Increased concentration of circulating acid glycosaminoglycans in chronic lymphocytic leukaemia and essential thrombocythaemia.
ABSTRACT To verify whether the increase in the number of circulating blood cells that synthesize glycosaminoglycans, B-lymphocytes or platelets, in proliferative disorders, may be associated with changes in the circulation of acid glycosaminoglycans, the serum and plasma concentrations of these polysaccharides have been measured in terms of their sugar components, following isolation and purification by chromatographic methods, in patients with chronic lymphocytic leukaemia or with essential thrombocythaemia and in healthy controls. In the patients, the concentrations of total circulating glycosaminoglycans and of both glucosamine-containing and galactosamine-containing serum glycosaminoglycans were significantly higher than in controls. These concentrations did not significantly correlate with the number of lymphocytes in patients with chronic lymphocytic leukaemia and of platelets in patients with essential thrombocythaemia. Analytical data suggest that excess glycosaminoglycans are mainly composed of chondroitin sulphate molecules and contain heparan sulphate structures.
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ABSTRACT: Heterogeneous heparan sulfate and chondroitin sulfate glycosaminoglycan (GAG) polysaccharides are important components of blood circulation. Changes in GAG quantity and structure in blood have been indicated in cancers and other human diseases. However, GAG quantities and structures have not been fully characterized due to lack of robust and sensitive analytical tools. To develop such tools, we isolated GAGs from serum and plasma. We employed liquid chromatography (LC) for GAG quantification and LC/mass spectrometry (MS) for GAG structural analysis. Twenty-four heparan and chondroitin sulfate motifs were identified, including linkage hexasaccharides, repeating disaccharide compositions, reducing, and non-reducing end mono-, di-, tri-, and tetrasaccharide structures. Disaccharides were detectable at picomolar level without radiolabeling or derivitization, so only a few ml of human and fetal bovine serum was required for this study. The detection of different reducing end structures distinct from GAG linkage hexasaccharides revealed that free GAG chains generated by GAG degradation enzymes co-existed with proteoglycans in serum. In addition, a novel sialic acid-modified linkage hexasaccharide was found conjugated to bikunin, the most abundant serum proteoglycan.Glycobiology insights. 02/2010; 2010(2):13-28.
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ABSTRACT: An improved isocratic high-performance liquid chromatography (HPLC) method for the analysis of l-(−)-fucose, d-(+)-galactosamine, d-(+)-glucosamine, d-(+)-galactose, obtained by hydrolysis of glycosaminoglycans (GAGs) and d-(+)-glucose and d-(+)-mannose is described. The presence in circulation of GAGs, acid polysaccharide sequences of alternate monosaccharide units, aminosugar and uronic acid (galactose in keratan sulfate), has been measured in terms of their sugar components. To evaluate concentration of these circulating sugars we considered blood samples obtained from healthy humans. Plasma or serum was filtered through weak anion-exchange Ecteola-cellulose either untreated or after mild alkaline treatment. GAGs adhering to resin were recovered by salt elution, and desalted on Bio-Gel P-2 resin. GAG fractionation by charge was carried out on a strong anion exchanger. GAG composition was evaluated in terms of galactose and aminosugars, measured in HPLC by the proposed procedure using anion-exchange resin and pulsed amperometric detection. The mobile phase consisted of 0.02 M NaOH and elution was carried out at flow-rate of 1.0 ml/min. The amperometric detector was set as follows: t1 (0.5 s), E1 (+0.1 V); t2 (0.09 s), E2 (+0.6 V); t3 (0.05 s), E3 (−0.6 V). The analysis required 14 min. Calibration standard curves for the six analytes were linear from 0.25 to 40 μM. RSD values for intra- and inter-day variabilities were ≤5.3% at concentrations between 0.25 and 40 μM. Accuracy, expressed as percentage error, ranged from −16 to 14%. The method was specific and sensitive with quantitation limits of 1 pmol for l-(−)-fucose, d-galactosamine and d-glucosamine, 3 pmol for d-(+)-galactose and d-(+)-glucose and 5 pmol for d-(+)-mannose. The results of the assay showed higher GAG concentrations in serum than in plasma.Journal of chromatography. B, Biomedical sciences and applications 01/2002;
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ABSTRACT: Hyaluronan (HA), a component of the extracellular matrix surrounding tumors, modulates tumor progression and the immune response. Dendritic cells (DC) may tolerize or stimulate immunity against cancer. In this report, we study the association between tumor progression, HA levels and DC activation in a lymphoma model. Mice injected with the cells with highest invasive capacity (LBR-) presented increased HA in serum and lymph nodes, and decreased DC activation in infiltrated lymph nodes and liver. These findings could be related to lack of an effective antitumor immune response and suggest that serum HA levels could have a prognostic value in hematological malignancies.Immunobiology 12/2011; 217(9):842-50. · 2.81 Impact Factor