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M Okamoto

Publications and source records attributed to M Okamoto.

At least 829 records · Page 46Linked to original sources

Localization of cytochrome P-450 in the colonic mucosa of 3-methylcholanthrene-pretreated and untreated rats. An immunohistochemical study.

Immunohistochemical localization of cytochrome P-450 in the colonic mucosa of 3-methylcholanthrene-pretreated and untreated rats was studied by indirect fluorescent antibody staining technique. A polyclonal antibody for cytochrome P-450MC purified from hepatic microsomes of 3-methylcholanthrene-pretreated rats was used for this experiment. A strong immunofluorescence was found to be localized in the cytoplasm of the surface epithelium of the mucosa in the colon of 3-methylcholanthrene-pretreated rats. A faint immunofluorescence was also observed in the epithelium of untreated rats. 7-Ethoxycoumarin O-deethylase activity of colonic microsomes was significantly enhanced by 3-methylcholanthrene-pretreatment in parallel with an increase in the intensity of immunostaining for cytochrome P-450MC in Western blotting analysis. This is the first report on the localization of cytochrome P-450 in the colonic mucosa.

7-Alkoxycoumarin O-Dealkylase↗

Biochemical switching device realizing McCulloch-Pitts type equation.

We analyzed cyclic enzyme systems, one of the best candidates for biochemical switching devices, especially focusing on their control mode against external perturbations. Since these systems have the reliability of ON-OFF types of operation (McCulloch-Pitts' neuronic equation), we shall present here the mechanical difference between these systems and electronic switching circuit, especially on the mnemonic mechanism of biochemical switching devices.

Computer Simulation↗

Immunohistochemical study of cytochrome P-450 11 beta-hydroxylase in human adrenal cortex with mineralo- and glucocorticoid excess.

Cytochrome P-450 specific for steroid 11 beta-hydroxylation (P-45011 beta) was immunohistochemically demonstrated in the adrenal glands of human, pig and bovine and of mineralo- and glucocorticoid excess using a specific monoclonal antibody against P-450 11 beta of bovine adrenocortical mitochondria. P-450 11 beta was present in all three cortical zones of the histologically normal adrenal glands of bovine, pig and human, particularly in the zona fasciculata (ZF) and reticularis (ZR). The P-450 11 beta immunoreactivity was intensive in cortical micronodules and inner ZF and ZR in Cushing's disease, and relatively intensive in the zona glomerulosa (ZG) and outer ZF in idiopathic hyperaldosteronism (IHA), corresponding to the sites of active steroidogenesis. In adenomas with Cushing's syndrome and primary aldosteronism, compact cells were generally stained well. In the adrenal glands attached to the adenomas, immunoreactivity was observed only focally in ZG cells but not in ZF and ZR cells.

Adenoma↗

Mechanics of percutaneous balloon valvotomy for mitral valvular stenosis.

The present study was performed to measure and calculate the mechanical force of percutaneous balloon valvotomy (PBV) for mitral valvular stenosis, using an equation pertaining to the mechanical force of the balloon needed to dilate the stenotic mitral valve. In case 1, the diameter of the mitral valve was enlarged by PBV from 1.43 cm to 2.40 cm and, in case 2, from 1.76 cm to 2.42 cm, with a mechanical force of 321 g and 436 g, respectively, following the equation.

Biomechanical Phenomena↗

Light scattering measurements of the gels of gelatin and acto heavy meromyosin solutions by sample rotation method.

The gelation processes of gelatin and acto heavy meromyosin solutions were investigated by a new method of light scattering. The sample was given a constant slow rotation around a vertical axis in a typical dynamic light-scattering setup. The measurements were made on solutions in which polystyrene latex spheres were added as the scattering probes. When a sample reached the gel state, the intensity of the light scattered from the sample fluctuated highly. The relative standard deviation of the intensity fluctuation has been shown to be a good measure of gelation. In addition, computer simulations of this scattering system were found to simulate well the experimental results.

Computer Simulation↗

Production of 19-oic-11-deoxycorticosterone from 19-oxo-11-deoxycorticosterone by cytochrome P-450(11)beta and nonenzymatic production of 19-nor-11-deoxycorticosterone from 19-oic-11-deoxycorticosterone.

19-Oxo-11-deoxycorticosterone was incubated with a cytochrome P-450(11)beta-reconstituted system, and the metabolites were analyzed by high performance liquid chromatography(HPLC). The main product found after chromatography was collected and treated with diazomethane. HPLC and 1H-NMR analysis of the methylated derivative indicated that it was 19-oic-11-deoxycorticosterone methyl ester. When 19-oic-11-deoxycorticosterone was stored at -20 degrees C for 1 month, it was spontaneously converted to other steroids. Structural analysis of the main degradation product indicated that it was 19-nor-11-deoxycorticosterone. These results suggest that the conversion of 19-oxo-11-deoxycorticosterone to 19-oic-11-deoxycorticosterone occurs through the P-450(11)beta-catalyzed reaction, and that the 19-oic-11-deoxycorticosterone thus formed is nonenzymatically converted into 19-nor-11-deoxycorticosterone.

Adrenal Cortex↗

Aldosterone biosynthesis in mitochondria of isolated zones of adrenal cortex.

An assumption that the aldosterone-synthesizing enzyme exists only in zona glomerulosa cells apparently contradicts our recent findings that a purified bovine adrenocortical cytochrome P-45011 beta catalyzes the aldosterone formation and the enzyme exists in both zones of the adrenal cortex. To gain more insight into the zone specificity of aldosterone production, the aldosterone-synthesizing activity of mitochondria prepared from the isolated zones of adrenal cortex of various animal species was investigated. The intact mitochondria from the bovine or porcine zonae fasciculata-reticularis could not produce aldosterone whereas those from the zona glomerulosa produced it at a significant rate. When the mitochondria from the zonae fasciculata-reticularis were solubilized by the addition of cholate, they produced aldosterone from corticosterone at a rate comparable to that of those from the zona glomerulosa. The presence of specific factor(s) in the zonae fasciculata-reticularis mitochondria inhibiting expression of the aldosterone synthetic activity is discussed. The mitochondria of the rat zonae fasciculata-reticularis could hardly catalyze aldosterone synthesis under the detergent-solubilized conditions, whereas those of the zona glomerulosa could. Immunoblot analysis revealed that the mitochondria of the zonae fasciculata-reticularis contained a protein of Mr 51,000 which was immunocrossreactive with a monoclonal antibody directed against P-45011 beta, whereas those of the zona glomerulosa contained two immunocrossreactive proteins of Mr 51,000 and 49,000. These results suggest that in the case of rat adrenal cortex, a specific aldosterone-synthesizing enzyme exists in the zona glomerulosa.

18-Hydroxycorticosterone↗

Inhibition of bovine adrenocortical mitochondrial cytochrome P-450(11)beta-mediated reactions by imidazole derivatives and mineralocorticoid analogs.

The effects of several imidazole antimycotic agents, an imidazole and several mineralocorticoid analogs on the cytochrome P-450(11)beta-catalyzed 11 beta-hydroxylation of 11-deoxycorticosterone and aldosterone synthesis were examined. Ketoconazole, clotrimazole, miconazole and etomidate were found to be potent inhibitors of the reactions, causing 50% inhibition of the 11 beta-hydroxylase activity at concentrations between 10(-8) and 10(-7) M. The potency of etomidate as to the inhibition of aldosterone- and 18-hydroxycorticosterone-production was found to be almost equal to that in the case of 11 beta-hydroxylation. Spironolactone and other newly synthesized mineralocorticoid analogs were also found to inhibit the cytochrome P-450(11)beta-mediated reactions. The ID50 values of these drugs for inhibition of the 11 beta-hydroxylase activity were almost equal to those in the case of the aldosterone- and 18-hydroxycorticosterone-biosynthetic activities. The results of kinetical studies indicated that one of the mineralocorticoid analogs, Compound 23-0586, acts as a competitive inhibitor for the cytochrome P-450(11)beta-mediated reactions.

Adrenal Cortex↗

Dose-response relationship for translocation induction in spermatogonia of the crab-eating monkey (Macaca fascicularis) by chronic gamma-ray-irradiation.

The induction of reciprocal translocations in spermatogonia of the crab-eating monkey (Macaca fascicularis) by chronic gamma-irradiation (1.8 x 10(-5) Gy/min, about 0.024 Gy/22 h/day) was examined. The frequencies of translocation per cell were 0.15% at 0.3 Gy, 0.27% at 1.0 Gy and 0.33% at 1.5 Gy. The dose-response relationship for translocation yield was a linear one with a regression coefficient (b) of 0.16 x 10(-2). When the slope (b) of the regression line was compared with that at a high dose rate (0.25 Gy/min, b = 1.79 x 10(-2), it was clear that the induction rate of translocations after chronic gamma-irradiation was only about one-tenth of that after high-dose-rate irradiation. Thus, there was evidence for a pronounced dose-rate effect in the crab-eating monkey.

Animals↗

Periarteritis of coronary arteries with severe eosinophilic infiltration. A new pathologic entity (eosinophilic periarteritis)?

A 40 year-old male presented symptoms of angina pectoris for about nine years and expired with symptoms of unstable angina, changing pattern at the terminal stage. At autopsy, both right and left coronary arteries of the subepicardial region were grayish white and elastic hard. Histologically, inflammatory infiltration was localized in adventitia of coronary arteries located in the subepicardial region. Inflammatory cells infiltrated into the adventitia were mostly eosinophiles. The medial smooth muscle cells were well preserved and the intima showed irregular thickening with fibrosis. Vascular obstruction or recanalization could not be observed. As a result of these findings, it was considered that these inflammatory changes of the coronary arteries could be termed eosinophilic periarteritis. These inflammatory changes could not be found in the intramural coronary arteries. Rather extensive fibrosis could be seen in the muscle layer centering about the posterior wall of the left ventricle. No findings of angiitis could be detected in the blood vessels except subepicardial coronary arteries.

Adult↗

Cervical lymph node metastasis in carcinoma of the tongue. Correlation between clinical and histopathological findings and metastasis.

The factors relating to lymph node metastasis occurring in association with carcinoma of the tongue have been investigated clinically and histopathologically by many authors. Nevertheless, accuracy in predicting lymph node metastasis has not yet been attained. The present study describes the results of analysis performed in an attempt to predict lymph node metastasis of carcinoma of the tongue. Ninety-eight patients with squamous cell carcinoma of the tongue who were treated at our institution were investigated. Among the 98 patients, lymph node metastases were histologically proven after neck dissection in 40 patients. For statistical analysis in the present study, the quantification theory Type II of Hayashi (1954) was employed for discriminatory analysis. According to this discriminatory analysis, the correlations between the clinical (age, tumour size, N-category) and histopathological (degree of keratinization, mitoses, structure, mode of invasion) findings and metastasis were assessed on the basis of the partial correlation coefficients; these coefficients for each finding were not greatly different from each other and 74 out of 98 patients (76%) could be differentiated between patients with metastasis and without metastasis accurately. The prediction of lymph node metastasis by such discriminatory analysis had significant validity.

Adult↗

Ammonia production from uric acid and its absorption from the caecum of the cockerel.

1. Experiments were done in vitro and in situ in the caeca to measure ammonia production from uric acid and its absorption. 2. When uric acid was introduced into a caecal sac containing mixed caecal micro-organisms 0.77 disappeared in 1 h, with the concomitant appearance of ammonia. 3. Amounts of ammonia produced from added [15N] uric acid in the caeca in situ after 30 min and in caecal medium in vitro after 10 h, were 0.28 and 0.25 respectively of the added 15N. 4. About 0.92 of the ammonia introduced into a caecal sac disappeared from the lumen fluid in 30 min. 5. It is concluded that ammonia is produced from uric acid by caecal micro-organisms and rapidly absorbed.

Ammonia↗

Binding and tyrosine kinase activities of the insulin receptor on Epstein-Barr virus transformed lymphocytes from patients with Werner's syndrome.

To assess the abnormalities of insulin receptor in patients with Werner's syndrome, we established Epstein-Barr virus transformed lymphocytes and studied the binding as well as kinase activities of insulin receptor. The insulin binding to both intact cells and WGA-purified insulin receptor preparations was within normal range. Autophosphorylation of the beta-subunit was not altered in patients with Werner's syndrome, and the receptors of these patients phosphorylated an exogenous substrate to a degree comparable to that of the normal participants. Taken together, these findings indicate that insulin resistance in Werner's syndrome likely is caused by a defect that cannot be detected by means used in the present study.

Adult↗

Clinical implications of pulmonary regurgitation in healthy individuals: detection by cross sectional pulsed Doppler echocardiography.

Pulsed Doppler echocardiography in healthy individuals often shows a disturbance of diastolic flow in the right ventricular outflow tract just below the pulmonary valve that suggests regurgitation. This disturbance of diastolic flow was studied in 50 healthy individuals and 40 patients with cardiopulmonary disease, some of whom had a pulmonary regurgitant murmur. Diastolic flow was disturbed in 39 of the 50 healthy individuals. In 32, cross sectional echocardiography gave a satisfactory image of the pulmonary valve. The characteristic Doppler signals usually lasted throughout diastole, were directed toward the right ventricular cavity, and gradually waned towards end diastole; they formed a spindle shaped area of abnormal signals that extended to within 10 mm of the coaptation of the pulmonary valve towards the right ventricular cavity and the pressure difference estimated from the signals by the modified Bernoulli equation seemed to be proportional to the normal retrograde transpulmonary pressure difference. In all 40 patients with cardiopulmonary disease, signals indicating pulmonary regurgitation were found whether or not a regurgitant murmur was present. When it was present, however, the spindle was longer than 20 mm and in patients with pulmonary hypertension the velocity of abnormal diastolic flow was higher than in healthy individuals. The Doppler signals registering disturbed flow in the healthy individuals resembled the signals caused by pulmonary regurgitation in the patients in terms of location, orientation, and configuration. These results show that healthy individuals usually have trivial pulmonary regurgitation. In practice the distance that the flow disturbance extends from the valve and estimated pressure difference across the valve are probably the most important variables for assessing the clinical significance of pulmonary valve regurgitation.

Adult↗

Change of insulin action with aging in conscious rats determined by euglycemic clamp.

To clarify the mechanism responsible for age-related changes in insulin action, the euglycemic clamp technique was performed with graded doses of insulin in conscious rats aged 2, 4, 10, and 20 mo. Insulin binding (IB) to muscle membranes was also studied. Maximal response of insulin-induced glucose disappearance rate (Rd) was decreased significantly between 2 and 4 mo of age. Dose-response curves shifted to the right progressively up to 20 mo of age. However, IB to the muscle membrane diminished between 1 and 4 mo of age without a decrease thereafter. When Rd was plotted against insulin bound to the membranes, the resulting curves shifted to the right with aging, suggesting a coupling defect between the binding and effector unit. In conclusion, insulin action alters in rats between 2 and 20 mo of age. The most pronounced impairment in IB and maximal response of insulin-induced Rd occurs during early life stage (through maturation) and then a coupling defect seems to be superimposed with further aging. However, we cannot exclude the possibility that these changes may be secondary to obesity or reduced physical activity, rather than aging per se.

Aging↗