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Biomedical subjects

M Ota

Publications and source records attributed to M Ota.

At least 271 records · Page 15Linked to original sources
PubMed1989

Effects of ryanodine and 9,21-didehydroryanodine on caffeine-induced contraction of rat and guinea pig aortae.

We compared the effects of ryanodine and 9,21-didehydroryanodine (DH-ryanodine), which are present in commercial preparations of 'ryanodine', on the contractions of rat and guinea pig aortae induced by 20 mM caffeine and tested the dependence of the action of each substance on external Ca2+. With the first protocol, the aortae were incubated with ryanodine or DH-ryanodine for 20 min in Ca2(+)-containing medium, and caffeine was added at 2 min incubation in Ca2(+)-free medium. With the second protocol, each substance was added when the external medium was changed to Ca2(+)-free medium, and 20 min later, caffeine was applied. Ryanodine and DH-ryanodine inhibited the caffeine-induced contractions in a similar way; i.e., with maximal effects at 3 microM and lesser effects at 10 microM. The potencies of inhibition by both substances were similar except that the effect of ryanodine at 1.5 microM was more potent than that of DH-ryanodine with the second protocol. The response by muscles previously loaded with Ca2+ to a second application of caffeine was more greatly inhibited by both compounds (use-dependent effect). The inhibition of the contraction due to the first or second application of caffeine was greater when either agent was applied in Ca2+-containing medium than in Ca2(+)-free medium. These results indicate that ryanodine and DH-ryanodine are similar in their effects on caffeine-induced Ca2+ release in vascular smooth muscle and that cellular Ca2+ levels may affect the action of ryanodine.

Alkaloids↗
PubMed1989

Intranuclear androgen-receptor complex binding sites of mouse submandibular gland.

Nuclear androgen-receptor binding sites in chromatin regions of mouse submandibular gland were studied. Cytosol androgen receptors prelabeled with [3H]androgen interacted with the crude nuclei in mouse submandibular gland and activation of the receptor was a prerequisite for this interaction. After in vivo administration of [3H]androgen to female mice, radioactivities were found in the nuclei purified from submandibular gland tissues and the [3H]androgen-labeled purified nuclei were further digested with micrococcal nuclease. The androgen receptor was found in solubilized, active chromatin fractions which contained mono- and dinucleosomes. By an in vitro exchange assay, endogenous androgen-receptor complexes associated with chromatin binding sites in intact males were found in the solubilized fraction after micrococcal nuclear digestion, whereas such complexes were not found in females. These results suggest that the androgen receptors translocated to the nuclei and became associated with chromatin and that this association occurred in transcriptionally active chromatin regions that were preferentially sensitive to micrococcal nuclease.

Animals↗
PubMed1989

[Normalizing method for determining the mode of crack-propagation in amalgams and composite resins].

A normalizing method for determining the mode of crack-propagation in a phase or component of a dental amalgam or composite resin was devised. The results calculated using the method were found to be satisfactory when the method was applied to the materials. Furthermore, details of the mode of crack-propagation in the materials which cannot be seen using pictures of the cross-section structure of the materials are revealed using the method.

Composite Resins↗
PubMed1989

Flow cytometric analysis of the Rh1 (Rho, D) antigen activity on red cells: various Rh blood group phenotypes including Du variants.

Rh1 (Rho, D) antigen activity has been analyzed by the use of the indirect immunofluorescence flow cytometry (FCM), and the Rh blood group genotypes were able to be successfully determined from the intensity of fluorescence detected in flow cytometry using the anti-D IgG that was fractionated in a Protein A Sepharose CL-4B column as the primary antibody. The relative amount of the fluorescein isothiocyanate (FITC) bound to the D (R1R1, CDe/CDe), the high grade Du (R2r',cDE/Cde), the low grade Du (K1r, CDue/cde), and the d (rr, cde/cde) red cells was estimated from the mean fluorescent intensity. The FITC-binding activity of the high grade Du and low grade Du was 83% and 21% that of D. The antigen-antibody complex density profile was analyzed by using the FITC-conjugated protein-A in place of the second antibody. Compared with the found results using anti-human globulin as the second antibody, this method was less sensitive but it still was able to demonstrate the different degrees of fluorescence according to the Rh genotypes. The present FCM method is both simple and useful for (1) measuring the relative amount of antigens, (2) for detecting the dosage effect and (3) for deferminins the blood group genotypes.

Erythrocytes↗
PubMed1989

[AE characteristics during compression test of high-copper amalgams and visible light-cured composite resins].

Differences in environmental strength of high-copper amalgams and photo-cured composite resins for posterior after storage in deionized water at 37 degrees C for 1 year were evaluated by examining compressive strength and acoustic emission (AE) characteristics. The results were as follows: Microfailure in amalgams and composite resins was detected from the onset of stress and their microfailures toward catastrophic failure occurred at 70 to 90% of their ultimate strength. One-year compressive strength for amalgam decreased to 80 to 98% of its 24-hour compressive strength and that for composite resin to 65 to 75%.

Composite Resins↗
PubMed1989

Characterization of lymphocytes in a patient with Wiskott-Aldrich syndrome: studies by fluorescence polarization.

Lymphocytes from a patient with Wiskott-Aldrich syndrome (WAS) were employed for a study of the intracellular viscosity and fluorescein permeability through the cell membrane by a fluorescence polarization spectrofluorometer, which was designed to calculate polarization value and permeable fluorescein intensity automatically. Fluorescein diacetate (FDA) was used as the indicator probe. The fluorogenic substrate is taken up by viable cells and converted to a fluorescent molecule, fluorescein, by intracellular esterase, where upon the fluorescein easily effluxes through the cell membrane. The response to stimulation with phytohemagglutinin (PHA) for 45 min led to a decreased polarization value (p-value) as compared to lymphocytes of healthy donors, and the fluorescein efflux through the cell membrane was greater than that of healthy donors. Fluorescein efflux from lymphocytes in the patient during 48 and 72 h incubation with or without PHA was markedly increased. In healthy donors, the degree of fluorescein permeability was not increased during the culture. These results indicate that intracellular viscosity of lymphocytes is altered in initial mitogenic stimulation, but that there was some abnormality in the fluorescein permeability properties through the cell membrane of lymphocytes in a patient with WAS.

Adolescent↗
PubMed1988

Nonactivated and activated glucocorticoid receptor complexes from human salivary gland adenocarcinoma cell line.

[3H]Triamcinolone acetonide glucocorticoid receptor complexes from human salivary gland adenocarcinoma cells (HSG cells) were shown to be activated with an accompanying decrease in molecular weight in intact cells, as analyzed by gel filtration, DEAE chromatography, the mini-column method and glycerol gradient centrifugation. Glucocorticoid receptor complexes consist of steroid-binding protein (or glucocorticoid receptor) and non-steroid-binding factors such as the heat-shock protein of molecular weight 90,000. To determine whether the steroid-binding protein decreases in molecular weight upon activation, affinity labeling of glucocorticoid receptor in intact cells by incubation with [3H]dexamethasone 21-mesylate, which forms a covalent complex with glucocorticoid receptor, was performed. Analysis by gel filtration and a mini-column method indicated that [3H]dexamethasone 21-mesylate-labeled receptor complexes can be activated under culture conditions at 37 degrees C. SDS-polyacrylamide gel electrophoresis of [3H]dexamethasone 21-mesylate-labeled steroid-binding protein resolved only one specific 92 kDa form. Furthermore, only one specific band at 92 kDa was detected in the nuclear fraction which was extracted from the cells incubated at 37 degrees C. These results suggest that there is no change in the molecular weight of steroid-binding protein of HSG cell glucocorticoid receptor complexes upon activation and that the molecular weight of nuclear-binding receptor does not change, although the molecular weight of activated glucocorticoid receptor complexes does decrease. Triamcinolone acetonide induced an inhibitory effect on DNA synthesis in HSG cells. Dexamethasone 21-mesylate exerted no such effect and blocked the action of triamcinolone acetonide on DNA synthesis. These results suggests that dexamethasone 21-mesylate acts as antagonist of glucocorticoid in HSG cells. The fact that dexamethasone 21-mesylate-labeled receptor complexes could be activated and could bind to DNA or nuclei as well as triamcinolone acetonide-labeled complexes suggests that dexamethasone 21-mesylate-labeled complexes can not induce specific gene expression after their binding to DNA.

Adenocarcinoma↗
PubMed1988

Detection of ABO blood group-active glycolipids extracted from red cell membrane and heat hematoma using TLC-immunostaining.

Glycolipids extracted from groups A, B, and O erythrocytes were developed on thin-layer plates; their ABO blood group antigenicities were detected by immunostaining method using avidin-biotin-complex (ABC). Among series of glycolipids of different flow rates, antigen-specific staining was observed in five bands from group A1 erythrocytes, four bands from group B, and two bands from group O. Monoclonal anti-A, -B, and -H antibodies specifically stained glycolipids from A1, B, and O erythrocytes, respectively. ABO blood grouping was possible from 5 g of epidural heat hematoma of a charred body by this method. ABC immunostaining on thin-layer chromatography is a useful and reliable method for ABO blood grouping in forensic practice.

ABO Blood-Group System↗
PubMed1988

Characterization of the nontransformed and transformed androgen receptor and heat shock protein 90 with high-performance hydrophobic-interaction chromatography.

The hydrophobicity of the nontransformed and transformed androgen receptor from rat submandibular gland and heat shock protein 90 (hsp90) from rat submandibular gland and liver was characterized by using high-performance hydrophobic-interaction chromatography on TSK gel Ether-5PW. In the absence of molybdate, cytosol [3H]R1881-androgen receptor complexes were mainly eluted in the 1.3 M region (Peak 1) with a small peak in the 0.8 M region (Peak 2) of a descending salt gradient (2 to 0 M) of ammonium sulfate. In the presence of molybdate, Peak 2 was predominant. When labeled-cytosol was applied after being heated at 25 degrees C for 30 min, a third peak (Peak 3) at around 0.64 M ammonium sulfate was newly observed. Peaks 2 and 3 were observed, while Peak 1 completely disappeared with the labeled-cytosol precipitated at 40% saturated ammonium sulfate. The Stokes radius of Peak 1 was 7 nm, and of Peak 2 was 8 nm. Both peaks were retained poorly by DNA-cellulose but bound rather well to DEAE-cellulose. These results suggest that these two peaks represent the nontransformed receptor, indicating that there are isoforms of the nontransformed androgen receptor which are distinguished by their hydrophobic properties and Stokes radii. Peak 3 had a Stokes radius of 5 nm and preferentially bound to DNA-cellulose, suggesting that this peak corresponds to the transformed receptor. These results indicated that the transformation of the androgen receptor accompanies the enrichment of the hydrophobicity of the receptor molecule. Hsp90 purified from rat livers and hsp90 in the cytosol both from livers and submandibular glands were eluted from Ether-5PW at 0.8 M ammonium sulfate, at almost the same position as Peak 2. This finding suggests that the enrichment of hydrophobicity on transformation is due to dissociation of hsp90 from the nontransformed androgen receptor.

Animals↗
PubMed1988

Ribonucleic acid association with androgen receptor from rat submandibular gland.

The transformed androgen receptor from rat submandibular gland converts to a faster sedimenting form (6-8S) on a glycerol gradient centrifugation after withdrawal of a transformation-inducing reagent (KCl or ATP). In this report, the association of cytosolic RNA with the transformed androgen receptor was investigated as a possible mechanism of molecular conversion of the androgen receptor. When the transformed and converted androgen receptors were treated with RNase A, these receptors sedimented at 4.5S in a low-salt glycerol gradient. Addition of RNA from rat submandibular gland to the RNase-Sepharose-treated transformed receptor caused a shift of receptor peak from 4.5S to 5.8S. RNA from rat submandibular gland, yeast RNA and E. coli rRNA inhibited DNA-cellulose binding of a RNase-treated transformed receptor in the absence of molybdate. These observations suggest that conversion from the transformed 4S androgen receptor to a 6-8S form resulted from the association of RNA(s) with the transformed receptor.

Animals↗
PubMed1988

Interaction of glucocorticoid receptor from rat liver with protamine and arginine.

The nontransformed glucocorticoid receptor (GR) from rat liver was found to bind to protamine-Sepharose and could be recovered by a salt gradient without a change in molecular configuration. The nontransformed GR also bound to arginine-Sepharose, but the transformed GR did not bind to either resin. Ligand-free GR interacted with both resins and was eluted without loss of its steroid binding ability. The bindings of GR to protamine- and arginine-Sepharose were saturable. The apparent dissociation constants of GR on protamine-Sepharose varied from 0.34 nM (-molybdate) to 0.68 nM (+ 10 mM molybdate) and those on arginine-Sepharose were 1.99 nM (-molybdate) and 0.65 nM (+ 10mM molybdate), respectively. The maximum binding capacity was achieved by arginine-Sepharose in the absence of molybdate. Higher salt concentrations (0.5 M NaCl) were required to elute GR from protamine-Sepharose than from arginine-Sepharose (approx 0.03 M NaCl). However, the effectiveness of several salts for the elution of GR was consistent in both resins as follows; MgCl2 = CaCl2 = Na2WO4 greater than (NH4)2SO4 = Na2MoO4 greater than arginine-HCl greater than lysine-HCl greater than KCI = NaCl. These results suggest that GR interacts with arginine residues in protamine. Chromatography using these resins resulted in 7-10-fold purification of occupied and unoccupied nontransformed GRs.

Animals↗
PubMed1988

Stability and transformation of the glucocorticoid receptor under acidic conditions.

The [3H]triamcinolone acetonide ([3H]TA)-binding ability of the rat liver glucocorticoid receptor (GR) was investigated under acidic conditions, ranging from pH 2 to 7.3. Both in the presence and absence of 10 mM molybdate, the [3H]TA-binding ability decreased below pH 6.5 and was almost completely lost below pH 5, pH 5.9 +/- 0.1 giving 50% [3H]TA-binding. The binding ability was recovered when the pH of the cytosol was reversed to 7.3 or the precipitate obtained on acidification was dissolved in a buffer of pH 7.3. Moreover, in the absence of molybdate, the [3H]TA-GR complexes formed at pH 7.3 remained unchanged until pH 5. Then they decreased, pH 3.9 +/- 0.1 giving 50% binding, and completely disappeared at pH 3. [3H]TA-binding activity recovered from the precipitate also decreased in a similar pH region (a 50% decrease in binding being observed at pH 4.2 +/- 0.04). These results suggest that rat liver GR is rather resistant under acidic conditions and that it exists in a peculiar state below pH 5.9 to approximately 4 as to its ligand binding property: unoccupied GR has no [3H]TA-binding ability but [3H]TA-GR complexes once formed at neutral pH do not dissociate. [3H]TA-GR complexes recovered from the precipitate at pH 5 had a Stokes radius of 7.5 nm, little DNA-cellulose-binding ability and sedimented at 8.6S on glycerol gradient centrifugation, indicating that the receptor existed in a nontransformed state. In addition, both occupied and unoccupied GR were transformed at about pH 4, their being 50% transformation. This transformation was accompanied by irreversible denaturation of the receptor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Effect of vasopressin on atrial natriuretic peptide release and renal function in dogs.

To assess the effect of arginine vasopressin (AVP) and 1-deamino-8-D-AVP (DDAVP) on atrial natriuretic peptide (ANP) release, renal water and electrolyte excretion, and cardiovascular function, AVP and DDAVP were administered at a dose of 10 ng.kg-1.min-1 iv for 30 min into anesthetized dogs receiving saline infusion at a rate of 1 ml.kg-1.min-1 (n = 12). In the control study, saline was infused alone (n = 6). AVP potentiated the plasma ANP response to an increase in plasma volume produced by saline infusion, increased mean arterial blood pressure (MAP), and exaggerated the natriuresis and kaliuresis. DDAVP did not potentiate the increase in plasma ANP but enhanced the natriuresis without any rise in MAP. Saline alone increased plasma ANP as well as sodium and potassium excretion with no changes in MAP. Inulin and p-aminohippuric acid clearances did not change during these studies. The results suggest that in hydrated dogs, AVP may increase ANP release and arterial blood pressure via the vasopressor activity of AVP and potentiate the natriuresis and kaliuresis, but the increased ANP may play little role in the natriuresis.

Animals↗
PubMed1988

Glucocorticoid regulates secretion of epidermal growth factor in the human salivary gland adenocarcinoma cell line.

The treatment of a human submandibular gland adenocarcinoma cell line (HSG cell line) for 48 h with triamcinolone acetonide (TA; 1-100 nmol/l) reduced the secretion of epidermal growth factor (EGF) in a closely related manner to a maximum of 66%. The reduction in the level of EGF secreted resulted in the suppression of DNA synthesis in the HSG cells to a similar extent. When the cells were incubated with TA and exogenous human EGF (hEGF), DNA synthesis was 1.7-fold higher than that without added hEGF. The removal of EGF by the addition of hEGF antibody reduced DNA synthesis in HSG cell cultures to the same extent as did TA. These results suggest that the growth inhibition of HSG cells by TA is due to the reduction in the amount of EGF secreted.

Adenocarcinoma↗