A modified protocol for isolation and purity evaluation of a staphylococcal acidic polysaccharide by chromatography and capillary electrophoresis

The extracellular slime of Staphylococcus epidermidis contains, amongst various macromolecules, an acidic polysaccharide (PS) of a molecular mass of 20 kDa with significant antigenic and biological properties. The isolation procedure used so far includes multiple fractionations in anion‐exchange chromatographic columns before its final purification by gel filtration chromatography. This protocol is laborious, time‐consuming and includes the risk of unnecessary loss of PS quantities. Because of the significance of this PS, a modified protocol resulting in an easier and quicker isolation procedure was developed. Furthermore, identification, purity, charge density and molecular integity of the isolated polysaccharide were evaluated by a reverse‐polarity capillary electrophoresis method. Copyright © 2010 John Wiley & Sons, Ltd.     

Preparation of New Sargassum fusiforme Polysaccharide Long-Chain Alkyl Group Nanomicelles and Their Antiviral Properties against ALV-J

Avian leukosis virus subgroup J (ALV-J) is an immunosuppressive virus which has caused heavy losses to the poultry breeding industry. Currently, there is no effective medicine to treat this virus. In our previous experiments, the low-molecular-weight Sargassum fusiforme polysaccharide (SFP) was proven to possess antiviral activity against ALV-J, but its function was limited to the virus adsorption stage. In order to improve the antiviral activity of the SFP, in this study, three new SFP long-chain alkyl group nanomicelles (SFP-C12M, SFP-C14M and SFP-C16M) were prepared. The nanomicelles were characterized according to their physical and chemical properties. The nanomicelles were characterized by particle size, zeta potential, polydispersity index, critical micelle concentration and morphology. The results showed the particle sizes of the three nanomicelles were all approximately 200 nm and SFP-C14M and SFP-C16M were more stable than SFP-C12M. The newly prepared nanomicelles exhibited a better anti-ALV-J activity than the SFP, with SFP-C16M exhibiting the best antiviral effects in both the virus adsorption stage and the replication stage. The results of the giant unilamellar vesicle exposure experiment demonstrated that the new virucidal effect of the nanomicelles might be caused by damage to the phospholipid membrane of ALV-J. This study provides a potential idea for ALV-J prevention and development of other antiviral drugs.     

Embedding magnetic nanoparticles into polysaccharide-based hydrogels for magnetically assisted bioseparation.

Based on the preparation of biocompatible polysaccharide-based hydrogels with stimuli-responsive properties by the copolymerization of maleilated carboxymethyl chitosan with N-isopropylacrylamide, novel magnetic hybrid hydrogels were fabricated by the in situ embedding of magnetic iron oxide nanoparticles into the porous hydrogel networks. Scanning electron microscopy (SEM) and thermogravimetric (TG) analyses showed that the size, morphology, and content of the iron oxide nanoparticles formed could be modulated by controlling the amount of maleilated carboxymethyl chitosan. As confirmed by X-ray diffractometry (XRD), equilibrium swelling ratio, and differential scanning calorimetry (DSC) measurements, the embedding process did not induce a phase change of the magnetic iron oxide nanoparticles, and the resultant hybrid hydrogels could retain the pH- and temperature-responsive characteristics of their hydrogel precursors. By investigating the partition coefficients of bovine serum albumin as a model protein, this magnetic hydrogel material was found to hold a potential application in magnetically assisted bioseparation.     

Structural properties and antioxidant activities of polysaccharide from fruit bodies of Pholiota nameko

A polysaccharide named PNP was extracted and purified from Pholiota nameko. The total sugar content of PNP was 95.29% and the molecular weight was 1.89 × 10³ kDa. The structural features of PNP were investigated by the combination of chemical and instrumental analysis such as UV spectrophotometer, specific rotation determination, FT-IR, methylisation analysis and Congo red. The results showed that the optical rotation of PNP was +120° and that it had a triple-helical structure. Besides, PNP was mainly composed of glucose and mannose at the molar ratio of 4.24:1.00. The backbone of PNP was composed of (1→3)-linked-Glc and (1→3)-linked-Man whereas the branches of (1→3,6)-linked- Glc, (1→3,6)-linked-Man and T- Glc. Consistenting with the results of UV–Vis spectra, FT-IR spectroscopy and ¹H NMR, indicated that PNP was a complex of polysaccharides and polyphenols. In vitro antioxidant results suggested that PNP was processed with certain scavenging capacity.     

A sensitive and specific HPGPC‐FD method for the study of pharmacokinetics and tissue distribution of Radix Ophiopogonis polysaccharide in rats

Interest in antimyocardial ischemic activity of a graminan‐type fructan with a weight average molecular weight of 4.8 kDa extracted from Radix Ophiopogonis (ROP) has necessitated the study of its pharmacokinetics and tissue distribution. For that, a simple HPGPC–FD method was developed for the sensitive and specific determination of FITC‐ROP (fluorescein–isothiocyanate‐labeled ROP) in plasma and rat tissues (heart, liver, spleen, lung, kidney, brain and stomach). The analyte was separated on a Shodex Sugar KS‐802 high‐performance gel column with 0.1 M phosphate buffer (pH 7.0) as mobile phase at a flow rate of 0.5 mL/min, and fluorescence detection at λ_ex 495 nm and λ_em 515 nm. The calibration curve for FITC‐ROP was linear over the range 0.25–20.0 or 50.0 μg/mL in all studied biosamples with correlation coefficients >0.995. The inter‐day and intra‐day precisions of analysis were not more than 10%, and assay accuracy ranged from 93 to 105% for plasma and from 89 to 108% for tissue homogenates. This method has been confirmed here to be suitable for the study of pharmacokinetics and tissue distribution of ROP and the achieved results are highly instructive for the further pharmaceutical development of ROP, suggesting the promising application of the method to the increasingly important carbohydrate‐based drugs. Copyright © 2009 John Wiley & Sons, Ltd.     

Levan polysaccharide from Erwinia herbicola protects osteoblast cells against lipopolysaccharide-triggered inflammation and oxidative stress through regulation of ChemR23 for prevention of osteoporosis

Osteoblast cell injury is a type of degenerative disorder characterized by osteolysis. Levan polysaccharide is an active component of Erwinia herbicola, which shows potential anti-inflammatory and antioxidant properties. However, the protective effects of levan on lipopolysaccharide (LPS)-induced inflammatory and oxidative stress injury in osteoblast cells as an in vitro model of osteolysis remain largely unknown. The present study aimed to explore the functions of levan in LPS-triggered inflammation and oxidative stress in osteoblast cells (MC3T3-E1). The protective effects of levan on LPS-induced inflammatory and oxidative stress responses in MC3T3-E1 cells were assessed by quantification of superoxide dismutase (SOD) and catalase (CAT) activity as well as enzyme-linked immunosorbent assay (ELISA) for determination of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Also, the key signaling pathway of ChemR23 was determined by qPCR analysis. Results showed that levan significantly alleviated LPS-induced deactivation of SOD and CAT activity. Levan also downregulated the expression of IL-6, TNF-α, and ChemR23 at mRNA level. These findings indicated that levan may protect MC3T3-E1 cells against LPS-triggered oxidative stress, and inflammation via regulation of ChemR23. This data may provide a potential basis for further clinical investigation of levan in the prevention and treatment of LPS-triggered bone resorption and osteolysis.     

Formation of Aromatic and Other Unsaturated End Groups in Carboxymethyl Cellulose During Hot Alkaline Treatment

Carboxymethyl cellulose (CMC, DS 0.58) was treated in solutions of sodium hydroxide (0.001–1 M) at 95 °C. The treated (1–12 h) CMC samples were purified by dialysis and analyzed by UV spectroscopy and by UV resonance Raman spectroscopy (UVRRS) with excitation at 244 nm. A UV absorption maximum at 265 nm and a UVRR signal at 1650 cm⁻¹ were indicative of formation of -conjugated aldehyde end groups in CMC through -elimination. Another strong UVRR band at 1610 cm⁻¹ gave evidence on conversion of some of the -conjugated aldehyde end groups to alkali stable aromatic structures.     

Thermosensitive Microparticles Based on Unsaturated Esters of some Poly- and Oligosaccharides: Preparation,Characterization, Drug Inclusion and Release

Summary: The paper concerns the preparation and characterization of hydrogel microparticles based on exopolysaccharide (Gellan, Xanthan) unsaturated derivatives and respectively on cyclodextrin as well as their application for some hydrosoluble and liposoluble drugs inclusion. In the first step the polysaccharide and cyclodextrin unsaturated esters (maleate, acrylate) were synthesized and their hydrogel forming capacity was tested using a grafting-crosslinking free-radical reaction with N-isopropyl acrylamide (NIPAm), at room temperature. For a better control of the crosslinking degree N,N' methylene-bis-acrylamide (BIS), replaced by cyclodextrin tri-acrylate (A-CD) in a few experiments, was used. The microparticles were obtained by using the method in w/o emulsion, in which the dispersed aqueous phase is the reaction mixture and the oil phase is hexane. The particles containing polysaccharide esters showed an average diameter around 100 µm when crosslinking was achieved with BIS. They were smaller than those crosslinked with A-CD, which are in the range of 200-300 µm; the particles based on Xanthan maleate were smaller than Gellan maleate based ones. Even much smaller particles (2-2.5 µm in diameter) were obtained by starting from A-CD grafted-crosslinked systems. The synthesized microparticles are able to include chloramphenicol, as well as progesterone; the drug is slowly released according to diffusion controlled kinetics. The application of these microparticles in emergency ophthalmic treatments is possible as a result of their thermal sensitivity; they can collapse and release the drug instantly when placed in contact with the human eye, at 37 °C.