrate schematically the hemodynamic changes induced by antihypertensive drugs.

Antihypertensive Agents↗

Effect of dietary sodium on the Na-K ATPase inhibitor in patients with essential hypertension.

To study the circulating humoral factor modifying transmembrane sodium transport, plasma was obtained from 12 patients with essential hypertension (EH) fed a high sodium diet (NaCl 15 to 17 g/d) for seven days and thereafter a low sodium diet (NaCl 2 to 3 g/d) for seven days. Ouabain-sensitive 86Rb+ influx into the red blood cells (RBC) obtained from a healthy subject, and incubated with the plasma obtained during the high sodium diet was significantly lower than that incubated with the plasma obtained during the low sodium diet (3.74 +/- 0.26 v 3.97 +/- 0.30 nmol/10(8) cells, P less than .05). The changes in mean blood pressure from the high to low sodium diet showed a significant positive correlation with the changes in the ouabain-sensitive Rb influx into RBC in the plasma from the high to low sodium diet. These results suggest that a humoral factor modifying the sodium pump might be altered by sodium balance in EH, especially in salt-sensitive hypertension.

Aldosterone↗

Effects of salt, prostaglandin, and captopril on vascular responsiveness in essential hypertension.

To examine the influence of dietary sodium, prostaglandin, and captopril on vascular reactivity, 12 patients with essential hypertension (EH) and seven normotensive subjects (NT) were given a high sodium diet and thereafter a low sodium diet, each for ten days. Indomethacin (IND) (150 mg/d) was administered during the last three days of each dietary period. Blood pressure and cardiac output (CO) (by impedance cardiography) were measured during the angiotensin II (Ang II) infusion before and after the IND treatment of each dietary period. In eight patients with EH, captopril 100 mg was given before Ang II infusion. EH patients were classified as either salt sensitive (SS) or nonsalt sensitive (NSS). The mean blood pressure (MBP) response to Ang II was significantly higher on high sodium intake than on low sodium intake in NSS and NT, but not in SS. IND significantly increased the MBP response to Ang II on low sodium intake in NSS and NT, but not in SS. IND significantly increased the TPR response to Ang II on low sodium intake, remarkably in NSS and NT compared with SS. Salt sensitivity (% decrease in MBP from high to low sodium intake) highly correlated with the increase in the TPR response to Ang II by IND on low sodium intake (r = -0.90). After captopril administration, IND still increased the MBP and TPR response to Ang II on low sodium intake. These results suggest that the modulation of the vascular responsiveness to Ang II by prostaglandins is altered by sodium balance and salt sensitivity in EH.

Angiotensin II↗

Increased sodium-calcium exchange in arterial smooth muscle of spontaneously hypertensive rats.

We compared sodium-calcium (Na-Ca) exchange in vascular smooth muscle between spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. Aortic rings of 11 SHR and 11 WKY rats aged 11-12 weeks were superfused with physiological saline, and isometric tension was measured. Systolic blood pressure was higher in SHR (174 +/- 12 mm Hg) than in WKY rats (132 +/- 4 mm Hg): 1) In the presence of 10 microM phentolamine, 10 microM verapamil, and 5 mM caffeine, reduction of ionized extracellular sodium concentration [( Na+]o) from normal (139.2 mM) to 1.2 mM (replaced by N-methyl-D-glucamine) caused an external Ca2+-dependent increase in tonic tension (calcium entry by Na-Ca exchange). The rate of increase was higher in SHR (35.4 +/- 3.9 mg/min) than in WKY rats (17.9 +/- 1.3 mg/min) (p less than 0.01). 2) In the presence of phentolamine, verapamil, and caffeine, relaxation from low-Na+ contraction was promoted by external calcium removal. The rate of relaxation was directly related to [Na+]o. The rates of relaxation at normal (139.2 mM) [Na+]o were higher in SHR than in WKY rats (p less than 0.05). The rates of relaxation at 1.2 mM [Na+]o (calcium extrusion by adenosine triphosphate-driven calcium pump) were not different between SHR (11.6 +/- 2.8 mg/min) and WKY rats (8.9 +/- 2.5 mg/min). The increase in the rates of relaxation from 1.2 mM to normal (139.2 mM) [Na+]o (calcium extrusion by Na-Ca exchange) was greater in SHR (34.9 +/- 6.6 mg/min) than in WKY rats (17.1 +/- 4.5 mg/min) (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Effect of recombinant human granulocyte colony-stimulating factor on white blood cell count and neutrophil functions in patients with malignant lymphoma].

We studied the in vivo effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on white blood cell (WBC) count and neutrophil functions in nine patients with malignant lymphoma. The WBC count and absolute neutrophil count were rapidly increased without a nadir phase after chemotherapy. Neutrophil alkaline phosphatase (NAP) scores also markedly increased following chemotherapy in all patients. Phagocytosis of India ink and nitroblue tetrazolium (NBT) reduction were revealed tend to be increased, but not exceeded significantly to normal range. RhG-CSF repaired neutrophil function in patients with decreasing that. Thus, rhG-CSF may be useful for prevention and treatment of infection after chemotherapy.

Adult↗

[Estimation of the intrarenal hemodynamics in patients with primary aldosteronism].

Intrarenal hemodynamics were estimated in patients with primary aldosteronism (PA) using the Gomez's equations and by analyzing their renal function curve. The study was performed in 6 patients with PA for 2 weeks; they were given a regular sodium diet (12-15 g/day) in the 1st week and a sodium restricted diet (1-3 g/day) in the 2nd week. Blood pressure and urinary sodium excretion (UNaV) were measured on the last three days of each stage. Glomerular filtration rate (GFR) and renal plasma flow rate (RPF) were also measured on the regular sodium diet. Afferent (RA) and efferent (RE) arteriole resistances, and glomerular hydrostatic pressure (PG) were calculated using Gomez's equations. UNaV was plotted on the ordinate as a function of mean blood pressure (MAP) on the x-axis. Assuming that the difference between MAP and the x-intercept of this renal function curve represents the effective filtration pressure, RA, PG and gross filtration coefficient of capillaries (KFG) were also calculated. GFR and RPF were 102 +/- 6, 469 +/- 27 ml/min, respectively. Estimated RA (6600 +/- 700 dyns.sec.cm-5) was markedly elevated, while RE (2500 +/- 100 dyns.sec.cm-5), PG (57 +/- 2 mmHg) and KFG (0.195 +/- 0.041 [ml/sec]/mmHg) remained normal. The intrarenal hemodynamic parameters obtained by analyzing the renal function curve were consistent with those by Gomez's equations. Recently, in DOCA-salt hypertension, an animal model analogous to human PA, PG has been reported to be increased, leading to renal damage. However, the increase in RA was as marked as seen in essential hypertension, and further the increase in PG was not observed in the patients of this study. Thus, it is not clear whether there is a meaningful relationship between glomerular hypertension and renal damage in PA as reported in DOCA-salt hypertension.

Adult↗