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Biomedical subjects

W Keil

Publications and source records attributed to W Keil.

At least 55 records · Page 3Linked to original sources
PubMed1985

The major oligosaccharides in the large subunit of the hemagglutinin from fowl plague virus, strain Dutch. Structure elucidation by one-dimensional and two-dimensional 1H nuclear magnetic resonance and by methylation analysis.

The N-glycosidically linked glycans in the large subunit (HA1) of the hemagglutinin from fowl plague virus, strain Dutch (containing about 15%, w/w, of carbohydrates), were liberated by alkaline hydrolysis, and were filtrated through Bio-Gel as the re-N-acetylated oligosaccharide alditols. One major fraction (90%, mol/mol) was obtained. It was subfractionated by concanavalin A affinity chromatography and was analyzed by methylation/capillary gas chromatography/mass fragmentography and especially by one-dimensional and two-dimensional 1H nuclear magnetic resonance. The major HA1 glycans, which are not sialylated, were thus found to comprise about 40%, 30% and 20% (mol/mol), respectively, of biantennary intersected, biantennary, and triantennary N-acetyllactosaminic ('complex') oligosaccharides. About two thirds of the internal GlcNAc residues in these glycans are substituted by Fuc(alpha 1----6), all the triantennary species carry the third Gal(beta 1----4)GlcNAc(beta 1----unit at the Man(alpha 1----6)-branch, and roughly one fourth of the N-acetyllactosamine units in the non-intersected biantennary oligosaccharides are incomplete.

Amino Acids↗
PubMed1985

Carbohydrates of influenza virus. Structural elucidation of the individual glycans of the FPV hemagglutinin by two-dimensional 1H n.m.r. and methylation analysis.

The structures of the oligosaccharides of the hemagglutinin of fowl plague virus [influenza A/FPV/Rostock/34 (H7N1)] have been elucidated by one- and two-dimensional 1H n.m.r. spectroscopy at 500 MHz and by microscale methylation analysis. N-Glycosidic oligosaccharides of the oligomannosidic (OM) and of the N-acetyllactosaminic type have been found, the latter type comprising biantennary structures, without (A) or with (E) bisecting N-acetylglucosamine, and triantennary (C) structures. Analysis of the tryptic and thermolytic glycopeptides of the hemagglutinin allowed the allocation of these oligosaccharides to the individual glycosylation sites. Each attachment site contained a unique set of oligosaccharides. Asn12 contains predominantly structures C and E which are highly fucosylated. Asn28 contains OM and A structures that lack fucose and sulfate. Asn123 shows A that has incomplete antennae but is highly fucosylated and sulfated. Asn149 has fucosylated A and E. Asn231 shows fucosylated A and E with incomplete antennae. Asn406 has OM oligosaccharides. Asn478 has A and E with little fucose. Localization of the oligosaccharides on the three-dimensional structure of the hemagglutinin revealed that the oligomannosidic glycans are attached to glycosylation sites at which the enzymes responsible for carbohydrate processing do not have proper access. These observations demonstrate that an important structural determinant for the oligosaccharide side chains is the structure of the glycoprotein itself. In addition, evidence was obtained that the rate of glycoprotein synthesis also has an influence on carbohydrate structure.

Glycopeptides↗
PubMed1984

A mutant of fowl plague virus (influenza A) with an altered glycosylation pattern in its hemagglutinin.

A temperature-sensitive mutant (ts 1/1) with a defect in the hemagglutinin (HA) gene, which was obtained by undiluted passage of fowl plague virus (FPV) at 33 degrees, is described. At 33 degrees proteolytic cleavage of the abnormal HA yielded an altered HA2 (XHA2) which migrated ahead of the NS1 protein and lacked the complex oligosaccharide side chain. At the nonpermissive temperature of 40 degrees, the migration of the HA of ts 1/1 from the rough endoplasmic reticulum (RER) via the Golgi apparatus to the cell surface was rate limiting for virus maturation. The HA was only slowly cleaved and migrated during polyacrylamide gel electrophoresis ahead of the HA of wild type FPV. Some revertants of ts 1/1 exhibited the same protein pattern as the mutant, others resembled wild type FPV, while one revertant gave rise to a mixture of HA2 and XHA2 at 40 degrees. These results suggest that (1) the loss of the complex oligosaccharide side chain is not responsible for the ts phenotype, (2) the mutation is presumably not at the site where the oligosaccharide side chain is linked to the protein backbone, and (3) ts 1/1 presumably carries a mutation located in RNA segment 4, which by pseudoreversion (suppressor mutation) in the same gene leads to different ts+ phenotypes.

Animals↗
PubMed1984

[The effect of electricity on the myoglobin content of the human heart and skeletal muscles].

The myoglobin content of heart muscles was examined by the indirect immunoperoxidase method in eight autopsy cases where death was due to electricity. Seven cases showed a considerable release of myoglobin from the myocardial fibers. In experimental comparison, the heart muscles and skeletal muscles from nine cases where death was natural were exposed to electricity, and considerable deletion of myoglobin was also demonstrated. We believe that the findings of ischemia or thermal effects due to electricity were the cause of the release of myoglobin. Because the demonstration of myoglobin is preferable to estimation of the damaged areas of muscles by hematoxylin-eosin staining, this examination may be more advantageous in forensic examination of death by electricity.

Adult↗
PubMed1984

[1st results with the Treponema pallidum hemagglutination (TPHA) test in blood stains].

Bloodstains were produced from probationers who had had syphilis at some time in their lives and from others whose anamnesis had no indication of syphilis. After storage the stain eluates underwent the treponema-pallidum-haemagglutination (TPHA) test, with the eluates' IgG content being adapted to a concentration adequate to the test conditions. The results received from the stain eluates of the previous syphilis patients corresponded in 85% of the cases with the serum findings of these probationers. It appears that the TPHA test can already provide clues as to the identity of an unknown stain producer at the beginning of the police investigation. Methodical parallels to dried-blood tests of syphilis as a clinical problem will be discussed.

Adult↗
PubMed1984

Carbohydrates of influenza virus. V. Oligosaccharides attached to individual glycosylation sites of the hemagglutinin of fowl plague virus.

The carbohydrate side chains of the hemagglutinin of fowl plague virus (A/FPV/Rostock/34 (H7N1] have been localized by a procedure involving fragmentation of the polypeptide with cyanogen bromide and various proteases. The positions of the fragments were determined by radioactive labeling of the sugars and of specific amino acids. Side chains of the complex type I are attached to asparagine residues 12, 28, 123, 149, and 478. A mannose-rich (type II) side chain is linked to asparagine 406. Asparagine 231 is not glycosylated. The side chains attached to asparagine residues 12, 123, 149, and 478 contain sulfate. Glycopeptides derived by Pronase digestion from the individual attachment sites have been analyzed by their affinity to concanavalin A and Lens culinaris agglutinin. The results indicate that each glycosylation site has a typical set of heterogeneous oligosaccharides. Comparison of the glycosylation patterns of the hemagglutinins of FPV and other influenza A viruses reveals that the glycosylation sites at asparagine residues 12, 28, and 478, which are located at the base of the spike, are highly conserved. Mannose-rich side chains appear to be located preferentially at interfaces between the three monomers of a spike or between the globular and fibrous domains of a monomer.

Acetylglucosaminidase↗
PubMed1982

[Haptoglobin type and isoantibodies (anti-A and anti-B)].

In 582 sera of blood donors of the groups A1, A2, B and 0 the Hp type as well as the anti-A or anti-B isoantibody titres respectively were determined. The frequency distribution of isoantibody titres in serum samples with different Hp-type were compared. As far as the numerical difference of distribution was concerned there was only one significant observation in the group of B alpha--titres of test persons with a different Hp-type. On the basis of these findings the following Hp-type/isoantibody relation can be established: low anti-A or anti-B titres respectively (approximately 1:8) will occur more frequently in persons with the Hp-type 1-1, higher titres (approximately 1:64) are to be found predominantly in persons with Hp2 genes. These findings are in accordance with other results, on the basis of which is was suggested that persons with Hp2 gene product have a higher immunogenic reactivity in comparison to type Hp 1-1.

ABO Blood-Group System↗
PubMed1982

[Prenatal AB0(H) blood group determination of amniotic fluid cells by mixed cell agglutination].

A modification of the mixed-cell agglutination reaction (MCAR) was used in prenatal AB0(H) blood typing of cells obtained from 41 samples of amniotic fluid. The findings are compared with results obtained from the use of the agglutination inhibition test in testing water-soluble AB0(H) substances of amniotic fluid based on the same samples. For prenatal AB0(H) typing, MCAR exhibited advantages over the agglutination inhibition test. Completion of data in this field will provide a good foundation for prenatal detection and identification of hereditary factors which can be linked with gene loci of the AB0 system.

ABO Blood-Group System↗
PubMed1979

Carbohydrates of influenza virus. III. Nature of oligosaccharide-protein linkage in viral glycoproteins.

Four different glycopeptides can be distinguished after pronase digestion of influenza A virus glycoproteins: Ia and Ib, containing N-acetylglucosamine, mannose, galactose, and fucose, and IIa and IIb, containing mannose and N-acetylglucosamine. All glycopeptides yielded N-acetylglucosaminyl-asparagine after mild acid hydrolysis. There was no evidence for O-glycosidic bonds. Thus, the carbohydrate complement is linked to the polypeptide exclusively by N-glycosidic linkages between N-acetylglucosamine and asparagine.

Acetylglucosamine↗
PubMed1977

Structural adaptations of lactate dehydrogenase isozymes.

The three-dimensional structures of dogfish M4 (muscle) and pig H4 (heart) lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) have been determined and correlated with the amino acid sequences of the dogfish M4, pig M4, pig H4, chicken M4, and chicken H4 lactate dehydrogenase isozymes. These results have been related to the known differences of physicochemical properties between the M and H lactate dehydrogenase isozymes. By far the largest structural alterations occur in the transition between the "apo" and "ternary complex" conformational states of the enzyme rather than between species or isozymes. The major catalytic difference can be explained by a replacement of alanine (in the M chain) with a glutamine (in the H chain) in the vicinity of the binding site of the coenzyme phosphates. The known immunological differentiation of the M and H isozymes is consistent with the differences in their amino acid sequences.

Adenosine↗