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Biomedical subjects

F Watanabe

Publications and source records attributed to F Watanabe.

At least 199 records · Page 11Linked to original sources
PubMed1989

Flow cytometric analysis of glucocorticoid receptor using monoclonal antibody and fluoresceinated ligand probes.

Conditions were established for single cell analysis of glucocorticoid receptor (GR) content by flow cytometry using several clones of a human leukemic cell line (CCRF-CEM). These included CEM-7A, 7R, C1, and ICR 27 Tk.3 cells which were examined both by standard [3H]dexamethasone radiometric binding and by two independent flow cytometry assays. The latter involved either mouse monoclonal antibody against GR (GR-MoAb) or fluoresceinated cortisol ligand probes. For CEM-7A, 7R, and C1 cells, there was a correlation between GR-MoAb and radiometrically defined GR values. However, clone ICR-27 Tk.3 with low [3H]dexamethasone binding exhibited the highest GR-MoAb fluorescence. The fluoresceinated cortisol assay correlated with dexamethasone binding values in all four clones. Thus, GR-MoAb identifies the total immunologically reactive GR present, while the fluoresceinated cortisol assay quantifies only the functionally intact GR in terms of its initial binding. Their combined use may reveal the cellular heterogeneity of GR expression and function also in human tumor samples, to which they have been successfully applied. When coupled with DNA counterstaining, GR expression can be related directly to frequently DNA-aneuploid tumor cells and cell cycle distribution.

Antibodies, Monoclonal↗
PubMed1989

Interaction between bacteriophage T4 coded gene 32 protein and poly(rA).

The cooperative binding of T4 gene 32 protein with polynucleotides, of which the quantitative aspects in the literature have not satisfied the requirements of thermodynamics, is studied by adopting a modified formula of the lattice theory. A moderate value is found for the cooperativity parameter (q approximately 200 at 0.2 M NaCl), which is weakly dependent on salt concentration. The cation effect on the binding suggests that the shielding of negative charges of the protein or a loose cation bridge between the bound protein molecules plays a role in the cooperative binding process.

In Vitro Techniques↗
PubMed1989

Regional effects of repetition time on NMR quantitation of water in normal and edematous lungs.

It is well known that pulmonary edema is, in general, spatially nonuniform. Since the NMR spin-lattice relaxation time (T1) is increased by lung edema, the spatial distribution of T1 will be nonuniform. When the repetition time (TR) is short relative to the T1 of edematous lung, lung water content will be underestimated and this underestimation will be spatially nonuniform as well. Therefore, technical artifacts which are a complex function of lung edema and its spatial distribution are expected. We compared overall and regional (topographic) lung water density measurements obtained from living rats (with normal or edematous lungs) using repetition times of 2.0 and 6.2 s (at a magnetic field of 1 T), to quantify this uneven T1 effect for normal and edematous lungs. NMR measurements at TR = 2.0 s underestimated whole lung water density (-rho H2O) TR = 6.2 s) by an average of 7.2% in normal rats and 22.5% in rats with pulmonary edema. Regional -rho H2O underestimation (%delta-rho H2O) varied from 2.2 to 8.8% (groups means) in normal lungs and from 7.3 to 30.8% in edematous lungs. As a result, the interquartile range (of the voxel distribution as a function of rho H2O) underestimated the spatial nonuniformity of lung water density by 28.0% in edematous lungs, likely because of greater loss of NMR signal from high-water-density, long-T1 lung regions. Both %delta-rho H2O and T1 were significantly correlated with -rho H2O at TR = 6.2 s.(ABSTRACT TRUNCATED AT 250 WORDS)

Body Water↗
PubMed1989

Factor XIIIa-containing cells and fibrosis in oral and maxillofacial lesions: an immunohistochemical study.

The distribution of subunit A of blood coagulation factor XIII (FXIIIa) was investigated by the avidin-biotin-peroxidase complex (ABC) method in various oral and maxillofacial tissues. These tissues were from normal tongue, gingiva, lip, and submandibular gland, and from Dilantin gingival hyperplasia (one case), pyogenic granuloma (three cases), peripheral fibroma (four cases), squamous cell carcinoma (seven cases), chronic sclerosing submandibular adenitis (two cases), and fibrous dysplasia of the mandibular bone (one case). The distribution of collagenous components was examined in the same tissues by means of the Sirius red F3BA method. By means of the ABC method, FXIIIa was detected in the cytoplasm of certain connective tissue cells in each of the tissues examined. These FXIIIa-containing cells were sparse in the normal tissues but evidently abundant in the fibrous connective tissue of inflammatory and neoplastic lesions. In the present study, the close relationship between the distribution of FXIIIa-containing cells and that of collagenous components is demonstrated. The role that FXIIIa-containing cells play in the process of fibrosis is discussed.

Collagen↗
PubMed1989

Site-specific mutagenesis of lysine-204, tyrosine-224, tyrosine-228, and histidine-307 of porcine kidney D-amino acid oxidase and the implications as to its catalytic function.

In order to evaluate the possible contributions of Lys-204, Tyr-224, Tyr-228, and His-307 in porcine kidney D-amino acid oxidase [EC 1.4.3.3] (DAO) to its catalytic function, we constructed four point mutant cDNAs encoding enzymes possessing Glu-204, Phe-224, Phe-228, and Leu-307 by oligonucleotide-directed in vitro mutagenesis. The four mutant cDNAs and the wild type cDNA could be expressed in vitro with similar efficiencies and about 200 ng of each enzyme protein was produced from 5 micrograms of the respective capped RNA. The electrophoretic mobilities of the in vitro synthesized mutant enzymes on SDS-polyacrylamide gel were almost identical with that of the wild type DAO, and the molecular weight was calculated to be 38,000. The Glu-204 and Phe-224 mutant DAOs showed comparable enzyme activities to that of the wild type enzyme, and were inhibited strongly by sodium benzoate, a potent competitive inhibitor of DAO. The kinetic parameters of the two mutant DAOs were also comparable to those of the wild type DAO. On the other hand, the Phe-228 and Leu-307 mutant DAOs showed no detectable activity. The results indicate that Tyr-228 and His-307 play important roles as to the constitution of the active site or participate in the reaction directly, while Lys-204 and Tyr-224 are not essential in the enzyme reaction.

Animals↗
PubMed1989

A mutagenic new iridoid in the water extract of catalpae fructus.

A mutagenic principle in the water extract from Catalpae Fructus (originated from Catalpa ovata G. DON) (Bignoniaceae) was isolated and characterized as a new iridoid named catalpin. The iridoid exhibited mutagenic activity towards Salmonella typhimurium strain TA100 in the presence and absence of rat liver homogenate (S9) mix in Ames' test.

Animals↗
PubMed1989

[Clinical use of castable ceramics (OCC) crown].

The new ceramics material (OCC) was developed for the purpose of aesthetic restration which can be fabricated by an usual lost wax process and it produces accurate marginal fit. OCC possesses mechanical, chemical, and physical properties. This article is discussed utility and security of OCC complete crown on the clinical application from Dec, 1986 to Jun, 1989. The results may be summarized as follows: 1) 2 of 69 cases were broken down in the period from Dec, 1986 to Jun, 1989. These reasons are caused by the lack of occlusal thickness and unsuitable fit of crown. 2) Although 4 of 69 cases are stained by tobacco tar and 5 of 69 cases are suffered from plaque accumulation, these staining and plaque accumulation are improved by plaque control. 3) An allergic reaction or an abnormality of oral tissues haven't been recognized during test period of clinical use. Consequently OCC crown showed itself about safety and usefulness.

Crowns↗
PubMed1989

[Establishment of a human skin squamous cell carcinoma cell line (SCC-1CB) in vitro and its characteristics].

A human skin tumor cell line SCC-1CB has been established in vitro. A specimen from a surgically excised tumor with moderately differentiated squamous cell carcinoma of skin from a Japanese male, aged 62, was transplanted subcutaneously into nude mice. After growth, a part of the tumor was passaged with a transplantation needle. At the fourth passage, part of the tumor was explanted into culture dishes. Cell cultures were maintained in Eagle's minimal essential medium supplemented with 10% fetal calf serum. A SCC cell line (SCC-1CB) was cloned and then established. Chromosome analysis revealed the human type with a mode number of 56. The cell line is tumorigenic in nude mice. Immunohistochemical staining revealed that keratin, cytokeratin and vimentin were positive and desmin was negative. It can be cryopreserved in liquid nitrogen. It has successfully maintained proliferation in continuous cell culture for over 2.5 years. The cells retain their original morphological appearance as determined by light and electron microscopy.

Animals↗
PubMed1989

[Intraoperative echocardiographic assessment of left ventricular muscle volume changes after intracardiac operation under cardiopulmonary bypass].

Cardiopulmonary bypass (CPB) and concomitant cardioplegic arrest (CA) may cause impairment of cardiac function with development of myocardial edema. There are few reports of intraoperative assessment of LV mass or muscle volume (MV) in patients, Fifteen patients (age; 0.5-68), 10 with congenital and 5 with valvular diseases, were studied by epicardial 2-dimensional and M-mode echocardiography. LVMV was obtained from M-mode study at pre- and post-CPB during surgery. Pre and post CPB LVMV-index (ml/m2) and percent change of LVMV were compared. CPB-time was 170 +/- 78 min (mean +/- SD), and CA time was 97 +/- 49 min. LVMV index increased significantly from 89 +/- 35 to 103 +/- 43 ml/m2 after CPB (p less than 0.01). Percent change ranging from -3 to +37% (14 +/- 12) correlated to CPB time (r = .81, p less than .01) and also to CA time (r = .62, p less than .05). These results indicated that CPB with CA caused acute increase in LVMV with a positive relation to its duration.

Adolescent↗
PubMed1989

[Activity of guanase in serum and liver function tests in HBe antigen-positive chronic hepatitis].

Forty-one patients with chronic hepatitis were divided into an HBe antigen-positive group (n = 13) and an HBe antigen-negative group (n = 28) to clarify the relationship between the presence of HBe antigen and liver function. In the HBe antigen-positive group, the activities of serum alanine aminotransferase (p less than 0.01), aspartate aminotransferase (p less than 0.05) and guanase (p less than 0.01) were significantly higher than those in the HBe antigen-negative group. The correlation coefficient between the HBe antigen titer and guanase activity was 0.528, which was higher than the corresponding values for alanine aminotransferase and aspartate aminotransferase. The determination of guanase activity in serum may be useful for evaluating the clinical severity of HBe antigen-positive chronic hepatitis.

Adolescent↗
PubMed1989

[Quantitative analysis of sulphur in the liquid of some dental cements].

The ceramometal restoration with porcelain margin have been clinically applied to provide accuracy and esthetics. But, we experienced the cases that the color of the labial marginal area of ceramometal restoration turned dark. Then, the crown was removed and observed. And it was found that the cement color had been changed black. As the result of the ultimate analysis of this cement by means of X ray microanalyzer, some elements; Al, P, S, Ca, Ni and Zn, were detected. As the inclusion of sulphur in commercially available cements was suspected the quantitative analysis of sulphur in the liquid of glass ionomer cement and polycarboxylate cement were done by means of inductively coupled plasma spectrometer. And following results were obtained that sulphur was included 5437 micrograms/ml in the liquid of glass ionomer cement, and 2147 micrograms/ml in the liquid of polycarboxylate cement, respectively. From the above, it is suspected that sulphur reacts on dental metal and causes tooth and gingival discoloration.

Dental Cements↗
PubMed1988

Effect of site-specific mutagenesis of tyrosine-55, methionine-110 and histidine-217 in porcine kidney D-amino acid oxidase on its catalytic function.

To assess the contributions of Tyr-55, Met-110 and His-217 in porcine kidney D-amino acid oxidase (EC 1.4.3.3, DAO) to its catalytic function, we constructed three mutant cDNAs coding for the enzymes possessing Phe-55, Leu-110 and Leu-217 by site-specific mutagenesis. The mutant and wild type cDNAs could be expressed in vitro with similar efficiency. The three mutant enzymes thus synthesized showed catalytic activities comparable to that of the wild type oxidase. It is concluded that Tyr-55, Met-110 and His-217 are not directly involved in the catalytic function.

Amino Acid Sequence↗
PubMed1988

Molecular cloning and sequence analysis of cDNA encoding human kidney D-amino acid oxidase.

cDNA clones encoding D-amino acid oxidase were isolated from a human kidney cDNA library by hybridization with cDNA for the pig enzyme. The cDNA insert of 2.0 kilobase pairs long provided coding information for a protein consisting of 347 amino acids. The molecular mass of the enzyme was calculated to be 39,410 Da. The amino acid sequence similarity between the pig and human enzymes is 84.4%, and among the active site residues proposed from chemical modification studies, methionine-110 of the pig enzyme was replaced by threonine. Northern blot analysis confirmed the expression of an mRNA of 2.0 kilobases encoding the D-amino acid oxidase in human kidney.

Amino Acid Sequence↗