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The sensitivity and specificity of the sheep erythrocyte - anti-sheep erythrocyte system to inhibition by pure cytolipin F has been studied with 5 antisera, in order to compare it with the rat erythrocyte-anti-rat lymphosarcoma system and its inhibition by pure cytolipin R. The cytolipin F - sheep erythrocyte system is much more sensitive than the cytolipin R - rat erythrocyte system, inhibition of hemolysis of 6 x 10(6) sheep cells being produced by 10 ng of cytolipin F (combined with a four-fold quantity of lecithin) compared with inhibition of hemolysis of 10(6) rat cells by 50 to 100 ng of cytolipin R (also combined with lecithin). Differences in sensitivity are attributed to the larger number of available cytolipin F determinants on sheep erythrocytes compared with cytolipin R determinants on rat erythrocytes. Studies of the auxiliary lipid enhancement of cytolipin F activity by galactocerebroside, lactosyl ceramide (cytolipin H), and lecithin are also reported.

Glial fibrillary acidic protein (GFAP) was studied in ependymomas by the three-step immunoperoxidase method and compared to results in astrocytomas and normal tissues. An order of reactivity for GFAP is presented. Diagnostic criteria, based on GFAP content, are proposed. In ependymomas GFAP-positive cells give rise to only some of the tumor cells, usually those forming tubules and perivascular arrangements. It is hypothesized that the same GFAP-positive cells may form tubules at their luminal poles and may produce perivascular arrangements at their other poles. The aberrant regulation associated with neoplastic transformation in glia is often, but not always, reflected in increased GFAP content. In both astrocytes and ependymal cells GFAP may have a similar function since in both cells the increase in GFAP appears to be related to the development of fibrillary processes.

Rabbit antisera to rat lymphosarcoma contain antibodies that are cytolytic for rat erythrocytes in the presence of complement. The reaction can be inhibited completely by pure cytolipin R showing that (a) immune hemolysis can be mediated through lipid determinants in the membrane, and (b) that cytolipin R determinants are present in the intact erythrocyte membrane and exposed on the surface. Optimal conditions for measurement of cytolysis in this system based on release of (51)Cr are described. Degrees of specificity of a number of different antilymphosarcoma sera are shown, based on inhibition of cytolysis by cytolipin R, cytolipin K, cytolipin H, cytolipin F (F-hapten), glucocerebroside, galactocerebroside, ceramide trisaccharide (cer-glu-gal-gal), and a mixed brain ganglioside preparation. The data suggest that cytolytic antibodies and agglutinating antibodies in these antisera are distinctive despite their common specificity for cytolipin R. Lymphosarcoma cells are more effective than erythrocytes in absorbing cytolytic antibodies.

An improved preparation of proteolipid from bovine brain white matter is described. The product obtained by repeated acetone precipitation is completely soluble in chloroform-methanol and has a fairly constant composition: 35% protein, 40% galactocerebroside, and about 25% phospholipid.

Cytolipin R, a ceramide tetrahexoside isolated from rat lymphosarcoma, was studied by sequential hydrolysis with specific glycosidases which revealed the anomeric configurations of the glycosidic bonds. Sugar linkages were established by combined gas-liquid chromatography and mass spectrometry of the partially methylated alditol acetates prepared after permethylation and hydrolysis of the intact lipid. Results indicated the structure of cytolipin R to be N-acetylgalactosaminyl(beta1-->3)galactosyl(alpha1-->3) galactosyl(beta1-->4)glucosyl ceramide. Cytolipin K (globoside I) differs in having a -galactosyl(alpha1-->4)galactosyl- internal linkage, and this difference must account for the immunological differences between cytolipin K and cytolipin R.

Ceramide lactoside [1-O-(galactosido-4-beta-glucosido)-2-N-acyl-sphingosine] was hydrolysed to ceramide glucoside and galactose by beta-galactosidase of rat brain. The reaction was not reversible, required cholate or taurocholate, had optimum pH5.0 and K(m) 2.2x10(-5)m. It was inhibited by gamma-galactonolactone and galactose as well as by ceramide, sphingosine and fatty acid. Ceramide lactoside could be degraded to ceramide, galactose and glucose by mixtures of rat-brain beta-galactosidase and ox-brain beta-glucosidase.

A method is described for determining the long-chain base content of glycosphingolipids after acid hydrolysis, using the new reagent fluorescamine. The reaction is sensitive and can be used to characterize or measure glycosphingolipids in quantities routinely separated by thin-layer chromatography.

The amount of time that monkeys (Macaca mulatta) slept was reduced after they were given p-chlorophenylalanine, a selective depletor of serotonin in animal tissues. The time spent in the rapid eye movement stage of sleep was unchanged, but the time in other sleep stages decreased. Seven regions of the brain had a 31 to 46 percent decrease in serotonin content; the concentration of cerebellar serotonin increased by 44 percent.