Polysaccharide-rich Fraction Research Articles

Macrophage stimulating polysaccharide purified from peels of grape (Vitis labrusca)

To characterize the macrophage stimulating polysaccharide in grape (Vitis labrusca) peels, the active crude polysaccharide (VL-3) has been fractionated from the hot-water extract of grape peels. A macrophage stimulating polysaccharide-rich fraction (VL-3IIb-1-1) was purified from VL-3 by 3 successive column chromatographies on DEAE-Sepharose CL-6B, Sepharose CL-6B, and Sephacryl S-300. VL-3IIb-1-1 was eluted as a single peak on high performance liquid chromatography (HPLC) and its molecular weight was estimated to be 194 kDa. VL-3IIb-1-1 consisted mainly of Ara and Gal in addition to uronic acid (GalA+GlcA) (molar ratio 1.00:0.81:0.72). Methylation analysis indicated that VL-3IIb-1-1 consisted mainly of terminal Araf, 4- or 5-linked Ara, 2,4-branched Rha, 6- or 3,4- or 3,6-branched Gal, and 3,4,6-branched Glc. Single radial gel diffusion also indicated that VL-3IIb-1-1 showed an intermediate reactivity with β-glucosyl-Yariv antigen. In addition, oral administration of VL-3IIb enhanced the stimulatory responses of macrophage stimulating activity ex vivo. Therefore, VL-3IIb-1-1 purified from grape peels is suggested to be pectic polysaccharide with arabino-3,6-galactan, and it is assumed that VL-3IIb-1-1 plays an important role for expression of its activity.

Physicochemical Properties and Antioxidant Capacity of 3 Polysaccharides from Green Tea, Oolong Tea, and Black Tea

Three polysaccharide-rich fractions named GTPS, OTPS, and BTPS were isolated from green tea, oolong tea, and black tea, respectively. Chemical characteristics, glycosidase inhibitory effects, and antioxidant properties of the 3 fractions were compared. Monosaccharides of GTPS were composed of D-rhamnose, L-arabinose, D-xylose, D-mannose, D-galactose, and D-glucose. But there were no xylose and mannose detected in OTPS and BTPS. The molecular weight distributions were decreased from 9.2 to 251.5 KDa to 3.8 to 32.7 KDa with the fermentation of the tea from green tea to black tea. BTPS showed the highest alpha-glucosidase inhibitory activity, antioxidant activities on hydroxyl radicals and DPPH radicals. The differences in antioxidant activities and glycosidase inhibitory properties among the 3 polysaccharide-rich fractions appeared to be related to differences in monosaccharide composition and molecular weight distribution of the polysaccharide. Diabetes mellitus (DM) is one of the primary threats to human health due to its increasing prevalence, chronic course, and disabling complications. Control of postprandial hyperglycemia and inhibition of oxidative stress are suggested to be important in the treatment of diabetes. Many efforts had been made to search for effective and safe alpha-glucosidase inhibitors and antioxidants from natural materials to develop a physiological functional food or lead compounds for curing diabetes. Coarse tea was used to cure diabetics in people in China and Japan. The hypoglycemic activity increased with the contents of polysaccharide in coarse tea. Many studies have focused on the hypoglycemic activities of tea polysaccharides, but little is known about the glycosidase inhibitory effects of tea polysaccharide. The aim of this study was to find a tea polysaccharide with the best potential for exploitation in curing diabetes.

Effect of Asparagus racemosus on sexual dysfunction in hyperglycemic male rats

Vajikaran rasayana (aphrodisiac) herbs of the Ayurvedic system of medicine have been advocated for improving failing sexual function in males (Thakur and Dixit, 2008). Diabetes mellitus is a major cause of male reproductive function, possibly due to an increased oxidative stress. Fructans and fructoligosaccharides (FOS), which are also considered as functional food components, have been reported to produce a beneficial effect against oxidative stress. The aqueous extract of the roots of Asparagus racemosus Willd. (Liliaceae) rich in 2→1 type FOS were evaluated for their efficacy against streptozotocin and alloxan induced diabetes leading to sexual dysfunction in rats. The behavioral analysis of rats was undertaken to observe the effect on mount, ejaculation and intromission latencies as well as frequencies, hesitation time and copulatory rate. It was observed that streptozotocin as well as alloxan induced hyperglycemic rats showed an overall reduced sexual performance. The deleterious effect was significantly ameliorated in animals treated with polysaccharide-rich fraction of A. racemosus. The study validates the traditional claim of using A. racemosus as an aphrodisiac herb for treating sexual dysfunction in males.

Polysaccharide-rich fraction of Agaricus brasiliensis enhances the candidacidal activity of murine macrophages

A polysaccharide-rich fraction (ATF) of medicinal mushroom Agaricus brasiliensis was evaluated on the candidacidal activity, H2O2 and nitric oxide (NO) production, and expression of mannose receptors by murine peritoneal macrophages. Mice received three intraperitoneal (i.p.) injections of ATF and after 48 h their peritoneal resident macrophages were assayed against Candida albicans yeast forms. The treatment increased fungicidal activity and it was associated with higher levels of H2O2, whereas NO production was not affected. We also found that the treatment enhances mannose receptor expression by peritoneal macrophages, which are involved in the attachment and phagocytosis of non-opsonized microorganisms. Treatment of animals with ATF was able to enhance the clearance of C. albicans during the first 6 h after the experimental i.p. infection. Our results suggest that this extract can increase host resistance against some infectious agents through the stimulation of microbicidal activity of macrophages.

Ameliorative Effect of Fructo-Oligosaccharide Rich Extract of Orchis latifolia Linn. on Sexual Dysfunction in Hyperglycemic Male Rats

Diabetes mellitus (DM) is one of the most prevalent modern degenerative disease that has been implicated for deleterious effects on male reproductive function possibly due to an increased oxidative stress. Fructans and fructooligosaccharides (FOS's) which are also considered as functional food components, have been reported to produce a benevolent effect against streptozotocin induced oxidative stress. The aqueous extract of Orchis latifolia rich in FOS's as well as phytosterols were evaluated for their efficacy against streptozotocin and alloxan induced sexual dysfunction. The behavioral analysis of rats was undertaken to observe the effect on mount, ejaculation and intromission latencies as well as frequencies, hesitation time and copulatory rate. It was observed that hyper- glycemia has an adverse effect on overall sexual behavior. The deleterious effect was significantly reduced in animals treated with polysaccharide rich fraction of O. latifolia. The study suggests that the diabetes induced sexual disability may be ameliorated by proper usage of herbal drugs.

Immunoactive Polysaccharide-Rich Fractions from Panax notoginseng

Panax notoginseng is a commonly used medicinal plant in south-western China. In a previous study, a sequential solubilisation of P. notoginseng high-molecular-weight (HMW) polymers using phenol-acetic acid-water, hot water, weak and strong alkali was performed to determine the structure of the component polysaccharides and proteins. The effects of these extracted HMW fractions on the human complement system, polymorphonuclear neutrophils (PMN) and peripheral blood mononuclear cells (PBMC) are reported here. Fr (1MKOH), which was extracted with 1 M KOH, showed the strongest complement-fixing activity and priming of reactive oxygen species (ROS) production by PMNs, as well as a mitogenic effect. Fr (1MKOH) was further fractionated by anion-exchange chromatography followed by gel-permeation chromatography. 1MD3-G2, the fraction most strongly bound to the DEAE anion-exchange column with a molecular weight of 1140 kDa, showed the highest complement-fixing activity. It is composed of acidic polysaccharides [including glucuronoarabinoxylan (GAX), homogalacturonan (HGA), rhamnogalacturonan I (RG I)], neutral polysaccharides (4-galactan and arabinan), and some protein.

280. Effect of Polysaccharides from Medicinal Herbs as Danger Signals: Implication for Regulating Immune Responses

Toll-like receptors (TLRs) are known to play an important role in binding of various pathogen associated molecular patterns (PAMPs) or non-PAMPs and signalizing the perception of stimuli from pathogens and that results in subsequent immune responses. Many reports have shown that plant polysaccharides can stimulate the immune system, but so far there is no clear evidence of molecular mechanism involved. We hypothesized that specific plant polysaccharides might mimic the functions of exogenous danger signals to mammalian immune cells and confer multiple effects in stimulating the immune cell systems via the TLR4-mediated NF-B signaling pathway. The main objective of our study is to find the potential immune modulators from plant extracts which act as Toll-like receptors (TLRs) ligands. In a primary screening, we identified few extracts enriched with polysaccharides significantly enhanced the transgenic NF-kB driven reporter gene activity (pNiFty-SEAP) in B-16 cell line. These extracts also showed increased IL-8 secretion in a monocytic cell line, THP-1. Further studies with DxI, a polysaccharide rich fraction from Dioscorea tuber, showed involvement of TLR4 in the secretion of pro-inflammatory cytokine (IL-8) and uric acid in primary monocytes. Treatment of THP-1 cells with polysaccharide rich fractions in combination with IL-4 resulted in up-regulation of DC-SIGN. Similarly, the treatment of human monocyte-derived immature DCs with DxI up-regulated many surface markers related to antigen-presentation. This suggested that polysaccharides may enhance the antigen-presenting activity of immune cells. These results thus reveal an apparently unrecognized strong interaction between the plant polysaccharides and the mammalian defense system.

Immunomodulating Polysaccharide Fractions of Menyanthes trifoliata L.

Looking for new plant sources of immunomodulating agents polysaccharide-rich fractions (PS) from Menyanthes trifoliata L. (Menyanthaceae) have been isolated. The herb of Menyanthes trifoliata L. was sequentially extracted with water, 0.1 M NaOH, 8% CH3COOH, and 1 M NaOH. After dialysis and resolution on Biogel P-10 four homogenic (B-4, B-5, C-4, D-5) and two nonhomogenic (A-3 and D-4) PS were isolated. About 0.5% of PS over 3500 Da were found in the dry plant material. They were characterized through chemical analysis, NMR and vibrational spectroscopy. Speciation analysis of chosen metal/metaloid elements was performed and an exceptionally high concentration of Se was found in PS of a pure water extract (A-3). The biological tests on the immunomodulating influence with human blood-derived lymphocytes and granulocytes revealed that two fractions, B-4 and B-5, were strong stimulators of immune cells, whereas fractions D-5 and A-3 were found as potent suppressive and anti-inflammatory agents. The applied isolation procedures led to the separation of active compounds into stimulatory and inhibitory fractions.

Effect of PSK and its Subfractions on Peripheral Blood Lymphocytes Mediated Cytotoxicity against Urinary Bladder Tumor Cells

Our previous studies have indicated that the protein-bound polysaccharide Kreha (PSK) enhances the cytotoxic activity of peripheral blood lymphocytes (PBL) against the T24 human urinary bladder tumor cell line in patients with bladder tumor. Since PSK consists of a mixture of various kinds of protein-bound polysaccharides, the present study was designed to examine which subfractions of PSK mediated the enhancement of cytotoxicity. When PSK was separated according to size, treatment of PBL with the 50kilodalton (kd) or less fraction killed T24 cells more efficiently than unfractionated PSK-treated PBL. The higher molecular weight fractions did not enhance killing above the control level. PSK was fractionated on a diethylaminoethyl (DEAE)-cellulose column to obtain a protein rich fraction that absorbed onto the column and a polysaccharide rich fraction that did not. PBL treated with the polysaccharide rich fraction were able to kill T24 cells more effectively than unfractionated PSK-treated PBL. The protein rich fraction had no effect on the killing. Further fractionation of the polysaccharide rich fraction was performed by differential precipitation with ammonium sulfate. PBL treated with the precipitated fraction at 70–80% saturation (PSK Fraction D) enhanced cytotoxicity equal to that of the polysaccharide rich fraction. Treatment of PBL with the other fractions did not augment the cytotoxicity. These enhancement by PSK fractions were observed in healthy donors and also in patients with bladder tumor. An increase of the proliferative response of PBL to PSK Fraction D as well as unfractionated PSK was observed. Treatment of PBL with PSK Fraction D had no effect on the proportion of PBL binding to T24 cells, thus suggesting a post-binding effect. The structure of PSK Fraction D as inferred from the results of methylation analysis was mainly an α-glucan. These results demonstrate that PSK mediated enhancement of cytotoxicity and proliferation of PBL may be largely due to an a-glucan of less than 50kd.

Anti-Complementary and Hypoglycemic Activities of the Glycans from the Seeds ofMalva verticillata

Seven polysaccharides and peptidoglycans obtained from the seeds of Malva verticillata were tested for anti-complementary activity. Remarkable activities were observed for MVS-I and MVS-IIA. The former is mainly composed of beta-1,3-linked D-glucan and of alpha-1,5-linked L-arabino-beta-3,6-branched D-galactan, and the latter is essentially alpha-1,5-linked L-arabino-beta-3,6-branched D-galactan. Further, considerable activities were observed for MVS-IIIA and MVS-IVA. Both glycans possess arabino-3,6-galactan moieties accompanying alpha-1,3-linked L-arabinopyranosyl, beta-1,4-linked D-xylosyl, and alpha-1,4-linked D-galacturonic acid units. In addition, the major neutral polysaccharide (MVS-I), the major peptidoglycan (MVS-V), and the polysaccharide-rich fraction (MVS-V-CH) obtained from MVS-V were tested for hypoglycemic activity. MVS-I especially showed remarkable activity, and MVS-V-CH also exhibited significant activity. From the viewpoint of structural features, the latter belongs to pectic substances.

Role of lipid A in the production of tumor necrosis factor and differences in antitumor activity between tumor necrosis factor and lipopolysaccharide.

The role of lipopolysaccharide (LPS) in the production of tumor necrosis factor (TNF) was examined. Alkaline treatment of LPS greatly reduced the TNF-producing activity of LPS, but TNF was produced when a large amount was injected. Free lipid A and lipid A-mouse serum albumin complex, which were prepared from the acid hydrolyzate, effectively induced TNF. However, the polysaccharide-rich fraction of the acid hydrolyzate was not capable of inducing TNF. Preincubation of LPS with polymixin B largely abrogated the TNF-producing activity of LPS. The differences in antitumor activity between TNF and LPS were also tested. TNF has a direct cytotoxicity against cancer cells in vitro but LPS does not. The activity of TNF was not inhibited by preincubation with polymixin B. Tumor necrosis in vivo was inhibited by preincubation of LPS with polymixin B but not by that of TNF. Galactosamine was found to induce susceptibility to the lethal effects of LPS, but did not influence the action of TNF. Lipid A is largely responsible for the TNF-inducing activity of LPS, but is not essential for the antitumor activity of TNF.

60 GRAM NEGATIVE SEPSIS OF INFANCY. MECHANISMS OF ACTIVATION OF THE COMPLEMENT SYSTEM

Bacterial lipopolysaccarides(LPS)from gram negative organisms are activators of the alternative and classical pathway of complement(C). This study was undertaken to evaluate if there is an relation between the activation initiated by LPS in vitro and the activation of C in patients with gram negative sepsis. Nine patients with bacteremia, endotoxemia(Limulus)and/or hipocomplementemia(CH50)were studied. Levels of Clq,C4,C3,C5,C3PA and properdine were measured. LPS from cultures were extracted by the procedure of Westphal.Lipid A(LA)and polysaccharide-rich fractions (PS)were obtained by acid hydrolisis. We examined the capacity of LPS, LA and PS to reduce hemolitic activity of NHS. In reaction mixtures with the activators we performed:measurements of Clq,C4,antigen B of C3,C5,C3PA and properdine hemolitic C4, mobility of C3,C3PA and properdine. Hypocomplementemia was present in 6 patients,the C profile suggests activation of both pathways in 5 and alternative in 1.Seven LPS reduced hemolitic capacity of NHS and breakdown C3.Different types of activators could be distinguished by C4 determinations and mobility of C3PA.Three LPS activate both pathways,2 alternative and 1 classical.LA initiate classical pathway and PS alternative.No correlation was found between the activation mechanism in vitro and the patients profile. Hypocomplementemia was observed particularly in severely compromised patients.

Biosynthesis of macromolecules in chloride cells in the gills of the common eel, Anguilla anguilla, adapting to sea water

1. 1. Rates of incorporation of( 3H)thymidine, ( 14C)leucine and ( 3H)glucoseamine into TCA-precipitable products were measured in chloride and respiratory epithelial cell populations in the gills of silver eels adapting to sea water. 2. 2. A transient wave of DNA synthesis occurs in both cell populations during early stages of adaptation. 3. 3. Chloride cells but not respiratory epithelial cells turn over faster in sea water than in fresh water. 4. 4. A polysaccharide-rich fraction appears to turn over faster in chloride cells in sea water. A glycoprotein fraction appears to turn over faster in respiratory epithelial cells in sea water.

Immune Adjuvancy of Lipopolysaccharide and a Nontoxic Hydrolytic Product Demonstrating Oscillating Effects with Time

Abstract Since the first reports of immune adjuvancy of endotoxic lipopolysaccharides (LPS) (1, 2), large numbers of such preparations have proven to be effective adjuvants for both soluble and cellular immunogens in a host of laboratory animals (for review, see Neter (3)). Kind and Johnson (4) as well as Franzl and McMaster (5) investigated the effect of time of LPS treatment on the primary immune response. In earlier investigations on radioprotective properties of LPS, we observed a unique time-dependent survival pattern of lethally irradiated mice pretreated with LPS (6). It was established that two optimal time intervals, peaking at -1 to -3 and -8 to -12 days, existed. A closely parallel time-dependent radioprotective profile of the nontoxic, polysaccharide-rich fraction (PS)3 of LPS could also be established. Quite recently a similar time-dependent, LPS-induced protection against TA3-Ha tumors in mice was also observed by us (7).

The purification of mycobacterial polysaccharides with concanavalin A.

Concanavalin A was demonstrated to react with 3 antigenic polysaccharide constituents of culture filtrates of Mycobacterium tuberculosis. Using direct precipitation and affinity chromatography methods, concanavalin A-reactive polysaccharides could be separated from concanavalin A-non-reactive polysaccharide in a partially purified, polysaccharide-rich fraction of mycobacterial culture filtrate. By the use of appropriate dissociation conditions, 2 concanavalin A-reactive polysaccharides of differing concanavalin affinities could be separated and purified. Four polysaccharides thus studied were identified with the United States-Japan mycobacterial antigen nomenclature. Antigen 1 was found to be a concanavalin A-reactive polysaccharide of high affinity. Antigen 2 was found to identify both with a concanavalin A-reactive polysaccharide of low affinity and with a concanavalin A-nonreactive polysaccharide. Antigen 3 was found to represent the large molecular weight polysaccharide II of Seibert and to react with...