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Biomedical subjects

N Takasu

Publications and source records attributed to N Takasu.

At least 127 records · Page 7Linked to original sources

Increase in colony stimulating factor (CSF) in serum and augmentation of CSF responsiveness of lymphoid mononuclear cells by acute Trypanosoma cruzi infection in mice.

Acute infection of Trypanosoma (T.) cruzi to C3H/HeN mice caused the induction of a higher level of serum colony stimulating factor (CSF) activity to support the proliferation of mouse bone marrow cells. The CSF activity reached a maximum 2 days after the infection and declined thereafter. Spleen cells of the T. cruzi-infected mice showed higher levels of responsiveness to CSF in L929-conditioned medium, mouse recombinant GM-CSF and infected mouse sera as compared with normal mouse spleen cells. The induction of CSF-responding cells became plateau 4 days after the infection and it decreased thereafter. In concomitant with the production of CSF activity in the infected mouse sera, large granular cells bearing high intensity of Mac-2 antigen increased in the infected mouse spleen. These cells were nylon nonadherent and displayed inhibitory effect on T cell response to Con A. These findings indicate that T. cruzi infection induces augmentation of in vivo CSF production, leading to the abnormal proliferation of CSF-responding cells and that augmented production of, and responsiveness to, CSF might be one of important mechanisms responsible for the induction of immune abnormalities in T. cruzi-infected mice.

Animals↗

Cytoplasmic pH in the action of insulin-like growth factor-I in cultured porcine thyroid cells.

The effects of insulin-like growth factor-I (IGF-I) on cytoplasmic pH (pHi) and [3H]thymidine incorporation were studied in primary cultures of porcine thyroid cells. IGF-I alkalinized thyroid cells and stimulated thymidine incorporation in a dose-dependent manner; the effects of IGF-I on alkalinization (the maximal rates of change of cytoplasmic pHi/min ((dpHi/dt)max)) and thymidine incorporation were observed at 2 ng/ml and were maximal at 100 ng/ml, with half-maximal stimulation at approximately 10 ng/ml. The results indicate that Na+/H+ exchange or cell alkalinization may function as a transmembrane signal transducer in the action of IGF-I on thyroid cell proliferation. Several mitogens and comitogens which activate sodium hydrogen exchange, including epidermal growth factor, platelet-derived growth factor and nerve growth factor, have been listed. Activation with IGF-I has not, however, been presented before. Thus the present study constitutes the first demonstration of IGF-I-stimulated activation of Na+/H+ exchange or cell alkalinization.

Amiloride↗

Early changes in fine structures of the aortic arch in hamsters fed a high cholesterol diet.

Intimal changes at the prelesional stage of atherosclerotic lesions were investigated ultrastructurally using hamsters fed a high cholesterol diet for 1 day to 1 month. Observations were restricted to the lesion-prone area in the aortic arch. The endothelial cells began to show well-developed rough endoplasmic reticulum and Golgi apparatus after a few days of the cholesterol diet. After three days, macrophages which contained a few lipid droplets were observed just below the endothelium. We found that the intimal smooth muscle cell formed a gap junction with the process of the medial smooth muscle cell. After a few weeks to 1 month on the diet, the intima became markedly thickened and filled with dense extracellular matrices. The intimal smooth muscle cells showed well-developed rough endoplasmic reticulum and Golgi apparatus with immature granules, suggestive of high secretory activity. The present study showed that endothelial ultrastructural changes, macrophage invasion, and medial smooth muscle cell migration are very early events occurring within a few days after cholesterol intake commences.

Animals↗

Elevated plasma atrial natriuretic peptide level in the early phase of microalbuminuria in patients with non-insulin-dependent diabetes mellitus.

Plasma atrial natriuretic peptide (ANP) levels were examined in 66 patients with non-insulin-dependent diabetes mellitus (NIDDM), and in 9 age-matched normal controls and 18 hypertensive controls. The diabetic patients were classified into three groups according to random urine albumin/creatinine ratio (ACR, mg/g or mg/88.4 nM); group 1 (normo-albuminuria, ACR less than 20, n = 34), group 2 (borderline microcalbuminuria, 20 less than or equal to ACR less than 100, n = 17) and group 3 (manifest microalbuminuria and macroalbuminuria, 100 less than or equal to ACR less than 2000, n = 15). Plasma ANP levels (pg per ml) were significantly elevated in group 2 (46.0 +/- 19.0 SD) when compared with either normal controls (23.8 +/- 14.2), group 1 (28.9 +/- 15.6) or group 3 (26.0 +/- 12.9). This increase in plasma ANP levels was not related to hypertension. Furthermore, plasma ANP levels correlated positively with log(ACR) among the patients with ACR under 100 (groups 1 and 2 combined, r = 0.4701, p less than 0.01). These results suggest that an elevated plasma ANP level in the early phase of microalbuminuria possibly plays a pathophysiological role in the development of nephropathy in NIDDM patients.

Albuminuria↗

Insulin-like growth factor I stimulates inositol phosphate accumulation, a rise in cytoplasmic free calcium, and proliferation in cultured porcine thyroid cells.

Insulin-like growth factor (IGF-I) stimulates thyroid cell proliferation. Using primary cultured porcine thyroid cells, we studied the intracellular pathways that mediate the action of IGF-I on thyroid cell proliferation. IGF-I stimulates inositol phosphate accumulation, a rise in cytoplasmic free calcium [( Ca2+]i), and cell proliferation. Exposure to IGF-I results in a time- and dose-dependent accumulation of inositol monophosphate, inositol bisphosphate, and inositol trisphosphate. IGF-I also increases [Ca2+]i, measured using fura-2, a fluorescent Ca2+ indicator; the IGF-I-induced [Ca2+]i response occurs immediately, reaches a maximum within 1 min, and then slowly declines. IGF-I stimulates thyroid cell proliferation, stimulates thymidine incorporation, and increases cell numbers. The IGF-I-induced inositol phosphate accumulation and [Ca2+]i response parallel thyroid cell proliferation in a dose-dependent manner; the maximal response is observed at a concentration of 100 ng/ml IGF-I, with half-maximal stimulation at approximately 10 ng/ml. Inositol phosphate accumulation and [Ca2+]i response after IGF-I stimulation may function as intracellular messengers for thyroid cell proliferation. This report may constitute the first demonstration of IGF-I-stimulated inositol phosphate accumulation and [Ca2+]i response in the cells.

Animals↗

Cytoplasmic pH in cultured porcine thyroid cells: alkalinization by epidermal growth factor.

We studied the effects of epidermal growth factor (EGF), thyroid-stimulating hormone (TSH) and amiloride on cytoplasmic pH (pHi) in cultured porcine thyroid cells. We used 2',7'-bis(2-carboxyethyl)-5- (and 6-)carboxyfluorescein (BCECF), an internalized fluorescent pH indicator, to measure pHi. EGF stimulated thyroid cell alkalinization and proliferation, which were blocked by amiloride. EGF-stimulated thyroid cell alkalinization depended on extracellular Na+ concentrations. EGF stimulation resulted in an activation of Na+/H+ exchange, which alkalinized the cells. The results indicated that Na+/H+ exchange or cell alkalinization might function as a transmembrane signal transducer in the action of EGF. In the present system, TSH did not stimulate alkalinization or proliferation.

Amiloride↗

Glucose raises cytosolic free calcium in the rat pancreatic islets.

Cytosolic free calcium [( Ca2+]i) was measured using fura 2 in the whole pancreatic islets obtained from male Wistar rats by collagenase dispersion. The pattern of change of [Ca2+]i in response to high glucose, potassium (K+) depolarization or the removal of extracellular calcium was compared with the temporal profile of insulin secretion. Twenty-nine mM glucose produced a gradual increase in [Ca2+]i with approximately 1.5 min of latency period. It remained elevated until the end of observation period (25 min) during which period the first phase of insulin secretion ceased and the second phase of secretion gradually increased. Depolarizing concentration of KCl also produced an elevation of [Ca2+]i, without detectable latency period, which lasted at a sustained level for the entire observation period (30 min). KCl caused a rapid increase of insulin secretion followed by a gradually decreasing level of secretion. Elevated [Ca2+]i and insulin secretion in response to high glucose returned to the basal level when external calcium was removed by the addition of EGTA. We conclude that high glucose and K+ depolarization raise [Ca2+]i in the pancreatic islet. However, the elevation of [Ca2+]i and insulin secretion are not always correlated in the later period of stimulation.

Animals↗

Age-related changes of thyroid function and immunologic abnormalities in patients with hyperthyroidism due to Graves' disease.

In order to clarify the reasons why hyperthyroidism due to Graves' disease is mild in aged patients compared to young patients, we examined the degree of hyperthyroidism and immune abnormalities in 371 untreated patients. The patients were divided into nine groups based on their age; groups A to I consisted of patients aged 0-9, 10-19, 20-29, 30-39, 40-49, 50-59, 60-69, 70-79, and 80-89 years, respectively. Serum thyroxine (T4), triiodo-thyronine (T3), and thyroglobulin levels were highest in group B (patients aged 10-19 years) and decreased progressively with advancing age. T3/T4 ratio was also highest in group B and decreased progressively with age. These data reconfirmed that the degree of hyperthyroidism is milder in older patients. The prevalence of positive microsomal and thyroglobulin antibodies was highest in group B (patients aged 10-19 years) and progressively lower in older groups, suggesting that an association with Hashimoto's thyroiditis is less prevalent in aged patients. We suggest that hyperthyroidism due to Graves' disease is mild in aged patients because responsiveness of the thyrocyte to the abnormal stimulator, thyrotropin-receptor antibody, may be reduced. Such reduced responsiveness may not be due to Hashimoto's thyroiditis, since the prevalence of antithyroid antibodies is lower in aged than in younger patients with hyperthyroidism due to Graves' disease.

Adolescent↗

A new method for predicting the outcome and survival period in paraquat poisoning.

1. To elucidate the importance of lung damage in the prognosis of paraquat poisoning, the respiratory index (RI: A-aDO2/PO2) was analysed in 51 patients with paraquat poisoning. 2. Progressive deterioration of the RI was observed in 43 non-survivors, but not in 8 survivors. In addition, the RI of non-survivors became greater than 1.5 in the course of their poisoning, while that of the survivors remained less than 1.5. 3. The level of the RI (i.e. whether it became greater than 1.5 or not) was found to be a good indicator for the prognosis for life. 4. The RI-time, defined as the time taken from ingestion for the RI to be greater than 1.5, was found to be a good indicator for predicting the survival period in fatal cases. 5. As a result, we conclude that it is important and useful to examine a series of RI and the RI-time in poisoned patients not only to provide proper respiratory care, but also to predict outcome and survival period.

Adolescent↗

A new variant of 17 alpha-hydroxylase deficiency with hyperaldosteronism in two Japanese sisters.

We present a report on two sisters who have 17 alpha-hydroxylase deficiency with hyperaldosteronism. They have hypertension and hypergonadotropic hypogonadism. The steroid profiles suggest that they have 17 alpha-hydroxylase deficiency. In contrast to the classical biochemical findings in 17 alpha-hydroxylase deficiency, both of these patients have hyperaldosteronism. Thus this report describes a new variant of 17 alpha-hydroxylase deficiency with hyperaldosteronism. Dexamethasone suppressed the mineralocorticoid excess, including aldosterone, and improved their hypertension. In the untreated state, ACTH, instead of the renin-angiotensin system, regulated plasma aldosterone levels, but during dexamethasone treatment the renin-angiotensin system regulated these levels.

Adolescent↗

A microtestplate-immunofluorescence assay for anti-Trypanosoma cruzi antibodies.

A new microassay method for the detection of anti-Trypanosoma (T.) antibodies was developed by using Terasaki's microtestplates, and T. cruzi or other parasites were immunofluorescently labeled with polyclonal or monoclonal anti-T. cruzi antibodies. The fluorescence intensity of immunofluorescently stained parasites was assessed by either visual observation or quantitative analysis using a microscopic spectrophotometer. Although the sensitivity of the microtestplate assay was nearly equal to that of the conventional glass slide assay or ELISA, this method is easier in the assay procedures and spares the amount of antibodies and the number of target parasites requisite to the assay.

Animals↗

Production and characterization of monoclonal antibodies against Trypanosoma cruzi-associated antigens.

Nine monoclonal antibodies (mAb) recognizing characteristic antigens in Trypanosoma (T.) cruzi were obtained from hybridomas which had been established by a fusion between mouse myeloma cells and the spleen cells of a mouse immune to the epimastigote form of T. cruzi (Tulahuen strain). Antigen specificities of these mAb were assessed by an indirect immunofluorescence (IIF) method using T. cruzi in different life cycles (amastigote and trypomastigote) and other members of the Trypanosomatidae. The mAb were classified into 3 groups from their reaction patterns to different parasites: 1) The strain specific mAb that reacted only with T. cruzi Tulahuen epimastigote, 2) the species specific mAb that reacted with all T. cruzi strains but not with other species of parasites, and 3) the mAb that were cross-reactive with other species of Trypanosomatidae. Most mAb were specific to epimastigote form of T. cruzi, but some reacted weakly with trypomastigote and amastigote form of the parasites. Immunoblotting and glycolipid analyses of the membrane fraction of homogenized parasites using the mAb identified at least 3 distinct antigenic molecules; those of protein nature having Mr. 43,000 and 58,000 and Mr. 43,000 and 62,000 and molecule(s) of glycolipid nature.

Animals↗

Cytoplasmic pH in cultured porcine thyroid cells: alkalinization by Ca2+.

The role of calcium in cytoplasmic pH (pHi) changes was studied using 2',7'-bis(2-carboxyethyl)-5-(and 6-)carboxyfluorescein, an internalized fluorescent pH indicator, in cultured porcine thyroid cells. The Ca2+ ionophores A23187 and ionomycin stimulated thyroid cell alkalinization. An increase in cytoplasmic free calcium resulted in activation of Na+/H+ exchange which alkalinized the cells.

Animals↗

Generation of hydrogen peroxide in cultured porcine thyroid cells: synergistic regulation by cytoplasmic free calcium and protein kinase C.

This study set out to elucidate the mechanism by which H2O2 generation is regulated in cultured porcine thyroid cells. We monitored continuously the effects of the calcium ionophore A23187 and the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) on H2O2 generation, using homovanillic acid and horseradish peroxidase. A23187 and TPA stimulated H2O2 generation. A23187 increased cytoplasmic free calcium and TPA activated protein kinase C. Generation of H2O2 is therefore regulated by cytoplasmic free calcium and protein kinase C. Exposure to A23187 or TPA augmented further the stimulation of H2O2 generation by TPA or A23187 respectively. Thus A23187 and TPA, by increasing cytoplasmic free calcium and activating protein kinase C respectively, synergistically activate H2O2 generation.

Animals↗

Three-dimensional studies of cytoskeletal organizations in cultured thyroid cells by quick-freezing and deep-etching method.

Isolated porcine thyroid cells were cultured on collagen gels (control group, TSH-stimulated group, and double-layered culture). They were split or cut to remove cytoplasmic soluble proteins for replica preparations. Some specimens were immunostained with anti-actin antibody or decorated with S1 myosin fragments to identify actin filaments. The basal cell membranes of thyroid cells of monolayer culture were in contact with collagen gels and the apical cell membranes faced the culture medium. Networks of actin filaments were attached to the cytoplasmic sides of the apical cell membranes, while intermediate filaments were localized along the basal ones. The thyroid-stimulating hormone (TSH) treatment induced the formation of microvilli only on the apical cell membranes and the accumulation of actin filaments under the apical cell membranes, indicating the apical-basal polarity of the cells. In double-layered culture, the primitive follicular lumens with microvilli appeared between two adjacent cells. The interaction of cell membranes with collagen gels is a determinant factor in the orientation of apical-basal polarity. Moreover, the TSH treatment and cell-cell contact further intensify the polarization through reorganizing the cytoskeletons.

Animals↗

[An undifferentiated carcinoma of the stomach--a report of 2 cases].

Two cases of an undifferentiated carcinoma of the stomach have been investigated by conventional histologic, immunohistochemical, and ultrastructural methods. The patients were a 73-year-old man and 65-year-old man. A postoperative histological examination of the resected stomach of each patient disclosed large areas of an undifferentiated carcinoma which were relatively well demarcated from areas of the adenocarcinoma. In the undifferentiated carcinomatous areas, the reticulin fiber stain was epithelial in pattern, and mucin staining proved negative. Similarly both Grimelius and Fontana-Masson staining also negative. In like manner immunostains using cytokeratin , vimentin , IgG, IgA, s-100 protein, and NSE were all negative, and an electron microscopic study showed no neurosecretory granules or mucin secretory granules. Based on these findings, the diagnosis of either a malignant lymphoma or small cell anaplastic carcinoma could be excluded, and thus an undifferentiated carcinoma was the determination.

Adenocarcinoma↗

Cytoplasmic pH in the action of epidermal growth factor (EGF) in cultured porcine thyroid cells.

We demonstrate measurement of cytoplasmic pH (pHi), using 2',7'-bis(2-carboxyethyl)-5 (and 6-) carboxyfluorescein (BCECF), and internalized fluorescent pHi indicator, in thyroid cells. Using cultured porcine thyroid cells, we studied the effects of epidermal growth factor (EGF) on pHi and [3H] thymidine incorporation; 10 nM EGF alkalinizes thyroid cells and stimulates thymidine incorporation. The results indicate that Na+/H+ exchange or cell alkalinization may function as a transmembrane signal transducer in the action of EGF in the thyroid cells.

Animals↗