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T Furuno

Publications and source records attributed to T Furuno.

At least 73 records · Page 4Linked to original sources
PubMed1996

Confocal fluorescence microscopy for antibodies against a highly conserved sequence in SH2 domains.

We have prepared monoclonal antibodies for a highly conserved sequence (GTFLVRESETTK) in SH2 domains. Mouse IgG1s (12E and 32D) prepared against a peptide-conjugated keyhole lympet hemocyanin specifically bound the antigenic peptide but not the carrier protein. Western blot analysis showed that one IgG1 (12E) recognized mainly 62 kDa proteins (possibly src-family tyrosine kinases) from triton X-100 extracts of RBL-2H3 cells and that another (32D) recognized mainly 32 and 110 kDa proteins. Confocal fluorescence microscopy showed that the SH2 domains had a diffuse cytoplasmic distribution and were not present in the nucleus. Following antigen stimulation, a markedly different cellular distribution was observed in the cells stained with 12E and 32D IgGs. 12E IgGs strongly stained the plasma membranes while 32D IgGs stained small granules in the cytoplasm. As 12E IgGs bound 62 kDa proteins on Western blotting, the results suggest that tyrosine kinases cluster along the plasma membranes and/or than conformational changes occur in the domains after antigen stimulation.

Amino Acid Sequence↗
PubMed1996

Surface expression of CD63 antigen (AD1 antigen) in P815 mastocytoma cells by transfected IgE receptors.

The surface expression CD63 antigen in rat basophilic leukemia cells (RBL-2H3) was observed after antigen stimulation by confocal fluorescence microscopy. The surface expression of CD63 antigen reflected the degranulation in RBL-2H3 cells. Then, we did the same experiments in P815 mastocytoma cells with transfected IgE receptors. The expression was observed in P815 cells with normal IgE receptors, but not in P815 variant cells with IgE receptors which were missing a C-terminal cytoplasmic domain of beta or gamma subunit. In addition, the expression in P815 cells with normal IgE receptors was mostly blocked by the pretreatment of herbimycin A. The results suggested that tyrosine phosphorylation of the C-terminal cytoplasmic domains of beta and gamma subunits was essential for degranulation.

Animals↗
PubMed1996

Modulatory effect of roxithromycin on human neutrophil function.

Neutrophils are thought to play a key role in tissue injury. We investigated the effect of roxithromycin, a 14-membered ring macrolide, on human neutrophil functions. The drug inhibited N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced superoxide (O2-) production and Ca2+ influx of human neutrophils. The inhibition was overcome by adding an inhibitor of cyclic AMP-dependent protein kinase (PKA), H-89. These results suggest that the drug affects O2- production and intracellular Ca2+ concentration of neutrophils via the action of PKA. Moreover, roxithromycin ameliorated endothelial cell injury induced by neutrophils, which may be, in part, due to the effect of the drug on neutrophils. Thus, roxithromycin may contribute to the treatment of diseases worsened by the excessive action of neutrophils.

Anti-Bacterial Agents↗
PubMed1996

[Two cases of Candida endocarditis associated with abdominal disease].

Two cases of Candida endocarditis are reported. The first case was of a 63-year-old man who had a positive blood culture for Candida albicans during treatment for liver abscess and early gastric cancer. He was transferred to our department, and aortic and tricuspid regurgitation due to Candida endocarditis was diagnosed. The patient was successfully treated with aortic valve replacement, tricuspid valve plasty and anti-fungal agents. The second case was of a 65-year-old man who complained of fever. Despite a diagnosis of common bile duct cancer and resection of the tumor, the fever persisted. He was transferred to our department and was diagnosed having aortic regurgitation due to Candida endocarditis, complicated by heart failure. Although intense medical therapy including antifungal agents, diuretics, catecholamines and digoxin was initiated, the patient died from multiple embolisms 9 days later. In the treatment of Candida endocarditis, early diagnosis and early decision-making for either surgical or medical therapy is indispensable. Although the prevalence of Candida endocarditis is low, the differentiation of this disease should be taken into account in febrile elderly patients with long-standing therapy with antibiotics.

Aged↗
PubMed1996

[An elderly patient with Leriche syndrome complicated by congestive heart failure and rhabdomyolysis].

A 64-year-old man, who had had bilateral intermittent claudication and leg pain for five years, was admitted because of sudden onset of severe leg pain, and acute respiratory failure. Laboratory data showed markedly elevated serum CK of 73,050 IU/L, and urinary myoglobin of 430,000 ng/ml. A diagnosis of rhabdomyolysis was made. Renal dialysis was required for the next two days because of acute renal failure. The aortogram was performed on the 32nd day and disclosed complete obstruction of the abdominal aorta immediately distal to the bilateral renal arteries. Good collateral flow was noted to the popliteal arteries. On the 49th day, the patient successfully underwent extra-anatomical bypass surgery (axillo-bifemoral bypass). The mechanism of rhabdomyolysis in this elderly patient was discussed.

Heart Failure↗
PubMed1995

An inhibitor of cyclic AMP-dependent protein kinase enhances the superoxide production of human neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine.

Intact human neutrophils produced superoxide (O2-) by the stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP) even when the extracellular Ca2+ was absent (0.56 +/- 0.13 nmol/min per 10(6) cells). The production by fMLP was enhanced more than twice in the presence of the extracellular Ca2+. Moreover, the O2- production by fMLP in the presence of extracellular Ca2+ was enhanced nearly three times by the treatment of cells with H-89, an inhibitor of cyclic AMP-dependent protein kinase (PKA). The enhancement was not observed when the extracellular Ca2+ was depleted from the reaction mixture. In addition, H-89 did not enhance fMLP-induced O2- production of electropermeabilized neutrophils in which the intracellular Ca2+ concentration was fixed to about 100 nM. These observations suggest that not only Ca2+ influx but the inhibition of PKA is necessary for the maximum O2- production by fMLP and that the O2- production is partially suppressed by the activation of PKA induced by fMLP.

Bucladesine↗
PubMed1995

Confocal fluorescence microscopy for studying thapsigargin-induced bivalent-cation entry into B cells.

We studied thapsigargin-induced bivalent-cation entry into antigen-specific B cells (TP67.21) with a confocal fluorescence microscope. Confocal fluorescence images of fluo-3-loaded B cells showed that thapsigargin-stimulated Ca2+ signals were transferred not only to the cytoplasm but also to the nucleus. In the absence of external Ca2+ ions, the free Ca2+ concentrations both in the cytosol and in the nucleus declined to basal levels by 5 min after addition of thapsigargin. However, subsequent addition of Ca2+ in the external medium made the fluo-3 (fura-2) fluorescence intensity rise, reflecting the fact that Ca2+ accumulated again in the nucleus as well as in the cytoplasm. Then, we added Ba2+ and Mn2+ instead of Ca2+, because Ba2+ and Mn2+ are known to enter via Ca2+ channels. The addition of Ba2+ and Mn2+ in the external medium quenched the fluo-3 fluorescence both in the nucleus and in the cytoplasm of B cells. This suggested the possibility that the increase in intranuclear Ca2+ after thapsigargin stimulation may come from the cytoplasm, not from the nuclear stores.

Aniline Compounds↗
PubMed1995

Case report: the first report of idiopathic hypereosinophilic syndrome involved with lung and middle ear.

It has been reported that various organs are involved in idiopathic hypereosinophilic syndrome. Frequently, the heart, lung, skin, and nervous system are involved. Involvement of the middle ear, however, has not yet been reported. In this article, the authors describe the first case of hypereosinophilic syndrome involving the lung and middle ear. A 39-year-old woman had a 4-month history of low grade fever, non-productive cough, and a feeling of fullness and hearing loss in both ears. Peripheral blood cell count showed eosinophilia. Bilateral tympanic cavities were obstructed with granulation tissue, and she was diagnosed as obliterative otitis media. The granulation tissue consisted of foamy histiocytes and eosinophils. Chest X-ray film and computed tomography showed patchy infiltrative shadow in the lung. Histologic examination of the open lung biopsied specimen showed alveolar spaces infiltrated by eosinophils. After treatment with 30 mg oral prednisolone daily, there was a rapid improvement in her clinical condition. Based on the clinical course and the histologic findings of this case, obliterative otitis media may be caused by eosinophilic infiltration and eosinophilic pneumonia.

Adult↗
PubMed1995

Serum cardiac troponin T in patients with acute myocardial infarction. Detection of coronary reperfusion and prediction of cardiac function.

Serum troponin T, a myocardial contractile protein, has been reported to be a sensitive marker for the diagnosis of acute myocardial infarction. However, there have been few reports on its ability to detect coronary reperfusion and to predict left ventricular function in the chronic stage. Twenty two patients (20 males and 2 females, 61 +/- 10 y.o.) with acute myocardial infarction were enrolled in this study. They were divided into 2 groups, one with successful reperfusion (group A: n = 13) and one without reperfusion (Group B: n = 9) and the serial changes of their serum troponin T levels were evaluated. Serum myosin light chain was measured in another group of patients with acute myocardial infarction without history of old myocardial infarction (group C: n = 8). The slope of the logarithm of serum troponin T on a time-value curve was calculated from the time of admission to the first peak within 24 hours of the onset of acute myocardial infarction. The correlation coefficient between the late peak of serum troponin T and the left ventricular ejection fraction in 11 patients with first Q wave acute myocardial infarction was compared with that between the serum myosin light chain peak and the left ventricular ejection fraction in group C. 1) The slope of the logarithm of serum troponin T on the time-value curve in group A was greater than that in group B (0.57 +/- 0.45 vs. 0.22 +/- 0.16) (p < 0.05). 2) There was a good correlation between the late peak level of serum troponin T (78 +/- 10 hours after the onset) and the left ventricular ejection fraction in 11 patients with first Q wave acute myocardial infarction (r = -0.84, p < 0.01), which was similar to that of the serum myosin light chain peak and the left ventricular ejection fraction (r = -0.72, p < 0.05). On the other hand, there was no correlation between the peak level of serum creatine phosphokinase and the left ventricular ejection fraction (r = -0.55, NS). The serum troponin T levels 24, 36, 48 and 60 hours after the onset also correlated well with the left ventricular ejection fraction (r = -0.65, -0.7, -0.65 and -0.89, respectively). We conclude that the serial measurement of serum troponin T in patients with acute myocardial infarction is useful in the evaluation of left ventricular function in the chronic stage and that it is a potential non-invasive predictor of coronary reperfusion.

Aged↗
PubMed1995

Mechanical measurements of single actomyosin motor force.

To elucidate the mechanism of force generation by actomyosin motor, a measuring system was constructed, in which an in vitro motility assay was combined with an optical trapping technique. An actin filament of several micron long was attached to a gelsolin-coated polystyrene bead, and was allowed to interact with a small number (approximately 1/1 micron actin filament) of rabbit skeletal heavy meromyosin (an active subfragment of myosin) molecules bound to a nitrocellulose-coated coverglass. The bead position was determined at 33-ms intervals. We measured the force generation event at relatively low (100-400 nM) ATP concentration so that the occurrence of individual force generation events could be detected with our time resolution. The actin-bound bead held in the optical trap moved in a stepwise manner in the direction of the actin filament only in the presence of ATP. At the trap strength of 0.3 pN/nm, the maximum size of the step was 11 nm, and the maximum force associated with the movement was 3.3 pN.

Actins↗
PubMed1994

Thapsigargin-induced nuclear calcium signals in rat basophilic leukaemia cells.

By a confocal fluorescence microscope with an argon-ion laser (488 nm) and a He-Cd laser (325 nm) we have studied thapsigargin-induced calcium signals in individual rat basophilic leukaemia (RBL-2H3) cells. In the presence or absence of external calcium ions, thapsigargin-induced calcium signals were transferred to the nucleus as well as to the cytoplasm of RBL-2H3 cells. The calcium signals were generally much stronger in the nucleus than in the cytoplasm. However, some of the RBL-2H3 cells had apparently reduced nuclear calcium signals. They had a basophil-like bilobed (multilobed) nucleus, although most RBL-2H3 cells had a mast-cell-like monolobed nucleus. In the cells with a bilobed nucleus, IgE-receptor-mediated calcium signals were neither transferred to the nucleus nor to the cytoplasm. The results gave a new insight into the understanding of the mechanism of the nuclear calcium signals in RBL-2H3 cells.

Animals↗
PubMed1994

Interaction between ganglioside-containing liposome and rat T-lymphocyte: confocal fluorescence microscopic study.

Interaction between rat T-lymphocytes and a ganglioside (GM3, GD3, GT1b, or GQ1b)-containing liposome was investigated in vitro. The direct stimulation of T cell by a ganglioside-containing liposome was followed by monitoring an increase in the intracellular calcium signal using a confocal laser fluorescence microscopic method. The GT1b- or GQ1b-containing liposome strongly stimulated the cell, while the GM3- or GD3-containing liposome showed much less effect. Free gangliosides did not stimulate the cell at all under the same condition. The extent of the stimulation depended on the surface density of GT1b on the liposome accompanied by a clear threshold concentration. The efficiency of this direct T cell stimulation with the ganglioside-containing liposome was closely related to the efficiency of the tumor growth suppression reported previously by ourselves.

Aniline Compounds↗
PubMed1994

Effects of herbimycin A and ST638 on Fc epsilon receptor-mediated histamine release and Ca2+ signals in rat basophilic leukemia (RBL-2H3) cells.

We examined the effect of the two protein tyrosine kinase inhibitors, alpha-cyano-3-ethoxy-4-hydroxy-5-phenylthiomethylcinnamide (ST638) and herbimycin A, on the activation processes of rat basophilic leukemia (RBL-2H3) cells by cross-linking of IgE receptors. RBL-2H3 cells sensitized with DNP-specific monoclonal IgE antibody were stimulated with multivalent antigen (DNP conjugate of bovine serum albumin). Analysis of phosphotyrosine-containing proteins in their lysates by SDS-PAGE and immunoblotting revealed that these two inhibitors efficiently inhibited the tyrosine phosphorylation of several proteins (32, 42, 56, 66, 72, 92, 150 kDa) including phospholipase C-gamma 1. The inhibitors also caused parallel inhibitions of the histamine release, the formation of inositol 1,4,5-trisphosphate, and the increase in cytosolic calcium ion concentration at the late sustained phase. A digital imaging fluorescence microscopic analysis of antigen-dependent calcium signals in individual cells showed that these two tyrosine kinase inhibitors inhibited the calcium influx from the external medium more powerfully than the mobilization of calcium ion from internal stores. In contrast, the inhibitors did not affect the increase in the cytosolic calcium ion concentration or the histamine release induced by the calcium ionophore A23187. Taken together, our results suggest that tyrosine phosphorylation following antigen stimulation regulates phosphatidylinositol hydrolysis and the influx of extracellular calcium.

Animals↗
PubMed1994

Tyrosine-specific protein kinase participates in the pathogenesis of acute immune complex alveolitis in rats.

Rabbit IgG antibodies against ovalbumin (OA) was injected intravenously into Wistar rats. When the animals were challenged with OA aerosolized by ultrasonic nebulization, acute lung injury occurred as reflected by increased recovery of bronchoalveolar cells, especially polymorphonuclear leukocytes (PMN) in bronchoalveolar lavage fluid (BALF). Lung morphology demonstrated cellular infiltration in the alveolar septa and intra-alveolar hemorrhage. When the rats were administered ST-638, a novel tyrosine kinase inhibitor, intraperitoneally prior to nebulization, the number of PMN in BALF decreased in a dose-dependent manner and superoxide anion (O2-)-producing activity in peripheral leukocytes was significantly suppressed. Furthermore, the reagent inhibited migration of human peripheral blood neutrophils induced by the chemotactic peptide f-met-leu-phe in vitro. These studies strongly indicate that tyrosine kinase plays an important role in immune complex-triggered neutrophil-related lung disorders, and the novel tyrosine kinase inhibitor ST-638 attenuates lung injury by preventing superoxide production and neutrophil migration.

Animals↗
PubMed1994

An initial signal of activation of rat peritoneal mast cells stimulated by Datura stramonium agglutinin: a confocal fluorescence microscopic analysis of intracellular calcium ion and cytoskeletal assembly.

A confocal fluorescence microscope using fluo-3 and 9-(dicyanovinyl)- julolidine (DCVJ) was used to study the mast cell activation by the N-acetyl glucosamine oligomer specific lectin Datura stramonium agglutinin (DSA) and inhibition by antagonist lectins having affinity to N-acetyl glucosamine (GlcNAc). DSA induced a transient increase in intracellular free calcium concentration ([Ca2+]i) followed by cytoskeletal disassembly and reassembly in rat peritoneal mast cells. These changes induced by DSA resulted in histamine release. The time course of fluorescence intensity in mast cells loaded with fluo-3- or DCVJ and activated by DSA resembled those activated by the basic polymer compound 48/80. Inhibition of [Ca2+]i rise by antagonist lectins was responsible for the inhibition of cytoskeletal assembly and the consequent histamine release induced by DSA. At the level of the individual cell, a mast cell stimulated by DSA responds in an all-or-none fashion. DSA possible induced intracellular calcium mobilization and cytoskeletal change by recognizing the GlcNAc-oligomer residues of specific glycoproteins of mast cells.

Agglutinins↗