Basic Exercises in Immunochemistry

Nowotny, Alois

doi:10.1007/978-3-662-00148-6

Cited by 183 publications

(123 citation statements)

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“…The tubes were centrifuged at 4 °C for plasma in a refrigerated centrifuge for 15 min at 1 000 g. Plasma was stored at -20 °C until being analyzed for vitamin A. Immunoglobulins (IgG1, IgG2, IgM) were determined by the single radial immunodiffusion procedure using a specific SRID kit (VMRD Inc, Pullman, WA, USA), and the standards and guidelines provided by the manufacturer. The method for the determination of KLH antibody titre was similar to one reported previously (Nowotny, 1969). The haemagglutinations were performed using KLHcoated sheep red blood cells prepared by the tanned red blood cell technique.…”

Section: Methodsmentioning

confidence: 99%

Effect of oral supplements of vitamin A on the plasma retinol levels in calves and their immunological unresponsiveness

Hidiroglou¹,

Markham²

1996

Reprod. Nutr. Dev.

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Section: Methodsmentioning

confidence: 99%

Effect of oral supplements of vitamin A on the plasma retinol levels in calves and their immunological unresponsiveness

Hidiroglou¹,

Markham²

1996

Reprod. Nutr. Dev.

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“…bovine-serum albumin as standard. Carbohydrate was determined by the phenol-sulfuric acid method (44), with Dextran as standard.…”

Section: Methodsmentioning

confidence: 99%

Molecular Complementarity of Yeast Glycoprotein Mating Factors

Crandall

Lawrence

Saunders

1974

Proc. Natl. Acad. Sci. U.S.A.

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Cell fusion between opposite mating types 5 and 21 of the yeast Hansenula wingei is initiated by a strong sexual agglutination reaction. The mating factors responsible for the specificity of cellular recognition are complementary glycoproteins which form a physical complex in vitro. The complex is assayed by recovery of agglutination activity of the multivalent 5-factor after the univalent 21-factor has been inactivated by treatment of the complex with alkali. The 5-factor 21-factor complex, purified on Sepharose 6B, is large (several million daltons) and heterogeneous. The three peaks of 5-factor activity contain a number of combining sites proportional to molecular size.In this paper, evidence is presented for complex formation in vitro between yeast glycoprotein mating factors. These cellwall factors determine the specificity of cellular recognition as the first step in mating. Thus, these factors may be considered "complementary" in the sense proposed by Emil Fisher (1) in his-analogy:... "dass Enzym und Glucosid wie Schloss und Schlussel" .... Shortly later, in 1913, Lillie (2) described an activity extracted from sea-urchin eggs that specifically agglutinated sperm cells and was therefore complementary to a sperm-surface component. These early ideas concerning preformed combining sites on macromolecules formed the basis of models proposed by Tyler (3) and Weiss (4) in which the specificity of cellular adhesion in tissue development was explained by the presence of complementary molecules on opposing cell surfaces which combined in a manner analogous to antigen-antibody complex formation. Subsequent studies of cell contact have been reviewed extensively (see refs. 5-16 and other papers in these volumes). In systems involving adhesion of identical cells, additional theoretical considerations and biophysical parameters have been proposed (17-21). Current research in cellular recognition is centered on the identification and characterization of cell-surface aggregation factors isolated from diverse systems (22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33). In general, the information for the specificity of aggregation factors is considered to reside in the primary sequence of the protein moiety. However, the role played by the carbohydrate moieties may be nontrivial since most of the aggregation factors studied thus far either interact with carbohydrates or glycoproteins on the cell surface or are glycoproteins themselves. The latter is true for the mating factors isolated from sexually agglutinative mating types of the yeast Hansenula wingei (34). This mating system has been reviewed (34), and criteria for determining specificity in other cell-aggregating systems were proposed (35). The hypothesis that the haploid mating factors are mutually repressed in the nonagglutinative diploid (36) has been supported by recent work in which conditions for the differential induction of each glycoprotein mating factor in the diploid were discovered (37).The mating factor from strain 5, called 5-factor (5f) (38) or ...

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“…The presence of fatty acids within lipid A seems to be essential for endotoxic activity of lipopolysaccharides, [9,15], since chemically [lo] It is not known, however, whether only the quantity or possibly the nature and type of linkage of fatty acids in lipidA determine the degree of biological activity of endotoxins. Comparison of the fatty acid composition in lipopolysaccharides derived from various genera of gram-negative bacteria may help to elucidate their significance for endotoxicity.…”

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confidence: 99%

Nature and Linkages of the Fatty Acids Present in Lipopolysaccharides from Vibrio metchnikovii and Vibrio parahemolyticus

Rietschel

Palin

Watson

1973

European Journal of Biochemistry

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The lipopolysaccharides of Vibrio metchnikovii and Vibrio parahemolyticus have been analyzed for their fatty acid composition. These fatty acids are probably bound to the hydroxyl and amino groups of glucosamine residues within the lipid A component of thc lipopolysaccharides. With regard to nature and to type of linkage of the fatty acids, both lipopolysaccharides are almost identical. Approximately equal amounts of myristic, palmitic and 3-lauroxylauric acid (3-0-lauroyl-3-hydroxylauric acid) are involved in cster linkages. I n addition, varying amounts of an unsaturated fatty acid are ester-bound. 3-Hydroxymyristic acid, whose hydroxyl group is free, is exclusively amide-bound. The similarities and differences regarding the nature and distribution of fatty acids in Vibrio lipopolysaccharides as compared to Salmonella lipopolysaccharides are discussed.Endotoxins (lipopolysaccharides) of many gramnegative bacteria (Salmonella, Escherichia and Xhigella) are built up according to a common structural principle : they consist of a heteropolysaccharide and a covalently bound lipid portion, the so called lipid A It is not known, however, whether only the quantity or possibly the nature and type of linkage of fatty acids in lipidA determine the degree of biological activity of endotoxins. Comparison of the fatty acid composition in lipopolysaccharides derived from various genera of gram-negative bacteria may help to elucidate their significance for endotoxicity.In the present study, the nature and linkages of fatty acids present in the lipopolysaccharides derived from two Vibrio strains, 8. metchnikovii and V . parahemolyticus, have been investigated. Lipopolysaccharides of both organisms are potent endotoxins, comparable in pyrogenic activity and lethal toxicity to endotoxins from Xalmmlla and Escherichia coli [12] (and Watson, unpublished results). It will be shown that in the lipid A component of both Vibrio lipopolysaccharides, about equal amounts of myristic, palmitic and 3-lauroxylauric acid are present in ester linkages and that 3-hydroxymyristic acid is exclusively involved in amide linkage. The fatty acids are probably linked to glucosamine residues. on trypticase soy broth (with 3O/,, NaCl for V. para- MATERIALS ANT) METHODS Bacteria and Lipopolysaccharides

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Basic Exercises in Immunochemistry

Cited by 183 publications

References 0 publications

Effect of oral supplements of vitamin A on the plasma retinol levels in calves and their immunological unresponsiveness

Effect of oral supplements of vitamin A on the plasma retinol levels in calves and their immunological unresponsiveness

Molecular Complementarity of Yeast Glycoprotein Mating Factors

Nature and Linkages of the Fatty Acids Present in Lipopolysaccharides from Vibrio metchnikovii and Vibrio parahemolyticus

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