Heteropolysaccharides isolated from liquid cultures of nine Tremella species contained 0.3 to 1.2% protein, 2.7 to 5% ash, 0.9 to 3.4% acetyl groups, 76.5 to 84.2% carbohydrates and trace amounts of starch. The polysaccharides in aqueous solution were slightly acidic (pH 5.1 to 5.6). They consisted of the following monomeric sugars: fucose, ribose, xylose, arabinose, mannose, galactose, glucose and glucuronic acid. The backbones of the polysaccharide structures consisted of alpha-(1-->3)-links while the side chains were beta-linked.
Arabinose ; Carbohydrates ; Fucose ; Galactose ; Glucose ; Glucuronic Acid ; Mannose ; Polysaccharides ; Ribose ; Starch ; Xylose

The present study was carried out to investigate the correlation with the sugar specificity of the apoptotic cells and the phagocytosis of the macrophage. Young rat thymocytes were isolated from the thymus and macrophages were isolated from the peritoneal cavity. Isolated thymocytes were treated with dexamethasone to induce apoptosis. After identified the apoptotic cells by TUNEL method, nine kinds of lectins (PSA, UEA I, GSL I B4, ECL, DBA, SBA, DSL, GSL II or WGA) were used to investigate the sugar specificity of the apoptotic thymocyte. And we carried out the phagocytosis assay using the lectin-inhibitory sugars. By TUNEL method, the most numbers of apoptotic thymocytes were observed in the cultured thymocytes treated with 10(-6)M of dexamethasone for 6 hours. From lectin histochemistry, apoptotic thymocyte showed positive reaction with PSA, UEA I, SBA, DSL and WGA, which indicate apoptotic cells have the sugar residues of alpha-D-mannose, alpha-L-fucose, terminal beta-N-acetylgalactosamine and internal beta-1, 4-N-acetylglucosamine oligomers. From phagocytosis assay using cultured thymocytes and macrophages, apoptotic thymocytes of positive reaction with TUNEL method were phagocytosed by macrophage. When the apoptotic thymocytes treated with the lectin-inhibitory sugars were mixed with the macrophages, it was observed that the phagocytosis of macrophages was reduced by methyl-alpha-mannopyranoside and alpha-L-fucose. Considering overall results, it can be assumed that the mannose and fucose may play an important role in the recognition of apoptotic thymocytes by macrophages.
Animals ; Apoptosis ; Carbohydrates ; Dexamethasone ; Fucose ; In Situ Nick-End Labeling ; Lectins ; Macrophages* ; Mannose ; Peritoneal Cavity ; Phagocytosis ; Rats ; Sensitivity and Specificity ; Thymocytes* ; Thymus Gland

OBJECTIVETo isolate and purify the polysaccharides from Radix Rehmanniae and analysis the monosaccharides composition.

METHODThe polysaccharides were extracted with hot water and precipitated by alcohol. Proteins in the precipitates were removed by TCA method. The products were further purified with column chromatography on Superdex 200 and Sephadex G100. The SRP I and SRP II were identified as homogeneous polysaccharide by HPLC, respectively, and then analyzed by GC after being hydrolysised.

RESULTTwo homogeneous polysaccharides (SRP I and SRP II) were obtained from Radix Rehmanniae.

CONCLUSIONSRP I contained rhamnose, arabinose, glucose and galactose in the percentage of 6.11%, 66.46%, 3.93% and 21.50%. SRP I was composed of rhamnose, fucose, mannose, galactose and fructose in the percentage of 21.82%, 24.47%, 10.48%, 29.94% and 13.29%.

Arabinose ; chemistry ; isolation & purification ; Chromatography, Gas ; methods ; Clinical Laboratory Techniques ; Drugs, Chinese Herbal ; analysis ; Fructose ; chemistry ; isolation & purification ; Fucose ; chemistry ; isolation & purification ; Galactose ; chemistry ; isolation & purification ; Glucose ; chemistry ; isolation & purification ; Mannose ; chemistry ; isolation & purification ; Monosaccharides ; chemistry ; isolation & purification ; Plant Extracts ; chemistry ; Polysaccharides ; chemistry ; isolation & purification ; Rhamnose ; chemistry ; isolation & purification ; Scrophulariaceae ; chemistry

Plants are known to be efficient hosts for the production of mammalian therapeutic proteins. However, plants produce complex N-glycans bearing β1,2-xylose and core α1,3-fucose residues, which are absent in mammals. The immunogenicity and allergenicity of plant-specific Nglycans is a key concern in mammalian therapy. In this study, we amplified the sequences of 2 plant-specific glycosyltransferases from Nicotiana tabacum L. cv Bright Yellow 2 (BY2), which is a well-established cell line widely used for the expression of therapeutic proteins. The expression of the endogenous xylosyltranferase (XylT) and fucosyltransferase (FucT) was downregulated by using RNA interference (RNAi) strategy. The xylosylated and core fucosylated N-glycans were significantly, but not completely, reduced in the glycoengineered lines. However, these RNAi-treated cell lines were stable and viable and did not exhibit any obvious phenotype. Therefore, this study may provide an effective and promising strategy to produce recombinant glycoproteins in BY2 cells with humanized N-glycoforms to avoid potential immunogenicity.
Amino Acid Sequence ; Blotting, Western ; Carbohydrate Sequence ; Cell Line ; Cloning, Molecular ; DNA, Complementary ; genetics ; Down-Regulation ; Epitopes ; genetics ; immunology ; Fucose ; metabolism ; Fucosyltransferases ; chemistry ; deficiency ; genetics ; immunology ; Glycoproteins ; chemistry ; genetics ; immunology ; Molecular Sequence Data ; Pentosyltransferases ; chemistry ; deficiency ; genetics ; immunology ; Polysaccharides ; chemistry ; immunology ; Protein Engineering ; methods ; RNA Interference ; Species Specificity ; Tobacco ; cytology ; genetics ; Xylose ; metabolism