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Medicinal plants used against malaria in Mali have previously been tested for their antiplasmodial activities using their organic solvent and water extracts. As the healers mainly use the water extracts for their treatments of malaria-patients, our aim was to study the water-soluble components from Malian plants used for treatment of malaria. Argemone mexicana (aerial parts), Sarcocephalus latifolius (root bark), Vitex doniana (leaves), and Malarial-5® (an improved traditional medicine (ITM) in tea) were the objects of our studies. Water extracts of these plants contained primarily polysaccharides. Due to this, the studies focused on the determination of the monosaccharide composition of the polymers present as well as assessing the immunomodulatory properties of the polysaccharide fractions isolated from these plants. Each plant material was extracted sequentially with dichloromethane, 80% ethanol and water at 100°C. The polysaccharides were obtained using gel filtration of the aqueous extracts and their monosaccharide compositions were determined using gas chromatography. Immunomodulatory effects were assessed using the complement fixation test and macrophage stimulation. All aqueous extracts from the four samples contained polysaccharides. The monosaccharide compositions vary between the plants. Arabinose, rhamnose, galactose, glucose and galacturonic acid were present in all samples, glucose being the main monomer. These polysaccharides showed complement fixing activity and induced nitrite oxide release from macrophages in a dose dependent manner. The polysaccharide fractions of A. mexicana (Am1) and V. doniana (Vd1) showed the most potent activities. These two fractions had an ICH50 of 2.4 and 6.3 μg/mL respectively in the complement fixation assay. The same two fractions induced a dose dependent release of nitrite oxide from macrophages. The results demonstrated that antimalarial plants contain polysaccharides with immunomodulatory properties. This preliminary work constitutes a new approach of antimalarial studies. Key words: Polysaccharides, immunomodulatory effects, antimalarial plants, Mali.

Based on previous studies about microflora regulation and immunity enhancement activities of polysaccharides from Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen (CPP), there is little study on intestinal mucosal immunity, which is a possible medium for contacting microflora and immunity. In the present study, the BALB/c mice were divided into five groups (eight mice in each group), including a normal group (Con), a model control group (Model), and model groups that were administered CPP (50, 100, 200 mg/kg/d) orally each day for seven days after intraperitoneal injection of 60 mg/kg BW/d cyclophosphamide (CP) for three days. CPP recovered the spleen index and restored the levels of IFN-&gamma, IL-2, IL-10, as well as serum IgG. In addition, it elevated ileum secretory immunoglobulin A (sIgA), the number of Lactobacillus and acetic acid content in cecum. These results indicated that CPP plays an important role in the protection against immunosuppression, especially mucosa immune damage, and the inhibition of pathogenic bacteria colonization, which could be considered a potential natural source of immunoregulator.

Based on previous studies about microflora regulation and immunity enhancement activities of polysaccharides from Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen (CPP), there is little study on intestinal mucosal immunity, which is a possible medium for contacting microflora and immunity. In the present study, the BALB/c mice were divided into five groups (eight mice in each group), including a normal group (Con), a model control group (Model), and model groups that were administered CPP (50, 100, 200 mg/kg/d) orally each day for seven days after intraperitoneal injection of 60 mg/kg BW/d cyclophosphamide (CP) for three days. CPP recovered the spleen index and restored the levels of IFN-γ, IL-2, IL-10, as well as serum IgG. In addition, it elevated ileum secretory immunoglobulin A (sIgA), the number of Lactobacillus and acetic acid content in cecum. These results indicated that CPP plays an important role in the protection against immunosuppression, especially mucosa immune damage, and the inhibition of pathogenic bacteria colonization, which could be considered a potential natural source of immunoregulator.

The inulin-type fructan was obtained by DEAE anion exchange chromatography from C. pilosula Nannf. var. modesta (Nannf.) L. T. Shen, after optimized extract condition, which was established by response surface methodology, designed using Box-Behnken factorial design and the optimum condition were: extracting 2.5 h with ratio of solvent to material 40 mL/g at 100 °C, twice. The maximum extraction yield was 20.6 ± 0.2%. It was confirmed as β-(2-1) linkage fructan, with terminal glucose, and with a degree of polymerization of 2–17 (DPav = 6), shown by the results of methanolysis, methylation, nuclear magnetic resonance and molecular weight determination. The prebiotic activity was proven on account of stimulation effect on Lactobacillus and pH reduction of medium in vitro. The results indicated that the inulin from C. pilosula could be used as a potential natural source of prebiotics.

Exo-polysaccharides were purified and characterized from the fermentation broth of Hypsizigus marmoreus, a popular edible mushroom consumed in Asia. Among them, B-I-I and B-II-I exhibited potent complement fixating activity, meanwhile, B-N-I, B-I-I, B-II-I and B-II-II exhibited significant macrophage stimulating activity. Molecular weights of the four exo-polysaccharides were determined to be 6.3, 120, 150 and 11 kDa respectively. Molecular characterisation showed that B-N-I is basically an α-1 → 4 glucan, with branches on C6; B-I-I is a heavily branched α-mannan with 1 → 2 linked main chain. B-II-I and B-II-II, have a backbone of rhamno-galacturonan with 1 → 2 linked l -rhamnose interspersed with 1 → 4 linked galacturonic acid. Structure–activity relationship analysis indicated that monosaccharide compositions, molecular weight, certain structural units (rhamno-galacturonan type I and arabinogalactan type II) are the principal factors responsible for potent complement fixating and macrophage-stimulating activities. Their immunomodulating activities may, at least partly, explain the health benefits of the mushroom.

Ethnopharmacological relevance Sutherlandia frutescens (syn. Lessertia frutescens) is an indigenous plant in Southern Africa and has been extensively studied from the ethnobotanical point of view. Amongst the various traditional uses, several illnesses involving the immune system have been reported. Due to some of the therapeutic effects observed, in relation to the traditional uses reported by the “khoi san” and “nama” people on cancer related illnesses, the plant has been given the local name kankerbos (cancerbush). Recently the plant has also been used amongst HIV/AIDS patients to stimulate the immune system. Materials and methods Leaves of Sutherlandia frutescens were extracted sequentially with ethanol, 50% ethanol/water, and water at 50 and 100 °C. The polysaccharides were extracted with water and fractionated by ion exchange chromatography and gel filtration to obtain enriched polysaccharide fractions. The bioactivities of the fractions were tested in the complement assay. Some of the fractions were treated with the enzyme pectinase, and the fragments thus produced were separated by gel filtration and their activities tested. Monosaccharide compositions and linkage analyses were determined for the relevant fractions. Results The leaves of Sutherlandia frutescens contain polysaccharides of the pectin type. Fractions from both the water extracts of 50 and 100 °C were bioactive. Fractions chosen for further studies showed that the fragment with the highest MW after the pectinase treatment had a substantially higher biological effect than the parent molecules. Based on a comparison of the different fractions it was concluded that galactose-rich regions were important for the bioactivity, these being of the AGII and AGI type, with the latter probably being more important than the former. Fragments rich in xylose also gave higher activity than those without it. Conclusions Our theory that the polysaccharides present in the leaves of Sutherlandia frutescens could be of importance as immunomodulating agents was confirmed. It was also shown that certain types of polysaccharides had a higher effect in the complement system than others. Thus both the water extracts obtained at 50 and 100 °C contain interesting biologically active polysaccharides.

Platycodon grandiflorus is a plant widely used in traditional Chinese medicine, of which polysaccharides are reported to be the main components responsible for its bio-functions. In this work, the inulin-type fructan (PGF) was obtained by DEAE anion exchange chromatography from the water extracted from P. grandifloras. Characterization was performed with methanolysis, methylation, and NMR and the results showed that PGF is a &beta, (2-1) linked fructan, with terminal glucose and with a degree of polymerization of 2&ndash, 10. In order to study its biofunctions, the prebiotic and immunomodulation properties were assayed. We found that PGF exhibited good prebiotic activity, as shown by a promotion on six strains of lactobacillus proliferation. Additionally, the PGF also displayed direct immunomodulation on intestinal epithelial cells and stimulated the expressions of anti-inflammatory factors. These results indicated that the inulin from P. grandiflorus is a potential natural source of prebiotics as well as a potential intestinal immunomodulator, which will be valuable for further studies and new applications.

Seaweeds waste is a main oceanic nonproductive material in related industries. In this study, it was utilized for incubation of Cantharellus sp., a popular edible mushroom. The micro particles obtained from the incubation were investigated for antioxidant activities. The results showed that the Cantharellus can grow on the seaweeds waste and produced dense mycelia. Meanwhile, antioxidant assays showed that the total antioxidant capacity and hydroxyl radical scavenging capacity of the micro particles obtained from Cantharellus incubation were significantly higher than those of the seaweeds waste. This is not only the first time to incubate Cantharellus sp. on seaweeds waste, but also the first study involved in antioxidant activities of fermented culture with Cantharellus sp.. It would be a beneficial pathway to recycle seaweeds waste with incubation of Cantharellus to produce potential natural antioxidants.

Single factor experiment and response surface methodology were conducted to optimize the production conditions for polysaccharide with higher antitumor activity from mushroom fermentation broth. Based on the results of single factor experiments, a central composite design was applied to the following optimization and second-order polynomial regression models were established. As the yield and antitumor activity of the polysaccharides were considered with equal weight, the optimal conditions were that ethanol concentration and pH of the broth was adjusted to 85.00% and 7.70 respectively, then precipitated at 12 °C for 1 h and dried the precipitates at 40 °C. The verification experiments completed under the optimal conditions gave the polysaccharides yield of 9.843 ± 0.217 g/l, and the cancer cell growth inhibitory rate of 83.69 ± 0.167% by the polysaccharides, which were in close agreement with values predicted by the models (RSD

The root bark, stem bark and leaves of Terminalia macroptera were sequentially extracted with ethanol, 50% ethanol-water, and 50 °C and 100 °C water using an accelerated solvent extractor. Ten bioactive purified polysaccharide fractions were obtained from those crude extracts after anion exchange chromatography and gel filtration. The polysaccharides and their native extracts were characterized with respect to molecular weight, chemical compositions and effects in the complement assay. The chemical compositions showed that the polysaccharides are of pectic nature. The results indicated that there was no great difference of the complement fixation activities in the crude extracts from the different plant parts when extracting with the accelerated solvent extraction system. The purified polysaccharide fractions 100WTSBH-I-I and 100WTRBH-I-I isolated from the 100 °C water extracts of stem and root bark respectively, showed the highest complement fixation activities. These two fractions have rhamnogalacturonan type I backbone, but only 100WTSBH-I-I contains side chains of both arabinogalactan type I and II. Based on the yield and activities of the fractions studied those from the root bark gave highest results, followed by those from leaves and stem bark. But in total, all plant materials are good sources for fractions containing bioactive polysaccharides.

The increasing commercial significance of natural polysaccharides for use in medicinal products is stimulating the development of efficient and easy scale-up techniques for polysaccharide purification. In this research, the crude polysaccharides from submerged cultivation broth of Hypsizigus marmoreus were purified using radial flow chromatography (RFC), and the antiproliferative activity of the purified fractions was evaluated in vitro. DEAE Sepharose CL-6B was selected to be packed in the RFC column based on its good resolution, physical stability, and low cost. Compared with axial flow chromatography (AFC), an efficient chromatographic process with significantly less time and buffer consumption but yielding higher polysaccharide recovery and resolution was established in RFC, which could clearly purify the crude polysaccharides into different fractions. An acceptable linear scale-up effect of RFC from 100 to 500 mL was successfully achieved without loss of resolution and enhancement of time consumption. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays in cell cultures indicated that the purified polysaccharide fractions possess moderate antiproliferative activities in three different human cancer cell lines, but have significantly lower cytotoxicity in normal human cell lines in vitro. Among the polysaccharide fractions, the main purified acidic fraction W-I could be considered as a novel potential antitumor agent candidate for several tumors, especially for human alveolar epithelial tumors. This research confirmed for the first time that RFC would be a new fast and efficient tool for purification of polysaccharides into different fractions, both at laboratory and commercial scales. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:872–878, 2014

The bark of Parkia biglobosa is used in traditional medicine to cure a wide range of illnesses. Polysaccharides were extracted from the bark with 50% ethanol–water, 50 °C and 100 °C water, and seven active fractions obtained by anion exchange chromatography and gel filtration. The complement fixation and macrophage stimulating activities of the different fractions were determined. The acidic fractions PBEII-I and PBEII-IV were the most active in the complement fixation assay, but the other fractions were also potent compared to the positive control BPII from Biophytum petersianum . Fractions PBEII-I and PBEII-IV were also the most potent fractions in stimulating macrophages to release nitric oxide. Structural studies showed that PBEII-I and PBEII-IV were pectic type polysaccharides, containing arabinogalactan type II structures. The observed differences in biological activities among the seven purified polysaccharide sub-fractions are probably due to differences in monosaccharide compositions, linkage types and molecular sizes.

In Malian traditional medicine the roots of Cochlospermum tinctorium are used in the treatment of gastric ulcer, but extending harvesting is causing a growing concern of a dramatic reduction in the wild plant population. In the present study cultivation of C. tinctorium is evaluated, and structural components and bioactive properties of crude water extracts and isolated polysaccharide fractions from roots of wild and cultivated C. tinctorium are compared. The crude water extracts were shown to contain starch, pectin- and inulin-type polysaccharides, in addition to phenolic substances and protein, while the isolated acidic polysaccharide fractions contained mainly monosaccharides typical for pectins. The monosaccharide compositions of the polysaccharide fractions from roots of wild versus cultivated plants were comparable, albeit the yields in the cultivated roots were lower. Furthermore, the crude extracts and isolated polysaccharide fractions from wild and cultivated roots exhibited similar complement fixating activities, but were not able to activate macrophages. The crude extracts from cultivated roots were also shown to be moderate radical scavengers. The present study has shown that roots of cultivated C. tinctorium contain the same types of bioactive polysaccharides as the wild roots. However, in order to utilize roots of cultivated C. tinctorium in traditional medicine the cultivation method should be improved.

Medicinal mushroom polysaccharides from submerged culture have lots of important biological activities, such as antitumor, immunomodulating, antioxidation and so on. In order to improve the biological activities of the polysaccharides, pigments impurities produced in the fermentation process have to be eliminated by a suitable technology before polysaccharides purification and application. 28 kinds of macro porous resins were investigated in this study and the results showed that macro porous resin OU-1 had satisfactory pigment elimination ability. After optimization by uniform design and verification experiments, the optimal condition for pigments adsorption by macro porous resin OU-1 was found. The optimized conditions were that the fermentation supernatant with pH ranging from 5.2 to 5.8 could be directly applied to pigments adsorption for 3.9 h at 40°C. At the optimized condition, about eighty percent of the pigment in fermentation supernatant could be eliminated, ninety-five percent of the polysaccharides in the fermentation supernatant could be recovered, and only about 41mg/ml of the resin (wet weight/v) was needed.