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Biomedical subjects

S Ohta

Publications and source records attributed to S Ohta.

At least 829 records · Page 46Linked to original sources

Induction of hyaluronic acid synthetase by estrogen in the mouse skin.

Hyaluronic acid synthetase activity was measured in male mouse skin following the topical application of estradiol in vivo. The enzyme activity increased in parallel with the hyaluronic acid content of the skin, and showed a similar response in the skin of ovariectomized female mice. The increase in enzyme activity was reduced by the anti-estrogen agents, tamoxifene citrate and clomiphene citrate, which block competitively the binding of estrogen to the estrogen receptor. The increase in hyaluronic acid synthetase activity was also reduced by topical application of cycloheximide or by subcutaneous injection of actinomycin D. The results suggest that the stimulation of hyaluronic acid synthesis in mouse skin in response to estrogen treatment is mediated through estrogen receptors and involves the induction of the enzyme hyaluronic acid synthetase.

Animals↗

Methanol dehydrogenase of Methylomonas J: purification, crystallization, and some properties.

A methanol dehydrogenase [EC 1.1.99.8] was purified and crystallized from methanol-grown Methylomonas J (formerly Pseudomonas sp. J), an obligate methylotroph. Its molecular weight was estimated to be 135,000 by gel chromatography. The sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis revealed two bands and their molecular weights were approximately 60,000 and 10,000. The enzyme was relatively stable at pH 6 and 10, and was unstable at pH 8. The enzyme activity was lost at pH 4.0; however, the prosthetic group was not liberated from the enzyme. Its isoelectric point was pH 9.3. The visible-ultraviolet absorption, fluorescence, CD, and ESR spectra were measured. The amino acid composition was analyzed after separation of the two components. Primary alcohols and formaldehyde served as substrates. Phenazine methosulfate (PMS) was an effective electron acceptor of the enzyme in the presence of either NH4Cl or methylamine. The enzyme was inhibited partially by metal chelators and completely by Mn2+ and Co2+.

Alcohol Oxidoreductases↗

Two cytochromes c of Methylomonas J.

Two kinds of c-type cytochromes, cytochrome c-551 (I), and cytochrome c-551 (II), were highly purified and crystallized from cell-free extract of methanol-grown Methylomonas J (formerly Pseudomonas sp. J) and their physiochemical and biochemical properties were studied. Cytochrome c-551 (I) had an absorption peak at 409 nm in the oxidized form and peaks at 417, 523, 551 nm, and a shoulder at 532 nm in the reduced form. The millimolar extinction coefficient of the alpha-peak of the reduced form was 25.3. The isoelectric point was at pH 5.3 and its standard redox potential was 0.29 V at pH 7.0. The molecular weight was estimated to be 16,000. Cytochrome c-551 (II) had absorption maxima at 409 nm in the oxidized form, and at 416, 521, and 551 nm in the reduced form. The millimolar extinction coefficient of the alpha-peak of the reduced form was 22.4. The isoelectric point was at pH 4.3 and its standard redox form was 22.4. The isoelectric point was at pH 4.3 and its standard redox potential was 0.24 V at pH 7.0. The molecular weight was estimated to be 12,500. The two cytochromes were reduced by methanol dehydrogenase [EC 1.1.99.8] of this bacterium, and formaldehyde was detected as an oxidation product. Ammonium chloride was not essential for reduction of the cytochromes. No significant reduction of the cytochromes was observed by methylamine dehydrogenase isolated from methylamine-grown cells or by 2,6-dichlorophenol-indophenol (DCPIP)-dependent aldehyde dehydrogenase of the methanol-grown cells. The reduced forms of the cytochromes were oxidized by blue copper protein of the methanol-grown cells.

Alcohol Oxidoreductases↗

Circumscribed myxoedema of lichen myxoedematosus as a sign of faulty formation of the proteoglycan macromolecule.

Using a new method devised by our laboratory, the ultrastructure of dermal glycosaminoglycan in an involved area of lichen myxoedematosus was examined. Although histochemical and biochemical studies have indicated simply an accumulated deposition of hyaluronic acid in the lesion, the glycosaminoglycan ultrastructure within it was distinctly different from that in normal skin. The glycosaminoglycan structure of normal skin was similar to the proteoglycan aggregate model described by Rosenberg (1975). As confirmed by the enzymatic digestion procedure, it represents the ultrastructure of hyaluronic acid bound to glycosaminoglycans such as dermatan sulphate or chondroitin sulphate. In contrast, hyaluronic acid filaments observed in lesions of lichen myxoedematosus contained no glycosaminoglycan subunits.

Adult↗

A modified technique of guinea pig testing to identify delayed hypersensitivity allergens.

A modified guinea pig testing technique was developed for the detection of weak allergens and allergenicity of materials unsuitable for testing by intradermal injection. This test involved the use of Freund's complete adjuvant to stimulate the immune system of the animal, and external application instead of intradermal injection of the test compound in the induction stage. The allergenicity of Sudan III, Brilliant Lake Red R and Sudan I was tested by this procedure. In the dose-effect study of Sudan I, the dose dependency of a positive reaction of the induction and challenge concentrations was recognized. The test was compared with three other guinea pig sensitization tests. The results obtained with this test correlated well with those obtained with the guinea pig maximization test.

Allergens↗

Galactosyltransferase activities in human sera of various diseases.

Recently galactosyltransferase has been noted as a new tumor marker. In this report galactosyltransferase activity was measured in sera of various diseases including malignant tumors and the levels of the enzyme activities were compared. The levels of the enzyme were higher in sera of malignant tumors and liver diseases than those in sera of other diseases. In malignant tumors little organ specificity was found with respect to the levels of galactosyltransferase activities. The levels of the enzyme were considered to have no relation to carcinoembryonic antigen or alpha-feto protein. On the other hand, in liver diseases the levels of the enzyme were correlated to the severity of liver damage. Therefore, it is reasonable to presume that the elevation of serum galactosyltransferase activity is caused chiefly by malignant tumors and liver damage.

Galactosyltransferases↗

[A simple method for enhancing interatrial right-to-left shunt in the diagnosis of small atrial septal defect using contrast echocardiography (author's transl)].

Commonly used contrast echocardiography is often inadequate to detect a small right-to-left (R--L) shunt in minimum atrial septal defect (ASD). We found that the diagnostic accuracy in ASD could be significantly improved by simply employing a Valsalva maneuver immediately followed by a Müller maneuver to enhance the R--L shunt. This procedure enabled contrast echo to be more clearly detected in both the left atrium and left ventricle, compared with conventional contrast echocardiography. The new technique was successful in diagnosing ASD in 5 patients with Qp/Qs ratio less than 2 determined by right heart cardiac catheterization. In conclusion, the diagnosis of a small ASD is greatly improved by simply employing this new procedure of a Valsalva-Müller maneuver during contrast echocardiography.

Adult↗

Intersubunit interactions in proton-translocating adenosine triphosphatase as revealed by hydrogen-exchange kinetics.

The rates of hydrogen-deuterium exchange in the peptide groups of the alpha and beta subunits and the alpha-beta subunit complex of proton-translocating adenosine triphosphatase from the thermophilic bacterium PS3 were examined. The exchange was found to be much slower in the isolated beta subunit than in the isolated alpha subunit. This has been taken as indicating that the structure of the beta subunit is tighter than that of the alpha subunit. Adenosine 5'-triphosphate (ATP) caused tightening of a relatively tight portion of the alpha subunit and of a relatively loose portion of the beta subunit. When the alpha and beta subunits are brought into contact, tightening of the alpha subunit, but not the beta subunit, occurs. The effect of ATP on the structure of the beta subunit is more pronounced in the presence of the alpha subunit than in its absence. These findings support the idea proposed previously that the alpha subunit has an allosteric site and the beta subunit a catalytic site and that the conformation of the beta subunit is controlled by the alpha subunit.

Adenosine Triphosphatases↗

The mechanism of estrogen-induced increase in hyaluronic acid biosynthesis, with special reference to estrogen receptor in the mouse skin.

The increase in hyaluronic acid and water contents induced by estradiol treatment in the mouse skin was dependent on dose and the number of treatments of estradiol. The anti-estrogen administered together with estradiol blocked the increase in hyaluronic acid content produced by treatment with estradiol alone. It was suggested that the anti-estrogen may act as an antagonist by competing for the cytoplasmic estrogen receptor on increase in hyaluronic acid synthesis. It was observed that the sensitivity in increase in hyaluronic acid biosynthesis by estradiol was related to the age of the mouse and the content of the cytoplasmic estrogen receptor in the mouse skin. It was suggested that there was a possible relationship between the increase in hyaluronic acid and cytoplasmic estrogen receptor in the mouse skin.

Animals↗

Neuronal depletion of cerebellum in late infantile metachromatic leukodystrophy.

The cerebellums of two children with late infantile metachromatic leukodystrophy were investigated by light and electron microscopy. These observations showed that the morphological abnormalities were comprised not only of severe demyelination and presence of metachromatic granules but also marked loss of Purkinje cells and granule cells. The depletion of Purkinje cells is considered to have resulted from demyelination of their myelinated axons. On the other hand, the reduction of granule cells is considered to be caused by the loss of Purkinje cells and presynaptic elements such as mossy fibers and glomerular collaterals of climbing fibers. The depletion of these neurons induced destruction of synaptic connection in the cerebellum.

Cerebellar Diseases↗

Nucleotide binding to isolated alpha and beta subunits of proton translocating adenosine triphosphatase studied with circular dichroism.

The catalytic and allosteric sites of proton translocating adenosine triphosphatase (ATPase) were studied by measuring the binding of nucleotides to the ATPase, and its alpha and beta subunits purified from thermophilic bacterium PS3, with a circular dichroic spectrometer. In contrast to mesophilic ATPases, this thermophilic enzmye contained no tightly bound nucleotides, and its subunits were stable after their purification. These properties were advantageous for analyzing both catalytic and allosteric sites. The former site showed rapid and loose binding, but the latter slow (t 1/2 = 1 h, for ADP) and tight binding. When a nucleotide was bound, the beta subunits showed a negative ellipticity at 275 nm corresponding to a tyrosyl residue, while the alpha subunits showed an ellipticity change corresponding to the absorption curve of the bound nucleotide. This difference enabled us to distinguish the binding sites in ATPase. At a low concentration, ADP selectively bound to alpha subunits in the ATPase, while at a high concentration, it bound to both subunits. This finding suggests that the tight binding sites are located in the alpha subunits. Although ADP and ATP bound to both the purified alpha and beta subunits, CTP did not bind to beta but only to alpha subunits, and ITP bound to beta but hardly to alpha. These nucleotide specificities also supported the idea that the catalytic sites are located in the beta subunits and the allosteric sites are located in the alpha subunits.

Adenosine Diphosphate↗