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Biomedical subjects

T Uda

Publications and source records attributed to T Uda.

At least 37 records · Page 2Linked to original sources
PubMed1994

Zinc hydroxide induced respiratory burst in rat neutrophils.

The effects of zinc hydroxide on the respiratory burst and phagocytosis by rat neutrophils were examined. Zinc hydroxide induced an increase in oxygen consumption and O2- production. Electronmicroscopy showed that neutrophils engulfed zinc hydroxide particles by phagocytosis. Pertussis toxin (0.25, 0.5, 1.0 micrograms/ml) and EGTA (1, 2, 5 mM) inhibited zinc hydroxide-induced O2- production in a dose-dependent manner. The inhibitors of protein kinase C, 1-(5-isoquinolinesulfonyl)-2-methyl-piperazine and N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide inhibited zinc hydroxide-induced O2- production with IC50 values ranging between 10 microM and 25 microM. The inhibitory study using an inhibitor of myosin light chain kinase, 1-(5-iodo-naphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine, showed IC50 values ranging from 5 microM to 10 microM. These findings indicate that zinc hydroxide induces respiratory burst and phagocytosis by rat neutrophils.

Animals↗
PubMed1993

Heavy-chain isoforms of non-muscle myosin in human tissues.

The heavy-chain isoforms of myosin in human non-muscle and smooth muscle tissues were analyzed by means of SDS/PAGE and using three distinct newly developed monoclonal anti-(human cerebrum myosin) Ig (HBM-1, HBM-3 and HBM-4). Purified cerebrum myosin contained three electrophoretic variants of non-muscle myosin heavy chain (NM1, NM2 and NM3, with apparent molecular masses of about 200, 198 and 196 kDa, respectively). Both NM1 and NM2 were recognizable by the brain-specific antibody HBM-1, while NM3 was recognizable by HBM-3. Each of the variants reacted with HBM-4 to a similar extent. Purified cerebellum myosin gave three electrophoretic variants of the heavy chain which were indistinguishable electrophoretically or immunologically from those of cerebrum myosin. Aortic myosin contained four electrophoretic variants, including the two smooth muscle myosin heavy chain isoforms and NM2-like and NM3-like heavy chains. Liver, platelet and kidney myosins contained a heavy chain very similar to NM3. Kidney myosin also contained a small fraction of an NM2-like electrophoretic variant. In addition, cerebrum, kidney, liver and platelet myosins appeared to contain minor, 194-kDa myosin heavy-chain-like polypeptide(s) (NM4). NM1, as well as NM2 and NM3, thus appear to be the brain-type and non-brain-type non-muscle myosin heavy-chain isoforms, respectively, and additional minor heavy-chain isoforms are also likely to be present in human tissues.

Animals↗
PubMed1993

Characterization of morphine-specific monoclonal antibodies showing minimal cross-reactivity with codeine.

Six murine monoclonal antibodies against morphine were produced using N-(4-aminobutyl)normorphine as a hapten. Most of the antibodies obtained distinguished the substituents at the 3 and 6 positions of morphine. This property of the antibodies led to a reduction in cross-reactivity with codeine, morphine-3-glucuronide (M-3-G) and morphine-6-glucuronide (M-6-G) to negligible levels. However, one of the antibodies distinguished the substituent only at the 3 position of morphine, which cross-reacted with M-6-G, naloxone and naltrexone. In the competitive inhibition enzyme-linked immunosorbent assay, morphine was detected at concentrations as low as circa 100 pg/ml.

Animals↗
PubMed1993

Human erythrocyte band 3 (EPB3) polymorphism: analysis of blood and bloodstains by an immunodetection method and frequency of the EPB3* Memphis variant.

The erythrocyte band 3 (EPB3) variant, band 3 Memphis (EPB3*Memphis), was detected by immunoblotting with a monoclonal antibody to the 41 kDa cytoplasmic N-terminal domain of band 3 without protease treatment of erythrocytes. EPB3*Memphis was also detected by immunoblotting from 3-month-old bloodstains subjected to alpha-chymotrypsin treatment. A population genetic study using this method indicated that the EPB3 variant would be useful for forensic work in Japan, since the frequency of this variant in Japanese (Wakayama prefecture) is relatively high (0.159).

Anion Exchange Protein 1, Erythrocyte↗
PubMed1993

ABO blood grouping of bloodstains by sandwich ELISA using monoclonal antibody specific for human red cell band 3.

ABO blood grouping of human bloodstains was performed by a sandwich ELISA using a species-specific monoclonal antibody to the amino-terminal cytoplasmic domain of human red cell membrane band 3. In a blind trial, all A, B and O bloodstains (a 1 cm long thread) and AB bloodstains (a 1.5 cm long thread) were accurately typed by this method. Even when bloodstains were contaminated by other body fluids (e.g., semen and saliva), only the ABO blood group epitopes on band 3 of the red cell membrane were detected. Thus, identification of human blood and ABO blood grouping of bloodstains which were contaminated by other body fluids could be simultaneously performed by this method.

ABO Blood-Group System↗
PubMed1992

Species-specific epitopes exist on the cytoplasmic amino-terminal domain of erythrocyte band 3 protein.

Monoclonal antibodies (P3-9H, P3-1F, P3-2H, P3-4A, and P3-4C) to human erythrocyte band 3 were produced using human erythrocyte membranes as the immunogen. All epitopes defined by these antibodies were found on the amino-terminal cytoplasmic domain of erythrocyte band 3. The antibodies crossreacted variously with erythrocyte band 3 of primates (chimpanzee, orangutan, Rhesus monkey, Japanese monkey, spider monkey, and capuchin monkey) in enzyme-linked immunosorbent assay. P3-9H did not crossreact with erythrocyte band 3 of any primate examined; P3-1F crossreacted only with that of chimpanzee; P3-2H crossreacted with erythrocyte band 3 of chimpanzee, spider monkey, and capuchin monkey; and P3-4A and P3-4C crossreacted with erythrocyte band 3 of all primates examined. These results suggest that evolutional changes in primates are accumulated in the amino-terminal cytoplasmic domain of band 3 and that species-specific epitopes exist on this domain.

Animals↗
PubMed1991

Clinical importance of coronary calcification detected by CT.

Coronary calcifications appear in advanced atheromatous lesions therefore fluoroscopy is useful for the detection of the atherosclerotic coronary artery disease (Aldrich et al., 1979). However, the detection of coronary calcification by fluoroscopy is difficult in the case of obesity or thick chest wall and also impaired by the background structures including bone and other intrathoracic calcifications. X-ray CT is more advantageous than fluoroscopy in the detection of coronary calcification. It can eliminate the interference from background structures and clearly demonstrates calcified sites of the coronary artery. Therefore, we investigated the clinical usefulness in the detection of coronary calcification with X-ray CT.

Adult↗
PubMed1990

Stimulation of Mn-superoxide dismutase expression by tumor necrosis factor-alpha: quantitative determination of Mn-SOD protein levels in TNF-resistant and sensitive cells by ELISA.

Marked increase in protein levels of Mn-superoxide dismutase (Mn-SOD) was found in TNF-resistant cell lines after treatment with Tumor Necrosis Factor (TNF). No such increase was observed in Cu,Zn-superoxide dismutase (Cu, Zn-SOD) protein in either TNF-resistant or sensitive cells. These results support the data that the Mn-SOD is one of the rescue proteins required for resistance to TNF cytotoxicity in these cell lines (Wong et al., Cell 58, 923-931, 1990). Mn-SOD was also responsive to TNF stimulation in KURAMOCHI, a human ovarian adenocarcinoma cell line. This may explain our previous result that Mn-SOD protein is highly expressed in epithelial ovarian cancer (Ishikawa et al. Cancer Res. 50, 2538-2542, 1990).

Adenocarcinoma↗
PubMed1990

Serum-manganese-superoxide dismutase: normal values and increased levels in patients with acute myocardial infarction and several malignant diseases determined by an enzyme-linked immunosorbent assay using a monoclonal antibody.

An enzyme-linked immunosorbent assay (ELISA) has been developed for human manganese-superoxide dismutase (Mn-SOD), using a specific monoclonal antibody raised against the purified enzyme. The Mn-SOD molecule comprises four identical sub-units and this permitted the development of a symmetrical assay, using the same monoclonal antibody as both capture and detector. The assay offers a specific, sensitive and convenient means of measuring immunoreactive Mn-SOD in human sera. Under optimum conditions, the sensitivity of the assay permits the detection of 2-200 ng of purified Mn-SOD from human liver. The mean serum Mn-SOD levels of normal healthy males and females were 99.8 +/- 24.8 (mean +/- SD) and 88.8 +/- 20.8 (mean +/- SD), respectively. A high level of the enzyme was found in the sera of patients with acute myocardial infarction as well as malignant diseases such as acute myeloid leukemia, primary hepatoma and gastric cancer. This is the first report of an ELISA using a monoclonal antibody specific for a distinct epitope of Mn-SOD.

Adult↗
PubMed1990

An immunohistochemical study of copper, zinc-containing superoxide dismutase detected by a monoclonal antibody gastric mucosa and gastric cancer.

The immunohistochemical localization of copper, zinc-superoxide dismutase (Cu,Zn-SOD) in human gastric mucosa and gastric cancer was studied using a monoclonal antibody. In gastric mucosa, parietal cells, pyloric glandular cells and foci of intestinal metaplasia showed positive staining in the cytoplasm and/or nucleus. The wide distribution of Cu, Zn-SOD in the gastric mucosa suggests cell function may be vulnerable to active oxygen species. In gastric cancer, 34 of 70 cases showed a positive reaction for Cu, Zn-SOD. There was a relationship between the grade of Cu,Zn-SOD immunoreactivity and the histological type of gastric cancer, well-differentiated types of gastric cancer being more frequently positive. The positive cases of poorly-differentiated adenocarcinoma were characterized by a pattern of diffusely infiltrative invasion. These results suggest that some types of gastric cancer are resistant to active oxygen species.

Adenocarcinoma↗
PubMed1990

Coronary artery calcification detected by CT: clinical significance and angiographic correlates.

Cardiac computed tomography (CT) to detect coronary calcification was performed on 161 patients undergoing coronary angiography for proven or suspected coronary artery disease. Among 108 patients in whom coronary calcifications was identified, 90% had significant coronary stenosis angiographically (greater than 75% stenosis), and 80% of 121 patients with significant coronary stenosis showed calcification by CT. The relationship between the calcification site and the significance in stenosis of each vessel was determined. Calcification was present in 133 arteries among 205 stenotic coronary arteries (sensitivity = 65%) as compared with 59 of 439 entire arteries with normal coronary angiograms (specificity = 87%). In the younger age group the sensitivity of calcification for stenosis of each coronary artery was lower and the specificity and predictive value were generally higher than those in the elderly group. These results demonstrate that CT is a valuable procedure for detecting coronary arterial disease, since this examination is easy to conduct, noninvasive, and widely applicable for screening a large population.

Adult↗
PubMed1989

Cytoplasmic orientation and two-domain structure of the multidrug transporter, P-glycoprotein, demonstrated with sequence-specific antibodies.

The predicted cytoplasmic orientation and two-domain structure of the multidrug efflux pump P-glycoprotein were demonstrated with sequence-specific antibodies. We synthesized peptides corresponding to amino acid residues, Glu393-Lys408 (anti-P) and Leu1206-Thr1226 (anti-C) in P-glycoprotein from human mdr1 cDNA and used these peptides to produce polyclonal antibodies. From the primary structure of P-glycoprotein, and anti-C antibody is expected to recognize another position, Leu561-Thr581, in the duplicate structure of P-glycoprotein, but anti-P recognizes only one site. These antibodies bind to multidrug-resistant cells (KB-C2) with permeabilized plasma membrane but do not bind to nonpermeabilized KB-C2 cells or parental KB cells, supporting the predicted cytoplasmic orientation of these sequences. With immunoblotting of the membrane fractions from KB-C2 cells, a major 140-kDa polypeptide of the P-glycoprotein was detected with both anti-P and anti-C. Two minor polypeptides with molecular mass of 95 and 55 kDa were also detected. When membrane vesicles were digested mildly with trypsin, the amount of these two polypeptides increased. Anti-P detected only the 95-kDa polypeptide, and anti-C detected both 95- and 55-kDa polypeptides. Achromobacter lyticus protease I (lysyl endopeptidase) and Staphylococcus aureus V8 protease also produced two polypeptides with similar molecular weights. Absorption into lectin-agarose beads and labeling with [3H]glucosamine indicated that the 95-kDa polypeptide was glycosylated but that the 55-kDa polypeptide was not. These two polypeptides as well as P-glycoprotein were photoaffinity-labeled with a calcium channel blocker, [3H]azidopine, but most of the label was found in the 55-kDa polypeptide. The yield of labeled fragments from membrane vesicles photolabeled after digestion with trypsin was similar to that from membrane vesicles digested with trypsin after photolabeling. These data indicate 1) that the 95-kDa polypeptide is the fragment corresponding to the amino-terminal half of P-glycoprotein containing sugar chains; 2) that the 55-kDa polypeptide is the carboxyl-terminal half which was mainly labeled with [3H]azidopine; and 3) that P-glycoprotein has a relatively rigid structure with a small number of protease-sensitive sites and its global structure is not destroyed by tryptic cleavage.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1989

Human serum immuno-reactive copper, zinc-superoxide dismutase assayed with an enzyme monoclonal immunosorbent in patients with digestive cancer.

Two types of monoclonal antibodies, N4 and N6, were raised against human copper, zinc-superoxide dismutase (Cu,Zn-SOD). An enzyme-linked immunosorbent assay for Cu,Zn-SOD was developed in sandwich method using these monoclonal antibodies. The measurable assay range was 1.0-100 micrograms/l and the coefficient of variation within and between series were 7.5% and 8.1%. Sera of 154 healthy control subjects displayed a low level (14 micrograms/l) of immunoreactive Cu,Zn-SOD with a logarithmic normal distribution. On the other hand, serum Cu,Zn-SOD levels were significantly elevated in patients with various digestive cancers.

Antibodies, Monoclonal↗
PubMed1989

Preparation of monoclonal antibodies against methamphetamine.

15 monoclonal antibodies against methamphetamine (MA) were obtained from hybridoma clones by the fusion of mouse 653 myeloma cells with the spleen cells of the immunized mice. The monoclonal antibodies exhibited different cross-reactivities against MA-related compounds and one antibody (MA-15 Ab) was highly specific against MA. A competitive inhibition enzyme-linked immunosorbent assay was established and the detection limit for MA was approximately 20 ng/ml.

Animals↗
PubMed1989

A monoclonal antibody against COOH-terminal peptide of human liver manganese superoxide dismutase.

Three stable hybridoma cell lines producing monoclonal antibodies specific for human liver manganese superoxide dismutase were established, and one monoclonal antibody, PG 11, was chosen for immunochemical studies. Immunoblotting demonstrated that the monoclonal antibody binds exclusively to the manganese superoxide dismutase. Immunohistochemical studies indicated that the enzyme is localized in the matrix of human liver mitochondria. To localize antibody-binding epitope, synthetic peptides of the NH2-terminal (residues 1-16) and COOH-terminal (residues 182-189, 190-196, and 182-196) parts of the enzyme were synthesized, and then their effects on the binding were studied using an enzyme-linked immunosorbent assay method. All of the above COOH-terminal peptides inhibited the binding whereas the NH2-terminal ones did not, indicating that PG 11 recognizes several peptides of COOH termini of manganese superoxide dismutase. This is the first report of monoclonal antibodies against human manganese superoxide dismutase with a distinct epitope and of the immunocytochemical demonstration of manganese superoxide dismutase.

Animals↗