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Biomedical subjects

T Kanda

Publications and source records attributed to T Kanda.

At least 289 records · Page 16Linked to original sources
PubMed1997

Slime formation as a marker of serious infection with methicillin-resistant Staphylococcus aureus.

Strains of methicillin-resistant Staphylococcus aureus (MRSA) are potential pathogens in patients undergoing major surgery and in patients who are immunocompromised. Some MRSA produce a viscous extracellular slime that may interfere with immune function. In this study, slime formation by MRSA was examined in cases of nosocomial infection by this pathogen at a single hospital. The antibacterial resistance, phage and enterotoxin characteristics of strains isolated from patients with fatal infections were determined. MRSA strains were classified as those associated with fatal infection; causing progressive infection; causing superficial infection; and those isolated from the nasal cavity of the health-care professionals as a control group. The incidence of slime formation was highest in the MRSA associated with fatal infection. The incidence of slime formation in the control group was significantly lower than that in the other groups. Results suggest that slime formation by MRSA may be associated with a more severe infection. Rapid identification of slime-forming MRSA may facilitate the initiation of appropriate treatment and improve the patient's prognosis.

Biomarkers↗
PubMed1997

Expression of interleukin-6 in the ventricles and coronary arteries of patients with myocardial infarction.

While an overproduction of interleukin-6 (IL-6) has been observed in patients with acute myocardial infarction (AMI), its clinical significance and localization in the ischemic myocardium have not been elucidated. We examined immunohistochemically the expression of IL-6 in 12 autopsied patients with AMI who had died within seven days of the infarction. Twenty sections of ischemic myocardium and nine of the coronary arteries involved were stained with anti-IL-6 and anti-atrial natriuretic peptide (ANP). The diameter of the myocardium was analyzed. The greatest expression of IL-6 in the infarcted myocardium occurred in patients who had died three to four days after the onset (2.7 +/- 0.4), as judged by a scheme for grading IL-6 expression. Patients who died within one to two days (1.0 +/- 0.3) or five to eight days (0.6 +/- 0.4) less frequently showed an overproduction of IL-6. The IL-6-positive myocardium co-expressed ANP and was significantly (p < 0.05) hypertrophied, when compared with the IL-6-negative myocardium. The diameter of IL-6-positive myocardial myocytes was significantly (p < 0.02) increased in patients who died within one to two days (1.6 +/- 0.2), three to four days (1.8 +/- 0.3), or five to eight days (2.0 +/- 0.2) after the AMI. The involved coronary arteries expressed IL-6 in the intimal and smooth muscle cells, as did atherosclerotic coronary arteries not involved in AMI. An overproduction of IL-6 was confirmed in the injured myocardium with hypertrophy in patients who died of AMI within seven days after onset. The hypertrophied injured myocardium co-expressed IL-6 and ANP. The expression of IL-6 in the myocardium in AMI appears to be associated with the mechanism of cardiac hypertrophy.

Adult↗
PubMed1997

Immunosuppressive drugs inhibit the production of interleukin-6 and interleukin-8 in cultured cardiac myxoma cells.

Cardiac myxoma cells produce large amounts of interleukin (IL)-6 and IL-8. To determine whether immunosuppressive agents could be used to treat cardiac myxoma, we tested the effects of dexamethasone and three of the newer second-generation immunosuppressive drugs, cyclosporin A, tacrolimus, and deoxyspergualin, on the production of IL-6 and IL-8 in these cells. Cultured cardiac myxoma cells were used as in vitro model of cardiac myxoma. Cells were tested for 24 hours with 10(-7) M dexamethasone, 10(-6) M cyclosporin A, 10(-8) M tacrolimus, and 10(-6) M 15-deoxyspergualin, with aliquots of conditioned medium being assayed for cytokine levels at 0, 6, 12, and 24 hours. Cardiac myxoma cells isolated from 4 patients all produced quantities of IL-6 and IL-8. The concentrations of IL-6 in the medium after 7 days in culture ranged from 79,000 to 2,740,000 pg/ml, and the concentrations of IL-8 ranged from 40,000 to 1,000,000 pg/ml. Exposure of cyclosporin A and dexamethasone almost completely inhibited the production of IL-6 and IL-8 after 24 hours of treatment. Tacrolimus inhibited the production of both cytokines by 55%, while 15-deoxyspergualin reduced IL-6 levels by 24% and IL-8 levels by 48% after separate 24 hour treatments. These results suggest that these newer immunosuppressive agents may be useful in reducing the production of IL-6 and IL-8 in patients with cardiac myxoma.

Aged↗
PubMed1997

Removing tRNA from a cell-free protein synthesis system for use in protein production.

The cell-free system for biosynthesis of proteins is becoming an important tool for protein engineering. In particular, introduction of the unnatural amino acids is achieved though cell-free protein synthesis with the use of chemically acylated tRNA that recognizes a specific codon. In the original method, however, it was difficult to control the system through changing tRNA composition, as the endogenous tRNAs are involved in the reaction. Thus, in the present study, we digested the tRNA within Escherichia coli S30 extract with resin-bound RNase A, and estimated the protein synthesis activity. It was revealed that this digestion process does not damage the activity, if a protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), is present in the digestion reaction.

Cell-Free System↗
PubMed1997

Reduced serum T3 level in a patient with nodular goiter and cardiac myxoma.

We report the unusual case of an 87-year-old woman with cardiac myxoma and adenomatous goiter. She exhibited slight elevations of serum interleukin-6 (IL-6), but levels of thyroid hormones such as T3, free T3 and free T4 were all abnormally low. Interleukin-6 may potentiate the alteration of thyroid metabolism.

Aged↗
PubMed1997

Activated protein C resistance assay as a screening test for thromboembolic disposition.

An activated partial thromboplastin time (APTT) based method was developed to determine the anticoagulant response in patient plasma to added purified activated protein C (APC). On the other hand, Bertina et al. (1994) reported that one point mutation of amino acid 506 in the factor V was observed in such APC resistance patients. APC resistance for blood samples with known abnormal findings on the coagulation test also may be determined. Coagmaster II for the APTT assay system (Sankyo, Tokyo, Japan) was employed. The APC resistance assay system was a useful reagent as a screening test for lupus anticoagulant (LA), in addition to APC resistance.

Calcium Chloride↗
PubMed1997

Two doses of artemether/mefloquine or artesunate/mefloquine combination for multidrug resistant falciparum Malaria.

Plasmodium falciparum in Southeast Asia is highly resistant to chloroquine, sulfadoxine/ pyrimethamine, quinine and even mefloquine. The use of two doses of short course artemether/mefloquine combination has been shown to be effective in a recent study. In the present study, we have assessed the efficacy of short course treatment with artesunate/mefloquine, in comparison with artemether/mefloquine in patients with multidrug resistant falciparum malaria. Ninety-nine Thai male patients who sought consultation at Makham Malaria Clinic, Chantaburi (eastern part of Thailand), were randomized to receive either the combination of artemether (150 and 100 mg; group A) or artesunate (150 and 100 mg; group B) with mefloquine (750 and 500 mg) at 24 hours apart. The follow-up was on days 1, 2, 7, 14, 21, 28, 35 and 42. Patients in both groups showed a rapid initial response to treatment; fever and parasite were cleared within 48 hours in 100 and 100% vs 91.8 and 96%, for group A vs B, respectively. All patients in group A had completed the 42 day-follow up; however, two patients in group B did not finish the 42-day follow-up. The cure rate was 100% in either group. No serious adverse effects were found. Artemether or artesunate with mefloquine given two doses at 24 hours apart can be used as effective alternative treatment regimens for multidrug resistant falciparum malaria.

Acute Disease↗
PubMed1996

Generation and characterization of anti-sulfoglucuronosyl paragloboside monoclonal antibody NGR50 and its immunoreactivity with peripheral nerve.

Sulfoglucuronosyl paragloboside (SGPG) is a member of the sulfated glucuronic acid-containing glycolipid (SGGL) family found primarily in peripheral nerves. These glycolipids contain the HNK-1 carbohydrate epitope and are recognized by monoclonal IgM from patients with chronic demyelinating neuropathy and paraproteinemia. Recent studies indicate that SGGLs may serve as ligands for selectins, amphoterin, and laminin, suggesting that these glycolipids may play an important role in cellular adhesion. To elucidate the biological function of these glycolipids, we produced a murine monoclonal antibody (mAb) and studied its antigenic specificity. Using an enzyme-linked immunosorbent assay (ELISA), we found that the mAb designated as NGR50 belonged to the IgG2a subclass, and that the minimal titer (2 SD above the mean optical density value of control) of this mAb was 1:640, with 20 ng of purified SGPG as the antigen. Thin-layer chromatography (TLC) immunoblotting revealed that this mAb reacted specifically with SGPG and sulfoglucuronosyl lactosaminyl paragloboside (SGLPG), which is a structural analogue of the former, but not with other glycolipids. Desulfated derivates of SGPG and SGLPG did not react with mAb NGR50. Western blot analysis showed crossreactivity with human myelin-associated glycoprotein (MAG), but not with rat MAG or rat glycoprotein P0. Unlike anti-HNK-1 monoclonal antibody, however, NGR50 reacted only weakly with several proteins in the 20-30-kD regions, including human P0, suggesting that mAb50 has a different fine specificity as an anti-HNK-1 antibody. Immunocytochemical study of rat sciatic nerve using mAb NGR50 revealed positive staining at the outer surface of the myelin sheath and Schwann cells, as well as in the intervening connective tissues. Faint staining was also visible at the axolemmal-myelin interface; however, compact myelin was not stained.

Animals↗
PubMed1996

Effect of bile on the intestinal bile-acid binding protein (I-BABP) expression. In vitro and in vivo studies.

Enterocytes actively transport bile acids from the ileal lumen to the portal blood. This physiological process greatly contributes to maintaining the bile acid homeostasis. However, little is known about the molecular mechanisms involved in this transport system. The effect of bile on gene expression of the intestinal bile-acid binding protein (I-BABP) expressed in the enterocytes was studied in vivo, using the by-pass method, and in vitro, using organ culture of ileum explants and Caco-2 cell line. The low cytosolic I-BABP concentration and I-BABP mRNA level found in diverted ileum was totally recovered when bile was added in the ileal lumen. Northern blot analysis of the ileal explants revealed a dose-dependent increase in the I-BABP mRNA in the presence of bile. In Caco-2 cells, the I-BABP transcript was dramatically increased in the presence of human bile while it was undetectable in the control cultures. These data offer the first evidence that biliary components regulate the I-BABP gene expressed in the enterocytes.

Animals↗
PubMed1996

Sensitization of Lewis rats with sulfoglucuronosyl paragloboside: electrophysiological and immunological studies of an animal model of peripheral neuropathy.

Antibodies against sulfoglucuronosyl glycosphingolipids (SGGLs) are known to be present in sera of patients with chronic polyneuropathy associated with IgM paraproteinemia. We recently studied rats sensitized with sulfoglucuronosyl paragloboside (SGPG), a major SGGL species, emulsified with keyhold limpet hemocyanin and Freund's adjuvant. The titer of the IgM class antibodies against SGPG increased up to 1:1,600, while that of the IgG class increased up to 1:800 2 weeks after sensitization. The antibodies showed a high degree of antigenic specificity; no cross-reactivity with other brain glycolipids could be detected. They, however, reacted with human myelin-associated glycoprotein (MAG) by Western blot analysis, but not with rat MAG. These animal models showed minor but clear clinical signs of neuropathy, consisting of mild tail muscle tone loss and walking disabilities. Electrophysiological examination of the sciatic nerves revealed nerve conduction abnormalities which consisted of conduction block and mild decrease in conduction velocity. Thus, our results support the concept that anti-SGPG antibodies may play an important pathogenetic role in this type of chronic neuropathy.

Action Potentials↗
PubMed1996

Synthesis and characterization of polymer-coated mixed-functional stationary phases with several different hydrophobic groups for direct analysis of biological samples by liquid chromatography.

Three types of polymer-coated mixed-functional (PCMF) silica column-packing materials for the direct analysis of biological liquids have been synthesized. These packing materials all have polyoxyethylene groups as their hydrophilic part but differ in their hydrophobic part (methyl groups for Me-POE, phenyl groups for Ph-POE, and octyl groups for Oc-POE). Retention characteristics with respect to several drug molecules, and protein recovery, were studied for each of the three types with different amounts of hydrophobic groups attached. Of the PCMFs prepared, Oc-POE showed the greatest overall retention, and Me-POE displayed the greatest protein recovery.

Anti-Bacterial Agents↗
PubMed1996

Semotiadil improves survival of rats with monocrotaline-induced pulmonary hypertension: comparison with diltiazem.

We compared the effects of semotiadil, a novel Ca2+ channel blocker, with those of diltiazem on survival and regression of right ventricular hypertrophy and media thickening of pulmonary arteries in a rat model of pulmonary hypertension. Pulmonary hypertension was induced by a single injection of monocrotaline (80 mg/kg). Four weeks later, after pulmonary hypertension was confirmed, oral administration of semotiadil (10, 30, or 100 mg/kg/day) or diltiazem (100 or 300 mg/kg/day) was initiated. The rats were observed for 3 weeks. Survival was significantly longer in the group that received semotiadil 100 mg/kg/day than in the groups treated with diltiazem 100 or 300 mg/kg/day. Media thickness and smooth muscle area in pulmonary arteries were significantly less in rats treated with semotiadil 100 mg/kg/day than in animals treated with diltiazem 100 mg/kg/day. The right ventricle to left ventricle mass ratio, right ventricular wall thickness, and right ventricular myocardial fiber diameter were equal in these two groups. Semotiadil 100 mg/kg/day improved the survival of rats, which responded with a significant regression of right ventricular hypertrophy and media thickening of pulmonary arteries in comparison with rats treated with diltiazem 100 or 300 mg/kg/day.

Animals↗
PubMed1996

Crystallization and preliminary X-ray study of a new crystal form of cytochrome c' from Rhodobacter capsulatus.

A new crystal form of diheme cytochrome c' from Rhodobacter capsulatus has been obtained and preliminary crystallographic experiments have been performed. The crystals belong to the space group P2(1)2(1)2 with unit-cell dimensions of a = 47.82, b = 72.59, c = 34.32 A. The assumption that an asymmetric unit of the crystal contains one half of the homodimer molecule indicates that the monomers in the dimeric molecule may be related by a crystallographic twofold axis. Crystals diffract up to 1.7 A resolution using the X-ray beam from synchrotron radiation, and 11 127 unique structure factors were obtained with an R(merge) of 7.1% from 52 922 indexed reflections. Structure analysis by means of molecular-replacement methods is now underway.

Journal Article↗
PubMed1996

Expression and localization of Lewis(x) glycolipids and GD1a ganglioside in human glioma cells.

We analysed the glycolipid composition of glioma cells (N-370 FG cells), which are derived from a culture of transformed human fetal glial cells. The neutral and acidic glycolipid fractions were isolated by column chromatography on DEAE-Sephadex and analysed by high-performance thin-layer chromatography (HPTLC). The neutral glycolipid fraction contained 1.6 micrograms of lipid-bound glucose/galactose per mg protein and consisted of GlcCer (11.4% of total neutral glycolipids), GalCer (21.5%), LacCer (21.4%), Gb4 (21.1%), and three unknown neutral glycolipids (23%). These unknown glycolipids were characterized as Lewis(x) (fucosylneolactonorpentaosyl ceramide; Le(x)), difucosylneolactonorhexaosyl ceramide (dimeric Le(x)), and neolactonorhexaosyl ceramide (nLc6) by an HPTLC-overlay method for glycolipids using specific mouse anti-glycolipid antibodies against glycolipid and/or liquid-secondary ion (LSI) mass spectrometry. The ganglioside fraction contained 0.6 micrograms of lipid-bound sialic acid per mg protein with GD1a as the predominant ganglioside species (83% of the total gangliosides) and GM3, GM2, and GM1 as minor components. Trace amounts of sialyl-Le(x) and the complex type of sialyl-Le(x) derivatives were also present. Immunocytochemical studies revealed that GD1a and GalCer were primarily localized on the surface of cell bodies. Interestingly, Le(x) glycolipids and sialyl-Le(x) were localized not only on the cell bodies but also on short cell processes. Especially, sialyl-Le(x) glycolipid was located on the tip of fine cellular processes. The unique localization of the Le(x) glycolipids suggests that they may be involved in cellular differentiation and initiation of cellular growth in this cell line.

Animals↗
PubMed1996

Positive and negative host factors for Sendai virus transcription and their organ distribution in rat.

In vitro mRNA synthesis by Sendai virus is almost entirely dependent on the addition of cellular proteins (positive host factors), one of which could be tubulin. In this study, we investigated the distribution of host factors in various rat organs. Extracts from the brain, thymus, heart, lung, testis, ovary, and uterus all supported in vitro Sendai virus transcription, among which the highest activity was obtained with the brain extract. On the other hand, little or no activity was detected in the liver, spleen, and kidney extracts. An inverse correlation between the apparent host factor activity to stimulate mRNA synthesis and RNase activity that hydrolyzes Sendai virus mRNAs was found, except in the liver extract. However, when a transcription initiation complex was isolated and subjected to RNA chain elongation reaction, all of the extracts including those from liver, spleen and kidney, were active. Immunoblotting showed that tubulin molecules were integrated in these initiation complexes, supporting the notion that tubulin is involved in the initiation complex formation. We also identified a transcription inhibitory activity without any detectable RNase activity in the liver extract. This negative host factor seemed to act on RNA chain elongation. It is likely that Sendai virus transcription is regulated by both positive and negative regulatory factors.

Animals↗