Water-soluble Crude Polysaccharide Research Articles

A novel flavonol-polysaccharide from Tamarix chinensis alleviates influenza A virus-induced acute lung injury. Evidences for its mechanism of action

BackgroundTamarix chinensis Lour. is a Chinese medicine used for treating inflammation-related diseases and its crude polysaccharides (MBAP90) exhibited significant anticomplement activities in vitro. PurposeTo obtain anticomplement homogenous polysaccharides from MBAP90 and explore its therapeutic effects and potential mechanism on influenza A virus (IAV)-induced acute lung injury (ALI). MethodsAnticomplement activity-guided fractionation of the water-soluble crude polysaccharides from the leaves and twigs of T. chinensis were performed by diethylaminoethyl-52 (DEAE-52) cellulose and gel permeation columns to yield a homogeneous polysaccharide MBAP-5, which was further characterized using ultra-high-performance liquid chromatography-ion trap tandem mass spectrometry (UPLC-IT-MS) and nuclear magnetic resonance (NMR) analysis. In vitro, the anticomplement activity of MBAP-5 through classical pathway was measured using a hemolytic test. The therapeutic effects of MBAP-5 on ALI were evaluated in H1N1-infected mice. H&E staining, enzyme linked immunosorbent assay (ELISA), immunohistochemistry, and western blot were used to systematically access lung histomorphology, inflammatory cytokines, degree of complement component 3c, 5aR, and 5b-9 (C3c, C5aR, and C5b-9) deposition, and inflammasome signaling pathway protein expressions in lung tissues. ResultsMBAP-5 was a novel flavonol-polysaccharide with the molecular weight (Mw) of 153.6 kDa. Its structure was characterized to process a backbone of →4)-α-D-GlcpA-(1→, →6)-α-D-Glcp-(1→, →3,4)-α-D-Glcp-(1→, →3,4,6)-α-D-Glcp-(1→, and →4,6)-β-D-Glcp-(1→, as well as branches of α-L-Araf-(1→ and β-D-Galp-(1→. Particularly, O-3 of →3,4,6)-α-D-Glcp-(1→ was substituted by quercetin. In vitro assay showed that MBAP-5 had a potent anticomplement activity with a CH50 value of 102 ± 4 µg/ml. Oral administration of MBAP-5 (50 and 100 mg/kg) effectively attenuated the H1N1-induced pulmonary injury in vivo by reducing pulmonary edema, virus replication, and inflammatory responses. Mechanistically, MBAP-5 inhibited the striking deposition and contents of complement activation products (C3c, C5aR, and C5b-9) in the lung. Toll-like receptor 4 (TLR4) /transcription factor nuclear factor κB (NF-κB) signaling pathway was constrained by MBAP-5 treatment. In addition, MBAP-5 could suppress activation of the inflammasome pathways, including Nod‐like receptor pyrin domain 3 (NLRP3), cysteinyl aspartate specific proteinase-1/12 (caspase-1/12), apoptosis‑associated speck‑like protein (ASC), gasdermin D (GSDMD), interleukin (IL)-1β, and IL-18 expressions. ConclusionsA novel flavonol-polysaccharide MBAP-5 isolated from T. chinensis demonstrated a therapeutic effect against ALI induced by IAV attack. The mechanism might be associated with inhibition of complement system and inflammasome pathways activation.

Properties of Yogurts Enriched with Crude Polysaccharides Extracted from Pleurotus ostreatus Cultivated Mushroom

Increasingly, consumers are looking for products with specific nutritional and health-promoting properties. The answer of the producers for this demand is fortified food. The raw material that can be used to enrich food is, among others, mushrooms. Crude water soluble polysaccharides (cWSP) were isolated from fruiting bodies of Pleurotus ostreatus (oyster) mushroom. Chemical analysis showed that they consisted mainly of carbohydrates (~61%), protein (~9%) and phenolics (~0.8%). The isolated cWSP were used to obtain enriched cow milk set yogurts. cWSP were added at the concentration of 0.1%, 0.2%, 0.3%, 0.4% and 0.5%, and milk containing no cWSP was prepared as the control. All of the variants were fermented via applying two commercially available culture starters. The addition of cWSP led to a drop in pH in the case of one starter culture. Also, the decline in total soluble solids (TSS) content was higher where cWSP was used for the enrichment. Texture profile analysis (TPA) revealed that parameters of hardness and gumminess increased along with the concentration of cWSP (reaching values approximately 7–8 times higher, compared to the control). A significant increase in syneresis level (proportional to cWSP concentration and ranging from ~10% to ~50%) was also observed after the fermentation. Fortifying milk with cWSP led to a slight increase in antioxidant capacity in FRAP assay (up to ~12%) and ABTS assay (up to ~23%). The results demonstrate that using cWSP to enrich set-type yogurts is fairly limited.

Optimized extraction of sulfated polysaccharide from brown seaweed Sargassum polycystum and its evaluation for anti-cancer and wound healing potential

• Optimized acidic hot water extraction of WSCSPs. • WSCSPs exhibit antioxidant property. • WSCSPs promote wound healing in RAW 264.7 cells. • WSCSPs is not cytotoxic to normal L6 myoblast cells. • WSCSPs induce death of HeLa cancer cells. In the present study, we optimized the extraction of water-soluble crude sulfated polysaccharide (WSCSP) from Sargassum polycystum and its biological activity was investigated. The WSCSP was extracted from S. polycystum by the acidic water extraction method. The Box-Behnken experimental design was used as a tool for the extraction and evaluated in variable conditions extraction temperature as 60 °C, extraction time duration as 60 min, pH 5.0 and acid hot water: algal ratio 1:50 (g/ml) on these conditions, respectively. The use of acid hot water extraction was efficient to obtain a high WSCSP yield (59.80 ± 2.52%) and further characterization and morphology were analyzed by Fourier-transform infrared spectroscopy (FTIR), Nuclear magnetic resonance spectroscopy (NMR), Thermogravimetry analysis (TGA), X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and the dynamic light scattering (DLS). WSCSP was shown to possess antioxidant activity, as evident by the DPPH assay. Moreover, WSCSP did not induce any significant cytotoxicity in L6 myoblast normal cell line. WSCSP also triggers wound healing in RAW 264.7 cells. WSCSP also induces significant death of HeLa (cervical cancer) cells, marking its cancer specificity. Hence WSCSP derived from Sargassum polycystum can be used as an anticancer candidate in future.

Anticancer and Antioxidant Activity of Water-Soluble Polysaccharides from Ganoderma aff. australe against Human Osteosarcoma Cells.

Wild mushrooms have gained great importance for being a source of biologically active compounds. In this work, we evaluate the anticancer and antioxidant activity of a water-soluble crude polysaccharide extract isolated from the fruiting bodies of the Ganoderma aff. australe (GACP). This mushroom was collected in San Mateo (Boyacá, Colombia) and identified based on macroscopic and microscopic characterization. GACP was characterized by UV-Vis spectroscopy, Fourier-transform infrared spectroscopy, high-performance liquid chromatography-diode array detector, and nuclear magnetic resonance. The antiradical and antioxidant activity were evaluated by different methods and its anticancer activity was verified in the osteosarcoma MG-63 human cell line. Chemical and spectroscopic analysis indicated that GACP consisted of β-D-Glcp-(1→, →3)-β-D-Glcp-(1→ and α-D-Glcp-(1→ residues. The results of the biological activity showed that GACP exhibited high antioxidant activity in the different methods and models studied. Moreover, the results showed that GACP impaired cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay) and cell proliferation (clonogenic assay) in a dose-response manner on MG-63 cells. The findings of this work promote the use of mushroom-derived compounds as anticancer and antioxidant agents for potential use in the pharmaceutical and food industries.

In vitro Assessment of Chemical and Pre-biotic Properties of Carboxymethylated Polysaccharides From Passiflora edulis Peel, Xylan, and Citrus Pectin.

This study aimed to determine the carboxymethylation effect of crude water-soluble polysaccharides of Passiflora edulis peel (WPEP), xylan (XY), and citrus pectin (CP). Their chemical and pre-biotic properties were also determined. The polysaccharides were carboxymethylated by reacting with chloroacetic acid and sodium hydroxide. The carboxymethylated and non-carboxymethylated polysaccharides were also used as pre-biotics to study the growth pattern of selected intestinal microflora. These polysaccharides substituted the glucose solution in culture media for culturing Lactobacillus brevis GIM1.773, Lactobacillus plantarum GIM1.19, Lactobacillus delbrueckii subsp. bulgaricus GIM1.155, and Streptococcus thermophilus GIM1.540. The results showed that the carboxymethylated polysaccharides c-XY, c-CP, and c-WPEP, had substitution degrees of 0.682, 0.437, and 0.439, respectively. The polysaccharides demonstrated resistance to digestion in the simulated human digestive models. The resistance to digestion was enhanced by carboxymethylation, especially the carboxymethylated CP and WPEP. The results also showed that the pre-biotic activities of the polysaccharides increased after carboxymethylation. The c-XY had a better pre-biotic effect than XY and the other carbohydrate samples. The findings suggested that carboxymethylated polysaccharides may be developed into novel pre-biotics and nutraceuticals that could promote growth of the probiotic strains.

Purification, Characterization and Immunomodulatory Activities of Polysaccharides fromMulberryLeaf Fermented withPhellinus igniarius

Phellinus igniariusis a rare and precious medicinal fungus, displaying an outstanding physiological effect, especially the immunomodulatory effects. Previous studies indicated that water-soluble crude polysaccharide (MPFP) was obtained from mulberry leaf fermented withPhellinus igniarius. In vitrocell assay revealed that MPFP showed higher immunomodulatory activity than that of mulberry leaves polysaccharide (MP) andPhellinus igniariusmycelial polysaccharide (PP). Therefore, in this study, structure and immunomodulatory activity of MPFP were measured, a novel polysaccharide named MPFP2-1 was separated through DEAE-52 cellulose column and SephadexG-100 gel-filtration chromatography. Monosaccharide composition analysis showed that MPFP2-1 was mainly composed of L-rhamnose and D-glucose with the molar ratio of 1.0:5.4. The average molecular weight was 50.3 kDa by high performance gel permeation chromatography (HPGPC). FT-IR spectrum showed that MPFP2-1 contained a characteristic absorption peak of polysaccharide. The NMR spectrum indicated MPFP2-1 contained 1 → 6 glucosidic bond.In vitroimmunomodulatory assay revealed that MPFP2-1 significantly enhanced the macrophages proliferation, stimulated the macrophages phagocytic capacity, as well as induced NO and TNF-a generation. We further discovered that MPFP2-1 stimulated iNOS and TNF-αprotein expression in RAW264.7 cells by western blotting. The results are in agreement with ELISA. All the results suggest that MPFP2-1 possesses potent immunomodulatory activity and could be taken forward as new products for medicines.

Structural characterization and osteogenic activity in vitro of novel polysaccharides from the rhizome of Polygonatum sibiricum.

Polygonatum sibiricum Red., a kind of edible and medicinal plant, has a long history of use in traditional Chinese medicine and as a nutritional food additive. Water-soluble crude polysaccharide (PS50) was obtained from P. sibiricum rhizome. After deproteinization and dialysis, PS50 was isolated and purified via DEAE-52 cellulose and Sephadex G-75 gel filtration chromatography to obtain two novel polysaccharides (PSP50-2-1 and PSP50-2-2). Chemical analysis and nuclear magnetic resonance (NMR) studies indicated that PSP50-2-1 was composed of β-d-Glcp-(1→, →2)-β-d-Galp-(1→, →2,6)-β-d-Galp-(1→, β-d-Fruf-(2→, and PSP50-2-2 was composed of β-d-Glcp-(1→, β-d-Galp-(1→, →2)-β-d-Galp-(1→, →6)-β-d-Galp-(1→, →2,6)-β-d-Galp-(1→, β-d-Fruf-(2→. The morphological analysis suggested that PSP50-2-1 and PSP50-2-2 both had a rough surface with cracks. Pharmacological studies showed that PSP50-2-2 at concentrations of 2.59 and 5.19 μM significantly promoted the differentiation and mineralization of MC3T3-E1 cells in vitro, which indicated that PSP50-2-2 had a function of osteogenic activity. Above all, this study provides evidence that PSP50-2-2 may be used as an anti-osteoporotic agent, with applications in health-care and pharmaceutical industries.

Exploration of bioactive prospects of a polysaccharide fraction from Termitomyces heimii against colorectal cancer and broad spectrum bacteria

Termitomyces heimii is a highly cherished wild edible mushroom in India for its pleasant delicacy, rich proximate compositions and profound ethno medicinal applications. A water soluble crude polysaccharide fraction (THP-I) was isolated from dried fruit bodies of T. heimii and purified by GPC. The 1H- NMR spectrum of THP-I exhibited anomeric protons within chemical shifts (δH) between 3.40 to 4.0, indicating that the compound possess –CH and –CH2 functional groups, which resonate with the oxygen atoms of the –OH group, confirmed it as a polysaccharide. THP-I exhibited the MIC values of 62.5 and 125 <mu>g/ml against Staphylococcus aureus and Shigella flexneri respectively. DPPH and FRAP assays showed effective radical scavenging potential by THP-I. In vitro cytotoxicity of THP-I was evaluated by MTT assay and showed notable cellular toxicity against HCT cells at a dose concentration 200 <mu>g/ml. It is first time reported herein that, THP-I has effectively diminished hyperplasia in 1, 2-dimethylhydrazine (DMH) induced colon cancer of Swiss albino rats along with sprouting of goblet cells. Through the present study it is evident that polysaccharide from T. heimii exhibited manifold bioactivities and therefore could be exploited further for its undiscovered potential applications.

Content Analysis, Cytotoxic, and Anti-metastasis Potential of Bioactive Polysaccharides from Green Alga Codium intricatum Okamura

Background: Codium intricatum, locally known as Pukpuklo, is a seasonal and edible green alga found in Ilocos Norte, Philippines. In this study, the biochemical content, cytotoxic and inhibitory potential against Matrix Metalloproteinase-1 (MMP-1) production of the polysaccharide-protein fractions from C. intricatum is first reported. MMPs are novel targets for therapeutic intervention with the potential to inhibit tumor growth, metastasis and invasion either on their own or in conjunction with cytotoxic treatments. Methods: Water-soluble Crude Polysaccharide (CP) and its fractions (CFs) from C. intricatum were isolated using hot water and ion-exchange chromatography and analyzed using different analytical techniques. Cytotoxicity against MCF-7 breast cancer cells and Human normal Dermal Fibroblasts (HnDFs) was determined by MTT assay. The MMP-1 inhibitory potential was tested in UVB exposed Human normal Dermal Fibroblast Cells. Results: CP and CFs afforded carbohydrates (2.07-16.1%), sulfates (1.81%- 9.9%), protein (0.05-2.7%), ash (&lt;77.2%), lipids (&lt;1.0%) and uronic acid (0.15- 4.49%). FT- IR and NMR spectra of CP and CFs exhibited absorption peaks comparable to sulfated galactans. Both CP and CFs significantly inhibited the growth of breast cancer (MCF-7) in a concentration-dependent manner (p&lt;0.05), induced proliferation of HnDFs, inhibited the production of Matrix metalloproteinase-1 (MMP-1) in UV-B induced HnDFs (p&lt;0.05). Conclusion: Results are highly suggestive that polysaccharide fractions from C. intricatum are bioactive molecules with cytotoxic and anti-metastasis potential.

Structural characterization and anticomplement activities of three acidic homogeneous polysaccharides from Artemisia annua.

Artemisia annua L. is a heat-clearing Chinese medicine and well-known for its antimalarial constituent, artemisinin. It has gained increasing attention for its anti-inflammatory and immunoregulatory activities. Interestingly, the crude polysaccahrides of A. annua exhibited potent anticomplement activity. This study was to isolate and characterize its anticomplement homogeneous polysaccharides from A. annua, and reveal the relationship between structures and anticomplement activities of the isolated polysaccharides. Water-soluble crude polysaccharides from the aerial parts of A. annua were extracted and fractionated by DEAE-cellulose and Sephacryl S-300 gel permeation chromatography. Homogeneity, molecular weight, monosaccharide composition, methylation and NMR analysis were performed to characterize the structures of homogeneous polysaccharides. Their anticomplement activities and targeting components in the complement activation cascade were evaluated by hemolytic assays. Three homogeneous polysaccharides (AAP01-1, AAP01-2 and AAP01-3) were obtained from A. annua. AAP01-1 was composed of seven monosaccharides, including mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose. AAP01-2 and AAP01-3 had similar monosaccharides with AAP01-1, except the absence of glucuronic acid. They were all branched acidic heteropolysaccharides with different contents of galacturonic acid (8%, 28% and 15% for AAP01-1, AAP01-2 and AAP01-3, respectively). AAP01-2 showed potent anticomplement activity with CH50 value of 0.360 ± 0.020 mg/mL through the classical pathway and AP50 value of 0.547 ± 0.033 mg/mL through the alternative pathway. AAP01-3 exhibited slightly weaker activity (CH50: 1.120 ± 0.052 mg/mL, AP50: 1.283 ± 0.061 mg/mL), while AAP01-1 was inactive. Moreover, AAP01-2 acted on C1q, C3, C4, C5 and C9 components and AAP01-3 interacted with C3, C4 and C5 components in the activation cascade of complement system. These results indicated that the relatively high contents of galacturonic acid were important for anticomplement activities of the polysaccharides from A. annua. The anticomplement polysaccharides are another kind of bioactive constituents conferring heat-clearing effects of A. annua.

Antitumor and Immunomodulatory Activity of Water-Soluble Crude Polysaccharide from Solid Fermentation of Phellinus igniarius and Mulberry Leaves in H22 Tumor-Bearing Mice

Antitumor and Immunomodulatory Activity of Water-Soluble Crude Polysaccharide from Solid Fermentation of<i> Phellinus igniarius</i> and <i>Mulberry</i> Leaves in H22 Tumor-Bearing Mice

Interactions between ascorbic acid and water soluble polysaccharide from the seeds of Plantago asiatica L.: Effects on polysaccharide physicochemical properties and stability

Viscous polysaccharide is one kind of main bioactive components in Plantago asiatica L. seeds. This study aims to investigate effects of various monosaccharides, disaccharides, especially ascorbic acid (VC), on apparent viscosity and physicochemical properties of crude water soluble polysaccharide from the seeds of P. asiatica L. (PLWEP-C), as well as its stability and degradation mechanism. Results showed that small molecules such as glucose, sucrose, maltose, and lactose had little effects on apparent viscosity of PLWEP-C, while VC could significantly reduce the viscosity. There was no obvious change in FT-IR spectra and monosaccharide composition of PLWEP-C after VC addition. Hydroxyl radicals were probably involved in the degradation of polysaccharide induced by VC, which was confirmed by glucose inhibition test. SEM observation showed that PLWEP-C was mainly in lamellar and filamentous appearance, and no great change was generated by VC. The solution stability of PLWEP-C can only be kept well, when VC was at low concentration and low temperature. In conclusion, VC could significantly reduce the viscosity of polysaccharide from the seeds of P. asiatica L., which was probably attributed to the reduction of molecular weight. Hydroxyl radicals participation were probably one of main reasons in the degradation process.

Isolation and Characterization of Water Soluble Polysaccharides from Isaria cicadae

Isaria cicadae Miquel is an important medicinal food fungus. The crude polysaccharides from I. cicadae were shown to significantly extend the lifespan of fruit flies in our previous study. To understand the underlying mechanism, the composition and structural characterization of polysaccharides from I. cicadae were determined. A water-soluble crude polysaccharide, obtained from a cultured Isaria cicadae by boiling-water extraction and ethanol precipitation, was fractionated by DEAE-Sepharose column chromatography. The isolate gave four polysaccharide fractions termed B-I-1, B-I-2, B-II-1 and B-II-2. The average ratio of Glu: Man: Gla in B-I-2 was 1.28:1.25:1, B-II-1 was 16.08:1:1.95, and B-II-2, was 1:2.43:2.61. B-I-1 was composed of Ara, Xyl and Glu in the ratio of 1:2.36:33.14. GC and FTIR spectra analysis combined with NMR spectroscopic analysis gave preliminary structures of the four polysaccharide fractions, which may provide an elucidation on the action mechanisms of - polysaccharide on fruit flies proliferation.

Polysaccharides production from soybean curd residue via Morchella esculenta.

To minimize the serious environmental pollution caused by food byproducts, soybean curd residue was used as the substrate to ferment polysaccharides by Morchella esculenta. Water-soluble crude polysaccharides (CMP) were purified by different gradient concentrations (30%, 60%, and 90%) of ethanol into three precipitation fractions. The experimental results indicated that the precipitated polysaccharides by the ethanol concentration of 60% (PMP) had the highest purity, containing rhamnose, arabinose, xylose, mannose, glucose, and galactose, with the molar ratio of 0.08:0.25:0.16:0.07:0.28:1.00. The physicochemical properties were revealed by SEM, AFM, FTIR, NMR, and Congo red test. Furthermore, the measurement of antioxidant activities in vitro demonstrated that PMP exhibited higher antioxidant capacities with the dose-dependent manner than CMP and polysaccharides from unfermented soybean curd (USP). Overall, the results suggested that PMP had attractive functional activity which could be potentially utilized in functional food industries. PRACTICAL APPLICATIONS: The polysaccharides from fungi sources are increasingly being used in functional food and pharmaceutical industry. In allusion to the problem of polysaccharides fermented by edible fungi, such as low yield, high cost, and hard to controllable. Soybean curd residue (SCR), a food waste, has been successfully used for the production of polysaccharides and graded ethanol was used to enhance polysaccharides production, lower the difficulty and homologous costs of purity. The results indicated that the purified polysaccharides displayed excellent antioxidant activities on scavenging DPPH and hydroxyl radicals. The outcomes of this work can provide technical support for comprehensive utilization of SCR and references for promotion and application of polysaccharides fermented from SCR via Morchella esculenta in food and medicine fields.

Isolation, purification, and characterization of novel polysaccharides from lotus (Nelumbo nucifera) leaves and their immunostimulatory effects.

In this study, a crude water-soluble polysaccharide (LLWP-C) was extracted from lotus leaves and further purified by size exclusion chromatography, to obtain the two main polysaccharides, LLWP-1 and LLWP-3. Physical and chemical analyses showed that they were homogeneous polysaccharides in β-type glycosidic linkage. LLWP-1 was devoid of helical conformation, had a molecular weight of 85.1 kDa and was mainly composed of Rha, Ara, Gal, Glu, and GalA in a molar ratio of 7.0:24.8:28.0:6.0:26.4. LLWP-3 showed a helical conformation, had a molecular weight of 12.5 kDa and consisted mainly of Rha, Ara, Gal, Glu, Man, and GalA in a molar ratio of 6.6:9.8:15.0:8.9:6.1:47.2. It was demonstrated that LLWP-C and both purified LLWP-1 and LLWP-3 could effectively enhance the proliferation, phagocytosis, nitric oxide (NO), and cytokine secretions by activating corresponding mRNA expression in macrophages, via MAPK and NF-κB pathways. LLWP-3 displayed the greatest immunostimulatory potential, followed by LLWP-1 and LLWP-C. These findings suggest that polysaccharides extracted from lotus leaf exert immunostimulatory activity that could be further investigated to develop functional foods and natural immunopotentiating therapeutic agents.

Molecular structures, chemical properties and biological activities of polysaccharide from Smilax glabra rhizome

The water-soluble crude polysaccharides, extracted from the rhizome of Smilax glabra, were fractionated using an anion exchange chromatography, yielding two fractions, F1 and F2. The crude and fractions (F1 and F2) mainly consisted of carbohydrates (66.7%–91.1%), proteins (7.30%–23.9%) and minor amount of sulfates (1.60%–9.40%). Glucose was the major monosaccharide unit of the polysaccharides with different levels of sugar constituents including galactose, arabinose, rhamnose and mannose. The molecular weight (Mw) of crude and fractions ranged from 32,102–6.3 × 103 g/mol. The crude and fractions could stimulate RAW264.7 cells to release nitric oxide and cytokines via up-regulation of their mRNA expression by the activation of NF-κB and MAPKs pathways. The related pattern recognition receptors (PRRs) on the surface of the cells appeared to be TLR2 and CR3. The GC–MS analysis revealed that the main backbone of the most immune-enhancing F2 was (1 → 4)-linked glucose and galactose chain with minor linkages of (1 → 6)-galactose, (1 → 3)-mannose, (1 → 2)-rhamnose and (1 → 5)-arabinose with some branches at C-3 and C-4 rhamnose, or C-6 galactose.

Water Soluble Antioxidative Crude Polysaccharide From Russula senecis Elicits TLR Modulated NF-κB Signaling Pathway and Pro-inflammatory Response in Murine Macrophages.

Russula senecis has recently been reported as a new addition to macrofungal flora of West Bengal. Besides, it also emerged as a seasonal health promoting nutrient to local ethnic people and enlisted for the first time as tribal food in our previous publication. In this context, the present work was designed to establish such usefulness scientifically and to meet the aim, crude polysaccharide, Rusenan, was prepared using conventional heated water reflux. Initially, the polymers were characterized to determine chemical composition and for that spectrophotometry along with Fourier-transform infrared spectroscopy (FT-IR), high-performance thin-layer chromatography (HPTLC), and gas chromatography-mass spectrometry (GC-MS) were performed. Analysis indicated that Rusenan was consisted mainly of carbohydrate conjugated with trace amount of protein. Furthermore, glucose was detected as the major monosaccharide (mainly in β-type glycosidic linkage) while other monomers were presented in the order of galactose > mannose > xylose > rhamnose. Conversely, antioxidant potential was determined following eight in vitro systems where the fraction evidenced strong superoxide, hydroxyl, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity, high affinity to Fe2+ as well as instant ability to donate electron with EC50 values ranging from 80 to 3885 μg/ml concentration. In addition, effect on murine macrophages was also investigated where the polysaccharide treatment increased cell proliferation, phagocytic activity, filopodia or lamellipodia formation, nitric oxide (NO) production and reactive oxygen species (ROS) synthesis. Thereafter, through reverse transcriptase polymerase chain reaction (RT-PCR) analysis, significant increase in the expression of Toll like receptor (TLR)-4, TLR-2 and nuclear factor kappa B (NF-κB) was observed; as a result alleviated level of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, IκB-α, and interferon (IFN)-γ were also noticed explaining definite immune-stimulatory activity of the fraction. Thus, overall finding suggests that R. senecis can be considered as a functional food and may be used in preparation of dietary supplement to enhance general health.

Isolation of polysaccharides from Malva aegyptiaca and evaluation of their antioxidant and antibacterial properties

This study investigated the antioxidant and antibacterial properties of crude water-soluble polysaccharides extracted from the mallow (Malva aegyptiaca) by precipitation with cetylpyridinium chloride (P1) or ethanol (P2). The Polysaccharides fractions were characterized by FTIR spectroscopy, a monosaccharide composition, and antioxidant and antibacterial activities. P1 showed the highest total sugars (81.2%) and sulfated groups (2.9%) contents but with the lowest total proteins content (8.7%). The infrared spectroscopic spectra exhibited the typical bands and peak characteristic of polysaccharides. Monosaccharides analysis revealed the dominance of galactose (40.3–43.7%) and glucuronic acid (25.9–30.9%). Interestingly, P1 displayed an important antioxidant activity as evaluated by the (Fe2+) chelating activity (IC50=1.15mg/ml), (Fe3+) reducing power (EC50=1.22mg/ml), β-carotene bleaching inhibition capacity (IC50=1.56mg/ml) and DPPH-radical scavenging activity (IC50=1.94mg/ml). Furthermore, P1 at 10mg/ml was highly active than P2 against several bacterial strains and especially the Gram-positive bacteria, where the inhibition rate ranged between 84.2 and 90.3%. Obtained results suggest that M. aegyptiaca polysaccharides precipitated by cetylpyridinium chloride could be used as a naural antioxidant and antibacterial agent.

Introducing a novel mushroom from mycophagy community with emphasis on biomedical potency.

Mushrooms have been prized by humankind as medicine and culinary wonder since antiquity. Though several species are ethnically valued; many prospective species are still being discovered. One such wild macrofungus has recently been discovered during subsequent field surveys in West Bengal, India which in turn exposed as a traditionally consumed popular myco-food. The collected taxon was found to be unique with regard to its morphological as well as genetical features. After detailed characterizations, the fungus was identified as a novel taxon belonging to the genus Russula (Russulaceae, Basidiomycota). Besides, the investigation was further extended in search of new functional ingredients and in this context, a water soluble crude polysaccharide rich extract (Rusalan) was isolated from dried basidiocarps. Accumulating evidences from GC-MS, HPTLC, FT-IR along with several spectrophotometric methods postulated that the fraction consisted mainly of carbohydrate in triple helical conformation, where glucose was the major monosaccharide mostly with β-type glycosidic linkage. Conversely, Rusalan showed pronounced antioxidant activity in six in vitro assay systems with EC50 value ranging from 190–1328 μg/ml concentration. The crude polysaccharide was also evaluated against six bacterial strains using microdilution method and the growth of Staphylococcus aureus and Bacillus subtilis were found to be inhibited effectively. In addition, immune-stimulatory assays demonstrated that Rusalan could evidently promote proliferation, induce phagocytosis, release NO, produce intracellular ROS and upregulate mRNA expression of iNOS, TNF-α, COX-2, as well as IL-6 genes in in mouse macrophage cells. Therefore, aim of the present study was not only to describe a new taxon to the world mycoflora but also to introduce a potent therapeutic agent that could be explored for food and pharmaceutical purposes. However, isolation of active component and in vivo studies need to be designed further.

FTIR characterization and antioxidant activity of water soluble crude polysaccharides of Sri Lankan marine algae

FTIR characterization and antioxidant activity of water soluble crude polysaccharides of Sri Lankan marine algae