PubMed Health⌕ Search

Biomedical subjects

K Tsujimura

Publications and source records attributed to K Tsujimura.

35 records · Page 2Linked to original sources
PubMed1994

Visualization and function of vimentin phosphorylation by cdc2 kinase during mitosis.

To investigate the role of intermediate filament (IF) protein phosphorylation by cdc2 kinase during mitosis, we developed a monoclonal antibody 4A4 recognizing Ser55-phosphorylated vimentin. Western blotting indicated that this antibody reacted with vimentin phosphorylated by cdc2 kinase but not with non-phosphorylated vimentin or with vimentin phosphorylated by other kinases such as cAMP-dependent protein kinase, protein kinase C, or Ca(2+)-calmodulin-dependent protein kinase II. Immunofluorescence and immunoelectron microscopy showed that vimentin Ser55 residues distributed in the entire cytoplasmic vimentin filament system are phosphorylated when the cells enter mitosis and dephosphorylated in cytokinesis. All cell lines examined showed a similar appearance of immunoreactivity with antibody 4A4. Fractionation of mitotic cell extracts on Mono-Q Sepharose revealed a single peak of vimentin Ser55 kinase activity, and the anti-p34cdc2 antibody reacted with the 34 kDa band in the kinase containing fractions. Vimentin Ser55 kinase activities were nil in the interphase cell extract. Immunofluorescent evidence using antibody 4A4 and biochemical analysis using vimentin Ser55 peptide showed that the degree of disassembly of vimentin filament of various cell types at early mitotic phase correlated well with the amount of mitotically activated cdc2 kinase.

Amino Acid Sequence↗
PubMed1994

Identification of phosphorylation sites on glial fibrillary acidic protein for cdc2 kinase and Ca(2+)-calmodulin-dependent protein kinase II.

We identified the phosphorylation sites of glial fibrillary acidic protein (GFAP) for cdc2 kinase and Ca(2+)-calmodulin (CaM)-dependent protein kinase II. GFAP was phosphorylated to approximately 0.2 mol of phosphate/mol of GFAP by cdc2 kinase, and this phosphorylation did not induce disassembly of the filament structure. On the other hand, GFAP was phosphorylated to approximately 1.9 mol of phosphate/mol of GFAP by Ca(2+)-CaM-dependent protein kinase II, and this phosphorylation did induce disassembly of the filament. Sequential analysis of the purified phosphopeptides revealed that only Ser8 on GFAP was phosphorylated by cdc2 kinase, whereas Ser13, Ser17, Ser34, and Ser389 on GFAP were phosphorylated by Ca(2+)-CaM-dependent protein kinase II.

Amino Acid Sequence↗
PubMed1994

Expression of vimentin in gastric cancer: a possible indicator for prognosis.

Expression of vimentin in gastric cancer was immunohistologically examined in 85 surgically resected specimens and evaluated in terms of various clinicopathological features. Vimentin expression was more pronounced in gastric cancers of more advanced type and in those exhibiting a higher degree of infiltrative growth, lymph node involvement and vascular invasion. These results indicate that vimentin expression by gastric cancer might be related to the invasion and metastatic process of cancer cells and could be useful as a prognostic indicator for patients with gastric cancer.

Aged↗
PubMed1993

Phosphorylation of synthetic vimentin peptides by cdc2 kinase.

Synthetic peptide representing the site Ser-41 in vimentin, Leu-Gly-Ser41-Ala42-Leu-Arg44-Arg-Arg-NH2, and its analogs in which Ala-42 was replaced by various amino acids were tested as substrates for cdc2 kinase. Among them, the analog containing sarcosine as well as proline was an excellent substrate. The result suggests that the N-substituted structure of proline immediately following the site is important for cdc2 kinase phosphorylation. Replacement of Ala-42 by polar amino acids, especially lysine, had negative effects on peptide phosphorylation. The peptides in this study were also assayed with another type of proline-directed protein kinase, tau protein kinase II. The substrate specificity differed essentially from that of cdc2 kinase.

Animals↗
PubMed1993

Impairment of cefdinir absorption by iron ion.

The effect of iron ion on the absorption of cefdinir, a new oral cephalosporin derivative, was evaluated in healthy male volunteers in a randomized three-way crossover study. The subjects received 200 mg cefdinir alone, 200 mg cefdinir and two tablets of iron ion concomitantly, and two tablets of iron ion preparation 3 hours after 200 mg cefdinir administration. The area under the concentration curve [AUC(0-12)] of cefdinir with concurrent iron was significantly smaller than that with cefdinir alone (mean +/- SD, 0.78 +/- 0.38 versus 10.3 +/- 1.35 micrograms.hr/ml). While there were no differences in AUC(0-3) between drug alone and drug with iron 3 hours later, the AUC(3-12) with delayed iron was significantly smaller than that of cefdinir alone (4.60 +/- 1.54 versus 8.03 +/- 1.72 micrograms.hr/ml). These findings suggest that the mechanism of interaction between cefdinir and iron ion preparation is the formation of a chelation complex and that this complex probably restricts gastrointestinal absorption.

Adult↗
PubMed1991

[Morphological and functional features of endodermal cells in rat yolk sac, with special reference to the fetal macrophage differentiation].

Morphological and functional features of the yolk sac endodermal cells with special reference to the fetal macrophage differentiation were investigated morphologically under the light and electron microscopes and immunologically with the antigen phenotypic analysis and the phagocytic activity-test, using the syngeneic DA rat-embryos from 8 to 16 days of gestation. Based on the staining property with toluidin blue and the ultrastructural features, the endodermal layer from day 8 to 16-yolk sacs has been known to consist of two kinds of cell type; 10% "clear" cells with clear cytoplasm and 90% "dark" cells with dark cytoplasm. Numerous primary lysosomes, phagolysosomes, lipid droplets and coated vesicles distributed preferentially in the supranuclear portion of endodermal cells. A broad intercellular space was found between "clear" cells and "dark" cells, indicating the loose intercellular binding. It was often found that "clear" cells tend to migrate from the endodermal layer into the mesenchymal layer, where the poor development of basement membrane was seen between them. Cells phagocytosing red blood cells, that resemble morphologically "clear" cells, were also observed in the fetal liver. At ten hours after latex-injection into the yolk sac cavity of 14 days embryos, some cells which phagocytosed latex beads in their cytoplasm were found in the endodermal layer, and also in the liver tissue and loose connective tissue of fetus. These cells were stained positively with monoclonal antibody Mar3 which recognizes preferentially rat-mononuclear phagocyte system. In vitro-latex uptake of separated endodermal cells was also demonstrated by the culture-study of endodermal cell suspension. The present findings indicate that the yolk sac-endodermal layer derived from the proximal endoderm consists of at least two kinds of cell-population with a great similarity to tissue macrophages in morphological and functional senses, and support the concept that some cell-populations of endodermal layer may migrate into fetal tissue and are closely related to the differentiation of fetal macrophages and their precursors.

Animals↗
PubMed1990

Comparative studies on FcR (FcRII, FcRIII, and FcR alpha) functions of murine B cells.

Distribution of FcR II, FcRIII, and FcR alpha on murine splenic B cells was examined by using FITC-labeled heat-aggregated IgG of each subclass and IgA. Almost 60 to 80% of B cells expressed both FcRII and FcRIII. However, FcR alpha was expressed on only a small proportion (6%) of B cells that co-expressed FcRII. By inhibition assays with the use of cold IgG of each subclass and IgA in addition to anti-FcRII mAb (2.4G2), it was found that IgG1, IgG2a, and IgG2b utilized the same receptor (FcRII), whereas IgG3 and IgA bound only to their unique receptors, FcRIII and FcR alpha, respectively. Immune complexes IC prepared by IgG1, IgG2a, IgG2b, and IgA anti-TNP mAb with TNP-coupled SRBC inhibited the polyclonal Ig secretion and proliferative responses of B cells stimulated with either IL-4 or LPS. The inhibition of B cell activation was associated with the blockade of the membrane depolarization. Moreover, IC prepared by these antibodies caused production of suppressive B cell factor (SBF) as is the case with rabbit IgG antibody to SRBC, and SBF thus prepared regulated antibody responses in an isotype-nonspecific manner. In contrast, no inhibition for these responses or production of SBF was attained by the IC of IgG3 antibody. We concluded that FcRII and FcR alpha mediates a suppressive signal for B cells by acting on the initial step of activation, whereas FcRIII lacks this activity.

Animals↗
PubMed1990

Metabolism of 7 beta-alkyl chenodeoxycholic acid analogs and their effect on cholesterol metabolism in hamsters.

The metabolism of 7-ethyl- and 7-propyl-chenodeoxycholic acids was studied in hamsters. Both bile acid analogs were absorbed efficiently by the intestine and secreted into the bile at rates similar to those of chenodeoxycholic acid. After intraduodenal administration into bile fistula hamsters, the 7-alkyl analogs were present in bile as the glycine and taurine conjugates. The glycine/taurine ratios were: chenodeoxycholic acid, 1.9; 7-ethyl analog, 0.3; and 7-propyl analog, 0.2. After oral administration, during a 21-day feeding experiment, the 14C-labeled analogs were recovered quantitatively in the feces. Chenodeoxycholic acid was largely 7-dehydroxylated to lithocholic acid in the intestinal tract. In contrast, the 7 alpha-hydroxy group of the 7-alkyl bile acids was completely resistant to bacterial action. 7-Ethyl-chenodeoxycholic acid was transformed in part to a compound tentatively identified as 7 alpha-hydroxy-3-oxo- 7 beta-ethyl-5 beta-cholanoic acid while 7-propyl-chenodeoxycholic acid was excreted unchanged. In the hamsters used, the 7-alkyl bile acid analogs did not inhibit the bacterial dehydroxylation of chenodeoxycholic acid. At the end of the 21-day feeding period, analysis of the gallbladder bile showed that 7-methyl-, 7-ethyl-, and 7-propyl-chenodeoxycholic acids accounted for 38, 31, and 12% of total bile acids, respectively. The 7-alkyl bile acids decreased cholesterol absorption; the 7-propyl analog caused significant decrease in serum and liver cholesterol concentration. These experiments demonstrate that the 7-ethyl- and 7-propyl chenodeoxycholic acids, just like the 7-methyl-analog, are absorbed by the intestine and participate in the enterohepatic circulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Identification of (22R)-3 alpha,7 alpha,12 alpha,22- and (23R)-3 alpha,7 alpha,12 alpha,23-tetrahydroxy-5 beta-cholestanoic acids in urine from a patient with Zellweger's syndrome.

The nature of two novel C27 bile acids present as the taurine conjugates in urine from a patient with Zellweger's syndrome was studied. Bile acids conjugated with taurine were isolated from unconjugated and glycine-conjugated bile acids by means of ion-exchange chromatography. After alkaline hydrolysis of the taurine conjugates, the hydrolysate was acidified and extracted with ether; the extract was again subjected to ion-exchange chromatography to separate neutral from acidic compounds. The neutral fraction, which consisted mainly of two steroidal lactones, was treated with lithium aluminum hydride, and the reduction products were identified as (22R)-5 beta-cholestane-3 alpha,7 alpha,12 alpha,22,26-pentol and (23R)-5 beta-cholestane-3 alpha,7 alpha,12 alpha,23,26-pentol by direct comparison of their gas-liquid chromatographic behaviors and mass spectral data with those of chemically synthesized authentic samples. Thus, the chemical structure of two native bile acids present in urine from a patient with Zellweger's syndrome should be formulated as (22R)-3 alpha,7 alpha,12 alpha,22-tetrahydroxy-5 beta-cholestanoic acid and (23R)-3 alpha,7 alpha,12 alpha,12 alpha,23-tetrahydroxy-5 beta-cholestanoic acid, respectively.

Cholestanols↗
PubMed1983

[Clinical trial of carmofur (HCFU) in the treatment of malignant ovarian cancer--The first report: combination therapy with endoxan, mitomycin C, and toyomycin].

A new fluoropyrimidine antitumor agent, carmofur (HCFU, Mifurol) was administered to patients with malignant ovarian tumor. Two of these patients revealed favorable results. The first patient was a 72-year-old female, who was diagnosed as having ovarian serous cystadenocarcinoma with metastatic omental tumor at exploratory laparotomy, its size was newborn child head size. She was started on with a combination chemotherapy of Mifurol (600 mg p.o. daily), Endoxan (4 mg/kg i.v. twice a week), Mitomycin C (0.04 mg/kg i.v. twice a week) and Toyomycin (0.01 mg/kg i.v. twice a week). After four weeks, this combination therapy brought her a complete response with disappearance of pleural effusion, ascites and metastatic tumor. The second case was a 39-year-old female, who underwent adnexectomy elsewhere which led to the discovery of Krukenberg tumor, and was referred to our hospital. After the first course of the same combination chemotherapy, second look operation was performed. Histological examination of the specimen obtained by metastatic tumor of uterosacral ligament showed the degeneration (grade II b-III, Oboshi and Shimozato) of cancer cell. It is suggested that this combination chemotherapy including Mifurol is effective and useful for the patients with ovarian carcinoma.

Adult↗
PubMedDate not supplied

Effects of sevoflurane and halothane on reperfusion-induced arrhythmia in the isolated rat heart.

The effects of sevoflurane and halothane on reperfusion-induced arrhythmia in the isolated rat heart were examined. Reperfusion-induced ventricular fibrillation (VF) occurred in all rat hearts, and the duration of VF was 151 +/- 11 (mean +/- SEM) seconds in the control group (no anesthetics administered). Sevoflurane changed neither incidence nor duration of VF at 1, 2, and 3 minimum alveolar concentrations (MAC) of anesthetic. Halothane altered neither the incidence nor duration of VF compared with the control group at 1 MAC; however, at 2 and 3 MAC, halothane significantly reduced the incidence and duration of VF (P < 0.05 at 1, 2, and 3 MAC halothane: respective incidences were 100%, 18%, and 55% and respective durations, 82 +/- 27 seconds, 20 +/- 6 seconds, and 13 +/- 9 seconds). In the isolated rat heart, we found that halothane has antiarrhythmic effects against reperfusion-induced arrhythmia at 2 and 3 MAC. Sevoflurane, however, did not have any antiarrhythmic effects against reperfusion-induced arrhythmia at 1, 2, or 3 MAC.

Animals↗