Isolation and structure elucidation of teleocidin B-1, B-2, B-3, and B-4.
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Biomedical subjects
Publications and source records attributed to Y Endo.
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A subpopulation of lymphocytes forming rosettes with autologous erythrocytes was studied on peripheral blood and thyroid tissues obtained from the patients with various thyroid diseases. The mean (+/-S.D.) percentage of autorosette-forming cells (ARFC) was 10.1(+/-5.5)% in the peripheral blood from patients with hyperthyroid Graves' disease, which was higher than that in normal subjects (5.6 +/- 2.8%), while the levels of ARFC in the peripheral blood from euthyroid patients with Graves' disease under treatment and Hashimoto's thyroiditis did not significantly differ from the normal level. The mean percentages of ARFC in the thyroid tissues from patients with Graves' disease and Hashimoto's thyroiditis were 14.7(+/-8.5) and 13.3(+/-7.8)%, respectively, which were higher than those in the peripheral blood from the same patients. Most of these cases with abnormally high levels of ARFC were accompanied with the abnormally low T cell to B cell ratios. The microscopic examination of the cytological materials from these patients showed an increased number of large stimulated lymphoid cells or lymphoblasts as compared with those who had few ARFC. These results suggest an increase in an activated T cell subset in the circulation and/or in the thyroid tissue, which is probably caused by active immune response to some stimuli.
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We have attempted to determine the carbohydrate moiety of human alphafetoprotein (AFP) produced by a yolk sac tumor. AFP was obtained from the cystic fluid of human yolk sac tumors grown in nude mice and was purified using an immunoadsorbent column coupled with monoclonal anti-AFP antibody. Then, the carbohydrate chain of the purified AFP was quantitatively released from the polypeptide chain. The resulting oligosaccharide was labeled and, by sequential exoglycosidase digestion in combination with methylation analysis and periodate oxidation, the structure was determined to be: Sia alpha 2----6Gal beta 1----4GlcNAc beta 1----2Man alpha 1----6 (GlcNAc beta 1----4) (Sia alpha 2----6Gal beta 1----4GlcNAc beta 1----2Man alpha 1----3) Man beta 1----4GlcNAc beta 1----4 (Fuc alpha 1----6) GlcNAcOT Compared with the known structure of the sugar, chain of human hepatic AFP, it was found that the sugar chain of yolk sac AFP contained an additional sugar, N-acetylglucosamine (bisect GlcNAc) linked to the beta-mannose. In the light of recent knowledge, this result indicates that the Concanavalin A (Con-A) binding site of the sugar chain is blocked by this GlcNAc in human yolk sac AFP. This fact forms the basis for the clinical use of the Con-A binding test to determine the origin of AFP in patients.
A heritable deficiency in α(2)-macroglobulin (α(2)M) was identified in a 61-year-old man with arterial thrombosis. Plasma α(2)M levels among the patient's symptom-free relatives consistently ranged from 43 to 55 percent of laboratory mean-normal values. The new α(2)M variant displayed retarded anodal immunoelectrophoretic mobility when studied in plasma and serum. The affected members of this lineage showed no evidence of acquired or inherited thrombotic or consumptive derangements involving other plasma proteins. The significance of a possible causal association between α(2)M deficiency and the predisposition to arterial thrombosis is considered. The uncomplicated use of streptokinase and urokinase to treat the reference patient's arterial thrombosis is described. Recommendations are made for the adoption of a descriptive nomenclature. The new familial deficiency is tentatively designated α(2) (+)-macroglobulin deficiency Irvine.
Three olivoretins, A, B and C (isolated from Streptoverticillium olivoreticuli), which are O-methylated teleocidin B isomers, were found to be biologically inactive. A fourth olivoretin, D, which has a free hydroxyl group and is identical to one of the four teleocidin B isomers, teleocidin B-4 (teleocidin B of Hirata) was biologically active. These findings indicate that the free primary hydroxyl group of teleocidin B isomers is necessary for activity. The effect on biological activity of the structural difference between des-O-methylolivoretin B (teleocidin B-1) and des-O-methylolivoretin C was also studied.
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Synthetic analogues (indolactams) related to the tumor promoter teleocidin were synthesized chemically. Of four indolactam-Vs lacking the monoterpenoid moiety of native teleocidin, (-)-indolactam-V bound to the 12-O-tetradecanoylphorbol-13-acetate receptor in cell membranes and induced both adhesion of HL-60 cells and ornithine decarboxylase activity in mouse skin, although its effects were weaker than those of teleocidin. (+)-Indolactam-V and two isomers of epi-indolactam-V showed no induction of ornithine decarboxylase. These results indicate that the S,S configuration of native teleocidin at the isopropyl residue and the hydroxymethyl group is necessary for activity.
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Congenital hypoplasia of the thumb is not a rare condition in clinical practice, but a familial occurrence of this condition is exceedingly rare. We report a family with familial congenital hypoplasia of the thumb. In three generations, five members were proved to be affected and another two were presumed to be affected. To our surprise, a younger sister of the propositus had tetralogy of Fallot and triphalangeal thumbs on both hands.
In a sandwich enzyme immunoassay (EIA) for human growth hormone (hGH) with anti-hGH Fab'-peroxidase conjugate, the effects of inorganic salts on serum interference were examined, and serum interference was eliminated by incubation of serum samples with anti-hGH IgG-coated polystyrene balls in the presence of 0.4 mol/l NaCl, avoiding the need for hGH-free serum. The sensitivity for hGH was 60 fg/tube or 3 ng/l of serum. No cross-reaction was observed with prolactin, chorionic gonadotropin or luteinizing, thyroid-stimulating and follicle-stimulating hormones. The coefficients of within-assay and between-assay variations were 2.8-6.5% and 4.8-8.7%, respectively. The regression equation and correlation coefficient to radioimmunoassay (RIA) were y(EIA) = 0.89x(RIA) + 0.11 and 0.98 (n = 100), respectively. hGH levels in normal male and female adult serum taken between 9:00 and 10:00 a.m. after overnight fasting and 1 h rest were 312 ng/l (range 53-940 ng/l; n = 10) and 662 ng/l (112-2195 ng/l; n = 13), respectively.
Various types of mitogenic substances, such as a Escherichia coli lipopolysaccharide (LPS), concanavalin A (Con A), pokeweed mitogen, polyI:polyC (a synthetic double-stranded RNA) and 12-O-tetradecanoylphorbol-13-acetate (a component of croton oil), induced histidine decarboxylase (HDC) in the liver, spleen and lung of mice at 4.5 hr after injection. Other inflammatory agents without mitogenic activity, such as zymosan, carrageenan, glycogen, D-galactosamine and N-acetyl-muramyl-L-alanyl-D-isoglutamine, did not induce the enzyme. Both LPS (a B-cell mitogen) and Con A (a T-cell mitogen) induced HDC also in nude mice that lack T-cells, indicating that T-cells are not required for HDC induction by mitogens. C3H/HeJ mice, which are LPS-low responder mice in various immunological tests, were quite a bit less responsive to LPS also in the HDC induction. These results show that mitogens with different properties can induce HDC as a common characteristic. On the basis of these results, the possible participation of macrophages in the process of HDC induction by mitogens was discussed.
The sequence of the 521 nucleotides at the 3' end of a rat 28 S rRNA gene was determined. The region encompasses the site of cleavage of 28 S rRNA by the cytotoxin alpha-sarcin. The toxin hydrolyzes a phosphodiester bond on the 3' side of a guanine residue 393 nucleotides from the 3' end. The alpha-sarcin domain is composed of a purine-rich sequence of 14 highly conserved nucleotides.
The administration of an Escherichia coli lipopolysaccharide (LPS), or an endotoxin into mice produced a variation in tissue serotonin (5HT) levels within 4.5 h. 5HT levels in the kidney and lung were decreased by the higher doses of LPS, but those in the liver and spleen were increased even by lower doses of the agent. The increase in liver 5HT was most marked. Such variations in 5HT levels were also produced by the administration of concanavalin A. In vitro experiments using extracts from livers of LPS-treated and non-treated mice indicated that there was no difference in the 5HT formation from 5-hydroxytryptophan between the two groups, but that 5HT formation from tryptophan was higher in the LPS-treated mice. The LPS-induced 5HT increase in liver was suppressed by p-chlorophenylalanine (an inhibitor of tryptophan hydroxylase), actinomycin D, cycloheximide and dexamethasone, but not by Ro 4-4602 (an inhibitor of aromatic amino acid decarboxylase), pargyline (an inhibitor of monoamine oxidase) and indomethacin. A possible mechanism of the 5HT increase in the liver is discussed on the basis of these results.
The ribonuclease activity of the cytotoxic protein alpha-sarcin has been characterized. When rat liver ribosomes or 60 S ribosomal subunits were the substrates, alpha-sarcin cleaved a single oligonucleotide of about 488 residues, the alpha-fragment, from the 3' end of 28 S rRNA. In contrast, 40 S ribosomal subunits were not affected by alpha-sarcin. The alpha-fragment was cleaved from 28 S rRNA in 80 S ribosomes when the concentration of alpha-sarcin was 3 x 10(-8) M and the toxin retained its specificity even when the concentration was 3 x 10(-5) M. The turnover number (kcat) for the reaction of alpha-sarcin with ribosomes was 55 min-1, establishing that the toxin acts catalytically. When total rRNA or 28 S rRNA was the substrate, alpha-sarcin caused extensive progressive digestion of the nucleic acids; however, no formation of the alpha-fragment occurred. The extent of the digestion of 28 S rRNA was related to the concentration of alpha-sarcin, but the amount of the toxin required was somewhat greater than that needed with ribosomes. Digestion of homopolynucleotides with alpha-sarcin indicated that the protein is specific for purines. When [32P]5 S rRNA was the substrate, alpha-sarcin cleaved on the 3' side of purines in both single- and double-stranded regions of the molecule. The results suggest that the unusual specificity of alpha-sarcin, in that it cleaves only one of more than 7000 phosphodiester bonds in the ribosome, is a property both of the cytotoxin and of the ribosome.