Per Falk

Globo-A--a new receptor specificity for attaching Escherichia coli.

Uropathogenic Escherichia coli strains designated as ONAP, based on their O negative A positive agglutination of human P1 erythrocytes, were shown to prefer the globo‐A glycolipid as a receptor structure. The dependence on both the A terminal and the globoseries chain was confirmed by agglutination of human AP1, but not A or OP1, erythrocytes and by binding to the globo‐A glycolipid on TLC plates. Neither Galαl→Ga1β nor the A trisaccharide GalNAcαl→ 3(Fucαl→2)Galβ alone functioned as receptors. The bacteria thus appeared to recognize an epitope resulting from the combination of the terminal and internal structures.

Fecal excretion of intestinal glycosphingolipids by newborns and young children

Glycosphingolipids were shown to persist in human fecal excretions from birth up 2 years of age. The pattern of glycosphingolipids was dependent on blood group and secretor status of the child and changed dramatically during the first months of life. Perinatally cerebroside, hematoside and blood group active fucolipids were dominating among fecal glycolipids. From the time of weaning lactosylceramide abruptly became and then persisted as a dominating glycolipid although cerebroside, complex gangliosides and blood group active fucolipids could still be detected in feces even at 2 years of age.

The mono- and difucosyl blood group B glycosphingolipids of rat large intestine differ in type of core saccharide

Two blood group B-active glycosphingolipids were isolated from rat large intestine and characterized by mass spectrometry, proton NMR spectroscopy and methylation analysis. The following structures were concluded: Gal alpha 1----3(Fuc alpha 1----2)Gal beta 1----3GlcNAc beta 1----3Gal beta 1----4Glc beta 1----1Cer and Gal alpha 1----3(Fuc alpha 1----2)Gal beta 1----4(Fuc alpha 1----3)GlcNAc beta 1----3Gal beta 1----4Glc beta 1----1Cer. The two glycolipids thus differ in their core saccharides (type 1 and type 2 chain, respectively) and therefore must have different pathways for biosynthesis.

An antigen present in rat adenocarcinoma and normal colon non-epithelial stroma is a novel Forssman-like glycolipid based on isoglobotetraosylceramide

A glycolipid with blood group A activity detected in the non-epithelial stroma of normal rat colon but not in epithelial cells (Hansson, G.C., Karlsson, K.-A., and Thurin, J. (1984) Biochim. Biophys. Acta 792, 281-292), was purified to homogeneity from normal rat colon and rat colon adenocarcinoma. Mass spectrometry and 1H-NMR spectroscopy of the intact permethylated derivative and gas chromatography after degradation revealed the structure GalNAc alpha 1----3GAINAc beta 1----3Gal alpha 1----3Gal beta 1----4Glc beta 1----1Cer, with the predominant ceramide containing sphingosine and non-hydroxylated 24:0 fatty acid. This identifies this glycolipid as a novel Forssman-like glycolipid, which is a tumor-associated antigen by definition, since it is not present in the normal rat large intestinal epithelium cells but in rat adenocarcinoma derived from these cells.

Enhancing effects of bile salts on the degradation of glycosphingolipids by glycosidases from bacteria of the human fecal flora.

Different concentrations of ionic and non-ionic detergents were examined for optimization of the in vitro degradations of intestinal glycosphingolipids by alpha- and beta-glycosidases from human fecal bacteria. In 5 mM Triton X-100 the enzymes hydrolyzed glycosphingolipids with lactoseries type 1 and 2 chains essentially to lactosylceramide (LacCer). In 5 mM sodium di- and trihydroxy bile salts lactosylceramide was degraded to glycosylceramide (GlcCer) in varying extent by enzymes from all five strains. The minimal bile salt concentrations for optimal 1,4-beta-galactosidase activities varied between 1 and 20 mM, i.e., close to or above the critical micellar concentrations (cmc). Dihydroxy bile salts were the most efficient in promoting conversion of LacCer to GlcCer at concentrations below 10 mM and conjugation with a taurine residue did not markedly lower the GlcCer yield. The optimal detergent concentrations for hydrolyses of the p-nitrophenyl (pnp) glycosides Gal beta 1-pnp and GalNAc alpha 1-pnp were approximately 0.05 mM for Triton X-100 and 0.5 mM for sodium taurodeoxycholate, i.e., clearly below their reported cmc values. Galabiosylceramide, globotria- and globotetraosylceramides, not degraded in the Triton X-100 micelles, were also resistant to hydrolysis using the sodium bile salts as detergents. In contrast, lactotetraosylceramide and isoglobotriaosylceramide were significantly more degraded by enzymes from a Ruminococcus gnavus strain and gangliotetraosylceramide by enzymes from a Bifidobacterium bifidum and a Bifidobacterium infantis strain using bile salt detergents. All strains but R. gnavus released terminal GalNAc from para-Forssman but not from the globotetraosylceramide or Forssman structures using 5 mM sodium deoxycholate as detergent. GM1 desialylation by two Ruminococcus torques strains and the R. gnavus and B. bifidum strains were enhanced under identical conditions. We conclude that the observed effects on glycosphingolipid hydrolyses reflects variations in the micellar presentation of the substrates. In addition, detergents seem to have a direct stimulating effect on the glycosidases, however at concentrations 10-100-times below the ones optimal for glycolipid degradations. These results with optimized bile salt concentrations, further support our previous observations that these five fecal bacterial strains produce enzymes with selected specificities towards glycosphingolipid core chains of the lactoseries type 1 and 2.

Intestinal sphingolipid excretion associated with feeding of phytohemagglutinin lectin (Phaseolus vulgaris) to germ-free and conventional rats

Intestinal sphingolipids of feces of germ-free and conventional rats were analyzed during the pair feeding of a complete defined diet containing phytohemagglutinin lectin (PHA) from red kidney beans (Phaseolus vulgaris) as 1% dietary protein in comparison to casein fed controls. Phytohemagglutinin in the diet increased the total fecal excretion of sphingomyelins (18-fold for germ-free and 20-fold for conventional rats), of non-acid glycosphingolipids (3.5-fold for germ-free and 9-fold for conventional rats) and also of the gangliosides (2.5-fold) for the germ-free rats compared to controls. For germ-free rats the increase of non-acid glycolipids was ascribed to an effect of the lectin strictly on the small intestinal mucosa, while for conventional rats an effect was seen also on the large intestinal mucosa. Increase of fecal gangliosides of germ-free rats was due mainly to an increased excretion ofN-acetylneuraminosyl-lactosylceramide, a ganglioside species restricted to epithelial cells of duodenum, of upper jejunum and of large intestines. The effects on glycolipid excretion observed in germ-free rats and the rather similar effects seen in conventional animals suggested that the influence of dietary PHA was due directly to effects elicited by PHA binding to the enterocyte brush border membrane and not to secondary effects induced by increase in the luminal microflora.