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Biomedical subjects

T Tsuji

Publications and source records attributed to T Tsuji.

At least 847 records · Page 47Linked to original sources

In vivo T1 measurements of canine brain for deep tissue temperature measurements with MRI.

The possibility of deep tissue temperature imaging with a magnetic resonance (MR) imager was studied in 30 mongrel dogs. The coronary view of the brain of a dog anesthetized with muscle relaxant and pentobarbital and ventilated automatically with air was taken with a special head coil by a 0.1 Tesla MR imager, Asahi Mark-J, driven by a spin warp pulse sequence. T1s in the imaging of the right and left white matter, grey matter, thalamus and caudate nucleus were measured with rectal temperature in group I (10 dogs) for a control study every 30 min until 120 min without intervention, in group II (10 dogs) for hyperthermia with extracorporeal bypass method, before heating, at 41 and 42 degrees C, in group III (10 dogs) for hypothermia, before cooling, at 35 and 30 degrees C and after rewarming, respectively. In groups I, II and III, the T1s at all sites except the white matter at 42 degrees C did not change significantly at any rectal temperature. T1 prolongation in the white matter at 42 degrees C may be due to edema, probably because of biological reaction to hyperthermia. The present study suggests that deep tissue temperature imaging in vivo by T1s measured with the MR imager may be impossible in spite of obvious evidence of T1 dependance on temperature in theoretical, in vitro and ex vivo studies already achieved.

Animals↗

[An immunohistochemical evaluation of Warthin's tumor using the monoclonal anti-human lung adenocarcinoma antibody KM-93 and the anti-human gastric carcinoma antibody KM-231].

An immunohistochemical identification of a human lung adenocarcinoma antigen and a gastric carcinoma antigen, detected by means of the monoclonal antibodies (MoAbs) KM-93 and KM-231, respectively, has been investigated in cases of a Warthin's tumor and in normal salivary glands. The ductal basal cells of the excretory duct epithelium in normal salivary glands are characterized by their positive staining to MoAb KM-231, whereas MoAb KM-231 staining is confined to basal tumor cells in cases of Warthin's tumor. Further, MoAb KM-93 staining is slightly positive when luminal tumor cells are present. These results suggest that the histogenesis of Warthin's basal tumor cells might originate in the excretory duct epithelium.

Adenocarcinoma↗

The effects of photosensitization by hematoporphyrin derivative on the protein content of cultured human lung cancer cells. A flow cytometric analysis.

In order to study the cytotoxic mechanisms of photodynamic therapy (PDT), the changes in the intracellular content of protein and DNA of cultured human lung cancer cells were examined by flow cytometry using the fluorescent dyes fluorescein isothiocyanate (FITC) and propidium iodide (PI). Immediately after PDT, the protein content increased while the DNA content showed little change. After more than 24 hours, the protein and DNA content were markedly decreased in lethally damaged cells; in sublethally damaged cells, however, the protein content gradually decreased to the level of that before PDT while the DNA content showed little change. These changes in the protein content may reflect the impairment of the plasma membrane function by PDT and its subsequent recovery in some cells.

DNA, Neoplasm↗

[A case of chronic progressive radiation myelopathy successfully treated with corticosteroid].

One patient who underwent radiation therapy for laryngeal cancer two years earlier developed chronic progressive radiation myelopathy and exhibited localized swelling of the cervical spinal cord at the C4 level and resultant blocking of the subarachnoid space on myelography and CT myelography. The condition responded markedly to the administration of corticosteroid except for persisting mild spasticity and sensory disturbances. On myelography and CT myelography half a year later, the cervical spinal cord showed no swelling. The corticosteroid therapy is considered to be useful for radiation myelopathy.

Aged↗

Charge heterogeneity of heat-labile enterotoxins from human enterotoxigenic Escherichia coli.

We purified heat-labile enterotoxins (LThs) from YT3, H-10407 and YT240 strains isolated from human diarrheal patients. These LThs were immunologically identical to each other. The molecular weights of their A and B subunits were also the same by means of SDS-polyacrylamide gel electrophoresis. However, the ionic charges of the molecular surfaces of these LThs were different as shown by polyacrylamide gel isoelectric focusing. Though the pI points of B subunits of the LThs were identical to each other, the pI points of A subunits were found to be different. These data suggest that the ionic charge differences among A subunits cause differences in holo LThs in their charge, and that there is heterogeneity among A subunits produced by strains of human enterotoxigenic Escherichia coli.

Bacterial Toxins↗

A single amino acid substitution in B subunit of Escherichia coli enterotoxin affects its oligomer formation.

We isolated a mutant strain of enterotoxigenic Escherichia coli by nitrosoguanidine mutagenesis, which produces an immunologically altered B subunit of heat-labile enterotoxin. This mutant B subunit was detected as a monomer on sodium dodecyl sulfate-polyacrylamide gel electrophoresis even without prior heating, suggesting a problem in oligomer formation. Furthermore, this mutant B subunit could not form holotoxin with the native A subunit, and the affinity to GM1-ganglioside receptor was 10-fold lower than that of the native B subunit. The amino acid sequence analysis of this mutant B subunit revealed only one amino acid substitution compared with the native B subunit, at the 64th position from the N terminus (valine instead of alanine). These data suggest that the alanine at position 64 from the N terminus is important for the native B subunit to form an oligomer structure and express its functions.

Alanine↗

Studies on the binding substances on human erythrocytes for the heat-labile enterotoxin isolated from chicken enterotoxigenic Escherichia coli.

To study the predominant binding substance for the heat-labile enterotoxin (LTc) isolated from chicken enterotoxigenic Escherichia coli, competitive binding assays were performed with neuraminidase-treated human type B erythrocytes and 125I-labeled B subunit of LTc (LTc-B). Of all inhibitors used, the ganglioside GM1 was the most effective in inhibiting the binding of 125I-labeled LTc-B to the erythrocytes. The other gangliosides used as inhibitors, gangliosides GD1b, GD1a, GM2, GT1b and GM3, were about 24, 166, 250, 440 and at least 440 times less reactive than ganglioside GM1, respectively. With glycoproteins as inhibitors, on the other hand, hog A + H, porcine thyroglobulin and bovine salivary mucin were over 10(4) times less potent. No inhibition was obtained by other mono-, di- and polysaccharides at the highest concentrations used. These findings suggest that the predominant binding substance on neuraminidase-treated human type B erythrocytes for the LTc-B is ganglioside GM1 and that the combining site of LTc-B may be specific for the terminal disaccharide (galactose-N-acetyl-D-galactosamine)-linked portion of ganglioside GM1.

Animals↗

Crystallization and preliminary X-ray diffraction study of the heat-labile enterotoxin B subunit produced by enterotoxigenic Escherichia coli.

Heat-labile enterotoxin LT produced by enterotoxigenic Escherichia coli is composed of A and B subunits. The A subunit is enzymatically active; whereas, through the action of the B subunit, the toxin binds to the receptor, a GM1 ganglioside present on the cell surface. Crystals of the LT-B subunit were formed at room temperature by vapor diffusion with polyethylene glycol in the presence of the non-ionic detergent beta-octylglucoside. The crystals were characterized by X-radiation as orthorhombic, space group P2(1)2(1)2(1), with unit cell dimensions of a = 224.1 A, b = 65.3 A, c = 118.4 A. They diffract X-rays to a resolution of at least 2.5 A and are stable to X-rays.

Bacterial Toxins↗

Continued high albumin production by multicellular spheroids of adult rat hepatocytes formed in the presence of liver-derived proteoglycans.

Adult rat hepatocytes formed floating multicellular spheroids, when they were cultured with proteoglycan fraction isolated from rat liver reticulin fibers. Cells in the spheroid showed only low growth activity. Albumin production by the spheroids increased up to 1.5 micrograms/micrograms DNA/day (180 micrograms/mg Protein/day) during the first 6 days and remained constant thereafter. In contrast, the albumin production by the monolayer markedly decreased after 4 days. The spheroid culture appears to be more suitable than the monolayer in studying differentiated functions of adult hepatocytes.

Albumins↗

[Effects of photodynamic therapy on cell kinetics of cultured human cancer cells in reference to its cytocidal effects].

Photodynamic therapy (PDT) for cancer is a relatively new technique that has been used clinically in the treatment of a variety of neoplasms. In this report, the effects of PDT on cell kinetics of the two cultured human lung cancer cell lines (PC-6, PC-9) were investigated in reference to its cytocidal effects. The cellular uptake of hematoporphyrin derivative (HpD) was dependent on external HpD concentration. And in the same external HpD concentration, PC-9 cells incorporated more HpD than did PC-6 cells. On the other hand, PC-6 cells were more susceptible to PDT than did PC-9 cells. This finding is explainable for the reason that PC-6 cells incorporate more HpD per unit protein content in single cells than do PC-9 cells. Analysis of DNA histogram obtained by flow cytometry showed that a shift in the histogram peak to hypodiploid occurred at severe cytocidal effect of PDT, and this finding expressed the cell killing characterized by cell lysis after PDT. At moderate cytocidal effect of PDT, a prolongation of G2/M phase of the cell cycle caused by the inhibition of protein synthesis was observed. And partial synchronization was ascertained. These findings indicate the possibility for synergistic antitumor effect of PDT in combination with other therapies.

Cell Cycle↗

Studies on the binding substances on human erythrocytes for the heat-labile enterotoxin isolated from porcine enterotoxigenic Escherichia coli.

The binding substance for the heat-labile enterotoxin (LTp) isolated from porcine enterotoxigenic Escherichia coli was studied by competitive binding assays. The binding of 125I-labeled LTp to neuraminidase-treated human type A erythrocytes was most effectively inhibited by ganglioside GM1 among inhibitors used. Mono-, di- and polysaccharides, glycoproteins and lectins were over 10(4)-times less potent inhibitors. Similar results were also obtained in competitive binding assays with 3H-labeled ganglioside GM1 and LTp-coupled Sepharose 4B. On the other hand, hemagglutination of neuraminidase-treated human type A erythrocytes by LTp was inhibited by methyl alpha-D-galactopyranoside, galactose, melibiose and some glycoproteins, but not effectively inhibited by ganglioside GM1 at the highest concentration used. Preincubation of LTp with an appropriate amount of ganglioside GM1 resulted in much higher hemagglutination than LTp alone. Although these findings show that there may be fundamental differences between interactions with ganglioside GM1 in hemagglutination compared to interactions with ganglioside GM1 in binding, the predominant binding substance for LTp on neuraminidase-treated human type A erythrocytes is suggested to be ganglioside GM1.

Animals↗

Production of monoclonal antibody to human IgG allotype G3M T.

An efficient method for the production of monoclonal anti-G3M T antibody is described. IgG3 protein of GM B3ST phenotype was isolated by affinity chromatography on Ricinus communis lectin I-agarose and used for immunization. A mouse hybridoma clone was obtained by fusion of popliteal lymph node cells and P3U1 myeloma cells. The antibody produced was tested for allotype specificity by hemagglutination inhibition and ELISA methods using 101 IgG-allotyping control sera. The antibody was neutralizable by all G3M T-positive sera and entirely nonneutralizable by G3M T-negative sera in the inhibition test, and reacted only with G3M T-positive IgG coats in the ELISA test. The results prove the antibody to be allotype-specific, and therefore practically establishes its monoclonality.

Antibodies, Monoclonal↗

Hemagglutinating activity of the B subunit(s) of the heat-labile enterotoxin isolated from chicken enterotoxigenic Escherichia coli.

The hemagglutinating activity of the B subunit(s) of the heat-labile enterotoxin (LTc-B) produced by chicken enterotoxigenic Escherichia coli was studied by hemagglutination and hemagglutination inhibition. No or weak hemagglutination of intact human erythrocytes was found by the LTc-B at the highest concentration used, whereas strong hemagglutination of both neuraminidase- and pronase-treated human erythrocytes was found. Enhancement in hemagglutination of treated human erythrocytes induced by the LTc-B was over 2 to 120-fold for type A and B erythrocytes and over 8-fold for type O erythrocytes, respectively. With intact and treated sheep erythrocytes, on the other hand, no hemagglutination was found by the LTc-B at the highest concentration used. Hemagglutination of pronase-treated human type B erythrocytes by the LTc-B was inhibited by methyl-alpha-D-galactopyranoside, galactose, melibiose, hog A + H, asialo-bovine salivary mucin and asialo-thyroglobulin among mono-, di- and polysaccharides and glycoproteins used as inhibitors. These results suggest that the LTc-B is a galactose-specific bacterial lectin.

Animals↗

Flow cytometric analysis of the kinetic effects of cisplatin on lung cancer cells.

The effects of cisplatin on the cell cycle and DNA synthesis of human lung adenocarcinoma cell line PC-9 were examined by flow cytometry. The cellular DNA content and the bromodeoxyuridine (BrdUrd) incorporation rate were measured simultaneously using a monoclonal anti-BrdUrd antibody. Following exposure to cisplatin (1.0 micrograms/ml) for 1 and 24 hr, the bivariate DNA/BrdUrd distributions revealed a delayed S-phase transit and an accumulation of cells in the G2M phase. The BrdUrd-linked green fluorescence intensity continued to decrease with the lapse of time. However, early- and mid-S-phase cells soon recovered DNA synthesis activity, and the former showed higher activity than the control cells. These findings suggested the vigorous DNA synthesis of cells in early S phase. However, for quantitative analysis of chemotherapeutic effects, some problems remained to be resolved regarding the condition for DNA denaturation and its alteration by the agents.

Adenocarcinoma↗

Type 4 chain H expression by bile ductules and hepatocytes in cirrhosis.

The monoclonal antibody MBr1 defines the blood group H determinant with beta 1----3N-acetylgalactosamine linkage (Fuc alpha 1----2Gal beta 1----3GalNAc----R) carried by type 3 or 4 backbone. The distribution of the antigen detected by this antibody was studied immunohistochemically in liver tissues. Although bile ducts with a diameter of more than about 100 microns normally expressed the MBr1-reactive antigen supranuclearly, smaller bile ducts and bile ductules did not express the antigen. In cirrhotic liver, proliferated bile ductules extensively expressed the MBr1-reactive antigen. In spite of the absence in normal liver cells, the antigen was expressed membranously in some cirrhotic liver cells. Under subcellular fractionation, MBr1 reactivity was almost exclusively recovered in the microsomal fraction. By HPTLC immunostaining, the major MBr1-reactive antigen was shown to be carried by type 4 chain H glycolipid (globo-H, Fuc alpha 1----2Gal beta 1----3GalNAc beta 1----3Gal alpha 1----4Gal beta 1----4Glc beta 1----1Cer). MBr1 reactive glycoprotein was not found. In conclusion, although type 4 chain H glycolipid is not expressed by normal bile ductules and liver cells, it is actively synthesized and expressed by proliferated bile ductules and some of the liver cells in cirrhosis in the absence of any neoplastic change.

Antibodies, Monoclonal↗

Detection of IgA protease from Haemophilus influenzae by immunoblotting.

IgA protease produced by various strains of Haemophilus influenzae can digest serum IgA and yield its fragments which can react with anti-IgA serum. We assayed IgA protease activity by detecting the digests of IgA by SDS-PAGE and immunoblotting. The digests were separated with SDS-PAGE, transferred to nitrocellulose membranes and detected with anti- (alpha chain of human IgA, its Fab and its Fc) immunoglobulin conjugated peroxidases. Using this method, we can determine which type of IgA protease is produced by various of H. influenzae strains. All the 20 strains isolated from respiratory tracts produced IgA protease.

Blotting, Western↗