Quantitation of 4-hydroxynonenal protein adducts.
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Acetylcysteine↗
Biomedical subjects
K Uchida
Publications and source records attributed to K Uchida.
A novel model of cutaneous candidiasis produced in prednisolone-treated guinea-pigs.
In an attempt to develop an animal model of cutaneous candidiasis useful for the pre-clinical evaluation of antifungal drugs, experimental cutaneous Candida albicans infections were produced in mature and immature guinea-pigs treated with prednisolone. The morbidity of this model in terms of the extent and the duration of the superficial infection was compared with that of two other reported models: those produced by alloxan treatment, or the use of occlusive dressings. Infected animals were also included which received neither of these treatments. The Candida infection continued steadily for 14 days or more in both mature and immature animal groups treated subcutaneously with prednisolone and appeared to be more suitable than that produced with alloxan treatment or under occlusive dressings. In prednisolone-induced infection, most of the micro-organisms were confined to the epidermis and there was no evidence of their dermal penetration during the 14 day experimental period. In view of these facts, we attempted a more quantitative method by counting viable Candida in order to evaluate antimycotics in a shorter therapy period during which animals would receive less exposure to the vehicle. These findings appear to indicate that the experimental model of cutaneous C. albicans infection produced in prednisolone-treated mature and immature guinea-pigs would be useful for studies on the therapeutic efficacy of antifungal agents against cutaneous candidiasis.
Administration, Topical↗
Reversible hyperkalemia during antihypertensive therapy in a hypertensive diabetic patient with latent hypoaldosteronism and mild renal failure.
A 66-year-old hypertensive diabetic patient with latent hypoaldosteronism and mild renal failure was treated by adding enalapril, an angiotensin converting enzyme inhibitor, to the furosemide and nifedipine regimen because of an insufficient antihypertensive response for 1 month. Seven days after enalapril addition, the blood pressure was significantly reduced, but frank hyperkalemia occurred with a marked rise in BUN and a slight increase in serum creatinine. Plasma renin activity (PRA) and plasma aldosterone (PA) values remained low before and during enalapril therapy. Transient treatment with sodium polystyrene sulfate after enalapril withdrawal improved the hyperkalemia and renal function, but PRA and PA levels were low. PA and its precursor steroids also responded poorly to graded angiotensin II infusion and rapid ACTH injection. Latent hypoaldosteronism probably predisposed this patient to frank hyperkalemia with progressive dehydration and slightly reduced renal function during antihypertensive therapy.
Aged↗
Enhancement of tumor proliferation by cyclosporine A in early phase of experimental hepatic metastasis.
The effect of cyclosporine A (CsA) on in vivo growth of hepatic metastasis was studied. Murine colon 38 tumor cells (1 x 10(5) were inoculated via the superior mesenteric vein. Mice were grouped depending on CsA dosage and time schedules: Group A: CsA 30 mg/kg body weight on the 7, 8 and 9th post tumor inoculation days by gavage; Group B: CsA 15 mg/kg body weight 30 min before tumor inoculation and 2 times more at 24 h intervals; Group C: CsA 30 mg/kg body weight at the same dose timing as Group B. Measurement of the diameter of the largest tumor serially by weekly laparotomy up to 4 weeks revealed that the tumor growth rates were significantly greater in Groups B and C than those in Group A or the control (without CsA). The mean tumor doubling times in the control, and Groups A, B and C were 2.2 +/- 1.3, 2.0 +/- 0.5, 1.5 +/- 0.4 and 1.3 +/- 0.8 days, respectively. The mean tumor numbers of hepatic metastasis were 13.2 +/- 8.3, 11.3 +/- 7.3, 19.4 +/- 8.7 and 19.6 +/- 6.8, respectively. Values of tumor proliferation index obtained by bromodeoxyuridine immunohistochemistry were 10.0 +/- 6.1%, 14.9 +/- 8.0%, 28.6 +/- 8.2% and 30.1 +/- 12.4%, respectively, with significant differences (Groups B and C vs. A or control, P < 0.05). In vitro MTT assay showed that cell viability rates were greater than 100% in the medium containing CsA concentrations of less than 10(-7) mol/liter. However, a cytostatic effect of CsA was apparent at higher concentrations. In contrast to the previous in vivo finding of a cytostatic effect of CsA on tumor cells, we found a cytoproliferative action when CsA was administered early in the course of metastatic tumor implantation in the liver. The mechanism of cytoproliferative effect of CsA is unknown but may involve multiple factors.
Animals↗
Direct sequencing of a novel DPB1 allele (DPB1*5101) of the heterozygote from the membrane of reverse dot blot analysis.
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Alleles↗
Aggregation of collagen exposed to UVA in the presence of riboflavin: a plausible role of tyrosine modification.
Riboflavin-sensitized photodynamic modification of collagen led to significant formation of cross-linked molecules. Sodium azide or 1,4-diazabicyclo(2,2,2)octane, which are known to be singlet oxygen quenchers, and catalase could not inhibit the modification. Surprisingly, the collagen modification was accelerated in the presence of superoxide dismutase. The aggregation was accompanied by the loss of tyrosine and histidine residues in the collagen. An inhibitory effect of dissolved oxygen on the modification of collagen was observed. Similarly, the loss of tyrosine residues in the irradiated collagen was inhibited in the presence of dissolved oxygen. Dityrosine formation was also observed with the loss of tyrosine. These results indicate that photodynamic modification of tyrosine probably contributes to the riboflavin-sensitized cross-linking of collagen through the formation of dityrosine.
Animals↗
[Detection of Mycobacterium avium, M. intracellulare and M. kansasii in sputum by two-step PCR].
A DNA amplification assay using PCR, which consists of amplification of genus specific mycobacterial 16S rRNA gene as the 1st step and reamplification of the amplicon with species specific primers as the next step, could detect M. avium, M. intracellular and M. kansasii in the sputum. The results with type or standard strains showed that M. avium PCR and M. intracellular PCR were highly specific for identification of each species but M. kansasii PCR detected M. gastri besides M. kansasii. Among 22 clinical samples which were positive by PCR, the 17 results were confirmed by culture. The PCR detected 27 (7.5%) of nontuberculous mycobacteria from 360 sputum and showed that the 27 of NTM consists of 9 M. avium, 8 M. intracellulare, 5 M. kansasii, 1 reacted both M. avium and M. intracellulare and 4 unidentified.
Base Sequence↗
Therapeutic efficacy of lanoconazole, a new imidazole antimycotic agent, for experimental cutaneous candidiasis in guinea pigs.
The therapeutic efficacy of 1% cream and 1% solution of lanoconazole, a new imidazole antimycotic agent, in the model of cutaneous candidiasis in prednisolone-treated guinea pigs was evaluated in comparison with that of comparable formulations of bifonazole. Each preparation was topically applied once a day for 3 consecutive days, starting on the fifth day postinfection, and quantitative culture study wsa conducted on the ninth day postinfection. Both formulations of lanoconazole were much more highly effective in terms of eradication of fungi than the bifonazole formulations.
Administration, Topical↗
Glomerular endothelial cells in culture express and secrete vascular endothelial growth factor.
Vascular endothelial growth factor (VEGF) is a specific growth factor for endothelial cells, and its abundant expression has been reported in kidney glomeruli. In this study, we focused on glomerular endothelial cells (GEN) as a possible source of VEGF secretion and sought to uncover a potential autocrine role of VEGF for GEN. Ribonuclease protection assay demonstrated VEGF mRNA expression in cultured GEN, and 46-kDa VEGF protein was detected in the conditioned medium by immunoblot analysis using polyclonal antibody raised against the NH2-terminal portion of VEGF. Removal of fetal bovine serum (FBS) from the culture medium for 2 h decreased VEGF mRNA abundance, which was restored by the readdition of FBS (10%) within 2 h. The effect of FBS was completely abolished by protein kinase inhibitor H-7 (10 microM), suggesting that FBS-stimulated VEGF mRNA induction involves activation of protein kinases. The treatment of GEN with 10(-7) M 12-O-tetradecanoylphorbol-13-acetate (TPA) increased the VEGF mRNA abundance fivefold, supporting the idea that VEGF expression is regulated by protein kinase C. [3H]thymidine incorporation into GEN treated with TPA (10(-7) M) was inhibited by neutralizing antibody for VEGF. Thus VEGF was identified as an autocrine growth factor for GEN in vitro. Its physiological role might be the regulation of GEN proliferation, and the induction of VEGF expression by FBS and TPA suggests its involvement in the response of glomerular capillary endothelial cells to injury in certain pathophysiological states.
Animals↗
Determination of glycated proteins in biological samples based on colorimetry of 2-keto-glucose released with hydrazine.
We developed a novel method for measuring glycated (glc) proteins in biological samples, based on the colorimetry of 2-keto-glucose which is released from the glc protein (ketoamine) on heating with hydrazine. The ketoamine-induced coloration remained constant at room temperature (25-27 degrees C) for 1 h. The method gave reliable precision and accuracy. However, high concentrations of serum pigments caused positive interference, suggesting that hemolytic or hyperbilirubinemic serum would give false-positive results. The concentration of glc protein in clinical serum samples measured by the present method (y) correlated well with those (fructosamine values, x) measured by the nitroblue tetrazolium-reducing method: y = 1.27x-1.69 (r = 0.92, n = 93). The concentrations (microM, mean +/- S.D.) of glc protein in sera from normal and diabetic subjects were 275 +/- 37 (n = 32) and 403 +/- 98 (n = 32), respectively, and the concentrations (nmol/mg hair, mean +/- S.D.) of glc protein in back hairs from non-diabetic and diabetic rats were 3.7 +/- 0.3 (n = 10) and 8.6 +/- 1.5 (n = 10), respectively. Thus, the technique gave reasonable concentrations of glc proteins in humans and rats with diabetes mellitus, indicating it to be reliable and diagnostically useful.
Adult↗
Regio- and stereoselective synthesis of carbocyclic 2',3'-dideoxy-3'-fluoro nucleosides as potential antiviral agents.
The synthesis and antiviral activity of racemic carbocyclic 2',3'-dideoxy-3'-fluoro nucleosides are reported. Carbocyclic 2',3'-dideoxy-3'-fluoro nucleosides were obtained from the 3-fluoro cyclopentane derivative 4, which was prepared by two methods. The SN2-displacement of the hydroxyl group of (+/-)-(1 beta, 2 alpha, 3 beta, 4 beta)-4-acetamido-2-fluoro-3-hydroxycyclopentylmethyl acetate (1) with Ph3P-I2 followed by tin hydride reduction afforded the 3-fluoroamino alcohol derivative 3. Alternatively, the protected fluoroamino alcohol 3 was prepared by regio- and stereoselective bromo-fluorination of cis-4 beta-acetamidocyclopent-2-enemethyl acetate (5) with hydrogen fluoride-pyridine/N-bromosuccinimide followed by tin hydride reduction to remove the bromine atom. Carbocyclic 2',3'-dideoxy-3'-fluoroguanosine (14) thus obtained was moderately active against herpes simplex virus in vitro.
Adenoviridae↗
Synthesis and antiallergic activity of novel azaazulene derivatives.
Various azaazulene derivatives were synthesized and their antiallergic activity was examined. The structure-activity relationship among various derivatives modified by introducing substituents at the 1-,2-, or 3-position of the azaazulene ring was investigated. The inhibitory activities on allergic histamine release of the compounds bearing a 5-tetrazolyl group at the 3-position were more potent than those of the corresponding compounds with other groups (CN, COOH, and CHO). The compounds substituted with amino, azide and carboxymethylamino groups at the 2-position showed strong inhibitory activity. The compounds with various phenylalkyl groups at the 1-position showed a greater activity than those with other substituents. Among the compounds with substituents at the 1-,2-, or 3-position of the azaazulene ring, 1-benzyl-7-isopropyl-3-(5-tetrazolyl)-1-azaazulen-2-one (18f) and 1-(4-fluorobenzyl)-7-isopropyl-3-(5-tetrazolyl)-1-azaazulen- 2-one (19c) had the most potent inhibitory activities on histamine release from mast cells and on passive cutaneous anaphylaxis (PCA) in rats after oral administration (ED50 = 0.56 and 0.58 mg/kg, respectively).
Animals↗
Regioselective cleavage reaction of the aromatic methylenedioxy ring. VI. Synthesis of phenothiazine analogues by using the cleavage reaction with sodium methoxide-thiols in dimethyl sulfoxide and evaluation of their biological activities.
The reactions of aromatic methylenedioxy compounds containing electron-withdrawing groups with sodium methoxide-thiols in dimethyl sulfoxide gave 3- and 4-hydroxybenzene derivatives in good yield by regioselective attack of the thiolate ions on the methylenedioxy ring. The formation mechanism and the reactivity of thiolate ions in the cleavage reaction of the methylenedioxy ring are discussed. Various biologically active compounds, 32a, 32d, 36b, 38b, 41b and 44-47, were prepared from the 4-hydroxybenzene derivatives and their Ca2+ antagonistic activities were evaluated. Among these compounds, 2-(2-bromophenylthiomethoxy)-10-(2-diethylaminoacetyl)-3- methoxyphenothiazine (46) showed the most potent Ca2+ antagonistic activity. Biological activity could be conveniently evaluated by measurement of the peak height of the vanadyl ion (+4 oxidation ion) signal produced by redox reaction between the phenothiazine derivatives and vanadate ion +5 oxidation ion) with ESR spectroscopy.
Animals↗
Formation of beta-galactosyl compounds of arabinosylcytosine in growing culture of Sporobolomyces singularis.
Two new compounds of 1-beta-D-arabinofuranosylcytosine (ara-C) were found to be produced in a high yield in a culture filtrate of Sporobolomyces singularis, when grown on a medium containing lactose and ara-C. The compounds I and II were obtained as white needle crystals from the culture filtrate by preparative paper chromatography, gel-filtration on Sephadex G-10 and Toyopearl HW-40S, lyophilization, and ethanol treatment. The compounds I and II were identified as 3'-O-(beta-D-galactopyranosyl)-ara-C and 3'-O-[beta-D-galactopyranosyl-(1-->4)-O-beta-D-galactopyranosyl]-ara-C, respectively, on the basis of the various experimental results, viz., elementary analyses, UV, IR, 1H-, and 13C-NMR spectra, and products by hydrolysis with alpha- and beta-galactosidases. Also, the yeast produced a large amount of 3'-O-beta-galactosyl compounds of adenosine and inosine in the culture filtrate when grown on a medium containing lactose and their ribonucleosides.
Carbohydrate Sequence↗
Mammary fibroadenoma in a young Holstein cow.
A mammary mass approximately 60 cm in diameter, which developed in the left udder of a 17-month-old Holstein cow, was removed surgically. The mass was well demarcated from the surrounding tissues and its cut surface was lobular in appearance. Microscopically, the mass consisted of a proliferation of well differentiated ducts and mucinous connective tissue. Immunohistochemically, the proliferative stroma contained two distinct cell types; vimentin-positive fibroblasts and spindle cells positive for vimentin and actin at the periphery of the ducts. The biological features of the present mammary tumor are thought to be generally compatible with those of mammary fibroadenoma reported in humans and dogs.
Animals↗
Human c-yes-1-related proto-oncogene in clinically normal dogs.
Human c-yes-1-related canine proto-oncogene in genomic DNAs from 21 clinically normal dogs was analyzed by Southern blot hybridization. The present study indicated that this proto-oncogene was well conserved in clinically normal dogs, however, there were structural changes in some dogs. These changes were different in each individuals and detected in low frequency.
Animals↗
Tumors in domestic animals examined during a ten-year period (1980 to 1989) at Miyazaki University.
During the ten years from 1980 to 1989 inclusive, a total of 468 (16.1%) tumors were found in 2,907 pathological samples from domestic animals, collected from Southern Kyushu, around Miyazaki City. In this study, canine tumors were collected most commonly (340/468 cases, 73%). In small animals, the skin and mammary gland were associated particularly with tumorigenetic hazards. In cattle, high incidence of leukemia and mesothelioma was found. These tendencies were almost the same as those we reported for the preceding ten-year period, although the number of cases of tumor was higher in this study.
Animals↗
Amplification of a c-yes-1-related oncogene in canine lymphoid leukemia.
Genomic DNAs from 4 canine spontaneous lymphoid neoplasms were examined by Southern blot hybridization using 7 kinds of oncogene probes. Analysis using a human c-yes-1 cDNA probe revealed amplification of the proto-oncogene in a DNA sample from a dog with lymphoid leukemia. Its degree was about 4- to 8-fold in comparison with a control DNA sample from a healthy dog. The genomic abnormality of canine c-yes-1-related oncogene may have a role in the tumorigenesis of this neoplasm, although the significance of the structural change in this oncogene remains unclear.
Animals↗