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Biomedical subjects

M Satake

Publications and source records attributed to M Satake.

At least 253 records · Page 14Linked to original sources

A novel HLA class II alloantigen detected by an alloantiserum TK2.

An alloantiserum, TK2, obtained from a kidney transplant patient, was investigated. TK2 reacted with B lymphocytes that had only one detectable DQ antigen. DQ "blank" homozygous cell lines were positive for TK2, but DQwl, DQw2, and DQw3 homozygous cell lines failed to react with TK2. Positive reactions for TK2 segregated with DQ "blank" haplotypes in the families studied. The reactivity of TK2 to B lymphocytes was strong, while its reactivity to monocytes was very weak. When TK2 was absorbed with DQ "blank" cells, it became unreactive to other lymphocytes. TK2 blocked the binding of an anti-DQ monoclonal antibody to DQ "blank" cells. It did not affect the binding of the antibody to DQw1 and DQw3 cells, nor did it affect the binding of an anti-DR monoclonal antibody to a DQ "blank" cell line. These findings suggest that TK2 can detect a new HLA class II alloantigen that corresponds to a DQ "blank" specificity.

Antibodies, Monoclonal↗

Isolation and characterization of a novel 2-aminoethylphosphonyl-glycosphingolipid from the sea hare, Aplysia kurodai.

A novel phosphonoglycosphingolipid named SGL-I' containing 1 mol of 2-aminoethylphosphonate residue was isolated from the skin of Aplysia kurodai using two silicic acid chromatography systems. Data obtained on methanolysis, permethylation, mild acid hydrolysis, and hydrogen fluoride treatment combined with thin-layer chromatography, gas liquid chromatography, gas chromatography-mass spectrometry, and proton magnetic resonance spectrometry showed that this glycolipid was 3-O-MeGal beta 1----3GalNAc alpha 1----3[6'-O-(2-aminoethylphosphonyl)Gal alpha 1----2]Gal beta 1----4Glc beta 1----1Ceramide. Palmitic acid, octadeca-4-sphingenine and anteiso-nonadeca-4-sphingenine are its major aliphatic components. The new glycolipid has essentially the same structure as another major phosphonoglycosphingolipid in the skin of Aplysia, SGL-II, that contains 2 mol of 2-aminoethylphosphonate residue, suggesting a metabolic relationship between the two.

Animals↗

Cooperation of middle and small T antigens of polyomavirus in transformation of established fibroblast and epithelial-like cell lines.

We have reported recently that small T antigen of polyomavirus stimulates the growth of NIH 3T3 cells beyond their saturation density and induces weak anchorage-independent growth (T. Noda, M. Satake, T. Robins, and Y. Ito, J. Virol. 60:105-113, 1986). We examined whether small T antigen would cooperate with middle T antigen in the in vitro transformation of NIH 3T3 (fibroblasts) and NRK-52E (epitheliallike) cells. The small-T-antigen gene, when cotransfected with the middle-T-antigen gene, had no additional effect on the efficiency or size of dense foci formation induced by the middle-T-antigen gene on a monolayer of NIH 3T3 cells. However, the small-T-antigen gene dramatically increased the rate of growth of NIH 3T3 cells transformed by middle T antigen in semisolid medium. Introduction of the small-T-antigen gene into middle-T-antigen-transformed cells did not disturb the integrated middle-T gene, alter expression of the middle-T gene, or enhance middle-T-antigen-associated tyrosine protein kinase activity. For NRK-52E cells, the expression of middle T antigen alone resulted in small, slow-growing foci on a monolayer. These cells did not show anchorage-independent growth, despite the fact that middle-T-antigen-associated tyrosine protein kinase activity was clearly detected in these cells. NRK-52E cells expressing both middle and small T antigens formed faster growing foci on a monolayer than middle-T-antigen-expressing cells did and grew in semisolid medium, even when the amounts of middle T antigen and its associated kinase activities were lower than those of middle-T-antigen-expressing cells. We conclude that small T antigen cooperates with middle T antigen in the in vitro transformation of established cell lines of fibroblast and epitheliallike cells, that it does not share the middle-T-antigen function even though they are structurally related, and that it has a significantly more important role in the transformation of NRK-52E cells than that of NIH 3T3 cells.

Animals↗

Mu-conotoxins share a common binding site with tetrodotoxin/saxitoxin on eel electroplax Na channels.

The binding characteristics of conotoxin GIIIA purified from the venom of a marine snail, Conus geographus, with regard to electroplax membranes from Electrophorus electricus were studied using a radiolabeled monopropionyl derivative of the toxin (3H-Pr-CGIIIA). 3H-Pr-CGIIIA bound specifically to a single class of saturable binding sites in electroplax membranes with a dissociation constant of 1.1 +/- 0.2 nM and a maximal binding capacity of 11 +/- 2 pmol/mg of protein. The latter value was similar to the number of specific binding sites (10 +/- 2 pmol/mg of protein) for 3H-lysine-tetrodotoxin (3H-Lys-TTX). Monopropionyl CGIIIA and CGIIIA had similar inhibitory effects on the binding of 3H-Lys-TTX (1 nM) to electroplax membranes with IC50 values of 3.5 and 0.9 nM, respectively. The association and dissociation of 3H-Pr-CGIIIA and electroplax membranes were much slower than those of 3H-Lys-TTX and the membranes. Mu-Conotoxins (CGIIIA and CGIIIB) and guanidinium toxins (TTX and saxitoxin) inhibited 3H-Pr-CGIIIA (1 nM) binding to electroplax membranes with IC50 values of 0.6, 1.1, 7.1, and 2.2 nM, respectively. However, several other kinds of neurotoxins and local anesthetics known to interact with Na channels did not affect 3H-Pr-CGIIIA binding. These findings indicate that mu-conotoxins must be classified in the same group of Na channel inhibitors as guanidinium toxins, since they competed with guanidinium toxins for binding sites on the Na channel. The peptide mu-conotoxins should be useful in studies on the functional and structural domains of Na channel proteins.

Aconitine↗

[Transplantation in cancer therapy].

In 1978, the new immunosuppressant, cyclosporin A, was accepted in clinical transplantation by Calne. Cyclosporin A eventually improved the results obtained after transplantation, after which it was widely accepted all over the world. Transplantation of several organs can be accepted as an effective form of treatment for patients with end-stage disease of each organ. Recently, transplantation has proved to be an excellent therapeutic method for cancer, especially in primary liver malignancy. Starzl has obtained good results in cases of fibrolamellar hepatomas. However, conservative surgical treatment should be used, whenever possible. Today, patients with unresectable liver tumor can expect long-term survival and even cure after liver transplantation with adjuvant therapeutic methods. Naturally, organ replacement is the most extensive surgical procedure for cancer treatment. However, early recurrence of tumors has often been observed following this therapy, possibly due to suppression. Adjuvant chemotherapy will thus be needed to improve the results of transplantation for malignancy.

Carcinoma, Hepatocellular↗

Analysis of donor criteria for the prediction of outcome in clinical liver transplantation.

The results of 219 orthotopic human liver transplants performed during 1985 at the University of Pittsburgh were reviewed to determine whether donor parameters could be used to predict the quality of early graft function. Multivariate discriminant analysis demonstrated that traditional parameters of donor assessment are unreliable predictors of poor graft function. Furthermore, 56% of the donors considered poor by conservative selection criteria produced livers with good early posttransplant function. Survival of recipients of primary allografts from donors rated poor was no different than survival of recipients of allografts from donors rated good.

Adolescent↗

Useful antiglobulin crossmatch test for DST-sensitized patients.

The AG-CDC tests were performed in parallel with C-CDC tests for studying sensitization of DST patients. Fourteen of 56 DST patients were positive by the AG-CDC tests after DST. Ten of 14 were also positive by both tests after DST. In 8 of 14 patients AG-CDC antibodies continued to be detectable even after C-CDC antibodies became undetectable as time passed and with plasmapheresis. Thus, AG-CDC tests are more sensitive compared to C-CDC tests and can extend the detection rate of DST-sensitized patients. Except for two DST patients (highly sensitized), 12 of 14 DST-sensitized patients were given transplants from specific blood donors at the time when their C-CDC tests became negative. One graft failed, another one had impaired function, and ten of them functioned. It should be noted that DST patients were transplanted successfully with or without plasmapheresis across a positive AG-CDC test if the titer was less than 1:4 at the time of transplant. All four DST recipients who developed antibodies detected only by AG-CDC tests after DST experienced severe accelerated rejections. In contrast, two of eight patients who were positive by both tests had "severe" accelerated rejections. The incidence of severe accelerated rejections was shown to be highly associated with the presence of AG-CDC antibodies. These facts suggest that AG-CDC antibodies may damage kidney grafts and result in severe accelerated rejections; and that AG-CDC tests are useful for predicting occurrence of severe accelerated rejections in DST recipients.

Blood Transfusion↗

Characterization of a diphosphonopentaosylceramide containing 3-O-methylgalactose from the skin of Aplysia kurodai (sea hare)

The complete structure is proposed for a ceramide (Cer), bis(2-aminoethylphosphono)-pentaoside, isolated from the skin of Aplysia kurodai. This new phosphonoglycosphingolipid was purified using two systems of column chromatography on silicic acid. The purity of the glycolipid was confirmed by thin-layer chromatography, analysis of its composition, and proton magnetic resonance spectrometry. The component carbohydrates were glucose, galactose, N-acetylgalactosamine, and 3-O-methylgalactose. Most (90%) of the fatty acid was palmitic acid and the major sphingosine bases were octadeca-4-sphingenine (51%) and anteisononadeca-4-sphingenine (38%). 2-Aminoethylphosphonyl-6-galactose was identified after its partial hydrolysis. From studies by methanolysis, permethylation, mild acid hydrolysis, hydrogen fluoride treatment, chromium trioxide oxidation combined with thin-layer chromatography, gas liquid chromatography, gas chromatography-mass spectrometry, and proton magnetic resonance spectrometry, the structure of the glycolipid was concluded to be 3-OMeGal beta 1----3GalNAc alpha 1----3[6'-O-(2-aminoethylphosphonyl)-Gal alpha 1----2](2-aminoethylphosphonyl----6)Gal beta 1----4Glc beta 1----1Cer.

Animals↗

Blockade of [3H]lysine-tetrodotoxin binding to sodium channel proteins by conotoxin GIII.

Conotoxin GIII from Conus geographus inhibited the binding of [3H]lysine-tetrodotoxin (4 nM) to electroplax membranes from Electrophorus electricus and to the rat brain P2 fraction with IC50 values of 13 nM and 7.9 microM, respectively. This inhibition observed with electroplax membranes was irreversible. These and physiological findings (Life Sci., 21 (1977) 1759-1770 suggest that conotoxin GIII inhibits Na channel activation by its interaction with the tetrodotoxin binding site of the Na channel. The differences in structures related to the activation of Na channels between the eel electroplax and the rat brain are indicated.

Animals↗

Spectrophotometric determination of copper in environmental samples by solid-liquid extraction of its 9, 10-phenanthrenequinone monoximate complex into molten naphthalene.

A fairly selective and sensitive spectrophotometric method has been developed for determination of copper after extraction of its 9, 10-phenanthrenequinone monoximate complex into molten naphthalene in the pH range of 6.1-8.4. At room temperature, the solid naphthalene containing the metal complex is separated by filtration, dissolved in dimethylformamide (DMF) and the absorbance measured at 470 nm against the reagent blank. Beer's law is obeyed in the concentration range, 0.0-9.6 micrograms of copper in 10ml of DMF. The molar absorptivity and sensitivity are 6.30 X 10(4) 1 mol-1 cm-1 and 0.001 micrograms cm-2, respectively. The interference of various ions has been studied and the method has been applied for the determination of copper in various standard reference materials, beers, wines, human hair and environmental samples.

Copper↗

HLA-DR2 and Dw2 in narcolepsy and in other disorders of excessive somnolence without cataplexy.

Studies on HLA antigens were conducted in several patient populations with the following findings: (a) All 135 Japanese narcoleptic patients, eight of whom were considered to have "symptomatic" narcolepsy, were found to be HLA-DR2 and HLA-DQw1 positive. All 17 members of a subgroup of the original population were also found to be HLA-Dw2-positive. It was concluded that HLA-DR2 is a prerequisite for the development of narcolepsy and that the diagnosis of narcolepsy can be excluded if HLA-DR2 or HLA-Dw2 is negative. The distinction between idiopathic and symptomatic narcolepsy needs to be reconsidered. (b) Haplotype studies in three families with narcoleptic members enabled detection of children at high risk for narcolepsy. (c) Of the 54 patients with disorders of excessive daytime sleepiness other than narcolepsy, those with essential hypersomnia (EHS) had a higher frequency of HLA-DR2; the others had a lower frequency. The DR2-positive EHS group could include members with an incomplete form of narcolepsy; the DR2-negative EHS group had disorders essentially different from narcolepsy, although both positive and negative groups manifested hypnagogic hallucinations, sleep paralysis, and sleep onset REMs. Two further studies were conducted in subgroups of the original narcoleptic population studied. In a subgroup of 30 patients who underwent lymphocyte subset studies, no T-cell abnormalities were detected; it is unlikely that an autoimmune mechanism is involved in the development of narcolepsy. In a subgroup of 33 narcoleptic patients, Southern's blot analysis of DNA using a DQ beta probe revealed three specific restriction fragments. Further studies are necessary to locate the DNA locus that carries the susceptibility gene for narcolepsy.

Alleles↗

A possible split of antigen DQw1 identified by antiserum T3516.

One of the human class II antigens DQw1, which was officially recognized at the 9th International Histocompatibility Workshop in 1984, has a comparatively broad spectrum. One multiparous woman's serum was confirmed to have an antibody which shows a reaction pattern of short DQw1 or a split of DQw1. Our analysis was performed with a population study, two-dimensional gel electrophoresis and blocking experiment by monoclonal antibodies.

Antibodies↗

Isolation and characterization of NIH 3T3 cells expressing polyomavirus small T antigen.

The polyomavirus small T-antigen gene, together with the polyomavirus promoter, was inserted into a retrovirus vector pGV16 which contains the Moloney sarcoma virus long terminal repeat and neomycin resistance gene driven by the simian virus 40 promoter. This expression vector, pGVST, was packaged into retrovirus particles by transfection of psi 2 cells which harbor packaging-defective murine retrovirus genome. NIH 3T3 cells were infected by this replication-defective retrovirus containing pGVST. Of the 15 G418-resistant cell clones, 8 express small T antigen at various levels as revealed by immunoprecipitation. A cellular protein with an apparent molecular weight of about 32,000 coprecipitates with small T antigen. Immunofluorescent staining shows that small T antigen is mainly present in the nuclei. Morphologically, cells expressing small T antigen are indistinguishable from parental NIH 3T3 cells and have a microfilament pattern similar to that in parental NIH 3T3 cells. Cells expressing small T antigen form a flat monolayer but continue to grow beyond the saturation density observed for parental NIH 3T3 cells and eventually come off the culture plate as a result of overconfluency. There is some correlation between the level of expression of small T antigen and the growth rate of the cells. Small T-antigen-expressing cells form small colonies in soft agar. However, the proportion of cells which form these small colonies is rather small. A clone of these cells tested did not form tumors in nude mice within 3 months after inoculation of 10(6) cells per animal. Thus, present studies establish that the small T antigen of polyomavirus is a second nucleus-localized transforming gene product of the virus (the first one being large T antigen) and by itself has a function which is to stimulate the growth of NIH 3T3 cells beyond their saturation density in monolayer culture.

Animals↗