Angelica gigas polysaccharide induces CR3-mediated macrophage activation and the cytotoxicity of natural killer cells against HCT-116 cells via NF-κB and MAPK signaling pathways.

Angelica gigas (A. gigas) is traditional medicinal herb that mainly exists in Korea and northeastern China. There have been relatively few studies conducted thus far on its polysaccharides and their bioactivities. We purified and described a novel water-soluble polysaccharide derived from A. gigas and investigated its immunoenhancing properties. The basic components of crude and purified polysaccharides (F1 and F2) were total sugar (41.07% - 70.55%), protein (1.12-10.33%), sulfate (2.9-5.5%), and uronic acids (0.5-31.05%) in total content. Our results demonstrated that the crude and fractions' molecular weights (Mw) varied from 42.2 to 285.2 × 103 g/mol. As the most effective polysaccharide, F2 significantly stimulated RAW264.7 cells to release nitric oxide (NO) and express several cytokines. Furthermore, F2 increased the expression of tumor necrosis factor-α (TNF-α), interferon-gamma (IFN-É£), natural killer cytotoxicity receptors (NKp44), and granzyme-B in NK-92 cells and enhanced the cytotoxicity against HCT-116 cells. In our experiments, we found that F2 stimulated RAW264.7 cells and NK-92 cells via MAPK and NF-κB pathways. The monosaccharide and methylation analysis of the high immunostimulant F2 polysaccharide findings revealed that the polysaccharide was primarily composed of 1 â†’ 4, 1 â†’ 6, 1 â†’ 3, 6, 1 â†’ 3 and 1 â†’ 3, 4, 6 galactopyranose residues, 1 â†’ 3 arabinofuranose residues, 1 â†’ 4 glucopyranose residues. These results demonstrated that the F2 polysaccharide of A. gigas which possesses potential immunostimulatory attributes, could be used to create a novel functional food.

Inhibitory effects of polysaccharides from Korean ginseng berries on LPS-induced RAW264.7 macrophages.

Polysaccharides isolated from Korean ginseng berries (GBPs) have shown beneficial effects such as immunomodulatory, anti-inflammatory, anti-cancer, and anti-diabetic properties. However, little is known about anti-inflammatory effects of GBPs. Thus, the purpose of this study was to investigate anti-inflammatory properties of four fractions of GBPs, namely GBP-C, GBP-F1, GBP-F2, and GBP-F3, in macrophages. Their toxicities and effects on NO production in RAW264.7 cells were assessed by culturing cells with various concentrations of GBPs and stimulating cells with LPS. Furthermore, expression levels of inflammatory mediators, cytokines, cell surface molecules, and immune signaling pathways were evaluated in LPS-stimulated macrophages using different fractions of GBPs at 450 µg/mL. These GBPs activated LPS-stimulated RAW264.7 cells to significantly reduce NO production. They suppressed the expression of mRNA and cell surface molecules via MAPK and NF-κB pathways. Collectively, results revealed that all four GBP fractions showed anti-inflammatory effects, with GBP-F1 having a more efficient anti-inflammatory effect than GBP-C, GBP-F2, and GBP-F3. The structure of GBP-F1 mainly consists of 1 → 3)- Araf, 1 → 4)- Glcp, and 1 → 6)-Galp glycosidic linkages. These results demonstrate that GBPs can be employed as alternative natural sources of anti-inflammatory agents.

Korean Ginseng Berry Polysaccharide Enhances Immunomodulation Activities of Peritoneal Macrophages in Mice with Cyclophosphamide-Induced Immunosuppression.

Korean ginseng (Panax ginseng C. A. Meyer), a member of the Araliaceae family, is known as a traditional medicinal plant to have a wide range of health properties. Polysaccharides constitute a major component of Korean ginseng, and its berries exhibit immune-modulating properties. The purpose of this study was to investigate the immune effects of crude polysaccharide (GBPC) extracted from Korean ginseng berry on peritoneal macrophages in mice with cyclophosphamide (CY)- induced immunosuppression. BALB/c mice were divided into eight groups: normal control, normal control + CY, levamisole + CY, ginseng + CY, and four concentrations of 50, 100, 250, and 500mg/kg BW/day of GBPC + CY. Mice were orally administered with samples for 10 days. Immunosuppression was established by treating mice with CY (80 mg/kg BW/day) through intraperitoneal injection on days 4 to 6. The immune function of peritoneal macrophages was then evaluated. Oral administration of 500mg/kg BW/day GBPC resulted in proliferation, NO production, and phagocytosis at 100%, 88%, and 91%, respectively, close to the levels of the normal group (100%) of peritoneal macrophages. In CY-treated mice, GBPC of 50-500 mg/kg BW/day also dose-dependently stimulated the proliferation, NO production, and phagocytosis at 56-100%, 47-88%, and 53-91%, respectively, with expression levels of immune-associated genes, such as iNOS, COX-2, IL-1ß, IL-6, and TNF-α, of about 0.32 to 2.87-fold, compared to those in the CY group. GBPC could be a potential immunomodulatory material to control peritoneal macrophages under an immunosuppressive condition.

An effective bio-inspired synthesis of platinum nanoparticles using Caulerpa sertularioides and investigating their antibacterial and antioxidant activities.

In this study, we report the synthesis of platinum nanoparticles (Cs-PtNPs) using an aqueous extract of Caulerpa sertularioides as a reducing agent. Cs-PtNPs were characterized by UV-Vis spectroscopy, fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), field emission electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDAX), high-resolution transmission electron microscopy (HR-TEM) and dynamic light scattering (DLS) analysis. Cs-PtNPs are spherical with a particle size of 6-22 nm. Cs-PtNPs have been shown to have highly effective antioxidant activities with 74% for DPPH, 63% for reducing power, and 59% for total antioxidant at 1 mg/ml, and results were compared with standard L-ascorbic acid. Furthermore, the Cs-PtNPs demonstrated excellent antibacterial activity against the Gram-negative bacteria, Vibrio parahaemolyticus with the highest zone of inhibition (18 mm) at 50 µg/ml. Moreover, Artemia nauplii showed less toxicity when treated with Cs-PtNPs at 150 µg/ml, indicating that the Cs-PtNPs are less toxic and environment friendly.

Supramolecular nanogels based on gelatin-cyclodextrin-stabilized silver nanocomposites with antibacterial and anticancer properties.

An effective method for reducing silver ions using gelatin (Gel) and 2-hydroxypropyl-ß-cyclodextrin (HPCD) hydrogels, which stabilize silver at various concentrations is described. The formation of AgNPs in solution, as well as Gel-HPCD nanogels, is confirmed by the surface plasmon resonance (SPR) band at 420-440 nm in the UV-Vis spectrum. The resulting Gel-HPCD and Gel-HPCD/AgNPs composites are characterized using various techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), X-ray diffractometry (XRD), and thermogravimetric analysis (TGA). SEM images showed that the porous structure and the AgNPs are homogeneously dispersed throughout the Gel-HPCD/AgNP composites network. The AgNPs in the Gel-HPCD/AgNPs composite is crystalline, with spherical particles having an average size of 7.0 ± 2.5 nm, as determined by TEM. The Gel-HPCD/AgNPs composites are strongly effective against both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria. The assembled antibacterial Gel-HPCD/AgNPs composites are also assessed for their cytotoxic and anticancer activities using HCT-116 cancer cells. The results suggest that Gel-HPCD/AgNPs composites could be used as effective therapeutics in the future in tissue engineering applications, as their bactericidal properties and low toxicity make them ideal for clinical use.

Immune-Enhancing Effects of Crude Polysaccharides from Korean Ginseng Berries on Spleens of Mice with Cyclophosphamide-Induced Immunosuppression.

Panax ginseng C. A. Meyer is well known as traditional herbal medicine, and ginseng berries are known to exhibit potential immune-enhancing functions. However, little is known about the in vivo immunomodulatory activity of Korean ginseng berries. In this study, crude Korean ginseng berries polysaccharides (GBP) were isolated and their immunomodulatory activities were investigated using cyclophosphamide (CY)-induced immunosuppressive BALB/c mice. In CY-treated mice, oral administration of GBP (50-500 mg/kg BW) remarkably increased their spleen sizes and spleen indices and activated NK cell activities. GBP also resulted in the proliferation of splenic lymphocytes (coordinating with ConA: plant mitogen which is known to stimulate T-cell or LPS: endotoxin which binds receptor complex in B cells to promote the secretion of pro-inflammatory cytokines) in a dose-dependent manner. In addition, GBP significantly stimulated mRNA expression levels of immune-associated genes including interleukin-1ß (IL-1ß), IL-2, IL-4, IL-6, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), toll-like receptor 4 (TLR-4), and cyclooxygenase-2 (COX-2) in CY-treated mice. These results indicate that GBP is involved in immune effects against CY-induced immunosuppression. Thus, GBP could be developed as an immunomodulation agent for medicinal or functional food application.

Intranasal Administration of Codium fragile Polysaccharide Elicits Anti-Cancer Immunity against Lewis Lung Carcinoma.

Natural polysaccharides have shown promising effects on the regulation of immunity in animals. In this study, we examined the immune stimulatory effect of intranasally administered Codium fragile polysaccharides (CFPs) in mice. Intranasal administration of CFPs in C57BL/6 mice induced the upregulation of surface activation marker expression in macrophages and dendritic cells (DCs) in the mediastinal lymph node (mLN) and the production of interleukin-6 (IL-6), IL-12p70, and tumor necrosis factor-α in bronchoalveolar lavage fluid. Moreover, the number of conventional DCs (cDCs) was increased in the mLNs by the upregulation of C-C motif chemokine receptor 7 expression, and subsets of cDCs were also activated following the intranasal administration of CFP. In addition, the intranasal administration of CFPs promoted the activation of natural killer (NK) and T cells in the mLNs, which produce pro-inflammatory cytokines and cytotoxic mediators. Finally, daily administration of CFPs inhibited the infiltration of Lewis lung carcinoma cells into the lungs, and the preventive effect of CFPs on tumor growth required NK and CD8 T cells. Furthermore, CFPs combined with anti-programmed cell death-ligand 1 (PD-L1) antibody (Ab) improved the therapeutic effect of anti-PD-L1 Ab against lung cancer. Therefore, these data demonstrated that the intranasal administration of CFP induced mucosal immunity against lung cancer.

Polysaccharide extracted from Taraxacum platycarpum root exerts immunomodulatory activity via MAPK and NF-κB pathways in RAW264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Taraxacum platycarpum Dahlst. (Korean dandelion) is a medicinal herb used in traditional medicine in Korea to treat various disease such as furuncles, mammitis, hepatitis, jaundice. Moreover, a decoction prepared from T. platycarpum leaves and stems is an effective treatment for cancer, glycosuria, liver disease, pleurodynia, and stomach problems. AIM OF THE STUDY: The main objective of this work was to study the composition and structural properties of polysaccharides (TPP) from Taraxacum platycarpum Dahlst. root and investigate the immunostimulatory activity on RAW264.7 cells. MATERIALS AND METHODS: TPP was extracted from T. platycarpum using hot water extraction, ethanol precipitation method and its fractionated using DEAE-Sepharose fast flow column. The composition, molecular weight, and structural characterization of TPP and its fractions were evaluated by various techniques. Further, the immunostimulatory activity of polysaccharides was tested on murine macrophage cell line RAW264.7 by various in vitro assays. The structure effect of TPP on RAW264.7 cells was studied by the removal of sulfate (desulfation) and protein (deproteinization) contents from TPP. RESULTS: We obtained three fractions namely TPP-1, TPP-2, and TPP-3 which mainly consisted of carbohydrates (75.55, 52.71, and 48.41%), sulfate (8.42, 15.19, and 27.67%), uronic acid (1.27, 6.56, and 4.39%), and protein (8.15, 24.85, and 9.73%). The average molecular weight of the fractions was 56.7, 108.2, and 132.3 × 103 g/mol, respectively. The polysaccharides activate the RAW264.7 cell to produce a significant amount of NO and upregulate the various mRNA expression by the activation of MAPK and NF-κB pathways via TLR4, TLR2, and CR3 receptors. The structurally modified deproteinated derivative (DP-TPP-2) more effectively decreases the NO production which means the protein content of TPP-2 mainly contributes to the RAW264.7 cells activation. The structure of DP-TPP-2 primarily consists of 1 â†’ 2)-Galp, 1 â†’ 6)-Glup, 1 â†’ 2) - Rhap, and 1 â†’ 5) - Arap glycosidic linkages. CONCLUSIONS: The present study demonstrated that the polysaccharide isolated from T. platycarpum shows admirable immunostimulatory by the activation of MAPK and NF-κB pathways through TLR4, TLR2, and CR3 receptors. The protein content of polysaccharides mainly contributes to the RAW264.7 cells activation. Our study results could be useful for developing a new immunostimulant agent.

Optimization of ultrasonic-assisted extraction of polysaccharides from purple glutinous rice bran (Oryza sativa L.) and their antioxidant activities.

Purple glutinous rice bran (Kum Doi Saket rice (KUM)) contains high content of edible polysaccharides and anthocyanins and has an excellent antioxidant activity. This research aimed to optimize the extraction of crude polysaccharides from defatted purple glutinous rice bran using an ultrasonic-assisted extraction (UAE) and compared with a hot water extraction (HWE). Results showed that optimal extraction condition was as follows: a defatted rice bran to water ratio of 1:20 w/v, extraction temperature and time of 70 °C for 20 min. Under the optimal extraction condition, the yield of polysaccharide of UAE (4%) was significantly higher than that obtained from the HWE (0.8%). Additionally, antioxidant activities of extracted polysaccharide including IC50 value DPPH, IC50 value ABTS, and FRAP value were 1.09 mg/mL, 2.80 mg/mL and 197 µM Fe2+/g, respectively. It is suggested that the UAE process is promising method to decrease the processing time and to enhance extracted polysaccharide yields by 4 times.

Inducing inflammatory response in RAW264.7 and NK-92 cells by an arabinogalactan isolated from Ferula gummosa via NF-κB and MAPK signaling pathways.

The polysaccharide isolated from F. gummosa (FGP) was found homogenous with a weight average molecular weight (Mw) of 50.0 × 103 g/mol and radius of gyration (Rg) of 105.3 nm. The FGP was an arabinogalactan with a backbone formed of →6)-ß-Galp-1→ residues having random branching points at C-3 extended with either ß-Galp-(1→3)-ß-Galp-(1→ or α-Araf-(1→ side chain residues. FGP exhibited proliferative effect on RAW264.7 cells and induced macrophages to exert proinflammatory response releasing NO and up-regulating the transcription of cytokines including TNF-α, IL-1ß, IL-6 and IL-12. The FGP induced NK-92 cells to up-regulate the expressions of TNF-α, IFN-γ, granzyme-B, perforin, NKG2D and FasL. The presence of p-NF- κB, p-ERK, p-JNK and p-p38 in RAW264.7 and NK-92 cells indicated their activation through NF-κB and MAPKs signaling pathways. These findings suggested that polysaccharides from F. gummosa are potent in boosting immune system and thus may be considered for further studies of biomedical applications.

Isolation, structural elucidation and immuno-stimulatory properties of polysaccharides from Cuminum cyminum.

The aim of the present study was to evaluate the effect of water-soluble polysaccharides from Cuminum cyminum to induce inflammatory response in immune cells and understand their underlying mechanisms. Weight average molecular weight (Mw) of polysaccharides varied between 191.4-512.2 × 103 g/mol. Polysaccharides induced RAW264.7 cells to release nitric oxide and express TNF-α, IL-1ß, IL-6 and IL-12 inflammatory cytokines. Polysaccharides activated NK-92 cells to produce TNF-α, IFN-γ, perforin, granzyme B, NKG2D and FasL. Activations of RAW264.7 and NK-92 cells were through NF-κB and MAPKs signal pathways indicated by the presence of phosphorylated NF-κB, ERK, JNK and p38 proteins. The polysaccharide structure was mainly constituted of →4)-Galp-(1→, →3)-Galp-(1→, →2)-Arap-(1→ and →2)-Arap-(1→ glycosidic linkages. Overall results suggested that polysaccharides from C. cyminum possessing lower MW and greater expanded conformation more effectively stimulate RAW264.7 and NK-92 cells and thus could be considered for further studies on their biomedical applications.

Co-immunomodulatory Activities of Anionic Macromolecules Extracted from Codium fragile with Red Ginseng Extract on Peritoneal Macrophage of Immune-Suppressed Mice.

In this study we investigated the immune effects of oral administration of anionic macromolecules extracted from Codium fragile (CFAM) and red ginseng extract mixture on the peritoneal macrophage cells in immune-suppressed mice. Cyclophosphamide (CY) induces the immune-suppressed condition. CY-treated mice were orally fed with different concentrations of CFAM supplemented with red ginseng extract and the peritoneal macrophages collected. CY treatment significantly decreased the immune activities of peritoneal macrophages, compared to the normal mice. The administration of CFAM mixed with red ginseng extract significantly boosted the viability of macrophage cells and nitric oxide production of peritoneal macrophages. Further, the oral administration of CFAM mixed with red ginseng extract up-regulated the expression of iNOS, COX-2, and TLR-4 as well as cytokines such as IL-1ß, IL-6, TNF-α, and IFN-γ more than the red ginseng-treated group. This study showed that CFAM enhanced the immune activity of red ginseng extract in the peritoneal macrophage cells of immune-suppressed mice. Furthermore, CFAM might be used as a co-stimulant of red ginseng extract through the regulation of macrophage cells for the enhancement of human health and immunity.

Isolation and structural characterization of sulfated polysaccharide from Spirulina platensis and its bioactive potential: In vitro antioxidant, antibacterial activity and Zebrafish growth and reproductive performance.

In this study, the sulfated polysaccharide (SPs) was isolated from Spirulina platensis. The isolated SPs contains carbohydrate, sulfate, protein and uronic acid at 38.7 ±â€¯0.30%, 21.3 ±â€¯0.87%, 7.1 ±â€¯0.15% and 7.9 ±â€¯0.4% respectively. The elemental analysis confirmed the presence of carbon (18.01 ±â€¯0.10%), hydrogen (1.83 ±â€¯0.02%) and nitrogen (3.43 ±â€¯0.01%). The monosaccharide composition and molecular weight of SPs were analyzed by high-performance liquid chromatography and size exclusion chromatography respectively. The monosaccharide composition analysis showed the existence of glucose, rhamnose, xylose, fucose, mannose, galactose and the molecular weight of SPs was 1016 kDa. Further, the characterization of SPs was done by UV-visible spectroscopy, X-ray diffraction, FT-IR, 1H NMR and 13C NMR analysis. The obtained SPs exhibited potent antioxidant activity in DPPH (76.45 ±â€¯0.49%), reducing power (absorbance: 1.3 ±â€¯0.02), hydrogen peroxide scavenging (66.3 ±â€¯1.16%), hydroxyl scavenging (68.6 ±â€¯3.2%), nitric oxide (81.36 ±â€¯1.85%) and total antioxidant (absorbance:1.66 ±â€¯0.02) activities at 5 mg/ml. In addition, SPs revealed the highest antibacterial efficacy against the pathogenic bacteria Vibrio vulnificus in disc diffusion, agar bioassay and protein leakage assays at 100 µg/ml. Furthermore, the supplementation of 2% SPs through a feed to the Danio rerio fish enhances the growth and reproductive performances. This finding confirmed that the isolated SPs from S. platensis possess pharmaceutical as well as nutritional properties.

Immune Enhancement Effects of Codium fragile Anionic Macromolecules Combined with Red Ginseng Extract in Immune-Suppressed Mice.

Codium fragile is an edible seaweed in Asian countries that has been used as a thrombolytic, anticoagulant, antioxidant, anti-inflammatory, and immune-stimulatory agent. Ginseng has also been known to maintain immune homeostasis and to regulate the immune system via enhancing resistance to diseases and microorganisms. In this study, anionic macromolecules extracted from C. fragile (CFAM) were orally administered with red ginseng extract (100 mg/kg body weight) to cyclophosphamide-induced immunosuppressed male BALB/c mice to investigate the immune-enhancing cooperative effect of Codium fragile and red ginseng. Our results showed that supplementing CFAM with red ginseng extract significantly increased spleen index, T- and B-cell proliferation, NK cell activity, and splenic lymphocyte immuneassociated gene expression compared to those with red ginseng alone, even though a high concentration of CFAM with red ginseng decreased immune biomarkers. These results suggest that CFAM can be used as a co-stimulant to enhance health and immunity in immunosuppressed conditions.

Codium fragile F2 sensitize colorectal cancer cells to TRAIL-induced apoptosis via c-FLIP ubiquitination.

This study demonstrates that combined treatment with subtoxic doses of Codium extracts (CE), a flavonoid found in many fruits and vegetables, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), induces apoptosis in TRAIL-resistant colorectal cancer (CRC) cells. Effective induction of apoptosis by combined treatment with CE and TRAIL was not blocked by Bcl-xL overexpression, which is known to confer resistance to various chemotherapeutic agents. While TRAIL-mediated proteolytic processing of procaspase-3 was partially blocked in various CRC cells treated with TRAIL alone, co-treatment with CE efficiently recovered TRAIL-induced caspase activation. We observed that CE treatment of CRC cells did not change the expression of anti-apoptotic proteins and pro-apoptotic proteins, including death receptors (DR4 and DR5). However, CE treatment markedly reduced the protein level of the short form of the cellular FLICE-inhibitory protein (c-FLIPS), an inhibitor of caspase-8, via proteasome-mediated degradation. Collectively, these observations show that CE recovers TRAIL sensitivity in various CRC cells via down-regulation of c-FLIPS.

Effect of sulfation and partial hydrolysis of polysaccharides from Polygonatum sibiricum on immune-enhancement.

The aqueous polysaccharide from Polygonatum sibiricum was extracted and fractionated using anion-exchange chromatography to obtain F1 fraction. The F1 was chemically sulfated and partially acid-hydrolyzed for the production of its over-sulfated (OS1,2,3) and hydrolyzed (HP1,2,3) derivatives, in which the sulfate content of OS1,2,3 was 7.5-17.1%, and the Mw of HP1,2,3 ranged from 18.2 × 103 to 57.3 × 103 g/mol. Considerable RAW264.7 cell activation was observed by HP1,2,3 with NO production of 34.9, 44.3 and 42.7 µM, respectively, as well as the mRNA expression of cytokines (IL-1ß, IL-6, IL-10 and IL-12). NK cell cytotoxicity against HT-29 cell was facilitated by OS1,2,3 treatment with the increased gene expressions of INF-γ, Granzyme-B, perforin, NKG2D, and FasL. RAW264.7 cells appeared to be activated via MR and TLR4 mediated signaling pathway, but CR3 and TRL2 might play a main role in stimulating NK cells. Overall, the present study suggests the potential application of polysaccharides from P. sibiricum in functional foods and pharmacological industries.

Extraction, characterization and immunomodulatory property of pectic polysaccharide from pomegranate peels: Enzymatic vs conventional approach.

Molecular characteristics, structural properties and immunostimulatory activities of polysaccharides from pomegranate peel were evaluated after 0.1 M HCl, Cellic CTec2 and buffer extractions. The isolated polysaccharides were mainly formed of neutral sugars (32.1%-51.1%) and uronic acids (19.9%-30.8%) as well as varying levels of proteins (15.0%-39.5%). Different levels of sugars including glucose (44.9%-68.1%), galactose (14.6%-19.4%), mannose (3.4%-18.1%), arabinose (3.1%-18.1%) and rhamnose (3.5-6.0%) constructed the structure of isolated polysaccharides. The average molecular weight (Mw) of polysaccharides differed notably and ranged from 422.5 × 103 to 18,631.8 × 103 g/mol. Polysaccharide molecules obtained using Cellic CTec2 enzyme induced RAW264.7 murine macrophage cells to release considerable amount of inflammatory mediators including nitric oxide, IL-1ß, TNF-α, IL-6 and IL-10. The polysaccharide stimulation of macrophage cells initiated through NF-κB and MAPKs signaling pathways and activation of p-NF-κB, p-JNK, p-ERK and p-p38 proteins. The most potent immunostimulatory polysaccharides were consisted of (1 → 3)-linked glucose, (1 → 6)-linked galactose, (1 → 4)-linked mannose and (1 → 4)-linked arabinose residues with branching points at C6 and C3. These results indicated that the lower molecular weight of polysaccharides isolated with Cellic CTec2 enzyme could be one of the major determinant structural characteristics in stimulating macrophage cells.

Water-soluble polysaccharides from Ulva intestinalis: Molecular properties, structural elucidation and immunomodulatory activities.

Water-soluble sulfated polysaccharides extracted from Ulva intestinalis and fractionated using DEAE Sepharose fast flow column to identify their molecular properties and macrophage cells stimulating activities. Crude and fractions (F1 and F2) were formed of neutral sugars (58.7-74.7%), sulfates (6.2-24.5%), uronic acids (4.9-5.9%) and proteins (3.2-10.4%). Different levels of sugar constituents including rhamnose (30.1-39.1%), glucose (39.0-48.4%), galactose (0.0-15.8%), xylose (8.5-11.3) and arabinose (0.0-5.1%). The molecular weight (Mw) of crude and fractionated polysaccharides ranged from 87.1 × 103 to 194.1 × 103 (g/mol). Crude polysaccharides were not toxic to RAW264.7 cells and fractions induced cell proliferation. Fraction F1 stimulated RAW264.7 cells to release considerable amounts of nitric oxide, IL-1ß, TNF-α, IL-6, IL-10 and IL-12 cytokines. The main backbone of the most immunostimulating polysaccharide (F1) was consisted of mixed linkages of (1 â†’ 2)-linked rhamnose and (1 â†’ 2)-linked glucose residues.

Structural characterization of sulfated arabinans extracted from Cladophora glomerata Kützing and their macrophage activation.

Water-soluble sulfated heteropolysaccharides were extracted from Cladophora glomerata Kützing and fractionated by ion-exchange chromatography, which yielded two subfractions, F1 and F2. The crude and fractionated polysaccharides (F1 and F2) mostly consisted of carbohydrates (62.8-74.5%) with various amounts of proteins (9.00-17.3%) and sulfates (16.5-23.5%), including different levels of arabinose (41.7-54.4%), galactose (13.5-39.0%), glucose (0.80-10.6%), xylose (6.84-13.4%), and rhamnose (0.20-2.83%). Based on the size exclusion chromatography (SEC) profiles, the crude and fractions mainly contained one peak with shoulders having molecular weight (Mw) ranges of 358-1,501 × 10(3). The F1 fraction stimulated RAW264.7 cells to produce considerable amounts of nitric oxide and cytokines compared to the crude and F2 fraction. The backbone of the most potent immunostimulating fraction (F1) was α-(1→4)-L-arabinopyranoside with galactose and xylose residues as branches at O-2 position, and sulfates mainly at O-2 position as well.

Characterization and immunomodulatory activities of polysaccharides from Spirogyra neglecta (Hassall) Kützing.

Sulfated polysaccharides (SP) isolated from freshwater green algae, Spirogyra neglecta (Hassall) Kützing, and fractionated SPs were examined to investigate their molecular characteristics and immunomodulatory activity. The crude and fractionated SPs (F1, F2, and F3) consisted mostly of carbohydrates (68.5-85.3%), uronic acids (3.2-4.9%), and sulfates (2.2-12.2%) with various amounts of proteins (2.6-17.1%). D-galactose (23.5-27.3%), D-glucose (11.5-24.8%), L-fucose (19.0-26.7%), and L-rhamnose (16.4-18.3%) were the major monosaccharide units of these SPs with different levels of L-arabinose (3.0-9.4%), D-xylose (4.6-9.8%), and D-mannose (0.4-2.3%). The SPs contained two sub-fractions with molecular weights (Mw) ranging from 164 × 10(3) to 1460 × 10(3) g/mol. The crude and fractionated SPs strongly stimulated murine macrophages, producing considerable amounts of nitric oxide and various cytokines via up-regulation of their mRNA expression by activation of nuclear factor-kappa B and mitogen-activated protein kinases pathways. The main backbone of the most immunoenhancing SP was (1→3)-L-Fucopyranoside, (1→4,6)-D-Glucopyranoside, and (1→4)-D-Galactopyranoside.