Cell surface ligands for rotavirus: mouse intestinal glycolipids and synthetic carbohydrate analogs.

Rotaviral binding to receptors on epithelial cells in the small intestine is thought to be a key event in the infection process and may be carbohydrate-mediated. Strain SA11 of rotavirus bound in vitro both to glycolipids isolated from mouse small intestine and to authentic glycolipids using thin layer chromatography overlay and microtiter well adsorption assays. Neutral mouse intestinal glycolipids which bound rotavirus were GA1 (Gal beta 1----3GalNAc beta 1---4Glc beta 1----4Glc beta 1----1-ceramide) and pentaosylceramides with terminal N-acetylgalactosamine, while acidic lipids which bound rotavirus included cholesterol 3-sulfate and two compounds termed bands 80 and 81. Digestion with ceramide glycanase suggested that bands 80 and 81 have lactosyl ceramide cores and an unidentified acidic moiety(s). No sialic-acid-containing glycolipids tested were active in viral binding. Band 81, which may have a ganglio core, bound rotavirus with greatest avidity, followed by GA1. Of authentic glycolipids assayed, only GA1 and GA2 (GalNAc beta 1----4Gal beta 1----4Glc beta 1----1-ceramide) displayed rotaviral binding. A phosphatidylethanolamide dipalmitoyl-containing neoglycolipid analog of GA2 bound rotavirus with avidity similar to native GA2. Substitution of beta 1----4-linked GlcNAc or beta 1----3-linked GalNAc for terminal GalNAc of GA2 neoglycolipid supported rotaviral binding, while other substitutions abrogated it. These findings suggest that a carbohydrate epitope similar to that of GA2 is sufficient for in vitro rotaviral binding, although binding may be enhanced by galactose and/or an acidic moiety in a secondary epitope.

Non-reducing terminal linkage position determination in intact and permethylated synthetic oligosaccharides having a penultimate amino sugar: fast atom bombardment ionization, collisional-induced dissociation and tandem mass spectrometry.

Certain linkage positions in oligosaccharides can be discerned by collision-activated dissociation mass spectrometry, rationalized by molecular modelling. Previous work on synthetic oligosaccharides has suggested that daughter ion patterns can distinguish among intact compounds which terminate in alpha-L-fucose and have a penultimate amino sugar. The current study indicates that these observations can be extended to oligosaccharides terminating in beta-D-galactose. In addition, we have observed that protonated, ammoniated and lithiated molecular ions all produce linkage-specific daughter ion spectra in these two sets of oligosaccharides. Sodiated molecular ions could be fragmented usefully under high collision energy offset conditions; potassiated ions were stable and not dissociable under conditions available in a triple-quadrupole instrument. We also show linkage discernment among the permethylated set of these six synthetic oligosaccharides. Methylated derivatives of this set of compounds give more useful product ions, including a 3-linkage specific ion. A novel relationship was noted by a plot of collision energy against (daughter ion/parent ion) ratio, which gave a unique slope for each of the non-reducing terminal linkage positions 3, 4 and 6 in the set of six compounds. The slope of this plot is related to the ability of each linkage position in the oligosaccharide to absorb collisional energy. Rotational freedom of the individual glycosidic linkage is hypothesized to play a role in this phenomenon.

Premature parturition in the California sea lion.

Twenty percent of the California sea lion pups born on San Miguel Island die due to premature parturition. Specimens collected from premature-partus animals resulted in recovery of a virus, San Miguel Sea Lion Virus, indistinguishable from Vesicular Exanthema of Swine Virus, and Leptospira pomona from some of the premature cows and pups. The age range of 10 females delivering healthy pups in June was 10-14 years. With one exception, the ages in 10 aborting females was 6-8 years. The p,p'-DDE levels of the premature parturient cows' blubber and liver were 7.6 and 4.8 times greater, respectively, than corresponding tissue concentrations in the full-term animals. Polychlorinated biphenyls residues were 4.4 and 3.8 times greater in aborting animals' blubber and liver than in the same tissues of full-term sea lions. Premature-partus females had tissue imbalances of mercury, selenium, cadmium and bromine. Pathology, parasitology, serum enzyme and hormone results are also presented. These data suggest an interrelationship of disease agents and environmental contaminants as the cause of premature parturition.