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Biomedical subjects

A Ota

Publications and source records attributed to A Ota.

At least 91 records · Page 5Linked to original sources
PubMed1988

Characterization of the altered oligosaccharide composition of the insulin receptor on neural-derived cells.

Typical insulin receptors are present on neuroblastoma cell lines. High affinity binding for insulin was present in membrane preparations from NG108 (a hybrid mouse neuroblastoma-rat glioma) as well as in membranes from SK-N-MC and SK-N-SH, two human neuroblastoma cell lines. Specific [125I]insulin binding was 24.4% for NG108, 16.9% for SK-N-MC and 5.2% for SK-N-SH at membrane protein concentrations of 0.4 mg/ml. IC50 for [125I]insulin binding was 3.4 nM in NG108 membrane preparations and 0.9 nM for SK-N-SH and 1.8 nM in SK-N-MC membranes. Apparent mol. wt. for the alpha subunits (identified by specific immunoprecipitation using the anti-insulin receptor antiserum B10) on SDS PAGE was 134 kDa for NG108; 124 kDa for SK-N-MC and 120 kDa for SK-N-SH. Neuraminidase digestion increased the mobility of the alpha subunit from both NG108 and SK-N-MC receptors to 120 kDa, whereas that from SK-N-SH were unaffected. Endoglycosidase H and endoglycosidase F digestions increased the mobility of the alpha subunits of all 3 cell lines to varying degrees, suggesting the presence of N-linked glycosylation. Insulin induced autophosphorylation of the insulin receptor beta subunit in WGA-purified membranes from all 3 cell lines. In addition, phosphorylation of a protein with an apparent mol. wt. 105 kDa was stimulated by insulin in WGA purified membranes from NG108. Tyrosine-specific kinase activity was present in the membranes from each cell line and was stimulated by insulin in a dose-dependent manner from 10(-9) to 10(-6) M. Proinsulin was about 100 times less potent in stimulating phosphorylation of the artificial substrate poly (Glu, Tyr)4:1 when compared to insulin in accordance with its lower binding affinity to the insulin receptor. Hexose transport was stimulated by insulin in all 3 cell lines. These results indicate that neuroblastoma cells contain specific insulin receptors and that they may be useful as models for studying the role of insulin in nervous tissue.

Binding, Competitive↗
PubMed1988

Insulin-related molecules and insulin effects in the sea urchin embryo.

Insulin, the polypeptide hormone secreted by the differentiated pancreas, may play a role in vertebrate development at prepancreatic stages. In an invertebrate embryo, the sea urchin Strongylocentrotus purpuratus, we now find that insulin modulates the levels of developmentally regulated mRNAs of different lineages (one ectoderm-specific, one mesoderm-specific, and one found in all cell types). Using indirect immunofluorescence, we have localized a molecule which shares antigenic determinants with mammalian insulin in the unfertilized egg as well as in the gut of pluteus larva sea urchins. In addition, Southern hybridization reveals high similarity between sea urchin DNA sequences and the human insulin receptor gene. Our results suggest the presence of an insulin/insulin receptor-related system in sea urchin development.

Animals↗
PubMed1988

Functional insulin-like growth factor I receptors are expressed by neural-derived continuous cell lines.

High affinity insulin-like growth factor I (IGF-I) receptors are expressed by two human neural derived cell lines, SK-N-SH and SK-N-MC. Specific [125I]IGF-I binding to crude membranes was 23.4% for SK-N-SH and 10.7% for SK-N-MC, with 50% inhibition of binding by unlabeled IGF-I between 0.6-0.7 nM. Scatchard analysis of crude membrane binding was linear, whereas Scatchard analysis after wheat germ agglutinin purification of the receptor became curvilinear. The IGF-I receptor alpha-subunits of SK-N-SH have an apparent Mr of 126K, whereas that for SK-N-MC is 132K. Despite these differences in alpha-subunit structure both cell lines demonstrate IGF-I-induced autophosphorylation of their own beta-subunits as well as specific IGF-I induced tyrosine kinase activity, suggesting normal coupling between the ligand-binding alpha-subunit and the tyrosine kinase-containing beta-subunit. Furthermore, IGF-I stimulated iododeoxyuridine uptake in both SK-N-SH and SK-N-MC in a dose-dependent manner, suggesting that these cells may be used to study the role of IGF-I action on neural tissues.

Animals↗
PubMed1988

Insulin-sensitive tyrosine kinase is increased in livers of adult obese Zucker rats: correction with prolonged fasting.

Adult obese Zucker rats (fa,fa) are hyperinsulinemic and insulin resistant. Specific insulin binding to crude membranes prepared from livers was 2.8% (per mg protein) in fatty animals compared with 7.9% in homozygous lean (Fa,Fa) and 9.0% in heterozygous lean (Fa,fa) animals. Insulin binding increased in liver membranes from fatty animals after a 72-h fast to 6.4%. The reduced insulin binding in livers from fatty rats was associated with elevated insulin-sensitive tyrosine kinase activity, which fell towards control values after the fast. The elevated tyrosine kinase activity was associated with an increased maximum velocity (Vmax) without a change in Michaelis-Menten constant (Km) for its substrates, ATP and poly(Glu,Tyr)4:1. These findings suggest that, in adult fatty rats, insulin-sensitive tyrosine kinase has increased intrinsic activity. Further, the effect of the prolonged fast on both insulin binding and kinase activity, suggest that in this model environmental factors, and not necessarily a genetic abnormality, may regulate liver insulin receptors and their kinase. Whether the inverse relationship of the kinase and insulin receptor number is the result of a compensatory mechanism remains to be elucidated.

Animals↗
PubMed1988

ATPase activity in cells of the yeast Saccharomycopsis fibuligera during mycelial growth.

Levels of ATPase activity were examined in the cells of Saccharomycopsis fibuligera during mycelial growth. The mycelial cells (M-form cells) grown under standing culture conditions exhibited a marked increase of ATPase activity after 3 days, showing a broad peak between 5 and 20 days and a maximum at 10 days. The dry weight/ml of the yeast-like cells (Y-form cells) grown under shaking culture conditions reached its peak after 3 days and remained constant throughout the 10 day period of observation. The dry weight/ml of the M-form cells under standing culture conditions exhibited its peak in 10-15 days and then gradually decreased. A very small amount of Y-form cells among the M-form cells under standing culture conditions, exhibited very low ATPase activity even in 10 days' incubation. In M-form cells, the particles increased in number during 20 days of incubation.

Adenosine Triphosphatases↗
PubMed1987

Insulin-like growth factor I receptors in neuronal and glial cells. Characterization and biological effects in primary culture.

Primary cultures of neuronal and glial cells from 1-day-old neonatal rats contain high affinity receptors for insulin-like growth factor I (IGF-I). The IC50 for displacement of 125I-IGF-I binding by unlabeled IGF-I was 3 nM for neuronal cells and 4 nM for glial cells. Unlabeled insulin was 20-50 times less potent. Apparent molecular mass of the alpha subunits of the IGF-I receptor was 125 kDa in neuronal and 135 kDa in glial cells. IGF-I induced autophosphorylation of the IGF-I receptor beta subunit in lectin-purified membrane preparations in a dose-dependent manner. The major phosphoamino acid of the beta subunit in both cell types was tyrosine in the IGF-I-stimulated state and serine in the basal state. Apparent molecular mass of the beta subunits of the IGF-I receptors was 91 kDa for neuronal and 95 kDa for glial cells. Tyrosine kinase activity of the IGF-I receptors was demonstrated by IGF-I-induced phosphorylation of the exogenous substrate poly(Glu, Tyr) 4:1 in both cell types. IGF-I had no effect on 2-deoxyglucose uptake in neuronal cells. In contrast, in glial cells, IGF-I stimulated 2-deoxyglucose uptake at very high doses, presumably acting via the insulin receptor. The effect of IGF-I as a neurotrophic growth factor in both neuronal and glial cells was demonstrated by its stimulation of [3H]thymidine incorporation. These findings suggest the IGF-I is an important growth factor in nervous tissue-derived cells.

Animals↗
PubMed1986

Properties of catalase activity in vegetative and sporulating cells of yeast Saccharomyces cerevisiae.

Properties of catalase activities have been examined in the intact cells of early stationary phase and cells 3 hr after transfer to sporulation medium in Saccharomyces cerevisiae. The catalase activities of the two cells had a broad optimal pH from 6 to 8. Catalase activity in the intact cells increased throughout a 4-hr period of the observation following transfer to sporulation medium. Almost all the catalase activity in vegetative cells was lost by the treatment at 60 degrees C for 10 min. Catalase activities of both cells were inhibited by KCN, NaN3, o-phenanthroline, and PCMB. The catalase activity of the vegetative cells was slightly more inhibited and inactivated than that of the sporulating cells by the inhibitors and by the treatment with HCl or NaOH.

Catalase↗
PubMed1986

Purification and properties of dipeptidase from Escherichia coli AJ005.

A Co2+-dependent dipeptidase from E. coli strain AJ005, a peptidase-deficient mutant, was purified with streptomycin sulfate, ammonium sulfate and DEAE-cellulose. The purified dipeptidase increased by about 106-fold in specific activity, with dilysine as a substrate. The dipeptidase cleaved dilysine to two lysines among the lysine homopolymers, the possibility remaining that it is active toward peptides other than dilysine, since it was investigated in the present study only for activity toward lysine homopolymers. Activity was inhibited 54% by 10(-3) M KCN and completely by 10(-3) M PCMB, EDTA and benzethonium chloride, but not at all by soybean trypsin inhibitors. 78% and 95% of its activity was lost with 30 minutes' treatment at 45 degrees C and 50 degrees C, respectively. The apparent Km value was 6.7 X 10(-4) M for dilysine. It is probable that the dipeptidase differs from dipeptidase DP.

Dipeptidases↗
PubMed1986

Clinical significance of serum bone Gla protein and urinary gamma-Gla as biochemical markers in primary hyperparathyroidism.

The serum bone Gla-protein (BGP) and urinary gamma-carboxyglutamic acid (gamma-Gla) levels were determined in patients with primary hyperparathyroidism (PHP). The mean serum BGP and urinary gamma-Gla levels were 18.6 +/- 2.34 ng/ml and 65.5 +/- 4.62 nmoles/mgCr, respectively, for the 11 patients with the skeletal type of PHP, 5.13 +/- 0.85 ng/ml and 45.2 +/- 1.33 nmoles/mgCr for the 4 with the chemical type, and 7.91 +/- 2.43 ng/ml and 43.2 +/- 3.47 nmoles/mgCr for the 5 with the renal type. Thus, patients with skeletal-type PHP had significantly higher serum BGP and urinary gamma-Gla levels than those with the other type of PHP. Serum BGP levels had significant positive correlations with serum Ca (r = 0.64, P less than 0.005), serum A1-p (r = 0.77, P less than 0.001) and serum PTH (r = 0.45, P less than 0.005). Urinary gamma-Gla levels also had significant positive correlations with serum Ca (r = 0.50, P less than 0.05), serum A1-p (r = 0.67, P less than 0.005), serum 1,25(OH)2D (r = 0.62, P less than 0.02), and serum BGP (r = 0.72, P less than 0.001). Mineral content in the left radius had significant negative correlations with serum BGP levels (r = -0.73, P less than 0.001) and urinary gamma-Gla levels (r = -0.59, P less than 0.01). As these data show, serum BGP and urinary gamma-Gla levels clearly reflect the abnormal bone metabolism and can therefore be useful biochemical markers in PHP.

1-Carboxyglutamic Acid↗
PubMed1986

Properties of glucose uptake in vegetative and sporulating cells of Saccharomyces cerevisiae.

The effect of digitonin, acetic acid, urea and ethanol treatment on the glucose uptake of vegetative cells and of sporulating cells (3 h after transfer to sporulation medium) was examined in Saccharomyces cerevisiae. Both glucose uptake activities decreased at a similar rate, and a slightly different rate, in treatment with various concentrations of digitonin and of acetic acid, respectively, at 25 degrees C for 10 min. The glucose uptake activity of the sporulating cells was much more stable to urea treatment than that of the vegetative cells; the activity decreased about 36% and 76% in the sporulating cells and the vegetative cells, respectively, under conditions of 2.5 M urea at 25 degrees C for 10 min. The glucose uptake activity of the vegetative cells was more stable to ethanol treatment than that of the sporulating cells; the activity decreased about 56% and 88% in the vegetative cells and the sporulating cells, respectively, in 25% ethanol at 25 degrees C for 10 min.

Acetates↗
PubMed1985

Properties of ATPase activity in intact vegetative cells and sporulating cells of yeast Saccharomyces cerevisiae.

The properties of ATPase activity were examined in the intact cells of yeast. The activity was stimulated by Mg2+, Mn2+ and Co2+. The activity was inhibited by NaN3 and by high concentrations of NaF, NaVO3 and PCMB. Optimal pH for the activity was approximately 8. The maximum value of the activity was obtained in the cells at the early stationary phase and it decreased in 3 hr after transfer to sporulation medium.

Adenosine Triphosphatases↗
PubMed1985

[The role of hypertension in apical hypertrophy].

Recent advances in echocardiography have revealed that hypertension causes several types of cardiac hypertrophy. We classified hypertensive patients by type of cardiac hypertrophy, and evaluated left ventricular function and the severity of hypertension. The subjects consisted of 257 hypertensive patients, 13 patients with cardiac hypertrophy, and 95 normotensive controls. The hypertensives were classified in four groups: no hypertrophy, concentric hypertrophy, asymmetrical septal hypertrophy (ASH), and asymmetrical apical hypertrophy (AAH). The normotensive patients with cardiac hypertrophy included nine with ASH and four with AAH. Cardiac functions in these patients were determined by echocardiography, RI-angiocardiography and cardiac catheterization. The results were as follows: Among 257 hypertensive patients, the incidence of concentric hypertrophy, ASH, and AAH was 53%, 10%, and 4%, respectively. In patients with AAH and hypertension, the hypertensive blood pressure levels and hypertensive organ involvements were mild. The blood pressures of most of these patients fell to the normal range after admission. The cardiac index and left ventricular systolic function (FS, mVCF, and ejection fraction) were significantly higher in AAH with hypertension than in the other hypertensive groups or in the normotensive controls. The hypertensive patients showed lower E-F slopes and higher A/E ratios than the normotensive controls, as well as the normotensive patients with ASH or AAH. Cardiac function did not differ appreciably between normotensive AAH and hypertensive AAH. Left ventricular dimension and left ventricular end-diastolic volume index were larger in AAH with hypertension. The total peripheral resistance of the hypertensive AAH was significantly lower than that of the hypertensives with concentric hypertrophy (p less than 0.01), though it was higher than that of the normotensive AAH (p less than 0.01). It was, therefore, concluded that mild hypertension observed in patients with AAH may be the result of regulatory mechanisms in the hyperkinetic states.

Adolescent↗
PubMed1984

Estimation of rat thermoregulatory ability based on body temperature response to heat.

A new simple but general estimation method for survival time in a hot environment is presented in this study. Even in heat-tolerant rats showing a triphasic heat response, an accurate estimation of survival time (ST) is possible. Rat groups, which included some heat-tolerant individuals, were exposed to 42.5 degrees C, 40% rh. Colonic temperature (Tco) was measured continuously by copper-constantan thermocouple. The ST (Y) of male and female rats were expressed as a linear function of time (X) until the Tco of 42.5 degrees C was reached: Y = 0.976X + 30.6 and Y = 0.968X + 31.6, respectively. A Tco of 42.5 degrees C at rest was just below the maximum survivable body temperature and above the steady-state equilibrium Tco levels during the second phase of the triphasic heat-response curve. Heat-tolerant individuals showed lower equilibrium temperatures than heat-intolerant rats. All 140 rats survived the Tco of 42.5 degrees C and lived for more than 8 wk, thus enabling them to be used for future experiments on thermoregulation. The heat survivors were able to reproduce, and their genetically controlled offspring could be used for thermoregulatory experiments.

Acclimatization↗