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At least 937 records · Page 52Linked to original sources
PubMed2005

Determination of saccharide content in pneumococcal polysaccharides and conjugate vaccines by GC-MSD.

A simple and sensitive gas chromatographic method was designed for quantitative analysis of Streptococcus pneumoniae capsular polysaccharides, activated polysaccharides, and polysaccharide conjugates. Pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F polysaccharide or conjugate were subjected to methanolysis in 3N hydrochloric acid in methanol followed by re-N-acetylation and trimethylsilylation. Derivatized samples were chromatographed and detected using gas chromatography with mass selective detector. Gas chromatographic results were compared with colorimetric values with agreement of 92 to 123% over the range of all samples tested. Monosaccharides released during methanolysis included hexoses, uronic acids, 6-deoxy-hexoses, amino sugars, and alditols. Quantitative recovery of monosaccharides was achieved for all serotypes by the use of a single methanolysis, derivatization, and chromatography procedure. Response factors generated from authentic monosaccharide standards were used for quantitation of pneumococcal polysaccharides and conjugates with confirmation of peak assignments by retention time and mass spectral analysis. This method allows saccharide quantitation in multivalent pneumococcal vaccine intermediates and final drug products with low-level detection (10 pg) and peak purity.

Carbohydrate Sequence↗
PubMed2005

Non-covalent interaction between procyanidins and apple cell wall material. Part III: Study on model polysaccharides.

The adsorption of condensed tannins (procyanidins) of varying degrees of polymerisation and percentage of galloylation on solid polysaccharides substrates was quantified using the Langmuir isotherms formulation. Pectins and xyloglucans, which are soluble polysaccharides, were first cross-linked by, respectively, dibromopropane and epichlorohydrin to obtain insoluble covalent gels. Cellulose and starch, being insoluble in the buffer solution at room temperature, were used as bought. Apparent affinity constants obtained for the pure polysaccharides were as follows: pectin>>xyloglucan>starch>cellulose. The apparent affinity constants increased with the molecular weight of the procyanidins, except with cellulose. Higher affinities were obtained with pectin, a polysaccharide having the ability to develop a gel-like network, forming hydrophobic pockets able to encapsulate procyanidins. Filamentous and globular polysaccharides, like cellulose and xyloglucan, bound procyanidins weakly. Higher apparent saturation levels were obtained for cellulose and xyloglucans, the arrangement of which would favour cooperativity and stacking. Pectin had lower saturation levels probably due to a steric hindrance effect.

Absorption↗
PubMed2006

Ca2+-transport in sea urchin unfertilized eggs: regulation by endogenous sulfated polysaccharides and K+.

Previous data from our laboratory showed that the reticulum of the sea cucumber smooth muscle body wall retains both a sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) and a sulfated polysaccharide. In this invertebrate, the transport of Ca(2+) by the SERCA is naturally inhibited by these endogenous sulfated polysaccharides. The inhibition is reverted by K(+) leading to an enhancement of the Ca(2+) transport rate. We now show that vesicles derived from the endoplasmic reticulum of unfertilized eggs from the sea urchin Arbacia lixula retain a SERCA that is able to transport Ca(2+) at the expense of ATP hydrolysis. As described for the sea cucumber SERCA isoform, the enzyme from the sea urchin is activated by K(+) but not by Li(+) and is inhibited by thapsigargin, a specific inhibitor of SERCA. A new sulfated polysaccharide was identified in the sea urchin eggs reticulum composed mainly by galactose, glucose, hexosamine and manose. After extraction and purification, this sulfated polysaccharide was able to inhibit the mammal SERCA isoform found in rabbit skeletal muscle and the inhibition is reversed by K(+). These data suggest that the regulation of the SERCA pump by K(+) and sulfated polysaccharides is not restricted to few marine invertebrates but is widespread.

Animals↗
PubMed2005

The response of normal human osteoblasts to anionic polysaccharide polyelectrolyte complexes.

Polyelectrolyte complexes (PEC) were prepared from chitosan as the polycation and several synthesized functional anion polysaccharides, and their effects on cell attachment, morphology, proliferation and differentiation were estimated using normal human osteoblasts (NHOst). After a 1-week incubation, PEC made from polysaccharides having carboxyl groups as polyanions showed low viability of NHOst on it although the NHOst on it showed an enhancement in their differentiation level. On the other hand, NHOst on PEC made from sulfated or phosphated polysaccharides showed similar attachment and morphology to those on the collagen-coated dish. When the number of NHOst was estimated after 1 week, the number on the PEC was ranged from 70% to 130% of those on the collagen-coated dish, indicating few effects of these PEC on cell proliferation. In addition, NHOst on PEC films made from sulfated polysaccharides differentiated to a level very similar to that observed on the collagen-coated dish, indicating that these PEC films maintain the normal potential of NHOst to both proliferate and differentiate. Measurement of gap junctional intercellular communication of NHOst on PEC revealed that PEC did not inhibit communication, suggesting that PEC films have few effects on cell homeostasis. Thus, PEC made from the sulfated polysaccharide may be a useful material as a new scaffold for bone regeneration.

Alkaline Phosphatase↗
PubMed2003

Characterisation of bacterial polysaccharides: steps towards single-molecular studies.

Techniques used in studies of polysaccharides, including chemical composition, linkage pattern, and higher order structures are in constant development. They provide information necessary for understanding of the polysaccharide properties and functions. Here, recent advancements in studies of the polysaccharides at the single-molecule level are highlighted. Over the last few years, single-molecule techniques such as force spectroscopy have improved in sensitivity and can today be used to detect forces in the pN range. In addition, these techniques can be used to investigate properties of single molecules close to physiological conditions. The challenges in the interpretation of the observations are aided by control experiments using well-characterised polysaccharides and by data provided by complementary methods. This field is expected to have increasing impact on the further advancement of the molecular understanding of the role of polysaccharides in various biological processes such as recognition and cell adhesion.

Biochemistry↗
PubMed2005

Challenges for the modern analytical ultracentrifuge analysis of polysaccharides.

This article reviews some of the recent advances in analytical ultracentrifugation and how these advances have impacted--and can impact--on our understanding of the size, shape through conformation modelling, interactions and charge properties of polysaccharides in solution, particularly when used in combination with other solution techniques and also imaging techniques. Specifically we look at (1) polysaccharide polydispersity and simple shape analysis by sedimentation velocity, and in particular using new approaches such as SEDFIT analysis; (2) polysaccharide molecular-weight analysis by sedimentation equilibrium and MSTAR analysis and how this complements analysis of size exclusion chromatography coupled to multi-angle laser light scattering; (3) polysaccharide conformation analysis using traditional procedures such as the Wales-van Holde ratio, power law or 'scaling' relations, more specialised treatments for rigid cylindrical structures, semi-flexible chains and worm-like coils and complications through draining effects; (4) Analysis of polysaccharide interactions and in particular complex formation phenomena, focusing on interesting applications in the areas of mucoadhesion and sedimentation fingerprinting; and (5) the possibilities for macromolecular charge and charge screening measurement.

Amylopectin↗
PubMed2004

Elucidation of polysaccharide origin in Ramalina peruviana symbiosis.

A structural elucidation of polysaccharides extracted from the aposymbiotically cultured mycobiont of the lichen Ramalina peruviana was carried out in order to determine whether the polysaccharides found previously in the symbiotic thalli are produced by the mycobiont or photobiont or both. The mycobiont isolate was cultivated on a solid malt-yeast extract-medium and the freeze-dried colonies were defatted and the polysaccharides extracted successively with hot water and aq. 2% KOH, each at 100 degrees C. The alkaline extract was obtained in much higher yield (31.5%) and submitted to a freeze-thawing treatment, giving rise to a precipitate (PK2) of a mixture of (1-->3),(1-->4)-alpha-glucan (1.2:1 ratio, nigeran) and a (1-->3)-beta-glucan (laminaran). The mother liquor was treated with Fehling solution to give a precipitate (galactomannan). This had a (1-->6)-linked alpha-d-mannopyranosyl main chain, substituted at O-4 and in small proportion at O-2,4 by beta-Galp units. All three polysaccharides have previously been found in the symbiotic thalli of R. peruviana, showing that these are produced by the fungus, without the participation of the Trebouxia photobiont. Surprisingly, isolichenan, a cold-water soluble (1-->3),(1-->4)-alpha-linked-glucan (3:1 ratio) was not found in the isolated mycobiont, despite being the main polysaccharide found in the thalli.

Ascomycota↗
PubMed2005

The structure-anticoagulant activity relationships of sulfated lacquer polysaccharide: effect of carboxyl group and position of sulfation.

Regiospecific oxidation of the primary hydroxyl groups in lacquer polysaccharide (LPL, Mw 6.85 x 10(4)) and its NaIO4 oxidation derivatives (LPLde) to C-6 carboxy groups was achieved with NaOCl in the presence of Tempo and NaBr. Sulfate groups were incorporated into the oxidated polysaccharides using Py.SO3 complex as a reagent. Reactivity of polysaccharide hydroxyl group was C-6 > C-2 > C-4. Sulfate groups were mainly linked to the second hydroxy at C-2 in the products. The results of APTT assay showed after incorporation of carboxyl groups into lacquer polysaccharides, the intrinsic coagulation pathway was promoted, and all sulfated polysaccharides had very weak anticoagulant activity within the scope of studied DS (0.39-1.11). These indicated that carboxyl groups and sulfate groups had the synergistic action. At the same time, the anticoagulant activity increased very slowly with the DS in the second hydroxy. This indicated that 6-O-SO3- in the side chains took an important role in the anticoagulant activity.

Anticoagulants↗
PubMed2004

Intracellular disposition of polysaccharides in rat liver parenchymal and nonparenchymal cells.

Binding and internalization of arabinogalactan, pullulan, dextran, and mannan were examined in rat liver parenchymal and nonparenchymal cells using 125I or fluorescein isothiocyanate (FITC) labeled polysaccharides. Binding and uptake of arabinogalactan and pullulan into parenchymal cells was inhibited by asialofetuin, indicating that the asialoglycoprotein receptor is involved in the intracellular disposition of arabinogalactan and pullulan. Uptake of 125I-labeled dextran to parenchymal cells was unchanged upon addition of excess unlabeled dextran, suggesting that dextran uptake occurs via fluid phase endocytosis. Of the polysaccharides tested, mannan showed the strongest specific association with liver nonparenchymal cells. FITC-labeled polysaccharides showed arabinogalactan and pullulan are internalized to liver parenchymal cells, whereas mannan is internalized to nonparenchymal cells. This study demonstrates that intracellular disposition of polysaccharides in the liver occurs via receptor-mediated endocytosis (RME), indicating that RME plays a role in the biodisposition of these polysaccharides as drug carriers.

Animals↗
PubMed2004

Ultrastructure of acidic polysaccharides from the cell walls of brown algae.

We have studied the ultrastructure of acidic polysaccharides from the cell walls of brown algae using a variety of electron microscopy techniques. Polysaccharides from Padina gymnospora present self assembled structures, forming trabecular patterns. Purified fractions constituted by alginic acid and sulfated fucan also form well-organized ultrastructures, but the pattern of organization varies depending on the polysaccharide species. Alginic acid presents sponge-like structures. Sulfated fucan exhibits particles with polygonal forms with a polycrystalline structure. These particles are in fact constituted by sulfated fucan molecules since they are recognized by a lectin specific for alpha-l-fucosyl residues. X-ray microanalysis reveal that S is a constituent element, as expected for sulfated groups. Finally, an exhaustive purified sulfated fucan shows the same ultrastructure formed by polygonal forms. Furthermore, elemental analyses of acidic polysaccharides indicate that they retain Zn, when algae were collected from a contaminated area. This observation is supported by direct quantification of heavy metal in the biomass and also in the solubilized polysaccharides compared with the algae from a non-contaminated site. We conclude that these molecules have specific ultrastructure and elemental composition; and act as metal binder for the nucleation and precipitation of heavy metals when the algae are exposed to a metal contaminated environment.

Alginates↗
PubMed2005

Comparison of three ELISA protocols to measure antibody responses elicited against serogroup C meningococcal polysaccharide in mouse, monkey and human sera.

In this study we compared the following ELISA protocols to measure antibody levels against serogroup C meningococcal polysaccharide: a traditional protocol using poly-L-Lysine mixed with the polysaccharide as coating antigen, a second protocol coating with a mixture of methylated human serum albumin with the C polysaccharide, a modified protocol coating with derivatized polysaccharide and a modification to the last one, specifically without adding ammonium thiocyanate to the sample buffer. Serum bactericidal activity of mouse, monkey and human sera were measured and correlation coefficients were calculated. For all serum types the modified ELISA protocol showed the highest correlation coefficients while the traditional protocol showed the lower ones. We demonstrated that the traditional protocol measures non-specific antibodies to the C polysaccharide, because no differences were detected between pre-immune and post-immune human sera (P>0.05), while the modified protocol detected the highest difference (P<0.01).

Animals↗
PubMed2005

Effect of polysaccharides from Opuntia ficus-indica (L.) cladodes on the healing of dermal wounds in the rat.

In traditional medicine extracts of polysaccharide-containing plants are widely employed for the treatment of skin and epithelium wounds and of mucous membrane irritation. The extracts of Opuntia ficus-indica cladodes are used in folk medicine for their antiulcer and wound-healing activities. The present study describes the wound-healing potential of two lyophilized polysaccharide extracts obtained from O. ficus-indica (L.) cladodes applied on large full-thickness wounds in the rat. When topically applied for 6 days, polysaccharides with a molecular weight (MW)>10(4)Da from O. ficus-indica cladodes induce a beneficial effect on cutaneous repair in this experimental model; in particular the topical application of O. ficus-indica extracts on skin lesions accelerates the reepithelization and remodelling phases, also by affecting cell-matrix interactions and by modulating laminin deposition. Furthermore, the wound-healing effect is more marked for polysaccharides with a MW ranging 10(4)-10(6)Da than for those with MW>10(6)Da, leading us to suppose that the fine structure of these polysaccharides and thus their particular hygroscopic, rheologic and viscoelastic properties may be essential for the wound-healing promoter activity observed.

Animals↗
PubMed2006

Antiviral property and mechanisms of a sulphated polysaccharide from the brown alga Sargassum patens against Herpes simplex virus type 1.

A sulphated polysaccharide (SP-2a) from the brown alga Sargassum patens (Kütz.) Agardh (Sargassaceae) was found to significantly inhibit the in vitro replication of both the acyclovir (ACV)-sensitive and -resistant strains of Herpes simplex virus type 1 (HSV-1), in dose-dependent manners, with 50% inhibitions occurring with 1.5-5.3 microg/ml of the polysaccharide. SP-2a exhibited extracellular virucidal activity only against the ACV-sensitive strains, but not the resistant strain, at the concentration of 100 microg/ml. The strongest antiviral activities against the different strains of HSV-1 were observed when this polysaccharide was present during and after adsorption of the virus to host cells. The inhibitory effect of SP-2a on virus adsorption occurred dose-dependently in all the HSV-1 strains tested, and the adsorption of the ACV-resistant DM2.1 strain was reduced by 81.9% (relative to control) with 4 microg/ml of the polysaccharide. This study clearly demonstrated that the antiviral mode of action of SP-2a is mediated mainly by inhibiting virus attachment to host cells, and this sulphated polysaccharide might have different modes of action against the ACV-sensitive and -resistant strains of HSV-1.

Acyclovir↗
PubMed2004

The symbiotic defect in a Sinorhizobium meliloti lipopolysaccharide mutant can be overcome by expression of other surface polysaccharides.

In this work we have examined the extent of functional complementation in symbiosis among different Sinorhizobium meliloti surface polysaccharides including lipopolysaccharide (LPS). We show that a symbiotic deficiency associated with an LPS defect can be reversed by appropriate expression of other surface polysaccharides such as galactoglucan (EPSII) and a particular form of capsular polysaccharide (KdoPS). It is noteworthy that, while succinoglycan EPSI and LPS cannot functionally substitute for each other, they can both be replaced by the same common set of polysaccharides (i.e., EPSII/KdoPS). The complex pattern of functional complementation in symbiosis among S. meliloti surface polysaccharides was shown to be different in Medicago truncatula compared to that previously reported for M. sativa.

Blotting, Southern↗
PubMed2003

Inhibition of DNA topoisomerases I and II, and growth inhibition of human cancer cell lines by a marine microalgal polysaccharide.

We have previously reported purification of an extracellular polysaccharide GA3P, D-galactan sulfate associated with L-(+)-lactic acid, produced by a toxic marine microalga Dinoflagellate Gymnodinium sp. A(3) (GA3), and induction thereby of apoptosis on human myeloid leukemia K562 cells. In the present report, we show that the GA3P is a potent inhibitor of DNA topoisomerase (topo) I and topo II, irrespective of the presence or absence of the lactate group. Dextran sulfate also showed similar level of inhibition of topo I and topo II. We also demonstrated that, unlike camptothecin (CPT) or teniposide (VM-26), the inhibition of topo I or topo II by the polysaccharide does not involve accumulation of DNA-topo I/II cleavable complexes, clearly showing that they are not topo poisons but catalytic inhibitors with dual activity. Furthermore, the polysaccharide, when added to the reaction mixture with CPT or VM-26, inhibited stabilization of cleavable complex induced by the latter compounds. In addition, when added to the reaction mixture after the formation of the cleavable complexes by topo poisons, CPT for topo I and VM-26 for topo II, either GA3P or dextran sulfate diminished the amount of the complexes already accumulated, i.e. reversal of the reaction. These results suggest that the polysaccharides bind to the enzymes with high affinities, and that, as for topo I/II inhibition, the GA3P shares a common mechanism with dextran sulfate. As examined in vitro with a human cancer cell line panel, GA3P exhibited significant cytotoxicity against a variety of cancer cells. These findings show that the polysaccharide GA3P would prove to be a potential anticancer chemotherapeutic agent with dual activity of topo I and topo II catalytic inhibition.

Antineoplastic Agents↗
PubMed2000

A wide diversity of sulfated polysaccharides are synthesized by different species of marine sponges.

Sulfated polysaccharides were extracted from four species of marine sponges by exhaustive papain digestion. These compounds were purified by anion-exchange and gel-filtration chromatography. Analysis of the purified polysaccharides revealed a species-specific variation in their chemical composition and also in their molecular masses. In the species Aplysina fulva we found a sulfated glucan with a glycogen-like structure. The other three species contained sulfated polysaccharides with variable proportions of galactose, fucose, arabinose and hexuronic acid and also with different degrees of sulfation. Although the complex nature of these polysaccharides did not allow complete structure determination, we detected the occurrence of 4-sulfated residues of fucose and arabinose in the species Dysidea fragilis. The biological role of these sulfated polysaccharides requires further investigation. They may be involved in the species-specific aggregation of sponge cells and/or in the structural integrity of sponge, resembling the proteoglycans of mammalian connective tissues.

Animals↗
PubMed1976

An extracellular fungal polysaccharide composed of 2-acetamido-2-deoxy-D-glucuronic acid residues.

The black yeast-like fungus NRRL YB-4163, now tentatively identified as Rhinocladiella elatior Mangenot, has been found to produce an extracellular microbial polysaccharide composed mainly of 2-acetamido-2-deoxy-D-glucuronic acid residues. Polysaccharide (PS) YB-4163, when isolated in good yield as the neutral potassium salt, dissolves readily in water to produce extremely viscous solutions, which form stable foams and emulsions. By depolymerizing PS YB-4163 with [14C]methanol-HCl, the polysaccharide can be both identified and quantitated radiochemically by determining the individual [14C]methyl glycosides after their separation by paper chromatography. When the methyl glycosides of PS YB-4163 were reduced with NaB3H4, only the methyl glycosides of 2-acetamido-2-deoxy-D-[6-3H]glucose were found. Analysis of the monosaccharide released from carboxyl-reduced PS YB-4163 by acid hydrolysis or methanolysis also showed 2-acetamido-2-deoxy-D-glucuronic acid to be the main constituent. Previously, the only polysaccharides known to be composed entirely or hexosaminuronic acid have been cellular products from pathogens. Of these, the antigenic polysaccharide (SPSA) from Staphylococcus aureus is composed entirely of 2-amino-2-deoxy-D-glucuronic acid, but its amino groups are substituted equally with acetyl and N-acetylalanyl groups. The specific optical rotation of PS YB-4163, [alpha]20D -75 degrees (c 0.5, water), is similar to that of SPSA (-91 degrees), and suggests beta-D-linkages that must be either (1 leads to 3) or (1 leads to 4).

Glucuronates↗
PubMed1985

Structural and conformational features of the Escherichia coli K92 capsular polysaccharide.

Escherichia coli K92 polysaccharide, an alternating (2----8)-alpha- and (2----9)-alpha-linked heteropolymer of sialic acid, undergoes lactonisation (10-20%) at low pH or on treatment with a water-soluble carbodi-imide (greater than 90%). Under the latter conditions, the formation of O-acylisourea is a minor pathway. The antigenicity of the K92 polysaccharide is unaffected by lactonisation of approximately 10% of the residues, but is abolished by lactonisation of approximately 90% of the residues. The unexpected, relative resistance of the K92 polysaccharide to periodate oxidation contrasts with the behaviour of the meningococcal non-O-acetylated (O-Ac-) C polysaccharide, a (2----9)-alpha-linked homopolymer of sialic acid. Two-dimensional 13C-1H-n.m.r. correlation spectroscopy has been used to assign 1H chemical shifts which have aided the interpretation of a resolution-enhanced one-dimensional spectrum. This has led to a determination of conformational features of the K92 polysaccharide in solution. The side-chain adopts a conformation such that H-7 and H-8 are gauche in the (2----8)-alpha-linked residues (1a or 1b), but antiperiplanar in the (2----9)-alpha-linked residues (2). Molecular correlation times have been calculated and some aspects of internal motion elucidated.

Carbohydrate Conformation↗