Antioxidant Activity Derived from Marine Green-Lipped Mussel Perna canaliculus Extracts in Mice.

This study investigates the antioxidant activities of lipid, protein, and carbohydrate extracts from the marine mollusk Perna canaliculus. Lipids were extracted using acetone, which was followed by protein extraction using the broad-spectrum enzyme Alcalase and then carbohydrate extraction using cetylpyridinium chloride. Eighty white BALB/c mice were divided into eight groups according to the administered extracts. Groups 1 and 5 were the control and toxin control groups, respectively. Groups 2, 3, and 4 were administered lipid, protein, and carbohydrate extracts, respectively. The other groups were administered P. canaliculus extracts as well as gentamicin and acetaminophen, known as ethanolic extracts, derived from Nerium oleander to induce oxidation stress. All groups showed significant improvements in body weight (p < 0.05). The lipid extract group showed a significant decrease in low-density lipoprotein cholesterol (p < 0.05) and a significant increase in high-density lipoprotein cholesterol (p < 0.05). After the toxin injection, all groups treated with P. canaliculus extracts showed increased antioxidant effects on hepatocytes (p < 0.05). The lipid extracts induced antioxidant effects to protect the kidney by increasing lipid peroxidation (p < 0.05) and catalase activities (p < 0.05). Also, protein extracts showed antioxidant effects by increasing glutathione and catalase levels significantly (p < 0.005). In conclusion, P. canaliculus extracts, especially lipids and proteins, have potent antioxidant activities that protect vital organs from oxidation stress.

Aloe vera and its two bioactive constituents in alleviation of diabetes -proteomic & mechanistic insights.

ETHNOPHARMACOLOGICAL RELEVANCE: Aloe barbadensis Miller, commonly known as Aloe vera has been used since time immemorial for treatment of various diseases such as cancer, inflammatory disorders, diabetes, wound healing etc. AIM: Diabetes mellitus is a complex disorder and understanding the molecular mechanisms involved is a key to identify different markers for early diagnosis of the disease. The proteomic approach offers a plethora of opportunities to identify markers and targets involved in pathogenesis of diabetes. The present study was undertaken to understand the mechanism of action of Aloe vera and its two constituents (Carbohydrates and Polypeptides) in the alleviation of diabetes in streptozotocin-induced diabetic rats through a proteomics approach. METHODS: Different groups of rats were fed with Aloe vera extract, carbohydrate fraction and peptide/polypeptide fraction for three weeks. The diabetic rats fed with Aloe vera and its two fractions restored the glucose and insulin levels to normal. The plasma of the rats was depleted with IgG and albumin and proteomic analysis was carried out. Apolipoproteins (dyslipidemia), complement factors (inflammatory pathways), zonulin (intestinal permeability), anti-oxidant related proteins were selected in this study as these are involved in the progression of diabetes. RESULTS: It was observed that Aloe vera extract is involved in the alleviation of diabetes through these pathways while the carbohydrate fraction alleviates diabetes through an anti-oxidant mechanism and glucose uptake while the polypeptide fraction alleviates diabetes through the restoration of intestinal permeability by reduced zonulin levels. CONCLUSION: The constituents of Aloe vera works different pathways involved in diabetes and the synergistic effect of these constituents make Aloe vera extract a prospective candidate, which can alleviate diabetes through regulation of the pathways involved in the progression of diabetes.

Apoplastic maize fructan exohydrolase Zm-6-FEH displays substrate specificity for levan and is induced by exposure to levan-producing bacteria.

Fructan exohydrolases (FEHs) are structurally related to cell wall invertases. While the latter are ubiquitous in higher plants, the role of FEHs in non-fructan species has remained enigmatic. To explore possible roles of FEHs in maize, a full length putative Zm-6-FEH-encoding cDNA was cloned displaying high sequence similarity with cell wall invertases. For functional characterization, Zm-6-FEH protein was expressed in Picha pastoris and in Nicotiana benthamiana leaves. Enzyme activity of recombinant Zm-6-FEH protein showed a strong preference for levan as substrate. Expression profiling in maize seedlings revealed higher transcript amounts in the more mature leaf parts as compared to the growth zone at the base of the leaf, in good correlation with FEH enzyme activities. Subcellular localization analysis indicated Zm-6-FEH location in the apoplast. Noteworthy, incubation of leaf discs with levan and co-incubation with high levan-producing bacteria selectively up-regulated transcript levels of Zm-6-FEH, accompanied by an increase of 6-FEH enzyme activity. In summary, the results indicate that Zm-6-FEH, a novel fructan exohydrolase of a non-fructan species, may have a role in plant defense against levan-producing bacteria.

Optimization of extraction with salicylic acid, rheological behavior and antiproliferative activity of pectin from Citrus sinensis peels.

A Box-Behnken design was used to optimize extraction temperature, extraction time and concentration of the salicylic acid to obtain a maximum polysaccharide yield from Citrus sinensis peels. The optimal settings were: extraction time 3 h, extraction temperature 80 °C and concentration of the salicylic acid 1.5%. Under these conditions, the experimental yield and uronic acid content were 11.74% and 66.9% respectively. Preliminary characterization was performed via FT-IR, SEC/MALS/VD/DRI and GC-MS after hydrolysis. SEC analysis showed that the extracted polysaccharide had a weight average molar mass of 350 kDa and an intrinsic viscosity of 640 mL/g. The GC-MS results revealed that the extracted polysaccharide was composed of arabinose 56.7%, galactose 17.8%, xylose 13.8%, rhamnose 5.1%, mannose 2.5% and glucose 1.5% suggested a rhamnogalacturonan pectin type I with a degree of esterification of 50.9% (IRTF). The flow curve and the dynamic frequency sweep were obtained at 10, 20, 30 and 40 g/L in water and at 30 g/L in presence of CaCl2 or NaCl at 1 mol/L. The solutions showed shear-thinning behavior fitted with Ostwald-De Waele model, except 10 g/L with a Newtonian behavior. The apparent viscosity and, the G' and G" moduli increase with PACO concentration in agreement with a slow-down of the dynamic chain. In the presence of CaCl2 or NaCl the reduction of electrostatic repulsions between pectin chains decreases the rheological parameters. The effect is less sensitive with CaCl2 due to intermolecular interactions. The antiproliferative activity of the extracted pectin on human Caco-2 and Hep-2 cells was very interesting with an IC50 1.4 and 1.8 µg/mL respectively.

Ultrasound Assisted Cascade Extraction of Oil, Vitamin E, and Saccharides from Roselle (Hibiscus Sabdariffa L.) Seeds.

Roselle seeds, a waste biomass of the roselle calyx processing industry, were utilized to recover valuable compounds of oil, vitamin E, and water-soluble saccharides. Firstly, ultrasound-assisted extraction (UAE) and conventional stirring extraction were conducted for saccharide extraction, and the advantage of UAE was confirmed. Secondly, oil, vitamin E, and saccharides extracted from Vietnamese roselle seeds by UAE were analyzed for the first time. Oil of tri-, di-, and mono-glycerides, fatty acids of linoleic-, oleic-, palmitic-, and stearic-acids, vitamin E of γ- and α-tocopherol, and saccharides of sucrose, raffinose, stachyose, etc. were identified, and the amounts of these components were compared with those in other country's roselle seeds. Thirdly, cascade extraction of oil, vitamin E, and saccharides by UAE was investigated with solvents of hexane, hexane:ethyl acetate binary solvent, and water. The results indicated that the order of using solvents was very important for high and selective extraction: the best order to recover oil (almost 100%), vitamin E (95.7%), and saccharides (86.2%) was hexane, and then water.

Extraction techniques and potential health benefits of bioactive compounds from marine molluscs: a review.

Marine molluscs and their bioactive compounds are of particular relevance to the growing pool of nutraceutical resources under global investigation. A number of extraction techniques have been developed to isolate bioactive compounds according to their chemical characterization, such as proteins, carbohydrates and lipids. We briefly reviewed those methods in general. Bioactive molecules are 'concealed' in the primary structures of tissue samples of molluscs as amino acids, lipids or carbohydrates which are released by mechanical and chemical processes. The major health benefits of extracts of molluscs include antioxidant, anticancer, and anti-infectious disease activities and cardiovascular protection, which have been reviewed in detail. This review provides a novel view into the efficacy of isolation techniques and subsequent bioactivity analysis of compounds under investigation. Future development in extraction-bioactivity has also been discussed.

Three-phase partitioning for the direct extraction and separation of bioactive exopolysaccharides from the cultured broth of Phellinus baumii.

In this study, three-phase partitioning (TPP) was used to directly extract and separate bioactive exopolysaccharides (EPSs) from a cultured broth of Phellinus baumii. The maximum extraction yield of EPS was 52.09% under the following optimal conditions: 20% (w/v) ammonium sulfate concentration, 1.0:1.5 (v/v) ratio of cultured broth to t­butanol, 30 min, and 35 °C. A multifrequency power ultrasound in a sequential mode coupled with TPP resulted in ~9.12% increment in extraction yield and ~80% reduction in extraction time compared with those of traditional TPP. The carbohydrate (88.21%) and uronic acid (3.37%) contents of partially purified EPS were higher than those of EPS-C obtained through conventional ethanol precipitation and separation methods. EPS and EPS-C exhibited similar preliminary structural characteristics and different monosaccharide compositions and molecular weights. The radical-scavenging abilities, antioxidant capacities, α­amylase and α­glycosidase inhibitory activities, and macrophage stimulation activities of EPS were also higher than those of EPS-C. Therefore, it could be concluded that TPP as a simple and green separation technique could be used to directly extract and separate bioactive EPS from the fermentation broths of mushrooms and other fungi.

Gas chromatographic-based techniques for the characterization of low molecular weight carbohydrates and phenylalkanoid glycosides of Sedum roseum root supplements.

An extensive characterization of low molecular weight carbohydrates (LMWC) and phenylalkanoid glycosides (PAG) of Sedum roseum root supplements has been carried out for the first time by gas chromatography coupled to mass spectrometry (GC-MS) and by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC × GC-ToF MS). Optimization of the required derivatization procedure for improved determination of PAG showed the combined use of trimethylsilylimidazole and trimethylchlorosilane as the most appropriate reagents. Up to 37 compounds were qualitative- and quantitatively characterized in different dietary supplements of S. roseum by GC-MS. In addition to the well-known rosin, rosarin, rosavin and salidroside, other carbohydrates, polyalcohols, acids, etc. were determined. Among them, several seven-carbon monosaccharides such as coriose and 2,7-anhydro-ß-d-altro-heptulose were detected for the first time in S. roseum root supplements. Sedoheptulose was found to be the most abundant compound (9-151 mg g-1), followed by rosiridin (20-81 mg g-1) and rosavin (11-56 mg g-1). The use of GC × GC-ToF MS allowed the detection and tentative assignation of 48 additional compounds mainly belonging to the phenylalkanoid glycoside, pentosyl-hexose and hexosyl-hexose families.

Marine Carbohydrate-Based Compounds with Medicinal Properties.

The oceans harbor a great diversity of organisms, and have been recognized as an important source of new compounds with nutritional and therapeutic potential. Among these compounds, carbohydrate-based compounds are of particular interest because they exhibit numerous biological functions associated with their chemical diversity. This gives rise to new substances for the development of bioactive products. Many are the known applications of substances with glycosidic domains obtained from marine species. This review covers the structural properties and the current findings on the antioxidant, anti-inflammatory, anticoagulant, antitumor and antimicrobial activities of medium and high molecular-weight carbohydrates or glycosylated compounds extracted from various marine organisms.

Bioactive potential and composition analysis of sulfated polysaccharide from Acanthophora spicifera (Vahl) Borgeson.

Marine seaweeds contain a valuable source of functional bioactive polysaccharide and it plays main role for effective anticancer activity. The structural feature of SPs was studied through FT-IR and 1H NMR spectra analysis. The isolated SPs from A. spicifera contain 63.3% of total sugar, 21.9% of total sulfate and 12.6% of total uranic acid was found. The active F2 fraction molecular weight of SP was found to be 420 kDa. The sugar was composed of galactose (73.5%), xylose (9.2%), mannose (1.9%) and arabinose (10.9%). Further the SP showed DPPH free radical scavenging activity of 55.55% at 150 µg/mL and reducing power activity of 91.3% at 125 µg/mL. In the present study, the purified sulfated polysaccharide (fraction F2) were extracted, purified and characterized for red seaweed and evaluated for their potential anticancer activity of in A549 cell lines under in vitro condition. These polysaccharide fractions exhibited potential apoptotic effects on A549 cell lines.

A polyacrylamide-based silica stationary phase for the separation of carbohydrates using alcohols as the weak eluent in hydrophilic interaction liquid chromatography.

A hydrophilic interaction liquid chromatography (HILIC) stationary phase was prepared by a two-step synthesis method, immobilizing polyacrylamide on silica sphere particles. The stationary phase (named PA, 5µm dia) was evaluated using a mixture of carbohydrates in HILIC mode and the column efficiency reached 121,000Nm-1. The retention behavior of carbohydrates on PA stationary phase was investigated with three different organic solvents (acetonitrile, ethanol and methanol) employed as the weak eluent. The strongest hydrophilicity of PA stationary phase was observed in both acetonitrile and methanol as the weak eluent, when compared with another two amide stationary phases. Attributing to its high hydrophilicity, three oligosaccharides (xylooligosaccharide, fructooligosaccharide and chitooligosaccharides) presented good retention on PA stationary phase using alcohols/water as mobile phase. Finally, PA stationary phase was successfully applied for the purification of galactooligosaccharides and saponins of Paris polyphylla. It is feasible to use safer and cheaper alcohols to replace acetonitrile as the weak eluent for green analysis and purification of polar compounds on PA stationary phase.

Mycochemical Characterization of Agaricus subrufescens considering Their Morphological and Physiological Stage of Maturity on the Traceability Process.

Agaricus subrufescens Peck is a basidiomycete with immunomodulatory compounds and antitumor activities. This research evaluated the mycochemical composition of A. subrufescens, considering their morphological and physiological stage of maturity, with a particular focus on the development of a traceability process for the formulation of new nutritional products based on fungal foods. The stipes contained a high amount of dry matter (10.33%), total carbohydrate (69.56%), available carbohydrate (63.89%), and energy value (363.97 kcal 100 g-1 DM). The pilei contained a high amount of moisture (90.66%), nitrogen (7.75%), protein (33.96%), ash (8.24), crude fat (2.44%), acid detergent fiber (16.75 g kg-1), neutral detergent fiber (41.82 g kg-1), hemicellulose (25.07 g kg-1), and lignin (9.77 g kg-1). Stipes with mature physiological stage had higher values of dry matter (10.50%), crude fiber (5.94%), total carbohydrate (72.82%), AC (66.88%), and energy value (364.91 kcal 100 g-1 DM). Pilei of the mushrooms in the immature physiological stage had higher values of P (36.83%), N (8.41%), and A (8.44%). Due to the differences between the mycochemical compositions of the morphological parts of mushrooms linked to their physiological stage of maturity, such characteristics have immense potential to be considered for a traceability process. This study can be used for the purpose of providing the consumer with more product diversity, optimizing bioactivities of composts, and allowing farmers an efficient and profitable use of the mushroom biomass.

Multienzyme decorated polysaccharide amplified electrogenerated chemiluminescence biosensor for cytosensing and cell surface carbohydrate profiling.

A novel ECL biosensor for cytosensing and cell surface carbohydrate expression evaluation was developed, by the integration of the peptide modified interface for highly specific carbohydrate recognition and sodium alginate loaded glucose oxidase as the signal probe with high signal amplification efficiency. A cysteine-terminated peptide self-assembled on the electrode through Au-S bond to construct a functional interface for cell capture, with decent biocompatibility and high affinity for the human breast cancer cell MCF-7. Concanavalin A lectin modified gold nanoparticles specifically recognized the cell surface carbohydrates and were absorbed on the electrode, followed by the immobilization of multiple glucose oxidase conjugated sodium alginate, which could remarkably increase the sensitivity of the biosensor with enhanced catalysis. The as-proposed ECL cytosensor was successfully applied for the detection of the MCF-7 tumor cells, whose glycans on the cell membranes are over-expressed. A low detection limit of 150cellsmL-1 was obtained, with a wide dynamic linear range from 5.0×102 to 5.0×105cellsmL-1. Due to the excellent sensitivity, stability and biocompatibility, the ECL biosensor would be promising in reliable diagnostics of glycan relevant biomarkers for cancer and other diseases.

Detection, purification and characterization of a lectin from freshwater green algae Spirogyra spp

ABSTRACT Freshwater algae are rich sources of structurally biologically active metabolites, such as fatty acids, steroids, carotenoids and polysaccharides. Among these metabolites, lectins stand out. Lectins are proteins or glycoproteins of non-immune origin which bind to carbohydrates or glycoconjugates, without changing ligand structure. Many studies have reported on the use of Spirogyra spp. as effective bioindicators of heavy metals; however, reports on Spirogyra molecular bioprospecting are quite limited. Therefore, this study aimed to detect, isolate, purify and characterize a lectin present in the freshwater green algae Spirogyra. Presence of the lectin protein in the extract was detected by hemagglutination assays. Subsequently, the protein extract was subjected to a sugar inhibition assay to identify the lectin-specific carbohydrate. Following this, the extract was applied to a guar gum column to afford the pure lectin. The lectin was inhibited by N-acetyl-glucosamine and N-acetyl-beta-D-mannose, but more strongly by D-galactose. The apparent molecular mass of the purified lectin was evaluated by Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Electrophoretic analysis revealed a single protein band with an apparent molecular mass of 56 kDa. Thus, it could be concluded that a lectin was purified from Spirogyra spp.

Systemic localization of seven major types of carbohydrates on cell membranes by dSTORM imaging.

Carbohydrates on the cell surface control intercellular interactions and play a vital role in various physiological processes. However, their systemic distribution patterns are poorly understood. Through the direct stochastic optical reconstruction microscopy (dSTORM) strategy, we systematically revealed that several types of representative carbohydrates are found in clustered states. Interestingly, the results from dual-color dSTORM imaging indicate that these carbohydrate clusters are prone to connect with one another and eventually form conjoined platforms where different functional glycoproteins aggregate (e.g., epidermal growth factor receptor, (EGFR) and band 3 protein). A thorough understanding of the ensemble distribution of carbohydrates on the cell surface paves the way for elucidating the structure-function relationship of cell membranes and the critical roles of carbohydrates in various physiological and pathological cell processes.

Effects of changes in chemical and structural characteristic of ammonia fibre expansion (AFEX) pretreated oil palm empty fruit bunch fibre on enzymatic saccharification and fermentability for biohydrogen.

Oil palm empty fruit bunch (OPEFB) fibre is widely available in Southeast Asian countries and found to have 60% (w/w) sugar components. OPEFB was pretreated using the ammonia fibre expansion (AFEX) method and characterised physically by the Fourier transform infrared spectroscopy, X-ray diffraction and scanning electron microscopy. The results show that there were significant structural changes in OPEFB after the pretreatment step, and the sugar yield after enzymatic hydrolysis using a cocktail of Cellic Ctec2® and Cellic Htec2® increased from 0.15gg(-1) of OPEFB in the raw untreated OPEFB sample to 0.53gg(-1) of OPEFB in AFEX-pretreated OPEFB (i.e. almost a fourfold increase in sugar conversion), which enhances the economic value of OPEFB. A biohydrogen fermentability test of this hydrolysate was carried out using a locally isolated bacterium, Enterobacter sp. KBH6958. The biohydrogen yield after 72h of fermentation was 1.68mol H2 per mol sugar. Butyrate, ethanol, and acetate were the major metabolites.

Different compounds are extracted with different time courses from fruits during microwave hydrodiffusion: examples and possible causes.

We set out to determine how nutrients diffuse during extraction, using fractional collection. The highest concentrations of sugars (195.5, 64.8 and 60.8g/L, respectively for grape, 'Najbolia' plum and apricot) were found for the earliest stages of extraction, with a decrease in concentration (to 41.4g/L, 48.2g/L and 1.7g/L, respectively) at the end of extraction process. Total polyphenols showed the same trends for plum and apricot (from 4.1g/L to 2.9g/L for 'Najbolia' plum, from 2.2 to 0.2g/L for apricot) but highest concentrations of total polyphenols (for grape and cherry) were obtained at fraction 5 or 6 (out of 7). Carotenoids from cherry tomato also had highest concentrations (at circa 25mg/L) almost at the end of extraction. For volatile molecules from sweet cherry, hexanal, 2-hexenal and linalool had their highest concentrations at fractions 3-4 (out of 7). Diffusion of nutrients depended on fruit destructuring, molecule solubility and localization of the compounds. Fruit size seemed unimportant.

Metabolic profiles and free radical scavenging activity of Cordyceps bassiana fruiting bodies according to developmental stage.

The metabolic profiles of Cordyceps bassiana according to fruiting body developmental stage were investigated using gas chromatography-mass spectrometry. We were able to detect 62 metabolites, including 48 metabolites from 70% methanol extracts and 14 metabolites from 100% n-hexane extracts. These metabolites were classified as alcohols, amino acids, organic acids, phosphoric acids, purine nucleosides and bases, sugars, saturated fatty acids, unsaturated fatty acids, or fatty amides. Significant changes in metabolite levels were found according to developmental stage. Relative levels of amino acids, purine nucleosides, and sugars were higher in development stage 3 than in the other stages. Among the amino acids, valine, isoleucine, lysine, histidine, glutamine, and aspartic acid, which are associated with ABC transporters and aminoacyl-tRNA biosynthesis, also showed higher levels in stage 3 samples. The free radical scavenging activities, which were significantly higher in stage 3 than in the other stages, showed a positive correlation with purine nucleoside metabolites such as adenosine, guanosine, and inosine. These results not only show metabolic profiles, but also suggest the metabolic pathways associated with fruiting body development stages in cultivated C. bassiana.

Structural glycomic analyses at high sensitivity: a decade of progress.

The field of glycomics has recently advanced in response to the urgent need for structural characterization and quantification of complex carbohydrates in biologically and medically important applications. The recent success of analytical glycobiology at high sensitivity reflects numerous advances in biomolecular mass spectrometry and its instrumentation, capillary and microchip separation techniques, and microchemical manipulations of carbohydrate reactivity. The multimethodological approach appears to be necessary to gain an in-depth understanding of very complex glycomes in different biological systems.