Improved antimelanogenesis and antioxidant effects of polysaccharide from Cuscuta chinensis Lam seeds after enzymatic hydrolysis

Cuscuta chinensis polysaccharide (CPS) was extracted using hot water and enzymatically hydrolyzed C. chinensis polysaccharide (ECPS) was produced by the mannase enzymatic hydrolysis process. The purpose of this research was to investigate the antimelanogenic activity of ECPS and CPS in B16F10 melanoma cells. The in vitro antioxidant activity was assessed by their ferric iron reducing power and DPPH free radical scavenging activities. The molecular mass distribution of polysaccharides was determined using SEC-MALLS-RI. CPS was successfully enzymatically degraded using mannase and the weighted average molecular weights of CPS and ECPS were 434.6 kDa and 211.7 kDa. The results of biological activity assays suggested that the enzymatically hydrolyzed polysaccharide had superior antimelanogenic activity and antioxidant effect than the original polysaccharide. ECPS exhibited antimelanogenic activity by down-regulating the expression of tyrosinase, MITF, and TRP-1 without cytotoxic effects in B16F10 melanoma cells. In conclusion, ECPS have the potential to become a skin whitening product.

Characterization and bioactivities of a novel polysaccharide obtained from Gracilariopsis lemaneiformis

ABSTRACT Gracilariopsis lemaneiformis is a type of red alga that contains seaweed polysaccharide agar. In this study, a novel non-agar seaweed polysaccharide fraction named GCP (short of crude polysaccharide obtained from Gracilariopsis lemaneiformis) was isolated from Gracilariopsis lemaneiformis. Structural analysis showed that GCP shows triple helical chain conformation when dissolved in water and has many branches and long side chains. Also, 1→3 linkage is the major linkage and the sugar structures are galactopyranose configurations linked by β-type glycosidic linkages. Two macromolecular substance fractions (GCP-1 and GCP-2) were purified by DEAE Sepharose Fast Flow column chromatography. Moreover, a splenocyte damage assay and splenocyte proliferation assay were used to analyse the bioactivities of GCP, GCP-1 and GCP-2. It was demonstrated that polysaccharides could protect splenocyte damaged by H2O2; GCP-2 shows a greatest protection rate, that is, 92.8%, which significantly enhanced the splenocyte proliferation, and GCP showed the highest proliferation rate, 9.30%. The results suggested that this type of novel non-agar polysaccharide displayed remarkable antioxidant and immunomodulatory activities and early alkali treatment could decrease the activities. It may represent a potential material for health food and clinical medicines.

Isolation of the intracellular and extracellular polysaccharides of Ganoderma neojaponicum (Imazeki) and characterization of their immunomodulatory properties

Background The role of polysaccharides isolated from the Ganoderma species of fungi in innate immunity has recently become a topic of research. Although some work has been conducted concerning Ganoderma lucidum, the characteristics of polysaccharides isolated from Ganoderma neojaponicum (Imazeki) as immunomodulatory agents are largely unknown. The aims for this study were to isolate and characterize the intracellular polysaccharides (IPSs) and extracellular polysaccharides (EPSs) of G. neojaponicum from STR reactor. Results The production of EPS and IPS was optimized on day 4 of the cultivation time in 2 L STR reactor based on the amount of biomass yield, total carbohydrate, β-glucan and a-glucan content. Further analysis, both the EPSs and IPSs showed the enhancement on proliferation and increment of phagocytosis activities of macrophage (RAW264.7) cell lines. Using an oral toxicity test, we also observed that 2000 mg/kg body weight/day dosage of dried G. neojaponicum mycelium does not cause any significant toxic effects on Sprague-Dawley rats in 14 d of administration. Conclusion The findings of this study indicate that the IPSs and EPSs of G. neojaponicum have the potential to be used as immunomodulating agents to stimulate the innate immune system for fighting infectious diseases. The polysaccharides from G. neojaponicum have to be further commercially explored as an alternative for medicinal Ganoderma variety of G. lucidum production.

Trichuris trichiura in a post-Colonial Brazilian mummy

Trichuris trichiura is a soil-transmitted helminth which is prevalent in warm, moist, tropical and subtropical regions of the world with poor sanitation. Heavy whipworm can result either in Trichuris dysenteric syndrome - especially in children - or in a chronic colitis. In heavy infections, worms can spread proximally and may cause ileitis. Here we provide first microscopic evidence for a T. trichiura adult worm embedded in the rectum of a post-Colonial Brazilian adult mummy. During Colonial and post-Colonial times, many European chroniclers described a parasitic disease named Maculo whose symptomatology coincides with heavy helminthiasis. Based on our findings and on comparison of ancient textual evidence with modern description of heavy whipworm, we feel confident in considering that the two syndromes are expressions of the same pathological condition.

Neutralising antibodies for Mayaro virus in Pantanal, Brazil

The Pantanal hosts diverse wildlife species and therefore is a hotspot for arbovirus studies in South America. A serosurvey for Mayaro virus (MAYV), eastern (EEEV), western (WEEV) and Venezuelan (VEEV) equine encephalitis viruses was conducted with 237 sheep, 87 free-ranging caimans and 748 equids, including 37 collected from a ranch where a neurologic disorder outbreak had been recently reported. Sera were tested for specific viral antibodies using plaque-reduction neutralisation test. From a total of 748 equids, of which 264 were immunised with vaccine composed of EEEV and WEEV and 484 had no history of immunisation, 10 (1.3%) were seropositive for MAYV and two (0.3%) for VEEV using criteria of a ≥ 4-fold antibody titre difference. Among the 484 equids without history of immunisation, 48 (9.9%) were seropositive for EEEV and four (0.8%) for WEEV using the same criteria. Among the sheep, five were sero- positive for equine encephalitis alphaviruses, with one (0.4%) for EEEV, one (0.4%) for WEEV and three (1.3%) for VEEV. Regarding free-ranging caimans, one (1.1%) and three (3.4%), respectively, had low titres for neutralising antibodies to VEEV and undetermined alphaviruses. The neurological disorder outbreak could not be linked to the alphaviruses tested. Our findings represent strong evidence that MAYV and all equine encephalitis alphaviruses circulated in the Pantanal.

Anti-fatigue effects of polysaccharides extracted from Portulaca oleracea L. in mice.

Portulaca oleracea L. has been used as a food and medicinal plant for thousands of years in China. Polysaccharides extracted from P. oleracea L. (POP) are its main bioactive compound and have multiple pharmacological activities. However, anti-fatigue effects of POP have not yet been tested. This study was designed to investigate the anti-fatigue effects of POP in mice using the rotarod and forced swimming tests. The mice were randomly divided into four groups, namely normal control group, low-dose POP supplementation group, medium-dose POP supplementation group and high-dose POP supplementation group. The normal control group received distilled water and the supplementation groups received different doses of POP (75, 150 and 300 mg/kg, respectively). The POP or distilled water was administered orally and daily for 30 day. After 30 days, the rotarod and forced swimming tests were performed and then several biochemical parameters related to fatigue were determined. The data showed that POP prolonged the riding times and exhaustive swimming times of mice, decreasing blood lactic acid and serum urea nitrogen levels, as well as increasing the liver and muscle glycogen contents. These results indicated that POP had the anti-fatigue effects.

Extraction and in vitro antioxidant activity of exopolysaccharide by Pleurotus eryngii SI-02

The extraction parameters for Pleurotus eryngii SI-02 exopolysaccharide (EPS) produced during submerged culture were optimized using response surface methodology (RSM). The optimum conditions for EPS extraction were predicted to be, precipitation time 20.24 h, ethanol concentration 89.62% and pH 8.17, and EPS production was estimated at 7.27 g/L. The actual yield of EPS under these conditions was 7.21 g/L. The in vitro antioxidant results of the EPS showed that the inhibition effects of EPS at a dosage of 400 mg/L on hydroxyl, superoxide anion and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals were 59.63 ± 3.72%, 38.69 ± 2.59%, and 66.36 ± 4.42%, respectively, which were 12.74 ± 1.03%, 8.01 ± 0.56%, and 12.19 ± 1.05% higher than that of butylated hydroxytoluene (BHT), respectively. The reducing power of EPS of P. eryngii SI-02 was 0.98 ± 0.05, 60.66 ± 5.14% higher than that of BHT. The results provide a reference for large-scale production of EPS by P. eryngii SI-02 in industrial fermentation and the EPS can be used as a potential antioxidant which enhances adaptive immune responses.

Extracellular polysaccharide production by a strain of Pleurotus djamor isolated in the south of Brazil and antitumor activity on Sarcoma 180

Polysaccharides with medicinal properties can be obtained from fruiting bodies, mycelium and culture broth of several fungus species. This work was carried out in batch culture using a stirred tank reactor with two different initial glucose concentrations (40-50 g/L) and pH values (3.0-4.0) to enhance extracellular polysaccharides production by Pleurotus djamor UNIVILLE 001 and evaluate antitumor effect of intraperitonial administration of Pleurotus djamor extract on sarcoma 180 animal model. According to factorial design, the low pH value (pH 3.0) led to a gain of 1.6 g/L on the extracellular polysaccharide concentration, while glucose concentration in the tested range had no significant effect on the concentration of polysaccharide. With 40 g/L initial glucose concentration and pH 3.0, it was observed that yield factor of extracellular polysaccharide on substrate (Y P/S = 0.072) and maximum extracellular polysaccharide productivity (Q Pmax = 11.26 mg/L.h) were about 188% and 321% respectively higher than those obtained in the experiment performed at pH 4.0. Under these conditions, the highest values of the yield factor of biomass on substrate (Y X/S = 0.24) and maximal biomass productivity (Q Xmax = 32.2 mg/L.h) were also reached. In tumor response study, mean tumor volume on the 21th day was 35.3 cm³ in untreated group and 1.6 cm³ in treated group (p = 0.05) with a tumor inhibition rate of 94%. These impressive results suggests an inhibitory effect of P.djamor extract on cancer cells.

Optimization of exopolysaccharides production from a novel strain of Ganoderma lucidum CAU5501 in submerged culture

This study aimed at optimizing the medium of a new Ganoderma lucidum strain CAU5501 to enhance the yield of exopolysaccharides (EPS) and mycelial growth. Firstly, the suitable level of glucose, magnesium, phosphate and C/N ratio was determined by single factor experiment. Subsequently, the optimum concentrations of these medium components were investigated using the orthogonal matrix method. The results indicated that the higher levels of EPS were correlated with the level of cell growth when glucose concentration was studied (data no show). The optimum medium for EPS yield was found to be 70 g/l glucose, 5 C/N ratio, 2.5 g/l KH2PO4, 0.75 g/l MgSO4·7H2O, and for mycelial growth was 50 g/l glucose, 5 C/N ratio, 1.5 g/l KH2PO4, 0.5 g/l MgSO4·7H2O. When cultivated in the obtained optimal media in 3 L shake flask, compared to the basal medium, the EPS yield increased markedly from 1.003 to 1.723 g/l, and the mycelium formation was also markedly improved from 2.028 to 7.235 g/l. Results obtained in this study are beneficial to further study for enhancing the production of Ganoderma lucidum polysaccharides in large scale commercialized production.

Antitumor properties of ganoderma lucidum polysaccharidesand terpenoids: [review]

Ganoderma lucidum is an edible medicinal mushroom withimmunomodulatory and antitumor properties, which are mainly attributed to polysaccharides and triterpenesthat can be isolated from mycelia, fruiting bodies and spores. G. lucidum has been used in a powdered form, asa medicinal beverage and a nutraceutical food (usually dried). In the present review we report some historicalfacts and the experimental evidence that polysaccharides and triterpenes obtained from this mushroom presentpotential antitumor activity. Direct effects on tumor cells include induction of apoptosis and interference in thecell cycle, whereas indirect effects are based on the modulation of immune response, usually impaired bycancer cells. Data indicate that G. lucidum can be used as a complementary tool for treatment of cancerpatients.

Phytochemical and biological studies of some polysaccharides isolated from aloe, tamarindus, opuntia and citrus

The aim of this study is the phytochemical and biological evaluation of polysaccharides isolated from some medicinal plants belonging to the Genera Aloe, Tamarindus, Opuntia and Citrus. Antihyperglycemic, anti-inflammatory, immunomodulatory and antioxidant of the polysaccharides were studied. Extraction, isolation and identification of the biologically active constituents have been carried out aiming at discovery of new active constituents from botanical origin which can be utilized in drug industry. The Polysaccharides from the plants under investigation were isolated using cold and hot extraction methods then hydrolysed, the polysaccharide hydrolysates were identified by Paper and HPLC chromatographic techniques. The isolated polysaccharides were tested for their anti-inflammatory, antioxidant, hypoglycemic and immunomodulating activities. The optimum parameters for the extraction of polysaccharides isolated from the seeds of Tamarindus indica Linn. Family Leguminosae were examined by using three factor statistical techniques. Maximum yield of polysaccharides were obtained from Tamarindus indica Linn, seeds [21%], which was increased by varying temperature and time [55%]. The results of the biological activities revealed significant antihyperglycemic, immunomodulating, anti-inflammatory and antioxidant activities with variable degrees. Chromatographic investigation of the polysaccharide hydrolysates revealed the presence of hexoses, pentoses and uronic acids with qualitative and quantitative variations in the chemical composition. Tamarindus indica Linn, seeds had the highest yield of polysaccharide and the highest significant biological activities

Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies

Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular studies.

Polysaccharide-rich fraction of Agaricus brasiliensis enhances the candidacidal activity of murine macrophages

A polysaccharide-rich fraction (ATF) of medicinal mushroom Agaricus brasiliensis was evaluated on the candidacidal activity, H2O2 and nitric oxide (NO) production, and expression of mannose receptors by murine peritoneal macrophages. Mice received three intraperitoneal (i.p.) injections of ATF and after 48 h their peritoneal resident macrophages were assayed against Candida albicans yeast forms. The treatment increased fungicidal activity and it was associated with higher levels of H2O2, whereas NO production was not affected. We also found that the treatment enhances mannose receptor expression by peritoneal macrophages, which are involved in the attachment and phagocytosis of non-opsonized microorganisms. Treatment of animals with ATF was able to enhance the clearance of C. albicans during the first 6 h after the experimental i.p. infection. Our results suggest that this extract can increase host resistance against some infectious agents through the stimulation of microbicidal activity of macrophages.

Isolation, purification and modification of sulfated polysaccharides from spirulina platensis and its antitumor activity

The water-soluble polysaccharide obtained from a blue-green alga Spirulina platensis by boiling water extraction and ethanol precipitation [118.25g kg[-1] on dry weight basis] was fractionated by DEAE-cellulose column chromatography, and purified by Sephacry-S300 giving two active fractions termed Ca-SPIIa and Ca-SPIIb. These fractions were mainly composed of glucose, galactose, rhamnose, fructose, glucouronic acid, mannose, xylose with the molar ratio [6.5: 3.3: 50.0: 38.5: 9.6: 1.5: 1.0] and [7.2: 3.5: 55.0: 40.2: 8.5: 2.0: 1.0] and sulfate percent [2.25% and 3.47%, respectively]. Calcium ion binding with the anionic part of a molecule was replaced with various metal cation and their inhibitory effects on Ehrlich Ascites Carcinoma Cells [EACC] was determined. The Ca-SPIIb was able to kill 95% of the tumor cells when they were incubated with 200microg. While, replacement of Ca[+2] with Na[+], Ag[+], Cu[+2] and H[+] reduced the activity

immunomedulatory effect of Ganoderma Lucidum polysachharide extract on BALB/c peritoneal macrophage function

Ganoderma Lucidum has been regarded as a natural immunomodulator. The exact carbohydrate epitope responsible for the immunomodulatory activity and its receptor have not been identified, but it seems likely that it is the receptor CR3 [complement receptor 3] which can bind to beta-glucan polysachharide. Because glucose-6-phosphate dehydrogenase [G6PD] activity has a critical role in the regulation of macrophage functions such as nitric oxide [NO] production, we assessed the immunomodulatory effect of GL-PS in BALB/c peritoneal macrophages. For this purpose, BALB/c mice peritoneal macrophages were isolated and treated with various concentrations of GL-PS [0.001, 0.01, 0.1, 1, 10, 100 microg/ml]. After 24 hours, the viability of treated macrophages was measured by MTT assay at 540 nm and the effective dose was determined to be 0.1 microg/ml. Then, macrophages were sonicated and special activity of G6PD was measured in the cell extracts by measuring the alterations in NADPH absorption at 339nm and protein concentration by Bradford method. Also, NO production was determined by use of Griess-reagent after 18 hours. Results of this study showed that 0.1 microg/ ml of GL-PS had the maximal effect on cell viability [stimulation Index] in comparison to other doses [0.05

Hypoglycemic activity of polysaccharide fractions containing ß-glucans from extracts of Rhynchelytrum repens (Willd.) C.E. Hubb., Poaceae

ß-Glucans are soluble fibers with physiological functions, such as interference with absorption of sugars and reduction of serum lipid levels. The objective of the present study was to analyze the distribution of ß-glucans in different tissues of the African grass species Rhynchelytrum repens and also to evaluate their hypoglycemic activity. Leaf blades, sheaths, stems, and young leaves of R. repens were submitted to extraction with 4 M KOH. Analysis of the fractions revealed the presence of arabinose, glucose, xylose, and traces of rhamnose and galactose. The presence of ß-glucan in these fractions was confirmed by hydrolyzing the polymers with endo-ß-glucanase from Bacillus subtilis, followed by HPLC analysis of the characteristic oligosaccharides produced. The 4 M KOH fractions from different tissues were subjected to gel permeation chromatography on Sepharose 4B, with separation of polysaccharides with different degrees of polymerization, the highest molecular mass (above 2000 kDa) being found in young leaves. The molecular mass of the leaf blade polymers was similar (250 kDa) to that of maize coleoptile ß-glucan used for comparison. The 4 M KOH fraction injected into rats with streptozotocin-induced diabetes showed hypoglycemic activity, reducing blood sugar to normal levels for approximately 24 h. This performance was better than that obtained with pure ß-glucan from barley, which decreased blood sugar levels for about 4 h. These results suggest that the activity of ß-glucans from R. repens is responsible for the use of this plant extract as a hypoglycemic drug in folk medicine.

Partial characterization and anticoagulant activity of a heterofucan from the brown seaweed Padina gymnospora

The brown algae Padina gymnospora contain different fucans. Powdered algae were submitted to proteolysis with the proteolytic enzyme maxataze. The first extract of the algae was constituted of polysaccharides contaminated with lipids, phenols, etc. Fractionation of the fucans with increasing concentrations of acetone produced fractions with different proportions of fucose, xylose, uronic acid, galactose, and sulfate. One of the fractions, precipitated with 50 percent acetone (v/v), contained an 18-kDa heterofucan (PF1), which was further purified by gel-permeation chromatography on Sephadex G-75 using 0.2 M acetic acid as eluent and characterized by agarose gel electrophoresis in 0.05 M 1,3 diaminopropane/acetate buffer at pH 9.0, methylation and nuclear magnetic resonance spectroscopy. Structural analysis indicates that this fucan has a central core consisting mainly of 3-ß-D-glucuronic acid 1-> or 4-ß-D-glucuronic acid 1 ->, substituted at C-2 with alpha-L-fucose or ß-D-xylose. Sulfate groups were only detected at C-3 of 4-alpha-L-fucose 1-> units. The anticoagulant activity of the PF1 (only 2.5-fold lesser than low molecular weight heparin) estimated by activated partial thromboplastin time was completely abolished upon desulfation by solvolysis in dimethyl sulfoxide, indicating that 3-O-sulfation at C-3 of 4-alpha-L-fucose 1-> units is responsible for the anticoagulant activity of the polymer.

Estudio químico de los polisacáridos presentes en Aloe vera L. y Aloe arborescens Miller cultivados en Cuba / Chemical study of the polysaccharides present in Aloe vera L. y Aloe arborescens Miller cultivated in Cuba

Se presentan los resultados del estudio químico preliminar de Aloe vera L. y Aloe arborescens Miller cultivados en Cuba. Los polisacáridos, a partir de las hojas secas, fueron extraídos con agua a ebullición, y se sometieron a fraccionamiento con bromuro de cetiltrimetil amonio y precipitación fraccionada. Se realizó hidrólisis ácida de las fracciones y detección de azúcares mediante cromatografía en papel. Para la cuantificación de los carbohidratos se utilizó el método del fenol-sulfúrico. Se encontró un mayor contenido de polisacáridos en el Aloe vera L. La hidrólisis ácida del crudo de polisacáridos demostró la presencia de manosa en el Aloe vera L. y glucosa para Aloe arborescens Miller, ambos como mayoritarios. Se encontró además que los polisacáridos ácidos en el Aloe vera L. están presentes en muy pequeña cantidad

Aislamiento de polisacáridos a partir de cordones umbilicales humanos / Isolation of polysaccharides from human umbilical cords

Se realizó el aislamiento y caracterización de polisacáridos obtenidos a partir de cordones umbilicales humanos mediante una modificación a la técnica de Danishevsky y Bella. Se trataron los cordones con solución de cloruro de sodio bajo condiciones de temperatura y agitación controlada y se precipitaron los polisacáridos mediante la adición de una disolución de bromuro de cetiltrimetilamonio al 1 porciento. Se resuspendió el sedimento obtenido en una solución de cloruro de sodio 0,4 mol/L eliminando algunas impurezas que no se solubilizan a esta fuerza iónica mediante centrifugación y se precipitó el producto de interés mediante la adición de etanol al sobrenadante, caracterizándose por métodos químicos. El producto puede ser empleado como materia prima para obtener un gel cicatrizante en la Industria Médico-Farmacéutica