epiglucan and Hemolysis

Beta-glucans are widely used in treatment, cosmetics, and the food industry. Glucans play a significant role in activation of the immune and antioxidant system and inhibiting tumor proliferation. In the current study the antitumor activities of new high and low molecular weight beta-glucan derived from oats were investigated in two human lung cancer cell line (A549, H69AR) and normal keratinocytes (HaCaT). The effect of high and low molecular weight beta-glucan from oat was evaluated by cellular viability assessment, lipid peroxidation and manganese superoxide dismutase evaluation and cytoskeleton visualisation. Additionally the level of red blood cells hemolysis was performed. Our results indicate strong anti-tumor properties of new beta-glucan from oat and at the same time no toxicity for normal cells.

Topics: Antineoplastic Agents; Antioxidants; Avena; beta-Glucans; Cell Survival; Hemolysis; Humans; Lipid Peroxidation; Lung Neoplasms; Male; Middle Aged; Molecular Weight; Neoplasms, Glandular and Epithelial; Tumor Cells, Cultured

Two novel magnetic agarose bead based assays have been developed to measure complement component C5 interaction with C3b and the Factor I Modules (FIMs) of C7. One innovation was to couple C3b onto the magnetic agarose bead using the alternative pathway C3 convertase, which resulted in a linkage of the ligand by a covalent ester bond. A second innovation was to employ nickel ion charged N,N,N'-tris(carboxymethyl)ethylene-diamine-magnetic agarose to capture recombinantly prepared C7 FIMs that were expressed with an oligo-histidine linker followed by an acidic domain that provided a spacer enabling the C7 modules exposure to C5. Detection was brought about by peroxidase coupled to C5. Both assays exhibited adequate statistics suitable for screening. As examples of the utility of these new methods, we chose to examine influence of natural products on C5 interaction. Fucoidan and β-glucans were observed to inhibit C3b-C5 interaction, and dextran sulfate was similarly active; however, rosmarinic acid had no measurable effect. In contrast only β-glucans from two species of macrofungi were able to interfere with interaction of C5 with the FIMs of C7.

Topics: beta-Glucans; Complement Activation; Complement C3-C5 Convertases; Complement C3b; Complement C5; Complement C7; Complement Hemolytic Activity Assay; Complement Inactivating Agents; Dextran Sulfate; Hemolysis; Humans; Immunologic Techniques; Iron; Magnetics; Polysaccharides; Protein Binding; Sepharose

(1→3)-β-d-glucan derived from Poria cocos hardly exhibits bioactivities. To extend its use, three types of (1→3)-β-d-glucan derivatives, which were sulfated (1→3)-β-d-glucan (S-P), carboxymethyl (1→3)-β-d-glucan (CMP) and carboxylmethyl (1→3)-β-d-glucan sulfate (S-CMP), were synthesized. Potential antioxidant activities of S-P, CMP and S-CMP were evaluated in vitro. The experiments of scavenging abilities of free radicals were carried out, such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide anion and hydroxyl. Deeply study of the derivatives' inhibitory effect for lipid peroxidation, DNA oxidative damage, erythrocyte hemolysis, and malondialdehyde (MDA) production were determined. And S-CMP significantly (P<0.01) increased the antioxidant activity of β-glucan. These results showed that multiple modifications of polysaccharides may bring the derivatives with excellent properties and various applications.

Topics: Animals; Antioxidants; beta-Glucans; DNA Damage; Erythrocytes; Free Radicals; Hemolysis; Hydrogen Peroxide; Lipid Peroxidation; Male; Malondialdehyde; Mice; Mitochondria, Liver; Mycelium; Oxidative Stress; Poria; Rats; Structure-Activity Relationship; Sulfates

A carboxymethylated-sulfated derivative of (1-->3)-beta-d-glucan (PCS3-II) extracted from Poria cocos was synthesized and coded as CS-PCS3-II. Results of infrared (IR) and Carbon-13 nuclear magnetic resonance spectroscopy ((13)C NMR) indicated that CS-PCS3-II contained carboxymethyl and sulfate groups with a degree of substitution (DS) of 1.05 and 0.36 respectively. By using size exclusion chromatography (SEC) combined with laser light scatting (LLS), the dependence of radius of gyration ((z)(1/2)) on the molecular weight (M(w)) for CS-PCS3-II was established as (z)(1/2) = 6.92 x 10(-2)M(w)(0.59) in 0.15M NaCl solution at 25 degrees C, suggesting that CS-PCS3-II existed as an extended flexible chain. CS-PCS3-II exhibited significantly higher inhibition ratio to Sarcoma 180 tumor in BALB/c mice than PCS3-II. Histological examination of tumor cells treated with CS-PCS3-II had signs of necrosis and apoptosis. It is postulated that introduction of the carboxymethyl and sulfate groups to PCS3-II increased its possible contact with the receptors of immune cells through hydrogen binding and electrostatic attraction, leading to a stronger immunological responses that resulted in inhibition of tumor cell proliferation. Moreover, there were significant increases in phagocyte and thymus indexes, spleen index, hemolytic activity as well as spleen antibody production and delayed type hypersensitivity (DTH), suggesting that CS-PCS3-II could significantly enhance immunpotentiation in mice.

Topics: Adjuvants, Immunologic; Animals; Antibiotics, Antineoplastic; beta-Glucans; Cell Line, Tumor; Erythrocytes; Hemolysis; Hypersensitivity, Delayed; Immunity, Cellular; Macrophages; Magnetic Resonance Spectroscopy; Male; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Phagocytosis; Poria; Proteoglycans; Sarcoma 180; Scattering, Radiation; Sheep; Spectrophotometry, Infrared; Spectroscopy, Fourier Transform Infrared; Spleen; Thymus Gland

A series of lipopeptide compounds co-produced during the fermentation of pneumocandin A0 (L-671,329) and related semisynthetic compounds were evaluated in vivo against Pneumocystis carinii pneumonia and systemic candidiasis. In addition, they were tested in vitro against a panel of pathogenic Candida species and in a Candida membrane 1,3-beta-D-glucan synthesis assay. The results of these studies demonstrate that pneumocandin A0 and pneumocandin B0 (L-688,786) are the most potent compounds when considering both antipneumocystis and anticandida activity. Other compounds in the series are selectively more potent against P. carinii or Candida albicans suggesting a diverging structure-activity relationship. Evaluation of these compounds for their ability to inhibit C. albicans 1,3-beta-D-glucan synthesis in vitro demonstrates that they inhibit this process. A positive correlation between 1,3-beta-D-glucan synthesis inhibition and in vitro antifungal activity was also demonstrated for some of the pneumocandins.

Topics: Animals; Anti-Bacterial Agents; Antifungal Agents; beta-Glucans; Candida albicans; Cell Membrane; Disease Models, Animal; Echinocandins; Erythrocytes; Glucans; Hemolysis; Humans; Mice; Microbial Sensitivity Tests; Mitosporic Fungi; Peptides; Peptides, Cyclic; Pneumocystis; Structure-Activity Relationship

Lei Wan, Poliporus mylittae Cook et Mass (Omphalia lapidescens Schraet) is a kind of fungus used in traditional Chinese medicine, as an anthelminthic. From Lei Wan, an active component designated as S-4001 had been isolated. Preliminary results indicate that S-4001 belongs to D, beta, 1-3 glucan with some 1-6 linkages. After administration of S-4001, significant antiinflammatory activity was found in various experimental animal models, including croton oil induced ear edema in mice and agar or yeast induced ankle swelling in rats. An inhibitory action on leucocyte migration induced by intraperitoneal injection of CMC in rats was also observed. The plasma content of corticosterone was significantly increased, but the content of ascorbic acid in the adrenals did not change in rats given S-4001. Apart from these actions, S-4001 showed a number of immunostimulating actions such as increasing the clearance of Congo red from mice blood and potentiating the immunohemolysis reaction in 615 mice.

Topics: Adjuvants, Immunologic; Animals; Anti-Inflammatory Agents, Non-Steroidal; Basidiomycota; Cell Migration Inhibition; Congo Red; Glucans; Hemolysis; Metabolic Clearance Rate; Mice; Polyporaceae; Rats

Topics: Anti-Inflammatory Agents; beta-Glucans; Drug Combinations; Glucans; Hemolysis; Humans

An antitumor (1----3)-beta-D-glucan with a number-average degree of polymerization (DP) of 540 from Alcaligenes faecalis var. myxogenes IFO 13140, and its lower molecular weight derivatives were found to activate the alternative pathway of complement (APC), as judged by hemolytic and immunoelectrophoretic analyses. Of the native and derivative (1----3)-beta-D-glucans measured, the smallest one that showed APC-activating ability was that with a DP of about 20. The effect of carboxymethylation of the (1----3)-beta-D-glucans with DPs of 49, 131 and 540 on their APC-activating ability was investigated. In any (1----3)-beta-D-glucan the ability was decreased with the increase of carboxymethyl substitution and was completely lost when about one carboxymethyl group per glucose residue was incorporated. In contrast, strong inhibitory ability against C1 hemolytic activity appeared on carboxymethylation.

Topics: Alcaligenes; Animals; beta-Glucans; Complement Activation; Complement C3; Complement Pathway, Alternative; Erythrocytes; Glucans; Guinea Pigs; Hemolysis; Immunoelectrophoresis; Kinetics; Molecular Weight; Sheep; Structure-Activity Relationship