Immunofluorescence analysis of cultured mammalian cell nucleus using four monoclonal antibodies against the nucleus.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to Y Kaneda.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
A case of intestinal myiasis caused by larvae of Sarcophaga peregrina (Japanese name: senchi-nikubae) is reported. The patient passed stools containing live fly larvae for 2 days, but was otherwise asymptomatic. The patient seems to have been infested accidentally as a result of having eaten "yakitori" in which the fly had deposited its larvae. This paper is the first report of intestinal myiasis caused by the 3rd instar of this species.
Explore the source record for details and available documents.
Two monoclonal antibodies reacted with the Trypanosoma cruzi-specific antigen of an apparent Mr 25,000 from all developmental forms (Tachibana et al. 1986). This T. cruzi-specific antigen was found at the plasma membrane by immunoperoxidase electron microscopy using the monoclonal antibodies TCF48 and TCF87. The TCF48 and TCF87-treated cells showed stain deposits at the plasma membrane clearly distinguishable from those in cells treated with a monoclonal antibody against a surface antigen. This suggests that the epitope(s) of the Mr 25,000 antigen is located on the inner surface or in the matrix of the plasma membrane. TCF48 and TCF87 also reacted with an antigen on the microtubules of the axoneme, but not with the subpellicular microtubules. These results suggest that the T. cruzi-specific Mr 25,000 antigen is common to both the plasma membrane and axoneme but it is not located at the subpellicular microtubules. Its identity and that of the surface antigen, Gp25 (Scharfstein et al. 1983) as well as its role in the pathogenicity of the parasite are discussed.
Two monoclonal antibodies (designated as TCF48 and TCF87 were raised against Trypanosoma cruzi, strain Tulahuen, Both antibodies reacted with all developmental forms of several different strains of Trypanosoma cruzi. The antibodies showed no detectable cross-reactivity with other species of Trypanosomatidae, so far examined. TCF48 and TCF87 were classified as immunoglobulin subclasses IgG1 and IgG2b, respectively. Apparent molecular weight of the corresponding antigen(s) to these monoclonal antibodies was 25,000 in amastigotes and epimastigotes, and 25,000 and 24,000 in trypomastigotes, as determined by the Western immunoblotting analysis. This antigen appeared to be located at the plasma membrane and the flagellum ofT. cruzi. However, no evidence supported the localization of the epitope(s) at the external surface of the live cell. Since this antigen reacted with the sera from the chronically infected mice, these monoclonal antibodies may be useful in the study of Chagas' disease.
Two species of glycoproteins from Leishmania braziliensis promastigotes of apparent molecular weights of 53,000 (glycoprotein 53) and 47,000 (glycoprotein 47) were localized. Four lectins with different sugar specificities bound to the blotting sheet to which the electrophoretically separated materials were transferred. Concanavalin A and Ricinus communis agglutinin bound to the band of glycoprotein 53 and the lectin from Dolichos biflorus bound to the band of glycoprotein 47. Wheat germ agglutinin bound to the bands of both glycoproteins. Histochemical examinations using fluorescence labeled lectins demonstrated that the glycoproteins 53 and 47 were located on the cell surface and in the cytoplasm of promastigotes, respectively. The results are consistent with the result of agglutination test.
The neutral and phospholipid content at each of the three morphological stages of the parasite, Trypanosoma cruzi, was analyzed by thin-layer chromatography. Total lipid fatty acid composition at each stage was analyzed by gas-liquid chromatography and the results were compared. Change in lipid composition at each stage was observed.
Twenty patients with benign prostatic hypertrophy were treated with Cernilton, 6 tablets a day for an average of 13.2 weeks. Subjective effectiveness was observed in the improvement of sense of residual urine (92%), retardation (86%), night frequency (85%), strain on urination (56%), protraction (53%) and forceless urinary stream (53%). The overall subjective effectiveness was 80% of patients, and the overall objective effectiveness was 54% of patients. Night frequency, residual urine volume and tidal urine volume were improved significantly. The overall effectiveness was 80%. No side effects were observed.
A procedure is described for the isolation of subcellular fractions from epimastigotes of Trypanosoma cruzi. The method could separate most of the nuclei, mitochondria and microsomes. The plasma membranes were purified by discontinuous density gradient centrifugation in alkaline buffer containing sucrose and magnesium. The yield of plasma membrane was 3.7 mg of protein per 10(9) epimastigotes, accounting for approximately 4.2% of the total cell proteins. The plasma membrane obtained from the 34-50% interface of sucrose density gradients was shown, by electron microscopy, to be completely free of other organella and was homogeneous according to enzymatic marker criteria. The specific activity of the 5'-nucleotidase and acid phosphatase were 96- and 5.5-fold, respectively, higher than that in the total cells, suggesting that the enzymes can be considered as good plasma membrane markers for T. cruzi. The results confirmed the possibility of the presence of membrane-bound acid phosphatase of T. cruzi.
Acid phosphatase activity in Trypanosoma cruzi was found to be located on the external surface of the plasma membranes. Both specific activity and activity per cell of this bound enzyme were significantly higher in the cells of amastigote (an intracellular form) than that of trypomastigote (a bloodstream form) and epimastigote (culture form). During the transformation of epimastigotes to amastigotes in vitro the activity of surface acid phosphatase was elevated concomitant with the increase in population of amastigotes. These results were interpreted as that the elevated enzyme activity is required for the intracellular parasitization of this organism or is a consequence of the morphological transformation.
Elongation factor 2 (EF-2), polypeptidyl -tRNA translocase, is an essential factor for protein synthesis in eukaryotic cells and Archebacteria . We isolated diphtheria toxin-resistant human primary embryo cells that contain EF-2 that cannot be ADP-ribosylated by diphtheria toxin and Pseudomonas exotoxin A (PA toxin). Somatic cell hybrids were constructed from mouse L cells and toxin-resistant human embryo cell mutants. Forty-one hybrid clones were isolated, of which 15 clones were resistant to PA toxin. Karyotypic analysis and isozyme studies revealed that there was an absolute correlation between human chromosome 19 and resistance to PA toxin in the hybrids. On subcloning of PA toxin-resistant hybrid cells, we obtained one PA toxin-resistant hybrid subclone containing human chromosome 19 as the only human chromosome. Furthermore, the resistance to PA toxin of hybrid cell strains was lost after infection with poliovirus, for which sensitivity is conferred by human chromosome 19. It was confirmed by using two-dimensional gel electrophoresis that PA toxin resistance in hybrid cells was caused by the presence of EF-2 resistant to ADP-ribosylation by fragment A of diphtheria toxin. These facts suggest that the gene encoding EF-2 is located on human chromosome 19.
Ehrlich ascites tumor cells were found to be very insensitive to diphtheria toxin. We formed 37 hybrids from Ehrlich tumor cells and diphtheria toxin-sensitive human fibroblasts. The effects of diphtheria toxin on protein synthesis in those hybrids were examined. The hybrids were divided into three groups on the basis of toxin sensitivity. Group A hybrids were as sensitive to diphtheria toxin as human fibroblasts, Group C were as resistant as Ehrlich tumor cells, and Group B had intermediate sensitivity. Group A hybrids had diphtheria toxin-binding sites but Group B and C had no detectable binding sites. Elongation factor-2 of all the hybrids was susceptible to ADP-ribosylation by fragment A of diphtheria toxin. Cells of Group A and B became more sensitive to CRM 45 (cross-reacting material 45 of diphtheria toxin) after they were exposed to low pH (pH = 4.5). The resistance of Group C to CRM 45 was not affected by the same treatment. Group A and B hybrids and human fibroblasts had similar sensitivities to a hybrid toxin composed of wheat germ agglutinin and fragment A of diphtheria toxin, but Group C and Ehrlich tumor cells were resistant to this hybrid toxin. All the hybrids and Ehrlich tumor cells were more sensitive to a hybrid toxin composed of wheat germ agglutinin and subunit A of ricin than were human fibroblasts. On subcloning of Group B hybrids, one Group C hybrid was obtained, but no Group A hybrid. These facts suggest that Ehrlich ascites tumor cells differ from human fibroblasts in the expression of a factor(s) that is involved in entry of fragment A of diphtheria toxin into the cytoplasm after the toxin binds to its surface receptors.
A 24-year-old man whose chief complaint was male infertility for 3 years was referred to our clinic. Round nodules approximately 2 cm in diameter were observed both in the left epididymis-head and the right para-epididymal head. These tumors were removed and they were confirmed to be papillary cystadenoma histopathologically. Only 6 cases of papillary cystadenoma including our case were collected in Japan. Pathologic lesions of this disease associated with Lindau's disease were also discussed.
A case of mixed infection of Salmonella (type C2) and Parasarcophaga crassipalpis (an intestinal myiasis) in a two-year-old girl is reported. A girl after dining in a restaurant developed typical Salmonella food-poisoning symptoms. Stool culture showed both Salmonella and fly larvae. This paper is to report a first case of this fly larvae infection and to review the recent trends of food-poisoning in Japan.
Explore the source record for details and available documents.