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Biomedical subjects

Y Umezawa

Publications and source records attributed to Y Umezawa.

At least 55 records · Page 3Linked to original sources
PubMed1999

Specific inhibition of TH2-type cytokine production from human peripheral T cells by terfenadine in vitro.

BACKGROUND: Cytokine imbalance is thought to be one of the causes for allergic diseases. The effect of anti-allergic drugs on cytokine production from T cells should be examined in a convenient way. OBJECTIVES: To study the in vitro effect of terfenadine, a prototype non-sedating H1 receptor antagonist, on cytokine production from activated T cells. METHODS: T cells were cultured in the presence of terfenadine on anti-CD3 mAb and anti-CD26 mAb-coated wells, anti-CD3 mAb and anti-CD28 mAb-coated wells, and anti-CD3 mAb wells with PMA. T-cell proliferation, along with the concentrations of interleukin (IL) -2, interferon (IFN) -gamma, IL-4, and IL-5 were measured. RESULTS: Terfenadine inhibited T-cell proliferation and IL-4 and IL-5 production under each costimulatory condition tested, whereas it had no effect on IL-2 and IFN-gamma production. CONCLUSIONS: These results indicate that terfenadine has a specific inhibitory effect on TH2-type cytokine production induced by several ways of costimulatory activation.

Cells, Cultured↗
PubMed1999

Non-linear analysis of the sleep EEG.

A sleep electroencephalogram was analyzed by non-linear analysis. The polysomnography of a healthy male subject was analyzed and the correlation dimensions calculated. The correlation dimensions decreased from the 'awake' stage to sleep stages 1-3 and increased during rapid eye movement (REM) sleep. These results were seen during each sleep cycle. In each sleep cycle, the correlation dimensions decrease for slow wave sleep, and increase for REM sleep.

Adult↗
PubMed1998

An assay method for evaluating chemical selectivity of agonists for insulin signaling pathways based on agonist-induced phosphorylation of a target peptide.

An optical method for evaluating the physiologically relevant agonist and antagonist selectivity of an insulin signaling pathway based on an insulin-dependent on/off switching of phosphorylation of a target peptide via insulin receptor is described. Insulin receptor serves as a binding for insulin and a given insulin receptor-binding peptide as a target for an insulin-receptor complex. Upon binding of insulin to its receptor, the insulin receptor undergoes autophosphorylation which enables the receptor to have a kinase activity and phosphorylate various substrates. The phosphorylated tyrosine in the substrate was measured with a monoclonal anti-phosphotyrosine antibody. As the target substrate for insulin receptor, a Y939 peptide consisting of 12 amino acid residues derived from insulin receptor substrate 1 (IRS-1) was used. The present assay method involves different sequential steps: (1) immobilization of a biotin-coupled Y939 peptide on an avidin coated 96-well plate via biotin-avidin complexation; (2) insulin-dependent phosphorylation of the Y939 peptide by the insulin receptor; (3) enzymatic reaction and absorptiometric assay of the phosphorylated Y939 peptide using the anti-phosphotyrosine antibody labeled with horseradish peroxidase. An insulin-dependent absorbance was observed for insulin concentrations from 1.0 x 10(-10) to 1.0 x 10(-7) M, and it leveled off. The observed absorbance was explained to be due to an increase in the phosphorylated Y939 peptide caused by insulin and its receptor complexation. No signal was, however, induced by both vanadyl and vanadate ions at concentrations up to 1.0 x 10(-4) M; these results and previous intact cell level data taken together led to the conclusion that these ions did not induce phosphorylation of the Y939 peptide. Upon addition of tyrphostin 25, a specific inhibitor for insulin receptor kinase activity, phosphorylation of the Y939 peptide in the presence of 1.0 microM insulin was competitively inhibited over 1.0 x 10(-4) M tyrphostin 25. The present system thus exhibited "physiologically more relevant" agonist and antagonist selectivity, the principle of which is based in part on the insulin signal transduction rather than simply relying on the binding assay. The potential use of the present method for evaluating the selectivity of a wide range of agonists and antagonists toward the insulin signaling pathways is discussed.

Amino Acids↗
PubMed1998

Individual pharmacodynamics assessed by antilymphocyte action predicts clinical cyclosporine efficacy in psoriasis.

BACKGROUND: Cyclosporine (INN, ciclosporin) use for psoriasis has been proposed and clinically examined. However, individual variation in cyclosporine efficacy is currently observed. To evaluate individual therapeutic potency of cyclosporine, pharmacodynamic approaches were performed with use of peripheral blood mononuclear cells (PBMCs) from patients with psoriasis. METHODS: Cyclosporine effects on PBMC-blastogenesis were examined in 33 patients with psoriasis. The drug concentration that gave 50% inhibition of mitogen-stimulated PBMC proliferation in vitro (IC50, in nanograms per milliliter) was evaluated in each patient. Cyclosporine was administered at an initial dose of 5 mg/kg/day, and the dose was tapered for 16 weeks to 3 mg/kg/day. The recovery rate in the psoriasis area and the severity index (PASI) 16 weeks after cyclosporine therapy began was measured. RESULTS: Cyclosporine IC50 values in 33 patients deviated widely, from 0.1 to 120.6 ng/ml. We classified these patients into two groups on the basis of their PBMC sensitivity to cyclosporine with use of the median cyclosporine IC50 (3.0 ng/ml) of these patients as the cutoff point. The PASI recovery rate after cyclosporine therapy in the patients with high sensitivity was significantly higher than that in the patients with low sensitivity (p < 0.0007). Moreover, a significant negative correlation between the IC50 and the PASI recovery rate was observed in these 33 patients (r = -0.73; p < 0.0001). Blood trough levels and side effects of cyclosporine were not significantly different between the two patient groups. CONCLUSIONS: The results showed that we could use PBMCs to pharmacodynamically predict the patients with a poor response to cyclosporine therapy. These patients may require larger doses of cyclosporine or alternative approaches to treatment. The patients with PBMCs sensitive to cyclosporine should be evaluated for treatment with smaller doses of the drug to avoid serious side effects.

Adult↗
PubMed1998

CH/pi interactions as demonstrated in the crystal structure of guanine-nucleotide binding proteins, Src homology-2 domains and human growth hormone in complex with their specific ligands.

The CH/pi interaction is a weak attractive molecular force occurring between CH groups and pi-systems. Possibility has been examined for the role of CH/pi interaction, by use of a computer program, in the crystallographic data of several guanine-nucleotide binding proteins, src homology-2 domains and human growth hormone complexed with their specific ligands. Short CH/pi contacts have been found in every case where cohesive forces are expected. Comparison of the structures of functionary related proteins has shown that mutation may occur but necessary CH/pi interactions are conserved. A considerable part of the non-polar interactions, broadly ascribed in the past to the van der Waals interaction or the so-called hydrophobic effect, has been suggested to be attributed to a more specific attractive force, the CH/pi interaction.

Animals↗
PubMed1998

CH/pi interactions in the crystal structure of class I MHC antigens and their complexes with peptides.

The crystal structure of class I major histocompatibility complex antigens (MHC) bound to their specific ligand peptides were analyzed, in the context of the CH/pi interaction, with use of a computer program CHPI. A number of short CH/Csp2 distances have been shown at the boundary of the heavy chain and beta2 microglobulin. These interactions are conserved between species, human versus murine. A number of contacts shorter than the conventional van der Waals distance have been disclosed between CH hydrogens and aromatic side-chain groups in the MHC/peptide complexes. The CH/pi interaction has been suggested to contribute to the specificity in the complex formation of class I MHC.

Amino Acid Sequence↗
PubMed1998

Activation of peripheral blood monocytes and macrophages in Kawasaki disease: ultrastructural and immunocytochemical investigation.

Monocytes/macrophages are considered to play an important role in the pathogenesis of Kawasaki disease (KD). However, the morphological and immunocytochemical features of the cells in acute KD have not been investigated. The ultrastructural and immunocytochemical characteristics of peripheral blood CD14+ monocytes/macrophages sorted by a magnetic cell sorter (MACS) during the course of KD were, therefore, studied to evaluate their role in the disease pathogenesis. Electron microscopy showed that CD14+ monocytes/macrophages from patients with acute KD had nuclei with complex shapes, apparent nucleoli and abundant intracytoplasmic granules, some of which were positive for acid phosphatase. The quantity of intracytoplasmic granules was correlated with disease severity, in terms of the duration of fever, maximum level of C-reactive protein and the presence of coronary artery lesions (CAL), suggesting that the monocytes/macrophages were activated and showed increased phagocytosis. Immunocytochemical staining of smears made from cell suspensions of sorted CD14+ monocytes/macrophages was carried out using a monoclonal antibody against tumor necrosis factor (TNF)-alpha. The cytoplasm of monocytes/macrophages from patients with acute KD was strongly positive in comparison to that of cells from control subjects, suggesting that intracytoplasmic granules secrete TNF-alpha.

Acid Phosphatase↗
PubMed1997

An optical method for evaluating ion selectivity for calcium signaling pathways in the cell.

A method for evaluating a physiologically relevant ion selectivity of Ca2+ signaling pathways in biological cells based on a Ca(2+)-dependent on/off switch for cellular processes via calmodulin (CaM) chemistry is described. CaM serves as a primary ion receptor for Ca2+ and a given CaM-binding peptide as a target for a CaM-Ca2+ complex. Upon accommodating four Ca2+ ions in its binding sites, CaM undergoes a conformational change to form a CaM-Ca(2+)-target peptide ternary complex. This Ca(2+)-induced selective binding of the Ca(2+)-CaM complex to the target peptide was monitored by a surface plasmon resonance (SPR) technique. As a target peptide, a 26-amino acid residue of M13 derived from skeletal muscle myosin light-chain kinase was used. The target peptide was covalently immobilized in the dextran matrix on top of gold, over which sample solutions containing Ca2+ and CaM were injected in a flow system. Ca(2+)-dependent SPR signals were observed for Ca2+ concentrations from 3.2 x 10(-8) to 1.1 x 10(-5) M and it leveled off. The observed SPR signals were explained as due to an increase in the refractive indexes caused by a Ca2+ ion-switched protein/ peptide interaction, i.e., Ca2+ ion to CaM and subsequent additional binding of the thus formed complex with immobilized M13. No SPR signals were however, induced by Mg2+, K+, and Li+ at concentrations as high as 1.0 x 10(-1) M; these results and previous spectroscopic data taken together conclude that these ions do not induce CaM/peptide interaction. Large changes in SPR signals were observed with a Sr2+ ion concentration over 5.1 x 10(-4) M; Sr2+ ion behaved in this case as a strong agonist toward the Ca(2+)-dependent on/off switch of CaM. The present system thus exhibited "physiologically more relevant" ion selectivity in that relevant metal ions could switch on the CaM/peptide or -protein interaction rather than merely be bound to CaM causing no further signal transduction. The potential use of this finding for more widely evaluating cation selectivity toward the Ca2+ signaling process was discussed.

Amino Acid Sequence↗
PubMed1997

Receptor based chemical sensing.

This mini-review describes our recent approach of mimicking transmembrane and intracellular signalings displayed by various receptors in biomembranes for the development of new sensing membranes. Several important modes of receptor signaling have been utilized exploiting bio and synthetic receptors; (i) Ca2+ signaling by calmodulin, (ii) active transport of target and relevant compounds displayed by Na+/D-glucose cotransporter and Na+,K(+)-ATPase, (iii) membrane permeability changes induced by glutamate receptor ion channel proteins and (iv) membrane potential changes induced by synthetic receptors. The newly designed sensing systems are demonstrated and discussed in terms of their novel mode of signal transduction, sensitivity and selectivity.

Biological Transport↗
PubMed1997

Immunosuppressant pharmacodynamics on lymphocytes from healthy subjects and patients with chronic renal failure, nephrosis, and psoriasis: possible implications for individual therapeutic efficacy.

BACKGROUND: In organ transplantation, patients with peripheral blood mononuclear cells (PBMCs) that exhibit resistance to cyclosporine (INN, ciclosporin) or glucocorticoids in vitro are refractory to therapy based on these drugs in vivo. However, detection or distribution of the resistant patients with immunologic disorders remains to be documented. METHODS: Drug sensitivity tests were performed with PBMCs from four subject groups: 69 healthy subjects, 100 patients with chronic renal failure, 38 patients with nephrosis, and 51 patients with psoriasis. The values for the concentration that produces 50% lymphocyte-mitosis inhibition (IC50) of the drugs on PBMC blastogenesis were estimated, and individual variations or group differences in the IC50 values were examined. RESULTS: The median cyclosporine IC50 values of the four subject groups were similar, but large individual deviations in the IC50 values were observed. Individual differences in prednisolone IC50 values were spread from 1 to 3500 ng/ml. When compared with healthy subjects, a significantly large number of the patients with chronic renal failure group exhibited low responses to prednisolone (p < 0.04). In contrast, no significant difference in the methylprednisolone IC50 was observed among the groups. Normal upper thresholds for IC50 values of these drugs were estimated from the mean + 2 standard deviations (SD) of the IC50 values of healthy PBMCs, and the patients with IC50 values above these levels were considered to be resistant. The incidence of resistant patients with nephrosis or psoriasis was similar to that of healthy subjects; however, the incidence of cyclosporine- or prednisolone-resistant subjects with chronic renal failure was significantly higher (p < 0.04). Significant correlations between PBMC sensitivity to cyclosporine in vitro and clinical efficacy of the drug in vivo were observed in renal transplant recipients and in patients with psoriasis. CONCLUSIONS: A large subset of patients with chronic renal failure showed PBMC resistance to cyclosporine and prednisolone. Hyperresistant patients have a high risk of being refractory to immunosuppressive therapy with one of these drugs. Alternative treatment should be considered according to the individual drug-sensitivity data.

Adult↗
PubMed1997

Electrochemical evaluation of chemical selectivity of glutamate receptor ion channel proteins with a multi-channel sensor.

A new method for evaluating chemical selectivity of agonists towards receptor ion channel proteins is proposed by using glutamate receptor (GluR) ion channel proteins and their agonists N-methyl-D-aspartic acid (NMDA), L-glutamate, and (2S, 3R, 4S) isomer of 2-(carboxycyclopropyl)glycine (L-CCG-IV). Integrated multi-channel currents, corresponding to the sum of total amount of ions passed through the multiple open channels, were used as a measure of agonists' selectivity to recognize ion channel proteins and induce channel currents. GluRs isolated from rat synaptic plasma membranes were incorporated into planar bilayer lipid membranes (BLMs) formed by the folding method. The empirical factors that affect the selectivity were demonstrated: (i) the number of GluRs incorporated into BLMs varied from one membrane to another; (ii) each BLM contained different subtypes of GluRs (NMDA and/or non-NMDA subtypes); and (iii) the magnitude of multi-channel responses induced by L-glutamate at negative applied potentials was larger than at positive potentials, while those by NMDA and L-CCG-IV were linearly related to applied potentials. The chemical selectivity among NMDA, L-glutamate and L-CCG-IV for NMDA subtype of GluRs was determined with each single BLM in which only NMDA subtype of GluRs was designed to be active by inhibiting the non-NMDA subtypes using a specific antagonist DNQX. The order of selectivity among the relevant agonists for the NMDA receptor subtype was found to be L-CCG-IV > L-glutamate > NMDA, which is consistent with the order of binding affinity of these agonists towards the same NMDA subtypes. The potential use of this approach for evaluating chemical selectivity towards non-NMDA receptor subtypes of GluRs and other receptor ion channel proteins is discussed.

Animals↗
PubMed1997

Different regulatory effects of pentoxifylline on human T cell activation pathways.

Pentoxifylline (PTX), a methylxanthine derivative, was examined for its effects on T cell proliferation and cytokine production stimulated by cross-linking anti-CD3 alone, anti-CD3 with PMA, anti-CD3 with anti-CD26, or anti-CD3 with anti-CD28 mAb, respectively. PTX at a 3.5 x 10(-5) M concentration significantly inhibited T cell proliferation and the production of tumor necrosis factor-alpha, interleukin-2, and interleukin-4. Moreover, this effect was selective for stimulation by cross-linking anti-CD3 with PMA, or anti-CD3 with anti-CD26, but not by cross-linking anti-CD3 with anti-CD28. These results suggest that the inhibitory effect of PTX on T cell activation involves the CD3 and CD26, but not the CD28 signal pathway.

Cell Survival↗
PubMed1997

Decrease in the concentrations of transforming growth factor-beta 1 in the sera of patients with Kawasaki disease.

Kawasaki disease (KD) is one of the most important forms of vasculitis, and is characterized by the initiation of a proinflammatory cytokine cascade. To further characterize the immunological profile of KD, we measured the serum levels of transforming growth factor-beta 1 (TGF-beta 1) as a regulatory cytokine. We determined the concentration of TGF-beta 1 in the sera of the patients with KD, anaphylactoid purpura (AP), and scarlet fever, using a sandwich enzyme linked immunosorbent assay. The serum levels of TGF-beta 1 were decreased in patients with KD, but not in patients with AP or scarlet fever during the acute stage. We found an inverse correlation between TGF-beta 1 and soluble tumor necrosis factor (TNF) receptor levels in KD patients during the acute and subacute stage. Decreased levels of TGF-beta 1, in particular to suppress TNF alpha (TNF-alpha) production, is an important part of the regulatory system of increased TNF-alpha production which cause vasculitis.

Adolescent↗
PubMed1995

A case of drug eruption caused by the crude drug Boi (Sinomenium stem/Sinomeni caulis et Rhizoma).

We report a case of drug eruption caused by the crude drug Boi. A 41-year-old female patient had been diagnosed with chronic rheumatoid arthritis in the department of internal medicine. After ingestion of a decoction of the crude drug Boi for the alleviation of arthralgia, a slight fever developed, which was followed by systemic edematous erythema with itching. HPLC showed that the main components of the crude drug Boi are sinomenine and magnoflorine. The results of patch tests were negative for all oral drugs that the patient had been taking. Oral ingestion tests showed that the patient showed positive reactions to the as-is Boi boiling-water decoction and 1/10-volume sinomenine. Based on this, the drug eruption was judged to be caused by sinomenine. It is considered the first time that the causative component of a drug eruption was confirmed by oral ingestion tests with components of a crude drug of Kampo medicine (Sino-Japanese traditional medicine).

Adult↗
PubMed1994

Construction of a human full-length cDNA bank.

We aimed to construct a full-length cDNA bank from an entire set of human genes and to analyze the function of a protein encoded by each cDNA. To achieve this purpose, a multifunctional phagemid shuttle vector, pKA1, was constructed for preparing a high-quality cDNA library composed of full-length cDNA clones which can be sequenced and expressed in vitro and in mammalian cells without subcloning the cDNA fragment into other vectors. Using this as a vector primer, we have prepared a prototype of the bank composed of full-length cDNAs encoding 236 human proteins whose amino acid sequences are identical or similar to known proteins. Most cDNAs contain a putative cap site sequence, some of which show a pyrimidine-rich conserved sequence exhibiting polymorphism. It was confirmed that the vector permits efficient in vitro translation, expression in mammalian cells and the preparation of nested deletion mutants.

Animals↗
PubMed1994

Pentoxifylline and intravenous gamma globulin combination therapy for acute Kawasaki disease.

We compared the efficacy of oral administration of pentoxifylline (PTX) and intravenous infusions of gamma globulin (IVGG) combination therapy with that of IVGG in reducing the frequency of coronary-artery lesions (CAL) in children with Kawasaki disease (KD), in a randomized trial. All patients with KD received acetylsalicylic acid (30 mg/kg per day), until the 30th day, after the onset of fever, followed by daily acetylsalicylic acid at a dose of 3-5 mg/kg per day there-after, and intravenous IVGG, 200 mg/kg per day, for 5 consecutive days. In addition, patients randomly assigned to PTX and IVGG combination therapy groups received oral PTX at a dosage of 10 mg/kg per day (low-dose) or 20 mg/kg per day (high-dose), in three divided doses until the 30th day. Patients with KD were all free from CAL prior to treatment. We assessed the presence of CAL by two-dimensional echocardiography which was also done prior to treatment and then twice a week after hospital admission. We detected CAL in 3 of 18 patients (16.7%) in the IVGG therapy group, as compared with 2 of 18 patients (11.1%) in the low-dose PTX and IVGG combination therapy group. There were no significant difference between the two groups. In the next study, we detected CAL in 3 of 21 patients (14.3%) in the IVGG therapy group, as compared with none of 22 patients (0%) in the high-dose PTX and IVGG combination therapy group (chi 2 = 6.4, P < 0.02). No adverse side-effects were observed in 79 patients with KD.(ABSTRACT TRUNCATED AT 250 WORDS)

Aspirin↗
PubMed1994

Determination of IgG subclasses of immunoglobulin preparations for intravenous injection and the problems involved.

The IgG subclasses of immunoglobulin for intravenous injection (IVIg) were investigated after various different types of treatment. The level of IgG2 was relatively low in preparations treated with pepsin, but by changing the clone producing anti-IgG2 antibody, the level approached that of normal human serum. Though undetectable in sulfonated immunoglobulin preparations, IgG3 was detected after reoxidation. When determining the IgG subclasses of IVIg, it must be kept in mind that the apparent value may subsequently become lower due to the depressed reaction of the antibody.

Antibodies, Viral↗