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Biomedical subjects

K Takeo

Publications and source records attributed to K Takeo.

At least 91 records · Page 5Linked to original sources
PubMed1990

The pharmacokinetic pattern of glycosylated human recombinant lymphotoxin (LT) in rats after intravenous administration.

We have examined the pharmacokinetics of glycosylated recombinant human lymphotoxin (LT) after intravenous bolus injection in rats and compared them with those of tumor necrosis factor (TNF) or LT species. The results are as follows. 1) The mean half-life of glycosylated LT in serum increases for each increase in dose, and the distribution volume (V) and total body clearance [(Cl (total)] tend to decrease for increase in dose. On the other hand, the half-life of TNF also increases for increase in dose, but the V tends to increase for increase in dose and Cl (total) does not change. 2) The glycosylated LT distributes to all organs so far tested except brain, and tends to accumulate to kidney more than other tissues at 6 h after the injection. 3) Nonglycosylated LT produced by E. coli and the glycosylated LT species carrying both N-type and mucin-type sugar moieties (25 kDa LT) have shorter half-lives and higher Cl (total)s than 23 kDa LT carrying N-type sugar moieties alone. The 21 kDa LT, the same species as 23 kDa LT except that it lacks 15 amino acid residues at the N-terminus, disappears much faster than 23 kDa LT and shows higher V and Cl (total). Thus, glycosylated LT shows nonlinear pharmacokinetics like TNF, but the deposition is quite different from that of TNF. The high serum concentration of glycosylated LT depends upon the presence of N-type sugar moieties, but not mucin-type sugar moieties. The N-terminal protein chain of LT also correlates with the serum concentration.

Amino Acids↗
PubMed1989

Ultrastructural alterations in Saccharomyces cerevisiae cells in association with elevated temperature-induced autolysis.

By means of the freeze-etching technique ultrastructural alterations in Saccharomyces cerevisiae cells undergoing autolysis at elevated temperature were studied. Wall surfaces of intact cells were smooth. During autolysis wall surfaces became rough with granules of 20-40 nm diameter. This alteration occurred after extensive disintegration of cytoplasmic organelles and after functional and ultrastructural impairments of the plasma membrane, but well before the rupture of the plasma membrane.

Autolysis↗
PubMed1989

Improved separation of alpha chains of collagen type I, type III, and type V by noninterrupted electrophoresis using thioglycolic acid as a negatively charged reducer.

Improved separation of alpha chains of collagen type I (alpha 1 [I]2 alpha 2[I]), type III(alpha 1[III]3), and type V (alpha 1[V]alpha 3[V])was achieved by noninterrupted sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a negatively charged reducer, thioglycolic acid. The thioglycolic acid, added to the running buffer of the cathodic reservoir, in the middle of electrophoresis quickly migrated in the gel anode, reducing interchain disulfide linkages in collagen type III and dissociating it into its alpha chain monomer, alpha 1[III], without an interruption of electrophoresis. The alpha chain, alpha 1[III], migrated more slowly than the alpha 1 [I] and alpha 2[I] chains of collagen type I, resulting in an excellent separation of alpha 1[III] from alpha 1[I]. The mobility of alpha 1[III] could be controlled by varying the time of thioglycolic acid addition to the running buffer. This enabled us not only to separate alpha 1[III] from alpha 1[I] and alpha 1[V], but also to precisely quantitate these alpha chains, even at low protein loading of mixed samples.

Collagen↗
PubMed1989

A simple method for determination of the concentration of anti-dextran IgG in antiserum by means of affinity electrophoresis.

A simple technique for determination of the concentration of anti-dextran IgG in antiserum using affinity electrophoresis is described. In the presence of an excess of intermediary molecular size dextran in the polyacrylamide gel, polyclonal anti-dextran IgG migrated in a single sharp band, separated from the nonspecific IgG fraction and other serum protein fractions. With this technique, 1-10 micrograms anti-dextran IgG in antisera can be determined within 3 h. We call the procedure ligand saturating affinity electrophoresis.

Animals↗
PubMed1989

Complete separation of anti-hapten antibodies by two-dimensional affinity electrophoresis.

A high resolution two-dimensional affinity electrophoresis has been developed, using capillary isoelectric focusing as the first electrophoresis and slab gel affinity electrophoresis as second electrophoresis. By this method 1-2 micrograms of anti-dinitrophenyl antibodies have been separated completely into several hundred homogeneous IgG spots. They are grouped into a number of families which are composed of several IgG spots of the same affinity to the hapten but of a different pI. It is suggested that each individual family is derived from one monoclonal antibody producing cell line.

Animals↗
PubMed1989

Studies on the heterogeneity of anti-hapten antibodies by means of two-dimensional affinity electrophoresis.

The molecular heterogeneity of rabbit anti-hapten antibodies has been investigated by two-dimensional affinity electrophoresis (2D-AEP). Anti-dansyl and anti-arsanilic diazo-antibodies were separated into several hundred IgG spots as in the case of anti-DNP antibodies. They were grouped into a number of monoclonal IgG families. At the beginning of immunization, IgG spots having low pI and low affinity were predominant but one or two weeks after immunization the IgG spots with high affinity and high pI increased. After the second or third immunization, the 2D-AEP patterns became stable and constant. Anti-arsanilic diazo and anti-DNP antibodies exhibited only weak cross-reactivity with other aromatic haptens. In contrast, anti-dansyl antibodies cross-reacted to a considerable degree with DNP-hapten. A few anti-dansyl IgG families which have cross-reactivity with DNP-hapten were separated. Their apparent dissociation constants to the haptens and their affinity ratios were calculated from their 2D-AEP patterns, according to the affinity theory.

Animals↗
PubMed1986

Synthesis of tetrasaccharides related to the antigenic determinants from the glycopeptidolipid antigens of serovars 9 and 25 in the Mycobacterium avium-M. intracellulare-M. scrofulaceum serocomplex.

The synthesis of the tetrasaccharides O-(2,3-di-O-methyl-alpha-L-fucopyranosyl)-(1----4)-O-(2,3- di-O-methyl-alpha-L-fucopyranosyl)-(1----3)-O-alpha-L-rhamnopyranosyl -(1----2)-6-deoxyl-L-talose (36) and O-(2-O-methyl-alpha-L-fucopyranosyl)-(1----4)-O-(2-O-methyl-alpha-L- fucopyranosyl)-(1----3)-O-alpha-L-rhamnopyranosyl-(1----2)-6-deoxy-L- talose (41) is described. The former and the latter structures, respectively, have been proposed as the carbohydrate chains in the polar glycopeptidolipid antigens of serovars 9 and 25 in the Mycobacterium avium-M. intracellulare-M. scrofulaceum serocomplex. Glycosylation of allyl 2,3-di-O-methyl-alpha-L-fucopyranoside with 4-O-benzoyl-2,3-di-O-methyl-alpha-L-fucopyranosyl chloride gave the alpha-linked disaccharide derivative, which was O-deallylated and converted into the corresponding disaccharide alpha-chloride. This was coupled with benzyl 3,4-di-O-benzyl-6-deoxy-2-O-(2,4-di-O-benzoyl-alpha-L-rhamnopyranosyl)- alpha-L-talopyranoside (32) to give a fully protected tetrasaccharide derivative, which was deprotected to furnish 36. Likewise, 3-O-benzyl-4-O-(3,4-di-O-acetyl-2-O-methyl-alpha-L- fucopyranosyl)-2-O-methyl-alpha-L-fucopyranosyl chloride, prepared by way of condensation of allyl 3-O-benzoyl-2-O-methyl-alpha-L-fucopyranoside with 2-O-methyl-3,4-di-O-(p-nitrobenzoyl)-alpha-L-fucopyranosyl bromide, reacted with 32, to provide, after removal of blocking groups, 41.

Epitopes↗
PubMed1986

Mode of cell growth of Malassezia (Pityrosporum) as revealed by using plasma membrane configurations as natural markers.

The mode of cell growth of Malassezia was studied by freeze-fracture using the plasma membrane configurations of this organism as natural markers. The plasma membrane of the mature cell bodies of M. pachydermatis had a ring swelling, and on each side of the ring, one set of straight and spiral grooves and circumvallate bulgings. The cell always divided at the ring swelling (M. pachydermatis) or depression (M. furfur), soon followed by budding there. A new set of similar configurations formed on the bud. In all the 12 strains of Malassezia studied, the spiral grooves in the mother and bud parts were both left-handed but opposite in the direction of elongation. By comparing distances between the spiral grooves in short and long buds and in mothers, the bud tip was suggested as the major, and adjacent regions as the minor, sites of wall growth. Some characterizations of the plasma membrane invaginations, especially in relation to the mode of cell growth, were also described.

Cell Membrane↗
PubMed1985

Microassay for proteins on nitrocellulose filter using protein dye-staining procedure.

A simple and rapid microassay for proteins utilizing the protein dye-staining procedure with a nitrocellulose (NC) filter is described. Proteins were directly bound to an NC filter using the "BIO DOT" microfiltration apparatus to ensure their uniformity. The proteins were then stained with a dye (Ponceau Red 3R or amido black 10B), and the optical density of the stained protein spots was directly measured by a densitometer. A good linearity between the optical density and the amount of protein was obtained in the range 0.05 to 10 micrograms. A larger number of samples (up to 96 samples) could be assayed within 1.5 h simultaneously. Contaminating chemicals, such as amino acids, sugars, reducing agents, chelating agents, tris(hydroxymethyl)aminomethane, deoxyribonucleic acid, and nucleotides, did not interfere with the assay. The reproducibility, pH dependency, and application of the assay to the quantitation of a small amount of proteins in body fluids are discussed.

Adult↗
PubMed1985

A correlation between mode of growth and regional ultrastructure of the plasma membrane of Schizosaccharomyces pombe as revealed by freeze-fracturing before and after filipin treatment.

The ultrastructure of the plasma membrane of Schizosaccharomyces pombe was studied by freeze-fracture using invaginations of the plasma membrane as natural markers and filipin-induced deformations as artificial markers. In accord with the mode of growth of this organism, ultrastructural aspects of the plasma membrane were related to the following ring zones: the growing pole, adjacent regions, proximal regions, the new cell pole, and the middle in dividing cells. The growing pole and adjacent regions had no or only a few invaginations. Filipin induced numerous deformations in these regions. By contrast, the proximal regions of the plasma membrane had several invaginations and resisted filipin-induced deformation. Concomitantly with commitment to cytokinesis, both the invaginations and the resistance to filipin-induced deformation disappeared in the middle. The results presented here strongly suggest the existence of two states of the plasma membrane of S. pombe, a fact which correlates well with the mode of growth of this organism.

Ascomycota↗
PubMed1985

Inhibition of polymorphonuclear leukocyte 5-lipoxygenase and platelet cyclooxygenase by alpha-(3,5-di-tert-butyl-4-hydroxybenzylidene)-gamma-butyrolacto ne (KME-4), a new anti-inflammatory drug.

The effect of alpha-(3,5-di-tert-butyl-4-hydroxybenzylidene)-gamma-butyrolactone (KME-4), a new anti-inflammatory compound, on arachidonate lipoxygenase and cyclooxygenase was investigated. KME-4 showed a dose-dependent inhibition of 5-lipoxygenase activity in both the cytosol (IC50: 0.85 microM) and ionophore A23187-stimulated cells (IC50: 11.5 microM) of guinea pig peritoneal polymorphonuclear leukocytes. KME-4 was also found to inhibit rabbit platelet cyclooxygenase (IC50: 0.44 microM), but had no inhibitory effect on platelet 12-lipoxygenase at concentrations up to 100 microM, whereas BW755C inhibited both enzymes in the same range of concentrations. The results indicate that KME-4 is a dual 5-lipoxygenase and cyclooxygenase inhibitor which is different from BW755C in affecting 12-lipoxygenase. These effects may provide information for understanding the pharmacological activity of KME-4.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗