ABSTRACT
Our study investigated two groups of adult patients with established diagnoses of primary myelofibrosis (21 patients) and myelodysplastic syndromes (MDS) (21 patients). The objective was to assess fetal hemoglobin (HbF) concentration and to investigate correlations with organomegaly and extramedullary hematopoiesis and with the level of anemia and blood transfusion requirement. In all patients, the diagnosis was confirmed by histopathological examination. Patients with myelofibrosis were investigated by ferrokinetics using 59Fe. The percentage of marrow sideroblasts was assessed in patients with refractory anemia with ringed sideroblasts. Increased values of HbF were found to occur both in patients with myelofibrosis and with MDS, although a higher incidence and higher concentrations were evident in patients with myelofibrosis. Statistically significant increases in HbF concentration were found when there was accompanying organomegaly, as compared to patients without this feature. The average HbF concentration in both groups of patients under study was twice as high in cases with as in those without marrow fibrosis. The difference was statistically significant. Increased HbF levels appear to correlate with extramedullary hematopoiesis. HbF concentration did not correlate with the level of anemia or with requirement for blood transfusion.
Subject(s)
Fetal Hemoglobin/analysis , Myelodysplastic Syndromes/diagnosis , Primary Myelofibrosis/diagnosis , Adult , Aged , Anemia, Sideroblastic/complications , Erythropoiesis , Female , Humans , Male , Middle Aged , Myelodysplastic Syndromes/pathology , Primary Myelofibrosis/pathologyABSTRACT
Band 3 and PAS-1 (a dimer of glycophorin A) from erythrocyte membranes of three children with congenital disorder of glycosylation, type Ia (CDG-Ia), aged 1 month, 3 years and 10 years respectively, were examined by a new technique that allowed determination of carbohydrate molar composition of glycoproteins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In CDG children a single N-glycan of band 3 glycoprotein was hypoglycosylated and its mannose content was normal or elevated. Glycophorin A which is the major carrier of erythrocyte sialic acid, was deficient in N-acetylgalactosamine, and sialic acid residues. This finding indicated a partial unglycosylation of O-glycans in glycophorin A. In keeping with the results of PAS-1 analysis, total sialic acid in erythrocyte membranes from CDG children was reduced to 40-56% of normal values. A possible molecular mechanism of hypo- and unglycosylation of band 3 and glycophorin A, respectively, in CDG is discussed.
Subject(s)
Anion Exchange Protein 1, Erythrocyte/chemistry , Congenital Disorders of Glycosylation/blood , Glycophorins/chemistry , Carbohydrate Sequence , Carbohydrates/analysis , Case-Control Studies , Child , Child, Preschool , Congenital Disorders of Glycosylation/classification , Congenital Disorders of Glycosylation/genetics , Electrophoresis, Polyacrylamide Gel/methods , Erythrocyte Membrane/chemistry , Glycosylation , Humans , Infant , Molecular Sequence Data , N-Acetylneuraminic Acid/analysis , Proteome , Sodium Dodecyl SulfateABSTRACT
Congenital dyserythropoietic anaemia type II (CDA II) is well known for glycosylation abnormalities affecting erythrocyte membrane glycoconjugates that encompass hypoglycosylation of band 3 glycoprotein and accumulation of glycosphingolipids: lactotriaosylceramides, neolactotriaosylceramide and polyglycosylceramides. These abnormalities were not observed in erythrocytes from patients with CDA of either type I or III. Recently, however, we have described a CDA type I patient in Poland with identical, though less pronounced, glycoconjugate abnormalities to those observed in patients with CDA type II. The abnormalities included partial unglycosylation of O-linked glycosylation sites in glycophorin A. These abnormalities are now reported in three Bedouin patients from Israel with CDA type I. In addition, the erythrocyte membranes of these patients exhibited highly increased globotetraosylceramide content. Glycoconjugate abnormalities were also present in erythrocyte membranes from three patients from Northern Sweden with CDA type III but they almost exclusively affected glycosphingolipids. In erythrocytes of all patients examined including one with CDA type II, polyglycosylceramides were significantly hypoglycosylated although, on a molar basis, their contents in erythrocyte membranes were increased. Thus, glycoconjugate abnormalities of varying intensity occur in erythrocyte membranes from all patients with CDA that were investigated.
Subject(s)
Anemia, Dyserythropoietic, Congenital/blood , Erythrocyte Membrane/metabolism , Glycoconjugates/metabolism , Anemia, Dyserythropoietic, Congenital/classification , Anion Exchange Protein 1, Erythrocyte/chemistry , Anion Exchange Protein 1, Erythrocyte/metabolism , Case-Control Studies , Glycoconjugates/chemistry , Glycophorins/chemistry , Glycophorins/metabolism , Glycosylation , HumansABSTRACT
Congenital dyserythropoietic anaemias (CDAs) are rare hereditary disorders characterized by ineffective erythropoiesis and multinuclearity of erythroblasts. Three main types of the disease have been described. Glycoconjugate abnormalities in erythrocyte membrane glycoconjugates, consisting of hypoglycosylation of band 3 and accumulation of certain glycosphingolipids including lactotriaosylceramide, neolactotriaosylceramide and polyglycosylceramides, have been described only in patients with CDA type II (CDA-II). We report on identical, although less pronounced, abnormalities in erythrocyte glycoconjugates from a patient with CDA-I. A low degree of hypoglycosylation of band 3 in our patient with CDA-I suggests that hypoglycosylation is not a cause, but, most probably, a consequence of dyserythropoiesis.
Subject(s)
Anemia, Dyserythropoietic, Congenital/metabolism , Erythrocyte Membrane/metabolism , Glycoconjugates/metabolism , Anemia, Dyserythropoietic, Congenital/classification , Anion Exchange Protein 1, Erythrocyte/metabolism , Carbohydrate Sequence , Female , Glycosphingolipids/metabolism , HumansABSTRACT
Glycophorins A from erythrocyte membranes of two patients with congenital dyserythropoietic anemia type I and type II (CDA type I and II) were analyzed for carbohydrate molar composition employing a modification of the recently published method that allowed simultaneous determination of carbohydrates and protein in electrophoretic bands of glycoproteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Zdebska & Koscielak, 1999, Anal Biochem., 275, 171-179). The modification involved a preliminary extraction of erythrocyte membranes with aqueous phenol, subsequent electrophoresis and analysis of the extracted glycophorins rather than electrophoresis and analysis of the glycophorin from intact erythrocyte membranes. The results showed a large deficit of N-acetylgalactosamine, galactose, and sialic acid residues in glycophorin A from patients with CDA type I and type II amounting to about 45% and 55%, respectively. The results strongly suggest that glycophorin A in these patients is partly unglycosylated with respect to O-linked glycans. In addition, glycophorin A from erythrocytes of a patient with CDA II but not CDA I exhibited a significant deficit of mannose and N-acetylglucosamine suggesting that its N-glycosylation site was also partly unglycosylated.
Subject(s)
Anemia, Dyserythropoietic, Congenital/blood , Glycophorins/chemistry , Anemia, Dyserythropoietic, Congenital/classification , Blood Group Antigens , Carbohydrates/analysis , Electrophoresis, Polyacrylamide Gel/methods , Erythrocyte Membrane/chemistry , Glycophorins/isolation & purification , Glycosylation , HumansABSTRACT
A method is described for determination of carbohydrate and protein contents of glycoproteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then electroblotted onto polyvinylidene difluoride (PVDF) membranes. Blots were stained, and appropriate pieces of PVDF membranes were excised, destained, and subjected to sequential hydrolysis with 0.2 M trifluoroacetic acid (TFA) for 1 h at 80 degrees C, then with 2 M TFA for 4 h at 100 degrees C, and finally with 6 M HCl at 100 degrees C for 24 h to release sialic acids, neutral sugars with hexosamines, and amino acids, respectively. In some instances preliminary methanolysis was used. Carbohydrates including sialic acids were quantitated by high pH anion exchange chromatography with pulsed amperometric detection. Protein content of the bands was determined as amino acids by the fluorescamine or ninhydrin method. In the calculation of results proper adjustments were made for small amounts of fucose released by hydrolysis with 0.2 M TFA at 80 degrees C, and for partial degradation of protein during hydrolysis with 2 M TFA at 100 degrees C. Recoveries of amino acids from hydrolysates of glycoproteins that had been electroblotted onto PVDF membranes equaled those of carbohydrates. This was possible because of preliminary hydrolysis of glycoproteins with TFA, as well as washing of wet, instead of dried, PVDF membranes after hydrolysis with 6 M HCl. The two modifications increased yields of amino acids by about 30%. The method was successfully applied to the determination of molar and weight percentage composition of human transferrin, band 3 protein, glycophorin A, and alpha(1)-acid glycoprotein. In each case the results obtained for directly hydrolyzed and electrophoresed/electroblotted glycoproteins were practically identical. We also determined the glucosamine content of band 4.1 protein of erythrocytes.
Subject(s)
Carbohydrates/analysis , Electrophoresis, Polyacrylamide Gel/methods , Glycoproteins/chemistry , Proteins/analysis , Humans , Sodium Dodecyl SulfateABSTRACT
We analyzed early and late results of surgical treatment of 100 consecutive children with Down's syndrome (DS) and congenital heart defect (CHD) who were operated on between 1990 and 1997. Fifty had common atrioventricular canal (CAVC), 24 ventricular septal defect, 8 the ostium primum atrial septal defect, 8 tetralogy of Fallot (TOF), 3 patent ductus arteriosus, 3 the ostium secundum atrial septal defect, and 4 CAVC coexisting with TOF. In 93 patients total correction was performed. The total death rate was 6%. Death in the CAVC group was 8%, but it decreased to 2.7% during the past 3 years. The children who were followed up (from 7 months to 6 years; mean, 39 months) are in NYHA class I or II. There were no reoperations. The postoperative course was complicated by pulmonary infections in 38% of patients, which converted to generalized infection in 10% and was the cause of death in 8% of patients. These results indicate that CHD in DS children can be repaired with a low death rate and low incidence of severe mitral atrioventricular valve regurgitation in the CAVC group. A high incidence of severe infections can influence the final results. Repair of CHD in infancy helps to eliminate problems connected with congestive heart failure and pulmonary hypertension.
Subject(s)
Down Syndrome/complications , Heart Defects, Congenital/complications , Heart Defects, Congenital/surgery , Adolescent , Body Weight , Cause of Death , Child , Child, Preschool , Follow-Up Studies , Heart Defects, Congenital/classification , Heart Defects, Congenital/mortality , Humans , Infant , Infections/etiology , Postoperative Complications/etiology , Survival Analysis , Treatment OutcomeABSTRACT
Leukemic leukocytes from 12 patients with acute myelogenous leukemia (AML) and two patients with chronic myelogenous leukemia (CML) were isolated by centrifugations in Percoll gradients, and examined for total carbohydrates. In leukemic leukocytes from 10 of these patients ceramide-bound carbohydrates were also determined. Protein-bound carbohydrates were calculated by subtraction of ceramide-bound carbohydrates from total carbohydrates. In all samples analysed the contents of total and protein-bound carbohydrates were much lower in leukemic leukocytes than in normal neutrophils, irrespective whether the results were expressed relative to protein, DNA, cell number or dry mass. For immature leukemic cells of M0-M1 phenotype differences up to 10-fold were observed. Contents of ceramide-bound carbohydrates, i.e. those of neutral and acidic glycosphingolipids (GSLs) were also low in leukemic cells. However, when GSL carbohydrates were calculated as percentage of total carbohydrates, GSLs in leukemic leukocytes were elevated in half of the AML patients but depressed in the other half. The results are discussed in the light of the hypothesis on GSL function by one of us (Koscielak J., 1986, Glycoconjugate J. 3, 95-108). According to one element of the hypothesis, during cell differentiation newly synthesized glycoproteins (GPs) that perform specific functions are added to house-keeping GPs that are present in plasma membranes of all types of cells. Thus, during differentiation, the GP content of the cell membrane should increase and that of the so called "membrane packing" glycosphingolipids should decrease.
Subject(s)
Carbohydrate Metabolism , Leukemia, Myeloid, Acute/blood , Leukocytes/metabolism , Carbohydrates/chemistry , Cell Transformation, Neoplastic , Humans , Leukemia, Myeloid, Acute/pathology , Leukocytes/pathology , Protein Binding , ProteinsABSTRACT
Activated blood platelets shed microparticles with procoagulant activity that probably participate in normal hemostasis. We have isolated spontaneously formed microparticles from human blood and analysed them for ultrastructure, antigenic profile, and biochemical composition. In transmission electron microscopy microparticles appeared as regular vesicles with a mean diameter of 300 nm (50-600 nm). In flow cytometry almost all microparticles reacted with fluorescein isothiocyanate (FITC) labeled antibody to platelet glycoprotein complex IIb-IIIa (GpIIb-IIIa) and with FITC-annexin V but only 40-50% of microparticles reacted with FITC-antibody to platelet glycoprotein Ib (GpIb). The latter result was confirmed by double labeling of microparticles with FITC-antibody to GpIIb-IIIa and phycoerythrin (PE) labeled antibody to GpIb. Large microparticles reacted better with anti-GpIb than the small ones. A decreased level of GpIb was also demonstrated by SDS/polyacrylamide gel electrophoresis of microparticles. Compositional studies indicated, that in terms of cholesterol and protein contents, microparticles resembled platelets rather than platelet membranes as previously thought. They are, however, deficient in certain components. Thus, in comparison to platelets, microparticles had reduced contents of sialic acid (by 56.4%), galactosamine (by 48.2%), glucosamine (by 22.4%), galactose by (11.8%) and fucose (by 21.6%). Mannose content was increased by 11.8%. Total phospholipids in microplatelets were lower by 17.8%. Glycerophospholipids only were affected with phosphatidylserine being decreased as much as by 43.2%. Neutral glycosphingolipids, gangliosides and ceramides in microparticles were reduced by half.
Subject(s)
Blood Platelets/physiology , Cytoplasmic Granules/metabolism , Platelet Activation , Platelet Glycoprotein GPIb-IX Complex/metabolism , Blood Platelets/cytology , Carbohydrate Metabolism , Ceramides/metabolism , Glycerophospholipids/metabolism , Glycosphingolipids/metabolism , HumansABSTRACT
Plasma membranes of rat platelets produced at normal platelet counts and during early recovery from immune-mediated thrombocytopenia were examined for the contents of carbohydrates, lipids and glycosphingolipids. Glucosylceramide, two monosialo-gangliosides and one disialo-ganglioside were found to be the major glycosphingolipids of platelets. During thrombocytopenia the contents of these glycosphingolipids as well as of ceramides were several fold elevated. Among carbohydrate constituents of platelets and platelet plasma membranes, glycogen content was increased and that of sialic acid decreased. These results are discussed in the light of literature data on relevant biochemical characteristics of megakaryocytes at different stages of maturation and on thrombopoiesis during acute experimental thrombocytopenia.
Subject(s)
Blood Platelets/metabolism , Ceramides/blood , Glycogen/blood , Glycosphingolipids/blood , N-Acetylneuraminic Acid/blood , Thrombocytopenia/blood , Animals , Cell Membrane/metabolism , Male , Rats , Rats, Wistar , Thrombocytopenia/immunologyABSTRACT
The activity of serum alpha-6-fucosyltransferase, a platelet derived enzyme, determined in sera of 22 normal individuals and 86 patients with various disorders was positively correlated with platelet counts. When the enzyme activity in 1 microliters serum was calculated per 1000 of platelets in blood (coefficient F/P) an inverse correlation became evident in that F/P was proportionally the higher the lower was platelet count in blood. The F/P values were in a good agreement with the results of direct assays of enzyme activities in isolated platelets. Neither granulocytes, lymphocytes nor red cells significantly contributed to serum enzyme activity though granulocytes enhanced the thrombin-induced enzyme release from platelets. In platelets separated by centrifugation in density gradients the enzyme was shown to be present in platelets of intermediate and high density but missing from the light ones. It is suggested that alpha-6-fucosyltransferase of platelets may be a marker of the ploidy level of megakaryocytes.
Subject(s)
Blood Platelets/enzymology , Fucosyltransferases/blood , Platelet Count , Blood Platelets/cytology , Centrifugation, Density Gradient , Female , Granulocytes/enzymology , Humans , Lymphocytes/enzymology , Male , Octoxynol/chemistry , Thrombin/pharmacology , Thrombocytopenia/enzymology , Thrombocytosis/enzymologyABSTRACT
Two ganglioside-associated protein components I and II have been isolated from crude ganglioside preparations of calf brain by DEAE-Sephadex ion-exchange chromatography. Both components exhibited binding capacity in aqueous media for gangliosides of the 'ganglio' series but not for neutral glycosphingolipids (polyglycosylceramides) and only a low capacity for sialosylparagloboside. Each protein bound individual gangliosides with different efficiency. Upon prolonged incubation of component I with gangliosides, complexes with high (30:1) and low (6:1) glycolipid/protein molar ratios were formed. The latter but not the former complex was able to penetrate Sephadex G-200 beads. Both components inhibited plating efficiency of cultured mouse N2a neuroblastoma cells. The molecular masses of components I and II were determined by SDS/PAGE to be 11-12 kDa and 28 kDa, respectively. Carbohydrates (fucose, mannose, galactose, N-acetylglucosamine, N-acetylgalactosamine, and some sialic acid) were found only in component II. When examined by reverse-phase HPLC each component separated into two major closely migrating peaks which were subsequently examined by Edman degradation. Amino acid sequences of the N-terminal portions of three of these peaks (one peak from component I and both peaks from component II) showed, as far as the sequences were established, identity with the sequence of ubiquitin. It is hypothesized that the proteins may be instrumental in intracellular trafficking of gangliosides.
Subject(s)
Brain Chemistry , Carrier Proteins/isolation & purification , Gangliosides/metabolism , Nerve Tissue Proteins/isolation & purification , Ubiquitins/chemistry , Amino Acid Sequence , Animals , Carbohydrate Sequence , Carbohydrates/analysis , Carrier Proteins/chemistry , Carrier Proteins/metabolism , Cattle , Cell Division/drug effects , Chromatography, High Pressure Liquid , Mice , Molecular Sequence Data , Molecular Weight , Nerve Tissue Proteins/chemistry , Nerve Tissue Proteins/metabolism , Peptide Fragments/chemistry , Sequence Homology, Amino Acid , Sphingomyelins/metabolism , Tumor Cells, CulturedABSTRACT
The authors present the results of surgical treatment obtained in 288 children with tetralogy of Fallot. In 127 children palliative procedures (pulmonary-systemic anastomoses) were employed, whereas 161 patients were subjected to total surgical corrections of the defect, performed in cardiopulmonary bypass. In 127 (79%) belonging to the latter group the correction was a single stage procedure, and in 34 (21%), it followed pulmonary-systemic anastomoses. Perioperative mortality rate in children subjected to single stage procedures was 12%, being lightly lower than the rate observed in patients subjected to two stage procedures. The analysis of operative results corrected for age revealed a higher mortality rate in children operated when less then two years of age (17%) in comparison to older patients (11%).
Subject(s)
Tetralogy of Fallot/surgery , Adolescent , Age Factors , Cardiopulmonary Bypass/methods , Cardiopulmonary Bypass/mortality , Child , Child, Preschool , Humans , Infant , Palliative Care , Reoperation , Survival RateABSTRACT
Human erythrocyte membranes which had been thoroughly extracted with organic solvents contained 20 nmol of fatty acids/mg dry wt. The major fatty acids were palmitic and stearic with their monoethenoic derivatives as minor constituents. No other fatty acids were detected. When solvent-extracted membranes were digested with Pronase about 90% of the original content of fatty acids was retained in the insoluble residue. Fatty acids were linked to membrane proteins through alkali-labile bonds of which 30% were of a thiol ester and the remainder of an O-ester type. This conclusion is based on differential liberation of fatty acids by hydroxylamine at pH 7.0 and pH 11.0. Two extracts of membranes enriched in peripheral proteins (bands 1, 2, 5 and 2.1, 4.1, 4.2, 6) were prepared and extracted with organic solvents but each contained about six times less fatty acids than the parent solvent-extracted membranes. Glycophorin A contains little if any covalently bound fatty acids. Anion transporter (band 3) contains about 1 mol of thiol ester of fatty acid. This accounts for about half of the thiol ester-linked fatty acids in the parent solvent-extracted membranes. Most of the O-ester-linked fatty acids are linked to an undisclosed membrane protein.
Subject(s)
Blood Proteins/metabolism , Carrier Proteins/metabolism , Erythrocyte Membrane/analysis , Fatty Acids/analysis , Membrane Proteins/metabolism , Anion Transport Proteins , Electrophoresis, Polyacrylamide Gel , Esters , Fatty Acids/metabolism , Humans , Hydrogen-Ion Concentration , Membrane Glycoproteins/analysis , Sulfhydryl Compounds/analysis , Sulfhydryl Compounds/metabolismABSTRACT
Previously we have shown that human platelets release alpha-6-L-fucosyltransferase (EC 2.4.1.68) during coagulation of blood [(1987) Glycoconjugate J. 4, 43-49]. Here we report that agonists which induce platelet aggregation bring about release of the enzyme. In quantitative terms the release of alpha-6-L-fucosyltransferase by washed, aggregated platelets was very similar to that occurring during blood coagulation.
Subject(s)
Blood Platelets/physiology , Fucosyltransferases/blood , Hexosyltransferases/blood , Platelet Aggregation , Adenosine Diphosphate/pharmacology , Blood Platelets/enzymology , Collagen/pharmacology , Epinephrine/pharmacology , Fucosyltransferases/metabolism , Humans , Ristocetin/pharmacology , Thrombin/physiologyABSTRACT
The composition and structure of neutral and acidic oligoglycosylceramides, polyglycosylceramides and polyglycosylpeptides were determined in erythrocyte membranes of two patients with congenital dyserythropoietic anaemia type II. In keeping with previous studies we found an elevated accumulation in CDA II erythrocytes of LacCer, Lc3Cer and nLc4Cer. Gb4Cer was elevated in erythrocytes of only one of the two patients tested. In addition we found a significant increase of 6IVNeuAcnLc4Cer ganglioside. Polyglycosylceramides were elevated 6-fold but they resembled those of cord erythrocytes with respect to complexity and the number of side chains. Polyglycosylpeptides of CDA II erythrocytes were decreased 7-fold. These glycopeptides were, however, heterogeneous with respect to branching pattern; the minor fraction was highly branched whereas the major one was more linear in structure. Both polyglycosylceramides and polyglycosylpeptides exhibited high I and i antigenicity. We postulate that the accumulation of glycolipids and underglycosylation of glycoproteins in CDA II membranes results from the prolongation of G1 and possibly M phases of the mitotic cycle of the erythroid cells in which glycolipids are preferentially synthesized.
Subject(s)
Anemia, Dyserythropoietic, Congenital/blood , Anemia, Hemolytic, Congenital/blood , Erythrocyte Membrane/analysis , Glycolipids/blood , Glycopeptides/blood , Adult , Carbohydrate Sequence , Gangliosides/analysis , Glycosphingolipids/blood , Humans , Male , Membrane Lipids/blood , Membrane Proteins/bloodSubject(s)
Aortic Coarctation/diagnosis , Adolescent , Aorta/pathology , Aortic Coarctation/pathology , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , MaleABSTRACT
The lipid content and composition of the enamel pellicle from caries-resistant (CR) and caries-susceptible (CS) subjects and their effect on its ability to retard the diffusion of lactic acid were investigated. Lipids accounted for 22.2 per cent of the dry weight of CR pellicle and 23.7 per cent of CS pellicle. The content of glycolipids in both groups was similar but CR pellicle contained 42 per cent less neutral lipids and 31 per cent less phospholipids. CR lipids had a higher content of cholesterol, cholesterol esters and sphingomyelin, whereas CS pellicle was richer in free fatty acids and phosphatidylethanolamine. Retardation of lactic-acid diffusion by CR pellicle was 45 per cent higher than by CS. Removal of lipids caused 50 per cent reduction in retardation by CR pellicle and 35 per cent by CS pellicle.
Subject(s)
Dental Caries Susceptibility , Dental Deposits/analysis , Dental Enamel/metabolism , Lactates/metabolism , Lipids/analysis , Adult , Dental Deposits/metabolism , Dental Pellicle , Fatty Acids/analysis , Female , Glycolipids/analysis , Humans , Lactic Acid , Male , Permeability , Phospholipids/analysisABSTRACT
Alkaline borohydride reductive cleavage of the mucin, purified from gastric aspirates of the secretors with blood group A, resulted in a heterogeneous population of neutral (79.7%) and acidic (20.3%) oligosaccharide alditols. Nine oligosaccharides (I-IX), ranging from 6 to 15 sugar units, have been purified from the neutral oligosaccharide fraction. Based on the results of immunological assays, sugar composition, degradation with specific exoglycosidases, and methylation analyses, we propose the following structures for these oligosaccharides: (sequence in text)