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Biomedical subjects

D M Marcus

Publications and source records attributed to D M Marcus.

At least 145 records · Page 8Linked to original sources

Gangliosides as markers for murine lymphocyte subpopulations.

Antibodies to GM1 ganglioside were used to study murine lymphocyte populations. In A, AKR, and BALB/c mice, anti-GM1 reacts with thymocytes and peripheral T cells. This reactivity of anti-GM1, studied by immunofluorescence, is independent of Thy-1 type and appears to be related to the reactivity of cross-reacting antibodies to asialo GM1 and GD1b, rather than GM1 itself. In addition, a subpopulation of lymphocytes reacting with anti-GM1 and anti-immunoglobulin has been found in approximately 26% of the peripheral lymphocytes of C3H mice, nude mice, and nude heterozygotes. This subpopulation is found in small numbers in A, AKR, and BALB/c mice. These studies demonstrate that antibodies to a chemically defined antigen can be used to identify T cells in many strains of mice and may delineate previously unrecognized lymphocyte subpopulations.

Animals↗

Abnormalities in the glycosphingolipid content of human Pk and p erythrocytes.

Erythrocytes of the rare Pk phenotype lack the blood group P antigen, and p erythrocytes lack both P and Pk antigens. On the basis of immunological data we suggested previously that the P and Pk antigens are the glycosphingolipids globoside and trihexosyl ceramide, respectively, and we have now confirmed these designations by chemical analysis of erythrocytes lacking these antigens. The Pk erythrocytes contain only traces of globoside and have a marked excess of trihexosyl ceramide in comparison with normal erythrocytes. The p erythrocytes lack globoside and trihexosyl ceramide and contain an excess of lactosyl ceramide and other complex glycolipids. Our analyses of normal erythrocytes also revealed complex gangliosides with the approximate chromatographic mobilities of GD1b and GT1, and several gangliosides containing N-acetylglucosamine.

Acetylglucosamine↗

Structure of the human erythrocyte blood group P1 glycosphingolipid.

A glycosphingolipid with blood group P1 activity was extracted from an acetone powder of human erythrocyte stroma with chloroform-methanol. It was purified by chromatography on columns of silicic acid and by preparative thin-layer chromatography of the fully acetylated and deacetylated glycolipid. The purified glycolipid contained galactose, N-acetylglucosamine, and glucose in a molar ratio of 3:1:1. Treatment of the P1 glycolipid with fig alpha-galactosidase released a single galactosyl residue and destroyed the blood group activity, and the alpha-galactosidase product had the same chromatographic mobility as paragloboside. Substitution sites on the neutral sugars of the P1 glycolipid and the alpha-galactosidase product were established by identification of methylated alditol acetates, and substitution on N-acetylglucosamine was determined by identification of methyl glycoside derivatives. The terminal nonreducing disaccharide of the P1 glycolipid is Gal(alpha, 1 leads to 4)Gal. N-Acetylglucosamine was identified as the next sugar in sequence by mass spectrometric analysis of the permethylated P1 glycolipid. On the assumption that the glucose residue is linked to ceramide, we propose the following structure for the P1 glycolipid: Gal(alpha, 1 leads to 4)Gal(beta, 1 leads to 4)Glc-NAc(beta, 1 leads to 4)Glc-Cer.

Blood Group Antigens↗

An immunochemical study of the human blood group P1, P, and PK glycosphingolipid antigens.

The erythrocyte PK and P blood group antigens have been identified as ceramide trihexoside (CTH), Gal-(alpha, 1 leads to 4)Gal(beta, 1 leads to 4)Glc-Cer, and globoside, GalN-Ac(beta, 1 leads to 3)Gal(alpha, 1 leads to 4)Gal(beta, 1 leads to 4)Glc-Cer, respectively, and the following structure has been proposed for the P1 antigen: Gal(alpha, 1 leads to 4)Gal(beta, 1 leads to 4)GlcNAc(beta, 1 leads to 3)Gal(beta, 1 leads to 4)Glc-Cer. Although the P1 and PK determinants have identical terminal disaccharides, CTH did not inhibit anti-P1. The P1 glycolipid and hydatid cyst glycoprotein inhibited the agglutination of P1K erythrocytes by anti-P1 and unabsorbed anti-P1PPK sera, but neither antigen inhibited a specific anti-PK serum. The P1 and PK glycolipids were equally effective in inhibiting the hemagglutinating activity of a lectin with alpha-galactosyl specificity obtained from ova of Salmo trutta. Anti-P sera were inhibited most effectively by human erythrocyte globoside, and to a lesser extent by Forssman glycolipid and rat kidney globoside. In the latter glycolipid the linkage between the internal galactosyl residues is alpha, 1 leads to 3, rather than alpha, 1 leads to 4, as in erythrocyte globoside. No cross-reactions between P and P1 or PK antigens were detected. New hypotheses are offered to explain the genetic regulation and biosynthesis of the P1, P, and PK antigens.

Antigens↗

13C NMR studies of the interaction of concanavalin A with saccharides.

The binding of alpha- and beta-methyl-D-glucopyranoside and beta-(o-iodo-pheryl)-D-glucopyranoside to concanavalin A has been studied by carbon-13 nuclear magnetic resonance techniques. The kinetics and binding orientations of these saccharides relative to the transition metal ion site in the protein have been determined.

Binding Sites↗

Measurement of serum ferritin by radioimmunoassay: results in normal individuals and patients with breast cancer.

Ferritins are iron-containing proteins found in normal tissues; they increase in concentration in many tumors and the blood of tumor-bearing individuals. We utilized a double-antibody radioimmunoassay for measurement of serum ferritin and defined the upper limit of normal as 146 ng/ml for women (mean 34 ng/ml) and 193 ng/ml for men (mean 93 ng/ml). Serum ferritin levels exceeded these limits in preoperative sera of 41% of women with mammary carcinoma (mean 199 ng/ml) and in 67% of women with locally recurrent or metastatic mammary carcinoma (mean 671 ng/ml). Individuals with hepatic inflammatory states are known to have high serum ferritin, and ferritin was increased in 43% of patients with hepatitis or cirrhosis (mean 364 ng/ml) and in 13% of patients with ulcerative colitis or gastroduodenal ulcers (mean 106 ng/ml). Measurement of serum ferritin may be useful in evaluation of patients with breast cancer and in monitoring their response to therapy.

Breast Neoplasms↗

Binding of 13 C-enriched -methyl-D-glucopyranoside to concanavalin A as studied by carbon magnetic resonance.

Binding of alpha-methyl-D-glucopyranoside, uniformly enriched with 14% (13)C, to zinc and manganese derivatives of concanavalin A at pH 5.6 was studied by pulsed Fourier transform carbon magnetic resonance techniques. Spin-lattice relaxation (T(1)) of the [(13)C]carbons of the sugar was measured in the absence and presence of the two transition metal derivatives of the protein. In the presence of the manganese derivative of concanavalin A, selective relaxation of the sugar carbons was observed. The values for T(1) reflect different distances between the carbons of the bound sugar and the manganese ion. Calculation of the distance between the manganese ion and each carbon of the sugar permit the 3-dimensional orientation of the bound sugar to be determined relative to the transition metal site in the protein. The results indicate that alpha-methyl-D-glucopyranoside binds to the protein in the Cl chair conformation with its 3- and 4- carbons closest to the manganese ion at a mean distance of 10 A.

Binding Sites↗