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T Hosoya

Publications and source records attributed to T Hosoya.

At least 253 records · Page 14Linked to original sources
PubMed1987

Proton and iodine-127 nuclear magnetic resonance studies on the binding of iodide by lactoperoxidase.

Interaction of an iodide ion with lactoperoxidase was studied by the use of 1H NMR, 127I NMR, and optical difference spectrum techniques. 1H NMR spectra demonstrated that a major broad hyperfine-shifted signal at about 60 ppm, which is ascribed to the heme peripheral methyl protons, was shifted toward high field by adding KI, indicating the binding of iodide to the active site of the enzyme; the dissociation constant was estimated to be 38 mM at pH 6.1. The binding was further detected by 127I NMR, showing no competition with cyanide. Both 1H NMR and 127I NMR revealed that the binding of iodide to the enzyme is facilitated by the protonation of an ionizable group with a pKa value of 6.0-6.8, which is presumably the distal histidyl residue. Optical difference spectra showed that the binding of an aromatic donor molecule to the enzyme is slightly but distinctly affected by adding KI. On the basis of these results, it was suggested that an iodide ion binds to lactoperoxidase outside the heme crevice but at the position close enough to interact with the distal histidyl residue which possibly mediates electron transport in the iodide oxidation reaction.

Horseradish Peroxidase↗
PubMed1987

In vivo effects of dexamethasone and cycloheximide on the phosphorylation of 110-kDa proteins and the protein kinase activities of rat liver nucleoli.

In vivo effects of dexamethasone and cycloheximide on the phosphorylation of nucleolar proteins and the activities of nucleolar protein kinases of rat livers were studied. Phosphorylation of nucleolar proteins was accomplished by incubation of isolated nucleoli with [gamma-32P]ATP at 37 degrees C for 10 min followed by electrophoretic separation and autoradiographic demonstration of phosphorylated proteins. Of several nucleolar phosphoproteins observed in the liver of adrenalectomized rats, the incorporation of 32P into nucleolar 110-kDa proteins was rapidly increased, reaching about 1.8-fold of the adrenalectomized level at 12 h after administration of dexamethasone. In addition, the activities of liver nucleolar protein kinases which were measured using casein or phosvitin as substrates were stimulated similarly by the administration of dexamethasone. It was found by DEAE-Sephadex column chromatography that nucleolar protein kinases were separated into NI and NII, the activities being increased about 35 and 80% by dexamethasone treatment (12 h), respectively, and that the enhancements were completely abolished by 1-h treatment of cycloheximide. The injection of cycloheximide also suppressed the hormone-induced enhancement of 32P incorporation into 110-kDa proteins but did not reduce the amount of the proteins at least until 2 h. Evidence was obtained indicating that the phosphorylation of the nucleolar 110-kDa proteins is accomplished primarily by protein kinase NII. These results suggest that continuous synthesis of protein(s) is necessary for glucocorticoid-induced enhancement of phosphorylation of 110-kDa proteins of liver nucleoli and that the nucleolar protein kinase NII is involved in the glucocorticoid-induced short-lived protein(s).

Adrenal Glands↗
PubMed1987

Proton nuclear magnetic resonance studies on the iodide binding by horseradish peroxidase.

Binding of an iodide ion to horseradish peroxidase was studied by following the hyperfine-shifted proton nuclear magnetic resonance signals of the enzyme. For the enzyme in an iodide-free solution, the spectra of hyperfine-shifted methyl region were only slightly affected by varying pH. In the presence of iodide (200 mM), however, both chemical shifts and line widths of the heme peripheral 1- and 8-methyl proton signals were markedly affected by the pH change from 7 to 4 and broadened at pH 4. From the change in peak heights of these signals at various concentrations of iodide, the dissociation constant of the iodide to the enzyme was calculated to be about 100 mM at pH 4.0. The peak derived from the proximal histidyl imidazole N epsilon-H proton was not perturbed by the addition of 200 mM iodide at pH 4.0 and 7.1. The rate of oxidation of iodide with hydrogen peroxide catalyzed by the enzyme was increased with decreasing pH, indicating the participation of an ionizable group with the pKa value of 4.0. Optical difference spectrum studies showed that iodide exerts no effect both at pH 4.0 and 7.4 on the binding affinity of resorcinol which is associated with the enzyme in the vicinity of the heme peripheral 8-CH3 group. These results suggest that an iodide ion binds to the enzyme at almost equal distance from the heme peripheral 1- and 8-methyl groups at the distal side of the heme and that the interaction becomes stronger in acidic medium with protonation of the ionizable group with the pKa value of 4.0.

Horseradish Peroxidase↗
PubMed1987

Relationship between RNA polymerase I activity and ornithine decarboxylase in rat liver tissues.

In order to examine the relationship between RNA polymerase I and ornithine decarboxylase (ODC), three lines of experiments were performed, with the following results. The glucocorticoid-induced increase of RNA polymerase I in rat liver nuclei was not abolished by administration of inhibitors of ODC synthesis and activity, namely 1,3-diaminopropane and 2-difluoromethylornithine respectively. Anti-ODC antibody did not cross-react with RNA polymerase I solubilized from rat liver nucleoli, indicating the absence of a common protein sequence in these enzymes. The ODC preparation which was treated with transglutaminase in the presence of putrescine could not stimulate the activity of RNA polymerase I in nuclei of liver and prostate. All these results suggest that the increases in ODC protein or activity are not a prerequisite to the increase in RNA polymerase I after hormonal or physiological stimuli, but rather that the increases in both enzymes are separate responses to the primary stimuli.

Animals↗
PubMed1987

Effects of glucocorticoids and cycloheximide upon transcription of ribosomal RNA gene in rat liver: application of quantitative S1 nuclease mapping analysis.

The change in the amount of pre-rRNA synthesized in rat liver was examined by S1 nuclease mapping analysis using the probe containing the 5' end of the rat 45S rRNA gene. It was found that the amount of pre-rRNA of adrenalectomized rats, which was about half of the normal level, was increased after administration of dexamethasone attaining to the normal level at 12 h, and that the hormone-induced pre-rRNA synthesis was completely abolished within 1 h after injection of cycloheximide. These results indicate that the glucocorticoids-stimulated initiation of rRNA transcription is mediated by the hormone-dependent short-lived protein(s).

Animals↗
PubMed1987

Change in the amount of rRNA precursor synthesized in rat ventral prostates after administration of androgen and cycloheximide: application of quantitative S1 nuclease mapping.

The change in amount of pre-rRNA was observed by the use of total cellular RNA isolated from rat ventral prostates and quantitative S1 nuclease mapping. The amount of pre-rRNA from castrated rats was about 20% of that of normal rats, and increased to the normal level 24h after administration of testosterone. Cycloheximide caused a rapid decrease in the amount of pre-rRNA in normal and testosterone-treated castrated rats, to about 20% and 10% of that of normal rats, respectively, 2h after the administration of the drug. These results support the view that the androgen-stimulated initiation of the rRNA gene transcription is mediated by a hormone-dependent short-lived protein(s).

Animals↗
PubMed1987

[A case of malignant melanoma, axillary lymph node metastasis proved by fine needle aspiration biopsy cytology].

We report the case of a 57-year-old man with a malignant melanoma in the subunguinal region of his right thumb associated with the palpable axillary lymph nodes. Its metastasis was confirmed by fine needle aspiration biopsy cytology. Prognosis if a malignant melanoma is protean by its stage. Thus information regarding the axillary metastasis is mandatory for deciding the therapeutic plan. Fine needle aspiration biopsy cytology is a useful technique for diagnosing lymph node metastasis.

Axilla↗
PubMed1987

[Clinico-pathological study of atypical adenomas of the thyroid].

A total of five cases of atypical adenomas of the thyroid were clinicopathologically examined to clarify their malignant potential and incidence. These adenomas showed both structural and cellular atypia when compared with follicular adenoma of the usual type, A follow-up study after surgery revealed neither metastasis nor postoperative recurrence. Therefore it has been concluded that an atypical adenoma of the thyroid can be regarded as a benign neoplasm.

Adenoma↗
PubMed1986

Nuclear magnetic resonance studies on the spatial relationship of aromatic donor molecules to the heme iron of horseradish peroxidase.

The geometry of hydrogen donor molecules bound to horseradish peroxidase was investigated using nuclear magnetic resonance techniques. Between resorcinol and 2-methoxy-4-methylphenol which showed different optical difference spectra, little difference was observed in the orientation of the molecules bound to horseradish peroxidase: the minimal distances between the enzyme iron and the protons of the phenol rings are in the range of 8.4-11.0 A. This situation was not greatly different for the third compound studied in this paper, benzhydroxamic acid, providing evidence against the view that its side chain coordinates to the heme iron. Furthermore, it was found that transferred nuclear Overhauser effect for the signals of these compounds was observable only when the heme peripheral 8-methyl proton signal was irradiated. These results, together with a hypothetical model of the enzyme structure obtained by computer-aided simulation procedures, suggest that the binding of these donor molecules and competitive inhibitors occur in the vicinity of the heme peripheral 8-methyl group, with hydrophobic interactions probably with Tyr-185 and with hydrogen bond with adjacent amino acid residues such as Arg-183.

Cresols↗
PubMed1986

Effects of glucocorticoid and cycloheximide on the activity and amount of RNA polymerase I in nuclei of rat liver.

The activity of the template-engaged form of RNA polymerase I from livers of adrenalectomized rats was about 50-60% of that of normal control rats, and increased about 2-fold at 6 h after the administration of dexamethasone. However, no change was found in the activity of the 'free' form of RNA polymerase I or the template-engaged form of RNA polymerase II. Immunochemical studies using guinea-pig anti-(RNA polymerase I) serum disclosed that the total number of RNA polymerase I molecules did not vary during the treatment with dexamethasone. Cycloheximide caused a rapid decrease in the template-engaged form of RNA polymerase I activity in normal rats and in dexamethasone-treated (6 h) adrenalectomized rats, to the value in adrenalectomized rats, but affected it only slightly in adrenalectomized rats. The elongation rate of rRNA-precursor synthesis in liver nuclei was not affected by a change in the concentration of circulating dexamethasone. From these results, it is concluded that about half the rRNA-precursor synthesis in rat liver is regulated by glucocorticoids, probably through the synthesis of short-lived protein(s) which may play a role in conversion of the 'dormant' form of RNA polymerase I into the 'engaged' form.

Adrenalectomy↗
PubMed1986

[Peroxidase activity of thyroid tissue in toxic diffuse goiter. Difference among thyroids administered antithyroid drugs and potassium iodide].

Peroxidase activity in thyroid tissue from 25 patients with Graves' disease was measured by Mini assay method (J. Biochem. 98, 637-647, 1985) employing guaiacol or iodide as a second substrate. The mean values of protein-based specific activity were 0.496 guaiacol unit/mg protein and 0.187 iodide unit/mg protein, reaching 16 fold and 28 fold those of normal thyroids, respectively. The mean value of ratio of iodide unit to guaiacol unit in each thyroid, 0.68, was also much higher than that of normal human thyroid, 0.16. No significant difference in peroxidase activity was observed between patients treated with methylmercaptoimidazole and those with propylthiouracil, but the activities of those groups were significantly higher than those of patients treated with potassium iodide, suggesting that inorganic iodine therapy plays some role in suppressing the synthesis of thyroid peroxidase in vivo.

Adolescent↗
PubMed1986

Fat in the normal cavernous sinus.

The cavernous sinuses of 17 patients examined by metrizamide CT cisternography with 1.5 mm-thick slices were reviewed. Most of the cavernous sinuses contained fatty components. Anterior lateral end and posterior end of the cavernous sinus are the common sites where the fat is visible.

Adipose Tissue↗
PubMed1986

Effect of androgen on ornithine decarboxylase activity in androgen-dependent mouse mammary tumor (Shionogi Carcinoma 115) and its androgen-independent subline (CS 2).

The activity of ornithine decarboxylase in androgen-dependent mouse mammary tumor (Shionogi Carcinoma 115) was reduced to 25% by castration of tumor-bearing mice and restored to the normal level 12 h after administration of testosterone or 5 alpha-dihydrotestosterone. Administration of estradiol-17 beta to the tumor-bearing castrated mice also stimulated the enzyme activity while progesterone and cortisol had little effect. On the other hand, the enzyme activity was affected by neither castration nor androgen injection to CS 2, which is a subline of SC 115 and completely independent of androgen for growth. The inhibition of ornithine decarboxylase activity in SC 115 by injecting alpha-difluoromethylornithine did not affect the enhancement of RNA polymerase I activity by androgen, showing independent elevation of the levels of the two enzymes by androgen.

Androgens↗
PubMed1986

Autoantibodies to thyroid peroxidase in patients with chronic thyroiditis: effect of antibody binding on enzyme activities.

Using thyroid peroxidase (TPO), which was purified from the thyroid of patients with Graves' disease, we attempted to determine whether sera from patients with chronic thyroiditis contained antibodies to the enzyme. When the binding was tested by ELISA, sera from patients with chronic thyroiditis revealed high binding activities to TPO. When TPO was incubated with IgG from sera followed by treatment with protein A-Sepharose and centrifugation, the remaining TPO activities in the supernatant fraction were lower in most of the patients, as compared to normal controls. Moreover, IgG purified by DEAE-cellulose chromatography from sera in patients interfered with the TPO activities. Titres of anti-TPO antibodies correlated well with those of anti-microsome antibodies. These results indicate the presence of autoantibodies to TPO in sera of most patients with chronic thyroiditis and that TPO may be one component of microsome antigen complexes recognized by the autoantibodies. Studies on the inhibition of TPO by IgG isolated from sera of patients using guaiacol and iodide assays revealed that at least three epitopes of TPO molecule were recognized by autoantibodies and that the antigenic determinants on TPO molecule recognized by autoantibodies could be heterogeneous in patients.

Adolescent↗