Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method.
TL;DR: A new method for the quantitive determination of sialic acids is described, which is about 50% more sensitive than the orcinol-hydrochloric acid method generally used and considerably lower with the resorcin reagent.
About: This article is published in Biochimica et Biophysica Acta.The article was published on 1957-01-01. It has received 2741 citations till now. The article focuses on the topics: Sialic acid & Resorcinol.
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TL;DR: It is shown that a protein with a glycosylphosphatidyl inositol (GPI) anchor can be recovered from lysates of epithelial cells in a low density, detergent-insoluble form, supporting the model proposed by Simons and colleagues for sorting of certain membrane proteins to the apical surface after intracellular association with glycosphingolipids.
2,970 citations
TL;DR: Chromatographic methods for the quantitative isolation and separation of human brain gangliosides are described and it is shown that predominantly the gangLiosides have a common basic structure : N-acyl-sphingosine-glucose-Galactose-galactoseN-acetylgalactosamine-gal actose to which one or more molecules of N-acetelneuraininic acid are bound.
Abstract: GANGLIOSIDES are defined as acylsphingosyl oligosaccharides containing sialic acid (KLENK, 1942). For several years brain gangliosides were considered to contain a common carbohydrate moiety (KLENK and LAUENSTEIN, 1953 ; BOGOCH, 1958). The occurrence of brain gangliosides with different carbohydrate moieties was suggested (SVENNERHOLM, 1956, 1957a), since different ganglioside preparations varied in their composition and behaviour on cellulose columns. KUHN and EGGE (1959) confirmed the chromatographic separation of brain gangliosides and showed that the slowmoving gangliosides had a considerably more complicated structure than the fastmoving ones. During the last two years reports have appeared from many laboratories on the complexity of brain gangliosides. SVENNEMOLM and RAAL (1961) isolated monoand disialogangliosides from human brains, KUHN, WIEGANDT and EGGE (1961) found one inonosialoganglioside, two disialogangliosides and one trisialoganglioside in human and calf brains, and DAIN, et al. (1962) isolated four components from ox brain with similar composition to those of KUHN et al. (1961). KLENK and GIELEN (19614 isolated the hexosamine-containing and the hexosaniinefree monosialogangliosides originally postulated by SVENNERHOLM (19576). In a recent communication (SVENNERHOLM, 1962) it was shown that predominantly the gangliosides have a common basic structure : N-acyl-sphingosine-glucose-galactoseN-acetylgalactosamine-galactose to which one or more molecules of N-acetylneuraininic acid are bound. In the present report chromatographic methods for the quantitative isolation and separation of human brain gangliosides are described. These methods have been applied to studies of the ganglioside fraction of normal foetal, infant and adult brains while several different gangliosides have been isolated and further characterized. A knowledge of the normal ganglioside pattern is important in the study of the metabolism of the gangliosides but it is also necessary for the detection of disturbances in ganglioside metabolism. In infantile amaurotic idiocy (SVENNERHOLM, 1962) for example, there are no abnormal gangliosides but excessive amounts of a ganglioside are present which normally constitute only a few per cent of the total ganglioside fraction. It is likely that in several other mental or neurological disorders there is a disturbance of the normal ganglioside pattern. As this can now be determined by simple chromatographic methods it will be quite easy to detect minor deviations from the normal ganglioside metabolism.
1,595 citations
TL;DR: The myelin prepared from 20‐day animals had the highest content of the first three constituents and the values of the four constituents were relatively constant per unit weight of myelin; the amounts of nucleic acid and ganglioside recovered in the myelin fractions increased with increasing age and myelin yield.
Abstract: — A procedure is described for the preparation from rat brain of myelin having the same degree of purity at all ages. Such a procedure is essential for the study of myelin composition during development. Microsomal contamination was successfully eliminated by adjusting the method to maintain a constant amount of brain per unit volume in the initial density gradient step, and by including two osmotic shocks and two low-speed centrifugation steps. Myelin prepared in this way from animals ranging from 15 days to 14months of age had a total ATPase activity of 0.3-2.0 μmol of Pi.h−1.mg−1 dry wt of myelin, representing 0.1-1.2 per cent recovery of the total homogenate activity; a Na+, K+- ATPase activity of 0.1-1.6 μfnol of Pi.h−1.mg−1 dry wt, representing 0.04-1.5 per cent recovery; a nucleic acid content of 0.2-0.7 per cent of myelin dry wt, representing 0.2-2.0 per cent recovery; and a ganglioside NANA content of 0.04-0.07 per cent of myelin dry wt. representing 0.2-4.6 per cent recovery. The myelin prepared from 20-day animals had the highest content of the first three constituents; otherwise the values of the four constituents were relatively constant per unit weight of myelin. The amounts of nucleic acid and ganglioside recovered in the myelin fractions increased with increasing age and myelin yield.
1,433 citations
Cites methods from "Quantitative estimation of sialic a..."
...Ganglioside N-acetylneuraminic acid was analyzed by the method of SVENNERHOLM (1957) as modified by MIETTINEN and TAKKI-LUUKKAINEN (1959)....
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TL;DR: This chapter discusses the procedures employed for the analysis of the sugar components of glycoproteins, and the hydrolytic release of the monosaccharides is also considered.
Abstract: Publisher Summary This chapter discusses the procedures employed for the analysis of the sugar components of glycoproteins. Most of the methods used for the estimation of the monosaccharides present in glycoproteins are colorimetric in nature. Because of the relatively low specificity of some of these color reactions, it is often important to apply them only after appropriate separation of the constituents. In many cases complete identification, preferably by chromatographic means, of all the sugar and amino acid constituents of a glycoprotein should precede any colorimetric analysis so that potentially interfering substances may be taken into account. The hydrolytic release of the monosaccharides is also considered in the chapter. The optimal conditions of acid hydrolysis for the monosaccharides vary substantially because of differences both in the stability of their glycosidic bonds and in their susceptibility to destruction during the hydrolysis. One of the prerequisites for an accurate analysis of the monosaccharide components of a polymer is finding hydrolysis conditions under which all of a given sugar is released and yet is not significantly destroyed.
1,231 citations
TL;DR: Results of the present study suggest that natural immunity to meningococcal disease is initiated, reinforced, and broadened by intermittent carriage of different strains ofMeningococci throughout life.
Abstract: Results of the present study suggest that natural immunity to meningococcal disease is initiated, reinforced, and broadened by intermittent carriage of different strains of meningococci throughout life. In young adults, carriage of meningococci in the nasopharynx is an efficient process of immune sensitization. 92% of carriers of serogroup B, C, or Bo meningococci were found to develop increased titers of serum bactericidal activity to their own meningococcal isolate, and 87% developed bactericidal activity to heterologous strains of pathogenic meningococci. The rise in bactericidal titer occurred within 2 wk of onset of the carrier state, and was accompanied by an increase in titer of specific IgG, IgM, and IgA antibodies to meningococci. In early childhood, when few children have antibodies to pathogenic meningococci, active immunization seems to occur as a result of carriage of atypical, nonpathogenic strains. Immunity to systemic meningococcal infection among infants in the neonatal period is associated with the passive transfer of IgG antibodies from mother to fetus. The antigenic determinants which initiate the immune response to meningococci include the group-specific C polysaccharide, cross-reactive antigens, and type-specific antigens.
1,028 citations
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TL;DR: To 1.0 cc.
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TL;DR: The method makes use of the blue color produced when fructose is dehydrated with HCl in the presence of diphenylamine, a reaction first observed by Ihl and Pechmann (4).
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