Isolated Polysaccharide Fractions Research Articles

Chemical characterization and gastroprotective effect of an isolated polysaccharide fraction from Bletilla striata against ethanol-induced acute gastric ulcer

The chemical characterization and protective role against ethanol-induced gastric ulcerated rats of a polysaccharide fraction from Bletilla striata (BSP) collected by ultrafiltration membrane approach were evaluated. This BSP faction was consisted of mannose and glucose at a molar ratio of 2.4:1 approximately, with a molecular weight of 146 KDa. FT-IR, NMR and XRD spectra indicated that BSP faction contained α-Man and β-Glc residues with low overall crystallinity. The polysaccharide exhibited significant scavenging activities of ABTS and FRAP, as well as non-toxicity against human gastric epithelial GES-1 cells. Oral administration with 100 mg/kg of BSP for 3 days continuously could significantly prevent the formation of ethanol-induced gastric mucosal lesion. It could also reduce the levels of pro-inflammatory cytokines, including TNF-α, IL-1β, IL-6, and IL-18, and MPO activity in gastric tissue. Additionally, the BSP faction exhibited antioxidant activity, increased the content of PEG2 as a defensive factor, and suppressed MAPK/NF-κB signaling pathway in gastric tissue. These results indicated that the gastroprotective activity of BSP faction could be attributed to the reduction of pro-inflammatory cytokines and oxidative stress and the inhibition of MAPK/NF-κB pathways. Our results provided substantial evidence that BSP could be a promising phytomedicine for gastric ulcer prevention.

Determination of the Inhibitory Activity of Ferula sinkiangensis Polysaccharides for Protein Tyrosine Phosphatase 1B

Polysaccharide fractions were sequentially extracted in 28.86 wt% yield from roots of Ferula sinkiangensis K. M. Shen. The polysaccharide fractions from F. sinkiangensis were identified as heteropolysaccharides and contained primarily the monosaccharides glucuronic and galacturonic acids, rhamnose, xylose, arabinose, galactose, and fructose. The isolated polysaccharide fractions at low concentrations exhibited in vitro inhibitory activity for protein-tyrosine phosphatase 1B. The degree of inhibition correlated with the galactose content in the polysaccharide fractions.

Isolation and Characterization of Pectic Polysaccharide Fraction fromIn VitroSuspension Culture ofFumaria officinalisL.

In the current study, an acidic polysaccharide from thein vitrosuspension culture ofFumaria officinalisL. was obtained by extraction with 0.8% (w/v) aqueous ammonium oxalate. The polysaccharide fraction mainly consisted of galacturonic acid (41.0%), followed by galactose (7.3%) and arabinose (5.6%). This suggests the presence of arabinogalactan side chains in the rhamnogalacturonan-I segment of the studied pectin, which was mainly built up by homogalacturonan segments. The pectin was evaluated as low-methyl-esterified (45.0%) with degree of acetylation 3.4%. The polymer fraction was consisted of different molecular weight populations in the range of 6–600 kDa. The high amount of 4-L-hydroxyproline (11.7% of total protein) and the specific positive reaction to Yariv’s phenylglycoside reagent indicated the presence of an arabinogalactan protein in the cell walls. The functional properties of the polysaccharide fraction were evaluated, as it possessed better water-holding capacity than oil-holding capacity. The studied pectin demonstrated significant foaming ability and promising emulsifying properties in a concentration 1%. Therefore, the isolated polysaccharide fraction could be successfully used as emulsifier and foaming agent in food products and pharmaceutical supplements.

Physicochemical properties of pectin from green jelly leaf (Cyclea barbata Miers)

The water extract of Green Jelly leaves (GJL) obtained by crushing the leaves in water (1:40) was capable of forming a gel at room temperature. The composition of GJL consisted mainly of carbohydrate (∼70w/w), protein (∼13% w/w) and minerals (∼6% w/w). The mineral portion consisted of mainly calcium (∼1.2% w/w), zinc (∼0.12% w/w) and magnesium (∼0.11% w/w). The isolated polysaccharide fraction (∼42.6% w/w) consisted of mainly galacturonic acid (∼35.8% w/w) and neutral sugars (∼6.8% w/w), with a weight-average molecular weight of ∼4.4×105g/mol. The results obtained by Fourier Transform Infra-Red (FTIR) showed that GJL polysaccharide fraction had a fairly similar FTIR fingerprint as the commercial low-methoxyl pectin (LMP). The degree of esterification of the polysaccharide changed drastically (from 97% to 10%) depending on the temperature used during the extraction process. The zeta potential of the extracted polysaccharide showed high negative charged as compared to the commercial LMP but close to sodium alginate. The study showed that the gelation was divalent cation-mediated and probably facilitated by the low degree of esterification which reduced steric hindrance from the methyl ester groups.

Submerged cultivation of Ganoderma lucidum and the effects of its polysaccharides on the production of human cytokines TNF-α, IL-12, IFN-γ, IL-2, IL-4, IL-10 and IL-17

An original strain of Ganoderma lucidum (W.Curt.:Fr.) Lloyd, MZKI G97 isolated from Slovenian habitats was grown by a submerged liquid substrate cultivation in a laboratory stirred tank reactor. Five fractions of extracellular and cell-wall polysaccharides were obtained by extraction, ethanol precipitation, and purification by ion-exchange, gel and affinity chromatography. The capacity of isolated polysaccharide fractions to induce innate inflammatory cytokines, and to modulate cytokine responses of activated lymphocytes was investigated. Human peripheral blood mononuclear cells (PBMC) were activated in vitro with polysaccharide fractions, in order to induce innate inflammatory cytokines: tumor necrosis factor alpha (TNF-α), interleukin (IL) 12 and interferon gamma (IFN-γ). For the immunomodulation capacity, polysaccharide fractions were cultured with ionomycine and phorbol myristate acetate (IONO+PMA) activated PBMC, and the concentrations of induced IL-2, IL-4, IFN-γ, IL-10 and IL-17 were measured. The results showed that polysaccharides from G. lucidum induced moderate to high amounts of innate inflammatory cytokines. Fungal cell-wall polysaccharides were stronger innate inflammatory cytokines inducers, while extracellular polysaccharides demonstrated a higher capacity to modulate cytokine responses of IONO+PMA induced production of IL-17. The results indicate that G. lucidum polysaccharides enhance Th1 response with high levels of IFN-γ and IL-2, and display low to no impact on IL-4 production. A similar pattern was observed at regulatory cytokine IL-10. All of the polysaccharide fractions tested induced IL-17 production at different concentration levels.

Strategy to identify and quantify polysaccharide gums in gelled food concentrates

A strategy for the unambiguous identification and selective quantification of xanthan gum and locust bean gum (LBG) in gelled food concentrates is presented. DNA detection by polymerase chain reaction (PCR) showed to be a fast, sensitive, and selective method that can be used as a first screening tool in intact gelled food concentrates. An efficient isolation procedure is described removing components that may interfere with subsequent analyses. NMR spectroscopy enabled the direct identification of xanthan gum and the discrimination between different galactomannans in the isolated polysaccharide fraction. An enzymatic fingerprinting method using endo-β-mannanase, in addition to being used to differentiate between galactomannans, was developed into a selective, quantitative method for LBG, whereas monosaccharide analysis was used to quantify xanthan gum. Recoveries for xanthan gum and LBG were 87% and 70%, respectively, with in-between day relative standard deviations below 20% for xanthan gum and below 10% for LBG.

Inhibition of Helicobacter pylori adhesion to human gastric adenocarcinoma epithelial cells by aqueous extracts and pectic polysaccharides from the roots of Cochlospermum tinctorium A. Rich. and Vernonia kotschyana Sch. Bip. ex Walp

In Malian traditional medicine infusions of the roots of Vernonia kotschyana or Cochlospermum tinctorium in water are used for treating gastric ulcer. Helicobacter pylori is known to play a major role in gastric ulcer development, and it was of interest to evaluate a potential anti-adhesive activity towards H. pylori by crude water extracts and isolated polysaccharide fractions from the roots of V. kotschyana and C. tinctorium. The inhibitory effects were examined by an in vitro flow cytometric assay using human gastric adenocarcinoma epithelial cells, where fluorescent-labeled H. pylori were pre-treated with the test fractions. The crude extract Ctw50 from C. tinctorium, containing a mixture of inulin, pectic polysaccharides, phenols and protein, led to a 43% reduction of bacterial attachment. The isolated pectic type fractions CtwA1 and CtwA2 from C. tinctorium, and Vko-I from V. kotschyana resulted in approximately 30% inhibition of H. pylori adhesion. These fractions consist of rhamnogalacturonan backbones with side chains of arabinogalactans and/or arabinans. The low degree of uronic acids in the fractions compared to anti-adhesive polysaccharides reported previously, suggests that the neutral side chains might play a role in the binding of bacterial adhesins. The fraction Vko-III.1 from V. kotschyana consisting mainly of galacturonic acid resulted only in a 19% inhibition of H. pylori adhesion. The anti-adhesive properties shown by the crude water extracts and isolated polysaccharide fractions in the present study might partly explain the anti-ulcer activities by the roots of V. kotschyana and C. tinctorium.

Non-cellulosic polysaccharides from the leaves of small balsam (Impatiens parviflora DC.)

The leaves of the annual plant Impatiens parviflora DC., a herbal medicine in Asian countries and invasive in managed forests and natural environments in Central Europe, have the potential as a source of bioactive phenolic compounds and polysaccharides. Extractives accounted for ∼22% of the leaves, whereby, the methanolic extract contains mainly caffeic acid, ferulic acid, kaempferol, and quercetin derivatives, and 1,2,4-trihydroxynaphthalene-1-O-glucoside. From the pre-extracted leaves, non-cellulosic polysaccharides were isolated by a five-step extraction procedure using as extractants cold water, 0.05M EDTA, and DMSO in the first three steps, and 1% and 5% NaOH in the last two steps. The isolated polysaccharide fractions were characterized by chemical, physicochemical, and spectroscopic analyses (FTIR and 1H NMR). The first three fractions contained mainly pectin and the alkali-extracted ones methylglucuronoxylan and arabinogalactan. The suggested structural features were confirmed using HSQC NMR and COSY experiments for polysaccharides of the EDTA-fraction and the Pronase-treated 5% NaOH-fraction. The EDTA fraction comprised a pectin with low degree of methyl-esterification (DM, 37%) with few rhamnogalacturonan RG I segments bearing β-1,4-galactan as side chains. The alkali-extracted fraction comprised a degraded methylglucuronoxylan and type II 3,6-arabinogalactan in about equal amounts.

A comparison of bioactive aqueous extracts and polysaccharide fractions from roots of wild and cultivated Cochlospermum tinctorium A. Rich

In Malian traditional medicine the roots of Cochlospermum tinctorium are used in the treatment of gastric ulcer, but extending harvesting is causing a growing concern of a dramatic reduction in the wild plant population. In the present study cultivation of C. tinctorium is evaluated, and structural components and bioactive properties of crude water extracts and isolated polysaccharide fractions from roots of wild and cultivated C. tinctorium are compared. The crude water extracts were shown to contain starch, pectin- and inulin-type polysaccharides, in addition to phenolic substances and protein, while the isolated acidic polysaccharide fractions contained mainly monosaccharides typical for pectins. The monosaccharide compositions of the polysaccharide fractions from roots of wild versus cultivated plants were comparable, albeit the yields in the cultivated roots were lower. Furthermore, the crude extracts and isolated polysaccharide fractions from wild and cultivated roots exhibited similar complement fixating activities, but were not able to activate macrophages. The crude extracts from cultivated roots were also shown to be moderate radical scavengers. The present study has shown that roots of cultivated C. tinctorium contain the same types of bioactive polysaccharides as the wild roots. However, in order to utilize roots of cultivated C. tinctorium in traditional medicine the cultivation method should be improved.

Immuno-modulation effect of sulphated polysaccharide (porphyran) from Porphyra vietnamensis

Our investigation explores the immuno-efficiency of sulphated polysaccharides enriched Porphyra vietnamenis. Isolated polysaccharide fraction (17.1–25.8%) was characterized by FTIR and NMR spectroscopy which showed the presence of typical linear backbone structure called as porphyran. Oral administration of porphyran (200–500mg/kg) evoked a significant (P≤0.05) increase in weight of the thymus, spleen and lymphoid organ cellularity. The total leucocyte and lymphocyte count was increased significantly (P<0.005). The increase in the percent neutrophil adhesion to nylon fibres as well as a dose-dependent increase in antibody titre values was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen and significant changes were observed in the formation of formazone crystals. It also prevented myelosuppression in cyclophosphamide drug treated rats. The results indicated that P. vietnamenis possesses potential immunomodulatory activity and has therapeutic potential for the prevention of autoimmune diseases.

Ginseng root water-extracted pectic polysaccharides originate from secretory cavities

A range of molecular probes for cell wall polysaccharides has been used to explore the structure and location of water-extracted pectic polysaccharides occurring in fractions isolated from ginseng roots. The LM19 homogalacturonan (HG) epitope was abundant in an HG fraction and analysis of LM19 binding to a rhamnogalacturonan-I (RG-I) rich-fraction indicated that the LM19 epitope is sensitive to acetylation. A specific RG-I epitope (LM16), four arabinogalactan-protein (AGP) epitopes (LM2, LM14, JIM16, MAC207) and an extensin epitope (JIM20) were found to be abundant and co-located in several isolated polysaccharide fractions including an arabinogalactan fraction and two RG-I fractions. Detection of the RG-I, AGP and extensin epitopes identified in isolated polysaccharide fractions in sections of ginseng roots indicated that they were most abundant in secretory cavities found in the cortical regions of ginseng roots. In addition, the immunocytochemical study indicated that polysaccharide epitope masking is a widespread phenomenon in the primary cell walls of ginseng roots.

Polymer Networks Produced by Marine Diatoms in the Northern Adriatic Sea

Using high resolution molecular technique of atomic force microscopy, we address the extracellular polymer production of Adriatic diatom Cylindrotheca closterium analyzed at the single cell level and the supramolecular organization of gel phase isolated from the Northern Adriatic macroaggregates. Our results revealed that extracellular polysaccharides freshly produced by marine diatoms can self-assemble directly to form gel network characteristics of the macroscopic gel phase in the natural aquatorium. Based on the experiments performed with isolated polysaccharide fractions of C. closterium and of macroaggregates gel phase, we demonstrated that the polysaccharide self-assembly into gel network can proceed independent of any bacterial mediation or interaction with inorganic particles.

Solid-state cultivation of Grifola frondosa (Dicks: Fr) S.F. Gray biomass and immunostimulatory effects of fungal intra- and extracellular β-polysaccharides

Grifola frondosa strain GF3, was cultivated on solid-state substrate consisting of milled whole corn plant ( Zea mays) and olive press cake supplemented with mineral additives and olive oil. Maintenance of the moisture content in the solid substrate is of crucial importance. Moistures higher than 70% promote growth of G. frondosa mycelium and polysaccharide production. Four fractions of pure extracellular β- d-glucans with total mass 127.2 mg and four fractions of intracellular polysaccharides with total mass 47.2 mg were isolated. Polysaccharides were further separated by ion-exchange, gel and affinity chromatography. Isolated polysaccharide fractions from fungal mycelium proved to induce moderate amounts of TNF-α in PBMC cells in vitro. The extent of TNF-α induction was up to 322 pg mL −1 at a polysaccharide concentration of 200 μg mL −1 for the intracellular fraction. The TNF-α inducing activity is comparable to romurtide, which has been used as a supporting therapy in cancer patients treated with radiotherapy and/or chemotherapy.

IMMUNOSTIMULATING EFFECTS OF ACIDIC POLYSACCHARIDES EXTRACT OF PANAX GINSENG ON MACROPHAGE FUNCTION

ABSTRACTThe root of Panax ginseng C. A. Meyer is one of the most popular natural tonics in oriental countries. In this study, we have isolated polysaccharide fraction of Panax ginseng (ginsan) and examined its effect on the function of murine peritoneal macrophages. When macrophages were treated with ginsan, cytotoxic activity against B16 melanoma cells was significantly induced. In addition, the levels of cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6 and lnterferon-γ (IFN-γ) were increased and the production of reactive oxygen/nitrogen components such as nitric oxide (NO) and hydrogen peroxide (H2O2) was enhanced. Moreover, phagocytic activity was induced in ginsan-treated macrophages compared to the control. The expression of CD14 and l-Ab on murine peritoneal macrophages was increased by the treatment with ginsan, while the expression of CD11b was decreased. Taken together, these results suggest that ginsan has an immunopotentiating effects on macrophages and these abilities could be used clinically for the treatment of diseases such as cancer.

Effect of Cordyceps sinensis on the proliferation and differentiation of human leukemic U937 cells

Cordyceps sinensis is a herb medicine with antitumor activity capable of suppressing the growth of mouse Sarcoma 180 in vivo. In the present study, we have isolated polysaccharide fraction of Cordyceps sinensis (PSCS) and investigated its effect on the proliferation and differentiation of human leukemic U937 cells using an in vitro culture system. Our results showed that the conditioned medium from PSCS (10 μg ml )-stimulated blood mononuclear cells (PSCS-MNC-CM) had an activity that could significantly inhibit the proliferation of U937 cells resulting in a growth inhibition rate of 78–83%. Furthermore, PSCS-MNC-CM treatment induced about 50% of the cells differentiating into mature monocytes/macrophages expressing nonspecific esterase (NSE) activity and the surface antigens of CD 11b, CD 14, and CD 68. Yet, the differentiated U937 cells also had functions of phagocytosis and superoxide production. However, PSCS alone or normal MNC-CM had no such effects. The levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-1 were very low in normal MNC-CM, and they were greatly increased in MNC-CM prepared with PSCS stimulation. Antibody neutralization studies further revealed that the tumoricidal and differentiating effects of PSCS-MNC-CM were mainly derived from the elevated cytokines, especially IFN-γ and TNF-α. These two cytokines acted synergistically on inhibiting cell growth and inducing differentiation of the target U937 cells.

Activation of CD4-positive T cells by polysaccharide fractions isolated from the Cupressaceae Thuja occidentalis L. (Arborvitae).

Various mitogens have been described which act on different subpopulations of mouse and/or human peripheral lymphocytes. In recent years it has been mainly Japanese and German research teams that have isolated polysaccharide fractions with immunomodulative properties [1–3, 17]. In addition to fungal polysaccharides such as lentinan, schizophyllan, and PSK polysaccharide, which are basically in the experimental step of clinical use [6, 10, 13], the focus of attention has mainly been on plants belonging to the asteracean family, such as Echinacea purpurea and E. angustifolia, where an increase in granulocyte phagocytosis is observable. Thuja occidentalis L., a Cupressaceae, has hardly been investigated up till now.

Analysis of cell wall material from plant tissues: Extraction and purification

The alcohol-insoluble residue (AIR) of immature and mature runner beans contains co-precipitated cytoplasmic proteins, nucleic acids, starch and polyphenols, which contaminate the isolated polysaccharide fractions and their binding is sufficiently tenacious to resist complete extraction with the usual protein solubilizing reagents. Therefore, a method was developed for preparation of “cell wall material” from plant tissues in which the contamination with cytoplasmic constituents was minimal. Alternative solvents for cell disruption and protein extraction have been compared. The method depended for its success on the selective removal of the contaminants from fresh ball-milled tissue by sequential treatments with 1% aq. Na deoxycholate, PhOH-HOAc-H 2O followed by α-amylase digestion. Ball-milling the tissue ensured almost complete rupture of the cells and organelles and allowed the solvents to penetrate the sample fully and dissolve the cytoplasmic constituents. The purified “cell wall material” has protein contents varying from 2.5 to 5.5% depending on the type and maturity of the tissue. The residual proteins are resistant to pronase, rich in hydroxyproline and have the amino acid composition of purified cell wall proteins, showing that the wall preparations are relatively pure.

Influence of corneal epithelium on the incorporation of 35SO4 into stromal glycosaminoglycans

In vivo studies on 35SO4 incorporation into the polysaccharides of intact corneas and corneas lacking epithelium were performed. The isotope was injected into the anterior chamber and the activity measured on the isolated polysaccharide fractions at different times after administration. It was clearly demonstrated that the absence of epithelium strikingly decreased the sulfate incorporation in corneal polysaccharides.