Microarray Analysis of Paramylon, Isolated from Euglena Gracilis EOD-1, and Its Effects on Lipid Metabolism in the Ileum and Liver in Diet-Induced Obese Mice.

We previously showed that supplementation of a high fat diet with paramylon (PM) reduces the postprandial glucose rise, serum total and LDL cholesterol levels, and abdominal fat accumulation in mice. The purpose of this study was to explore the underlying mechanism of PM using microarray analysis. Male mice (C57BL/BL strain) were fed an experimental diet (50% fat energy) containing 5% PM isolated from Euglena gracilis EOD-1 for 12 weeks. After confirming that PM had an improving effect on lipid metabolism, we assessed ileal and hepatic mRNA expression using DNA microarray and subsequent analysis by gene ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The results suggested that dietary supplementation with PM resulted in decreased abdominal fat accumulation and serum LDL cholesterol concentrations via suppression of the digestion and absorption pathway in the ileum and activation of the hepatic PPAR signaling pathway. Postprandial glucose rise was reduced in mice fed PM, whereas changes in the glucose metabolism pathway were not detected in GO classification and KEGG pathway analysis. PM intake might enhance serum secretory immunoglobulin A concentrations via promotion of the immunoglobulin production pathway in the ileum.

Effect of Supplementing Seaweed Extracts to Pigs until d35 Post-Weaning on Performance and Aspects of Intestinal Health.

The objective of this study was to examine the effects of feeding laminarin (LAM) and fucoidan (FUC) enriched seaweed extracts up to d35 post-weaning on measures of animal performance, intestinal microbial and transcriptome profiles. 75 pigs were assigned to one of three groups: (1) basal diet; (2) basal diet + 250 ppm fucoidan; (3) basal diet + 300 ppm laminarin with 7 replicates per treatment group. Measures of performance were collected weekly and animals sacrificed on d35 post-weaning for the sampling of gastrointestinal tissue and digesta. Animal performance was similar between the basal group and the groups supplemented with FUC and LAM (P > 0.05). Pigs fed the basal diet had higher alpha diversity compared to both the LAM and FUC supplemented pigs (P < 0.05). Supplementation with LAM and FUC increased the production of butyric acid compared to basal fed pigs (P < 0.05). At genus level pigs fed the LAM supplemented diet had the greatest abundance of Faecalbacterium, Roseburia and the lowest Campylobacter of the three experimental treatments (P< 0.05). While neither extract had beneficial effects on animal performance, LAM supplementation had a positive influence on intestinal health through alterations in the gastrointestinal microbiome and increased butyrate production.

Using Flammulina velutipes derived chitin-glucan nanofibrils to stabilize palm oil emulsion:A novel food grade Pickering emulsifier.

We herein report chitin-glucan nanofibrils from edible mushroom Flammulina velutipes (CGNFs) as a novel stabilizer for palm oil Pickering emulsion (o/w, 30:70, v:v). Generally, these CGNFs being composed of glucose and glucosamine, are threadlike with 4.9 ± 1.2 nm wide and 222.6 ± 91.9 nm long. They were easily absorbed on the oil-water interface to form a compact layer around the oil droplets referring to Pickering emulsion. This emulsion presented shear-thinning and gel-like behaviors, wherein CGNFs concentration had a profound influence on the emulsion volume, droplet size, and stabilization index. Moreover, CGNFs showed an ability to stabilize the emulsion with a minimum of surface coverage approximately 30%. It indicated that moderate concentration of NaCl improved the emulsification effect, and the emulsion were stable in a large range of pH. These CGNFs are easy to prepare, eco-friendly and sustainable, which provides a potential for large-scale application of Pickering emulsion in food and nutraceuticals fields.

Structures and anti-atherosclerotic effects of 1,6-α-glucans from Fructus Corni.

In this study, two homogeneous polysaccharides (PFC-1 and PFC-2) having anti-atherosclerotic activity were isolated from Fructus Corni. PFC-1 and PFC-2 were 1,6-α-glucans with the molecular weight of 4.4 kDa and 82.0 kDa, respectively. In the in vitro experiments, PFC-1 and PFC-2 showed significant inhibitory effects on the cholesterol accumulation in RAW264.7 macrophages induced by oxidized low-density lipoproteins (ox-LDL), and the inhibitory rate of PFC-2 was 81.62%. Apolipoprotein E-deficient (ApoE-/-) mice fed high-fat diet (HFD) were used to evaluate the anti-atherosclerotic effects of PFC-2 in vivo. The aortic root lipid area decreased by 55.01% in the PFC-2-administered group as compared to the model group. PFC-2 decreased the levels of serum low-density lipoprotein cholesterol, total cholesterol, triglycerides, and malondialdehyde, increased the superoxide dismutase activity, and reduced the contents of lipid and macrophages in the aortic sinus plaque in ApoE-/- mice fed with HFD. Furthermore, PFC-2 markedly inhibited the expression of type A1 scavenger receptor (SR-A1) and cluster of differentiation 36 (CD36) in ox-LDL-treated macrophages. Taken together, 1,6-α-glucans from Fructus Corni showed significant anti-atherogenic effect, and the mechanism is related to enhanced antioxidant activity of the ApoE-/- mice and down-regulated the expression of SR-A1 and CD36 proteins in macrophages.

Isolation, purification, structure characterization of a novel glucan from Huangshui, a byproduct of Chinese Baijiu, and its immunomodulatory activity in LPS-stimulated THP-1 cells.

Huangshui (HS) has attracted considerable attention for the utilization of aroma compounds and microorganisms containing presently. However, little work has been done on biological polysaccharides in HS. In this study, a novel water-eluted HS polysaccharide, HSP-W, was identified as an α-D-glucan with a Mw of 166.00 kDa. It was consisted of a 1,4 linked α-D-Glcp backbone with the substitution at O-6 with 1,6-linked α-D-Glcp residue and non-reducing terminal of ß-Glc-1→ through monosaccharide composition, IR, methylation, and NMR analyses. SEM, AFM, and particle size distribution measurements showed that HSP-W had relatively clustered spherical shape with different sizes from nanoscale to micrometer. Additionally, HSP-W significantly induced NO and ROS production as well as the release of IL-1ß, IFN-γ, TNF-α, and IL-6, and upregulated pinocytic and phagocytic capacities of THP-1 cells. Meanwhile, HSP-W treatment markedly enhanced mRNA and protein expressions of these cytokines. HS probably has potential application as an immunostimulatory agent.

Chitosan-glucan complex hollow fibers reinforced collagen wound dressing embedded with aloe vera. II. Multifunctional properties to promote cutaneous wound healing.

This study presents an innovative multifunctional system in fabricating new functional wound dressing (FWD) products that could be used for skin regeneration, especially in cases of infected chronic wounds and ulcers. The innovation is based on the extraction, characterization, and application of collagen (CO)/chitosan-glucan complex hollow fibers (CSGC)/aloe vera (AV) as a novel FWS. For the first time, specific hollow fibers were extracted with controlled inner (500-900 nm)/outer (2-3 µm) diameters from mycelium of Schizophyllum commune. Further on, research and evaluation of morphology, hydrolytic stability, and swelling characteristics of CO/CSGC@AV were carried out. The obtained FWS showed high hydrolytic stability with enhanced swelling characteristics compared to native collagen. The hemostatic effect of FWS increased significantly in the presence of CSGC, compared to native CO and displayed excellent biocompatibility which was tested by using normal human dermal fibroblast (NHDF). The FWS showed high antibacterial activity against different types of bacteria (positive/negative grams). From in vivo measurements, the novel FWS increased the percentage of wound closure after one week of treatment. All these results imply that the new CO/CSGC@AV-FWD has the potential for clinical skin regeneration and applying for controlled drug release.

Prosopis juliflora as a new cosmetic ingredient: Development and clinical evaluation of a bioactive moisturizing and anti-aging innovative solid core.

Prosopis juliflora is an invasive plant distributed throughout the world and presents metabolites of interest for cosmetology. The aim of this work was to develop a new polysaccharide-based ingredient from P. juliflora and analyze its application in a solid core formulation that upon contact with water instantly forms a gel to improve moisturizing and anti-aging skin properties. Purified extracts by gel chromatography were characterized by NMR and LC-DAD-MS-MS. The in vitro and in vivo safety, antioxidant activity, formulation development and clinical evaluation were performed. The extract was characterized as containing an α-glucan and phenolics. It was non-cytotoxic, non-phototoxic and no skin reactions were observed in vivo. Antioxidant activity were present through different mechanisms. Clinical evaluation reinforced the potential of P. juliflora in skin hydration and microrelief improvement. This innovative form proved to be a prototype of a new product and the first study of an α-glucan as a cosmetic ingredient.

Chitosan-glucan complex hollow fibers reinforced collagen wound dressing embedded with aloe vera. Part I: Preparation and characterization.

Collagen (CO)/chitosan-glucan complex (CSGC) hollow fibers encapsulated aloe vera (AV) dressing scaffold (CO/CSGC@AV) were fabricated for the first time by the freeze-dried process. Extraction process, morphology, mechanical properties, pore size, porosity, swelling ability, and degradation behavior of composites scaffold were investigated. CSGC hollow fibers were extracted from mycelium of Schizophyllum commune CSGC hollow fiber exhibited inner diameter of (600 ±â€¯250 nm) and outer fiber diameter of (2.5 ±â€¯0.5 µm). The results of swelling and hydrolytic degradation studies demonstrated that the physicochemical of CO/CSGC@AV was significantly enhanced by CSGC in a concentration-dependent manner. The mechanical property of the CO/CSGC@AV was improved after encapsulated AV into CSGC hollow fibers compared with native CO. The pore size and porosity of the CO/CSGC@AV were slightly decreased in the presence of AV. All these results suggested that the new dressing scaffold has a potential for clinical skin regeneration, particularly for infected chronic wounds and ulcers.

Immunomodulating Effects Exerted by Glucans Extracted from the King Oyster Culinary-Medicinal Mushroom Pleurotus eryngii (Agaricomycetes) Grown in Substrates Containing Various Concentrations of Olive Mill Waste.

We have recently demonstrated that we could enhance glucan content in Pleurotus eryngii following cultivation of the mushrooms on a substrate containing different concentrations of olive mill solid waste (OMSW). These changes are directly related to the content of OMSW in the growing substrate. Using dextran sulfate sodium (DSS)-in-flammatory bowel disease (IBD) mice model, we measured the colonic inflammatory response to the different glucan preparations. We found that the histology damaging score (HDS) resulting from DSS treatment reach a value of 11.8 ± 2.3 were efficiently downregulated by treatment with the fungal extracted glucans. Glucans extracted from stalks cultivated at 20% OMSW downregulated to a HDS value of 6.4 ± 0.5 whereas those cultivated at 80% OMSW showed the strongest effects (5.5 ± 0.6). Similar downregulatory effects were obtained for expression of various intestinal cytokines. All tested glucans were equally effective in regulating the number of CD14/CD16 monocytes from 18.2 ± 2.7% for DSS to 6.4 ± 2.0 for DSS + glucans extracted from stalks cultivated at 50% OMSW. We tested the effect of glucans on lipopolysaccharide-induced production of TNF-α, which demonstrated that stalk-derived glucans were more effective than caps-derived glucans. Isolated glucans competed with anti-Dectin-1 and anti-CR3 antibodies, indicating that they contain ß-glucans recognized by these receptors. In conclusion, the most effective glucans in ameliorating IBD-associated symptoms induced by DSS treatment in mice were glucan extracts prepared from the stalk of P. eryngii grown at higher concentrations of OMSW. We conclude that these stress-induced growing conditions may be helpful in selecting more effective glucans derived from edible mushrooms.

Peyer's patch-immunomodulating glucans from sugar cane enhance protective immunity through stimulation of the hemopoietic system.

The aim of the present study was chemical clarification of in vitro Peyer's patch-immunomodulating polysaccharides in sugar cane molasses, and evaluation of in vivo modulating activity on immune function of T lymphocytes in Peyer's patches and on microenvironment of hemopoietic system. Five kinds of glucans, comprising of dextranase-sensitive and activity-related d-glucosyl moieties, were purified as in vitro Peyer's patch-immunomodulating polysaccharides from the molasses. Oral administration of a glucan-enriched subfraction induced IL-2 and GM-CSF-producing T lymphocytes in Peyer's patches, resulting in enhancement of IL-6 production in a hemopoietic microenvironment to boost neutrophil numbers in the peripheral blood stream. Oral administration of purified glucan or glucan-enrich sub-fraction of sugar cane reduced the number of Plasmodium berghei- or P. yoelii-infected erythrocytes in a murine infection model, using polysaccharide alone or via co-administration with the antimalarial drug, artesunate. These results suggested that Peyer's patch-immunomodulating glucans enhanced protective immunity through axis of Peyer's patches-hemopoietic system.

Isolation, purification, characterization and bioactivities of a glucan from the root of Pueraria lobata.

The root of Pueraria lobata is considered to be a medicinal and edible herb for the treatment of diabetes, and it has a long history of application in China. To explore the constituents responsible for the anti-hyperglycemic activities of P. lobata, a water-soluble polysaccharide (PL70-1-1) was isolated and purified by using a DEAE-Cellulose 52 anion exchange column and a Sephacryl S-100 gel filtration column. Its molecular weight (2584 Da) was determined by high performance gel permeation chromatography (HPGPC). Its structure was deduced by Fourier transform-infrared spectroscopy (FT-IR), monosaccharide composition analysis, gas chromatography coupled with mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR). It was deduced that PL70-1-1 was a glucan, and its main chain consisted of (1→)-linked ß-d-glucose, (1→4)-linked α-d-glucose, (1→4, 6)-linked ß-d-glucose, and (1→3)-linked α-d-glucose, and the branch chain consisted of (1→)-linked ß-d-glucose. The results of scanning electron microscopy showed that PL70-1-1 had a needle-like shape, and the surface had a scaly texture. The Congo red experiment showed that PL70-1-1 did not have a triple-helix structure. In addition, PL70 and PL70-1 displayed selective inhibitory effects on α-amylase and α-glucosidase in vitro. PL70 had remarkable α-glucosidase inhibitory activity. However, PL70-1-1 exhibited outstanding α-amylase inhibitory activity, with an IC50 of 3.945 µM in vitro. This indicated that its activity was 417 times higher than the positive control acarbose. PL70-1-1 may be beneficial as an α-amylase inhibitor, reducing the postprandial blood glucose level and treating type 2 diabetes.

Extraction, selenylation modification and antitumor activity of the glucan from Castanea mollissima Blume.

The present study aimed to characterize the glucan from C. mollissima Blume fruits and its selenium derivative, then investigate their antitumor activity in vitro. A glucan, designated as CPA, was firstly isolated from the fruits of C. mollissima Blume. Structure analysis indicated that CPA was a linear 1,6-α-D-glucan with the average molecular weight about 2.0 × 103 kDa. The selenylation modification derivative of CPA (sCPA), exhibited a stronger antiproliferative effect on tumor cells than CPA in vitro. CPA and sCPA could induce HeLa cells apoptosis and decrease mitochondrial membrane potential. sCPA could also arrest HeLa cells in S phase, promote reactive oxygen species generation and activate caspase-3 activity in HeLa cells. These results manifest that CPA and sCPA inhibit the proliferation of HeLa cells via different mechanisms, which is meaningful for their potential use as antitumor drugs.

Immune Enhancing Activity of ß-(1,3)-Glucan Isolated from Genus Agrobacterium in Bone-Marrow Derived Macrophages and Mice Splenocytes.

An effective method for activating macrophages and deriving a Th1 immune response could be used to improve the defenses of hosts. In this study, we investigated the immunomodulation effect and the related signaling mechanism of [Formula: see text]-(1,3)-glucan, isolated from the Agrobacterium species. Here, we found that [Formula: see text]-(1,3)-glucan predominantly induced the tumor necrosis factor (TNF)-[Formula: see text], interleukin (IL)-1[Formula: see text], IL-6, IL-12p70, and nitric oxide, which was dependent on mitogen-activated protein kinases (MAPK) and nuclear factor (NF)-[Formula: see text]B signaling. Additionally, [Formula: see text]-(1,3)-glucan treatment significantly up-regulated the expression of the co-stimulatory molecules CD80 and CD86, and also significantly increased the expression of iNOS and Dectin-1, which is a transmembrane protein that binds [Formula: see text]-glucan and associates with macrophage activation. Importantly, the splenic T cells co-cultured with [Formula: see text]-(1,3)-glucan-treated macrophages produced the a Th1 cytokine profile that includes high levels of IFN-[Formula: see text], but not IL-4 (Th2 cytokine), indicating that [Formula: see text]-(1,3)-glucan contributes to Th1 polarization of the immune response. Taken together, our results suggest that [Formula: see text]-(1,3)-glucan isolated from Agrobacterium species can induce macrophage activation through the MAPK and NF-[Formula: see text]B signaling pathway, as well as Th1 polarization.

Production of pullulan from raw potato starch hydrolysates by a new strain of Auerobasidium pullulans.

In the present study, hydrolysis of potato starch with marine cold-adapted α-amylase and pullulan production from the hydrolysates by a new strain of Auerobasidium pullulans isolated from sea mud were conducted. The hydrolysis conditions were optimized as follows: reaction time 2h, pH 6.5, temperature 20°C, and α-amylase amount 12 U/g. Under these optimum hydrolysis conditions, the DE value of the potato starch hydrolysates reached to 49.56. The potato starch hydrolysates consist of glucose, maltose, isomaltose, maltotriose, and trace of other maltooligosaccharides with degree of polymerization ranged 4-7. The maximum production of pullulan at 96 h from the hydrolysate of potato starch was 36.17 g/L, which was higher than those obtained from glucose (22.07 g/L, p<0.05) and sucrose (31.42 g/L, p<0.05). Analysis of the high performance liquid chromatography of the hydrolysates of the pullulan product with pullulanase indicated that the main composition is maltotriose, thus confirming the pullulan structure of this pullulan product.

Extraction and separation of polysaccharides from Laminaria japonica by size-exclusion chromatography.

A large number of studies have suggested that polysaccharides, such as fucoidan and laminarin, in various seaweeds have significant biological properties. A different distribution of molecular weights is a prominent sign of many polysaccharides. Therefore, a simple, fast and reliable high-performance size-exclusion chromatography (HPSEC) method was proposed to separate fucoidan and laminarin from Laminaria japonica. After evaluating the different separation conditions for HPSEC, such as the type of mobile phase and flow rate, an acid extraction method was established and optimized by a systematic investigation of the influencing factors. Under the optimal conditions, 169.2 and 383.8 mg g(-1) of fucoidan and laminarin, respectively, were extracted. This method is suitable for the extraction and separation of polysaccharides with good reproducibility of the retention time, acceptable linearity, small relative standard deviation and low detection limits.

Nanometric organisation in blends of gellan/xyloglucan hydrogels.

Mixtures of gellan gum (GL) and a xyloglucan (XGJ) extracted from Hymenaea courbaril seeds were prepared in a solution of 0.15 mol L(-1) NaCl. Rheology measurements revealed that 2.4 g L(-1) pure GL formed a brittle hydrogel, and GL-XGJ blends showed improved pseudoplastic character with higher XGJ contents. SAXS analyses showed that the Rg dimensions ranged from 1.3 to 4.9 nm, with larger values occurring as the amount of XGJ increased, and diffusion tests indicated that better diffusion of methylene blue dye was obtained in the network with a higher XGJ content. AFM topographic images of the films deposited onto mica revealed fewer heterogeneous surfaces with increased XGJ contents. The water contact angle revealed more hydrophobic character on all of the films, and the wettability decreased with increasing amounts of XGJ. Therefore, the demonstrated benefit of using XGJ blends is the production of a soft material with improved interface properties.

A glucan isolated from flowers of Lonicera japonica Thunb. inhibits aggregation and neurotoxicity of Aß42.

Inhibition of Aß aggregation and attenuation of its cytotoxicity are considered to valuable therapeutics for Alzheimer's disease (AD). Here, a glucan named as LJW0F2 was purified from flowers of Lonicera japonica Thunb. Using monosaccharides composition analysis, methylation analysis, IR and NMR spectroscopy, this polysaccharide was elucidated to be an α-D-(1→4)-glucan with an α-(1→4) linked branch attached to the C-6 position. Its inhibitory effect on Aß42 aggregation was measured by fluorescence spectroscopic analysis with thioflavine T (ThT) and atomic force microscopy (AFM). We showed that polysaccharide LJW0F2 could inhibit Aß42 aggregation in a dose-dependent-manner. Besides, LJW0F2 could attenuate the cytotoxicity induced by Aß42 aggregation in SH-SY5Y neuroblastoma cells. To the best of our knowledge, this was the first report that the exogenous plant-derived polysaccharide might block Aß42 aggregation directly and reduce its toxicity in SH-SY5Y cells.

Profiling the main cell wall polysaccharides of grapevine leaves using high-throughput and fractionation methods.

Vitis species include Vitis vinifera, the domesticated grapevine, used for wine and grape agricultural production and considered the world's most important fruit crop. A cell wall preparation, isolated from fully expanded photosynthetically active leaves, was fractionated via chemical and enzymatic reagents; and the various extracts obtained were assayed using high-throughput cell wall profiling tools according to a previously optimized and validated workflow. The bulk of the homogalacturonan-rich pectin present was efficiently extracted using CDTA treatment, whereas over half of the grapevine leaf cell wall consisted of vascular veins, comprised of xylans and cellulose. The main hemicellulose component was found to be xyloglucan and an enzymatic oligosaccharide fingerprinting approach was used to analyze the grapevine leaf xyloglucan fraction. When Paenibacillus sp. xyloglucanase was applied the main subunits released were XXFG and XLFG; whereas the less-specific Trichoderma reesei EGII was also able to release the XXXG motif as well as other oligomers likely of mannan and xylan origin. This latter enzyme would thus be useful to screen for xyloglucan, xylan and mannan-linked cell wall alterations in laboratory and field grapevine populations. This methodology is well-suited for high-throughput cell wall profiling of grapevine mutant and transgenic plants for investigating the range of biological processes, specifically plant disease studies and plant-pathogen interactions, where the cell wall plays a crucial role.

Isolation and characterization of a glucan-type polysaccharide from the red pine mushroom, Lactarius deliciosus (Higher Basidiomycetes).

A novel glucan-type polysaccharide has been isolated from the fruiting bodies of the edible mushroom Lactarius deliciosus. Two successive extractions (cold aqueous extraction at 25°C and hot aqueous extraction at 100°C) were performed, and the same polysaccharide was obtained in both fractions. The purity of the polysaccharide was evaluated by size exclusion chromatography. The SEC chromatogram showed a unique peak with a molecular weight of approximately 150 kDa. Analysis of the chain composition revealed that the polysaccharide was composed of glucose. Methylation analysis and NMR experiments showed that the glucan-type carbohydrate contained a main chain consisting of α-(1→3)-Glcp units with α-(1→4)-linked branches every sixth glucose residue.

Characterisation of cell wall polysaccharides from rapeseed (Brassica napus) meal.

To enable structural characteristics of individual cell wall polysaccharides from rapeseed (Brassica napus) meal (RSM) to be studied, polysaccharide fractions were sequentially extracted. Fractions were analysed for their carbohydrate (linkage) composition and polysaccharide structures were also studied by enzymatic fingerprinting. The RSM fractions analysed contained pectic polysaccharides: homogalacturonan in which 60% of the galacturonic acid residues are methyl-esterified, arabinan branched at the O-2 position and arabinogalactan mainly type II. This differs from characteristics previously reported for Brassica campestris meal, another rapeseed cultivar. Also, in the alkali extracts hemicelluloses were analysed as xyloglucan both of the XXGG- and XXXG-type decorated with galactosyl, fucosyl and arabinosyl residues, and as xylan with O-methyl-uronic acid attached. The final residue after extraction still contained xyloglucan and remaining (pectic) polysaccharides next to cellulose, showing that the cell wall matrix of RSM is very strongly interconnected.