Alcohol Precipitation Research Articles

Green biorefinery for sugar beet pulp valorisation: Microwave hydrothermal processing for pectooligosaccharides recovery and biobutanol production

The complete valorisation of sugar beet pulp (SBP) through pectooligosaccharides (POS) extraction and biobutanol production was studied. A central composite experimental design was planned to optimise the time and temperature of the hydrothermal microwave-assisted extraction. 59.7% of the POS present in the SBP were recovered, and a prehydrolysate with 31.1 g POS/L was obtained under optimal conditions (165 °C and 12 min). The prehydrolysate was further precipitated by ethanol. One-step and stepwise alcohol precipitation were evaluated. The prehydrolysate:ethanol ratio was increased from 1:1–1:3 in the stepwise precipitation, whereas a ratio of 1:2 was used in the one-step precipitation. The pectin fractions differed from each other in their chemical composition. The molecular weight of the pectin fractions ranged from 474 to 523 kDa, and the degree of esterification of all of them was higher than 50%, so they can be considered high methoxyl pectins. On the other hand, the pretreated solid enriched in glucan was subjected to enzymatic hydrolysis with cellulases and fermented by Clostridium beijerinckii, producing 8.3 g butanol/L after 48 h of fermentation (53 kg butanol/t SBP). As this research has demonstrated, SBP is a suitable feedstock for a green biorefinery approach. The fractionation of SBP by hydrothermal treatment avoids using acid and alkaline solvents and allows high POS and biobutanol production yields.

Analysis of Fecal Microbiota Dynamics in Lupus-Prone Mice using a Simple, Cost-Effective DNA Isolation Method.

Gut microbiota has an important role in educating the immune system. This relationship is extremely important for understanding autoimmune diseases that are not only driven by genetic factors, but also environmental factors that can trigger the onset and/or worsen the disease course. A previously published study on the dynamics of the gut microbiota in lupus-prone MRL/lpr female mice showed how changes of the gut microbiota can alter disease progression. Here, a protocol is described for extracting representative samples from the gut microbiota for studies of autoimmunity. Microbiota samples are collected from the anus and processed, from which the DNA is extracted using a phenol-chloroform method and purified by alcohol precipitation. After PCR is performed, purified amplicons are sequenced using a Next Generation Sequencing platform at Argonne National Laboratory. Finally, the 16S ribosomal RNA gene sequencing data is analyzed. As an example, data obtained from gut microbiota comparisons of MRL/lpr mice with or without CX3CR1 are shown. Results showed significant differences in genera containing pathogenic bacteria such as those in the phylum Proteobacteria, as well as the genus Bifidobacterium, which is considered part of the healthy commensal microbiota. In summary, this simple, cost-effective DNA isolation method is reliable and can help the investigation of gut microbiota changes associated with autoimmune diseases.

Research on batch-to-batch quality control of compound Danshen extract based on physical characterization and multivariate statistical analysis

Physical attributes of Chinese herbal extracts are determined by their chemical components, and the physical and chemical attributes jointly affect the preparation process performance and the final product quality. Therefore, in order to improve the quality control of Chinese herbal extracts, we should comprehensively study the batch-to-batch consistency of physical and chemical attributes as well as the correlations between them. This paper first explored the physical attributes affecting the preparation process performance of the compound Danshen extract and developed a method for characterizing the texture attributes. With such main chemical components as water, phenolic acids, saponins, and saccharides and texture, rheology, and other physical attributes taken into consideration, the batch-to-batch quality fluctuation of products from different production lines and time was analyzed by principal components analysis(PCA). Finally, the correlation and partial least squares(PLS) analysis was conducted, and the regression equation was established. The fitting result of the PLS model for dynamic viscosity was satisfying(R~2Y=0.857, Q~2=0.793), suggesting that the chemical components could be adjusted by the component transfer rate in the extraction process, the impurity removal rate in the alcohol precipitation process, and the water retention rate of the concentration process to meet the control of the extract dynamic viscosity. This study clarified the correlations between physical and chemical attributes of the compound Danshen extract and established a method for controlling its physical attributes based on process regulation, which would provide reference for improving the quality control of Chinese herbal extracts.

Synergistic antitumor effects of polysaccharides and anthocyanins from Lycium ruthenicum Murr. on human colorectal carcinoma LoVo cells and the molecular mechanism.

The antitumor effects of Lycium ruthenicum Murr. polysaccharides (LRPS) and Lycium ruthenicum Murr. anthocyanins (LRAC) were comprehensively investigated in this study. LPRS was obtained by water extraction and alcohol precipitation and further purified using diethylaminoethyl cellulose (DEAE‐Cellulose) and Sephadex G‐75 columns. High‐performance liquid chromatography (HPLC) and Fourier transform‐infrared (FT‐IR) spectroscopy were used to characterize the purified LRPS. The results showed that the purified LRPS contained heteropolysaccharides, mainly composed of arabinose, galactose, and glucose with weight percentage of 41.2%, 33.6%, and 10.8%, respectively. More importantly, LRPS (500 μg/ml) and LRAC (80 μg/ml) failed to impede the proliferation of tumor cells when applied solely (48 h incubation), yet remarkable antineoplastic effects were found once they were applied altogether, since the LoVo cells, a typical human colorectal carcinoma cell line, were significantly inhibited by the mixture of LRPS (150 μg/ml) and LRAC (20 μg/ml) (LRPS&AC) in 24 h. The antineoplastic activity resulted from the combination of both LRPS and LRAC (LRPS&AC), by means of blocking the cell cycle at the G0–G1 phase and inducing LoVo cell apoptosis via reactive oxygen species (ROS)‐dependent pathway. The inhibitory effects of LRPS&AC were specific to the tumor cells, without imposing on the proliferation of normal cells. Western blotting revealed that the antitumor effect was related to the mitochondria‐mediated apoptosis launched by the cross‐action of PI3K/Akt (phosphatidylinositol 3‐kinase/protein kinase B) and JAK2/STAT3 (janus kinase 2/signal transduction and activator of transcription 3) signaling pathways. These findings for the first time reveal the synergistic antitumor effects of LRPS&AC and the related mechanisms, which enable Lycium ruthenicum Murr. to serve as a natural source to develop therapeutic reagents and functional foods with antineoplastic properties.

Polysaccharide-Rich Fractions from Ganoderma resinaceum (Ganodermataceae) as Chemopreventive Agents in N-Diethylnitrosamine-Induced Hepatocellular Carcinoma in Wistar Rats.

Hepatocellular carcinoma (HCC) is one of the most common and lethal diseases worldwide. Its treatment remains ineffective and the prognosis remains severe, thus favoring the emergence of a preventive approach. Mushroom-derived polysaccharides offer great opportunities because of their less toxicity and bioactivities. The present study aimed to investigate the chemopreventive effects of water-soluble polysaccharides from Ganoderma resinaceum on HCC. Two G. resinaceum polysaccharide-rich fractions (GRP I and GRP II) were obtained following hot water and alcohol precipitation. Their proteins, phenol compounds, and total neutral sugar content were assayed. The in vitro antiproliferative effect was assessed in MDA-MB 231, Hela, and HepG2 using the MTT assay. Further, for the in vivo study, seven groups of nine rats each received N-diethylnitrosamine (100 mg/kg BW), vehicle (NaCl 0.9%), doxorubicin (10 mg/kg BW), or G. resinaceum polysaccharides (125 and 250 mg/kg BW). Liver cancer initiation and progression was assessed by evaluating histomorphology of liver section, hepatic injury markers, hematology, cytokines/chemokines levels, and stress oxidative markers. GRP II presented higher protein and sugar and lower phenol compound content than GRP I. GRP exhibited CC50 of 340 and 261.7 in HepG2 cells after 48 h. Moreover, GRP I and GRP II (125 and 250 mg/kg) prevented the alteration of the histoarchitecture of the liver induced by the DEN. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alpha-fetoprotein (AFP), proinflammatory cytokines (G-CSF, IFNγ, and TNFα), and chemokines (eotaxin and fractalkine) levels were significantly decreased in the GRP I- and GRP II-treated groups, while anti-inflammatory cytokines (IL-10 and IL-12p70) levels were increased. The antioxidant defense was also stimulated by reducing malondialdehyde (MDA) and nitric oxide (NO2) levels, increasing catalase (CAT) and superoxide dismutase (SOD) activities, and reducing glutathione (GSH) levels. Our results indicate that GRP I exhibits chemopreventive effects by inhibiting cell proliferation and restoring liver architecture, antioxidant enzymes, and cytokines/chemokines balance.

Structural characterization of a novel polysaccharide from sweet corncob that inhibits glycosylase in STZ-induced diabetic rats : Structural characterization of a novel polysaccharide.

In the current study, we extracted a polysaccharide from sweet corncob and evaluated its hypoglycemic function. After collection in water, alcohol precipitation, and purification by DEAE-52 and Sephadex G-100 columns, we obtained a polysaccharide (SCP50) that was composed primarily of mannose and glucose (9.73:190.27), with a molecular weight of 9280.33Da. We demonstrated that SCP50 exhibited significant inhibition of α-glucosidase activity, with an IC50 of 4.866mg/mL, Km of 1.297 × 10-3, and Vmax of 0.076mol/L·min-1 in vitro. We also observed that SCP50 markedly attenuated disaccharidase (maltase, sucrase, and lactase) activity in a rat model of T2DM. We conclude that SCP50 exerts a hypoglycemic effect via inhibition of intestinal glycosylase. These results thus provide new insight into the hypoglycemic action underlying sweet corncob polysaccharide's effects.

Anti-inflammatory effects of a SERP 30 polysaccharide from the residue of Sarcandra glabra against lipopolysaccharide-induced acute respiratory distress syndrome in mice

Ethnopharmacological relevanceSarcandra glabra (Thunb.) Nakai, a valuable dietetic Chinese herb, is still widely used today. Multiple ingredients of S. glabra with a variety of activities such as anti-inflammatory, antiviral, and antitumor were studied. However, the Sarcandra glabra (Thunb.) Nakai polysaccharide hasn’t been reported for its anti-inflammatory effect. Aim of the studyIn this study, the anti-inflammatory activity of Sarcandra glabra (Thunb.) Nakai polysaccharide was assessed in LPS-induced ARDS mice. Materials and methodsA polysaccharide coded as SERP 30 was obtained by water extraction, alcohol precipitation, and gel filtration. After the physicochemical properties determination and structural characterization, LPS induced-mice ARDS model was used to evaluate the anti-inflammatory and associated antioxidant activities of SERP 30. H&E staining was used to observe the seriousness of lung injury in mice. The ELISA method was used to measure the expression of inflammatory factors (TNF-α and IL-6) in the serum of the mice. The TBA method and the WST-1 method were used to evaluate the oxidative stress injury. Immunohistochemistry was used to distinguish the expression of metalloproteinase-9 (MMP-9), heparinase (HPA), syndecan-1, and decorin in ARDS-mice lung tissue. Western blotting was used to confirm the expression of related proteins in mouse lung tissue. ResultsSERP 30 had a potential role in improving lung damage, reducing inflammation, and preventing oxidative stress. Moreover, SERP 30 significantly attenuated the damage to the endothelial glycocalyx and maintained the integrity of the glycocalyx. The western blotting result implied that the main anti-inflammatory mechanism is directed towards NF-κB and MAPK signaling pathways with inhibiting the activation of associated proteins. ConclusionThis research provides a theoretical basis for treating ARDS by using a byproduct from food resource.

Exploration of nanofiltration conditions for separation of alcohol precipitation liquid during preparation of Reduning Injection by Box-Behnken design combined with flux attenuation

This study employed Box-Behnken design combined with flux attenuation to explore the nanofiltration conditions for separation of alcohol precipitation liquid during the preparation of Reduning Injection and discussed the applicability of nanofiltration in the separation of the liquid with high-concentration ethanol. The effects of nanofiltration molecular weight cut-off(MWCO) and pH on the rejection of chlorogenic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid were consistent with the principles of pore size sieving and charge effect, respectively. The rejection of the three phenolic acids was reduced by concentration polarization effect caused by trans-membrane pressure(TMP). The swelling of membrane surface decreased the pore size and membrane flux for effective separation. Chlorogenic acid and 4,5-dicaffeoylquinic acid were more sensitive to pH and ethanol concentration than 3,5-dicaffeoylquinic acid. A certain correlation existed between the compound structure and the separation factors of nanofiltration, and the separation rules were associated with the comprehensive effect of charge effect, pore size sieving, concentration polarization, steric hindrance and so on.

Extraction of Pectin from Citrus Peels - A Review

Pectin is complex polysaccharides contains 1, 4-linked x and β galactosyluronic acid residues extracted using alcohol precipitation method from citrus peels. The color of pectin from orange peel was pale yellow. Pectin is soluble in hot and cold alkaline water. The pectin is polysaccharide use as a stabilizer in foods. The pectin was extracted from various citrus fruit peels and its optimization by varying pH using hydrochloric acid and citric acid. Pectin can be used to improve the mouth feel and the pulp stability in juice-based drinks and as a stabilizer in acidic protein beverages. Extraction of pectin from citrus fruit peels like Orange peels, sweet orange and lemon gives higher yield of pectin. % Yield and rate of extraction for pectin from orange and sweet lemon (Mosambi) peels depends on parameters such as pH, temperature, solvent, time of extraction. Extraction carried out at pH 1.5 at 60 min with 60,70 & 80 OC temperature. The higher yield of pectin at pH value 1.5 and contact time 60 min. The extraction of pectin such as Citric Acid, Hydrochloric Acid, Sulphuric Acid, Nitric Acid and oxalic acid (C2H2O4). The time of extraction increased to an extreme there is less effect on yield of pectin reported and also decreased from maximum level due to thermal degradation of the extracted pectin.

Exopolysaccharides from the Energy Microalga Strain Botryococcus braunii: Purification, Characterization, and Antioxidant Activity.

Botryococcus braunii, a prestigious energy microalga, has recently received widespread attention because it can secrete large amounts of exopolysaccharides (EPS) with potential applications in food, cosmetics, and nutraceuticals. Unfortunately, the insufficiency of research on the bioactivity and structure–activity relationship of B. braunii EPS has impeded the downstream applications. In the present study, alcohol precipitation, deproteinization, and DEAE-cellulose column chromatography were used to extract and purify B. braunii SCS-1905 EPS. It was found that B. braunii SCS-1905 EPS were high-molecular-weight heteropolysaccharides containing uronic acid (7.43–8.83%), protein (2.30–4.04%), and sulfate groups (1.52–1.95%). Additionally, the EPS primarily comprised galactose (52.34–54.12%), glucose (34.60–35.53%), arabinose (9.41–10.32%), and minor amounts of fucose (1.80–1.99%), with the presence of a pyranose ring linked by a β-configurational glycosidic bond. Notably, the antioxidant activity of crude exopolysaccharides (CEPS) was stronger, and the half maximal inhibitory concentration (IC50) for ABTS and hydroxyl radicals was significantly lower than that of deproteinized exopolysaccharides (DEPS). Overall, this study indicated a potential application of B. braunii SCS-1905 EPS as a natural antioxidant. In summary, B. braunii EPS could be used as a potential feedstock for the production of antioxidant health foods.

Hypouricemic effect of gallic acid, a bioactive compound fromSonneratia apetalaleaves and branches, on hyperuricemic mice

As a tropical medicinal plant, Sonneratia apetala is mainly distributed in the southeast coastal areas of China. Recently, the hypouricemic effect of Sonneratia apetala leaves and branches (SAL) has been reported, but the active compound and its mechanism are unclear. Thus, this study aims to explore the effective fraction of SAL and the mechanism of its active compound on uric acid formation and excretion. SAL was extracted with ethyl acetate and concentrated to obtain solvent-free extracts (SAL-EA). The remains fraction (SAL-E) and the supernatant fraction (SAL-S) of SAL resulting from water extraction and alcohol precipitation were collected and dried. The effects of different fractions were explored on hyperuricemic mice. SAL-S showed excellent activities in decreasing the levels of uric acid (UA), blood urea nitrogen (BUN), and creatinine (CRE) in serum and in attenuating kidney damage. Then, the active compound gallic acid (GA) identified by HPLC was assayed for its mechanism of regulating uric acid metabolism in hyperuricemic mice. The hypouricemic effect of GA was probably associated with the downregulation of URAT1 and GLUT9, upregulation of ABCG2 and decreased activities of adenosine deaminase (ADA) and xanthine oxidase (XOD). Moreover, GA suppressed the level of MDA, IL-6, IL-1β, TNF-α, TGF-β1, COX-2 and cystatin-C (Cys-C), and enhanced the activities of SOD, GSH-Px, CAT, and Na+-K+-ATPase (NKA) in the kidneys. These results indicated that GA protects against hyperuricemia-induced kidney injury via suppressing oxidative stress and inflammation as well as decreasing the serum levels of UA by regulating urate transporters.

Anticancer and apoptotic effects of a polysaccharide extract isolated from Lactarius chrysorrheus Fr. in HepG2 and PANC-1 cell lines

Mushrooms are widely used in many cultures for nutritional and health benefits. The Lactarius species is found in the Aegean region of Turkey. Lactarius chrysorrheus Fr. is a wild mushroom that contains a milky juice. In this study, we investigated the in vitro cytotoxic potential and apoptotic effect of the polysaccharide extract prepared from L. chrysorrheus by water extraction and alcohol precipitation using the tetrazolium MTT dye assay, annexin V staining, wound healing and colony formation, and qRT-PCR. The molecular weights of three peaks observed in HPLC chromatograms were calculated as 1869.9, 3043.92 and 16821.47 Da. The extract exhibited cytotoxic activity at 72 h, with an IC50 value of 296.42 ?g/mL in HepG2 and 444.43 ?g/mL in PANC-1 cells; the extract that was tested on the normal HEK293 cell line exhibited no cytotoxicity. Further, L. chrysorrheus upregulated the expression of CASPASE 3 and CASPASE 9 while downregulating B-cell lymphoma 2 (BCL-2) and B-cell lymphoma-extra large (Bcl-xL) genes, and inhibited cell migration and colony formation in HepG2 and PANC-1 cells. This study provides new insight into the use of the polysaccharide from L. chrysorrheus in the development of novel anticancer agents.

Structural characterization and in vitro hypoglycaemic activity of glucomannan from Anemarrhena asphodeloides Bunge.

A new polysaccharide (AABP-2B) was obtained from Anemarrhena asphodeloides Bunge after purification by gradient alcohol precipitation and DEAE-52 cellulose column chromatography. AABP-2B was confirmed to be a homogeneous polysaccharide with a molecular weight of 5800 Da and was composed of mannose and glucose at a molar ratio of 7.2 : 2.8. Structural analysis demonstrated that the backbone of AABP-2B was mainly composed of 4)-β-D-Manp-(1, 4,6)-β-D-Glcp-(1 and 3,6)-β-D-Manp-(1. The hypoglycaemic effect of AABP-2B was evaluated by its inhibition of α-glucosidase activities and insulin resistance in a HepG2 cell model. The results showed that AABP-2B displayed α-glucosidase inhibitory activities and could significantly improve glucose consumption by activating the IRS-1/PI3K/Akt signalling pathway in insulin-resistant HepG2 cells. Hence, AABP-2B may have potential as a functional food or medicine for diabetes therapy.

СРАВНИТЕЛЬНАЯ ОЦЕНКА РАНОЗАЖИВЛЯЮЩЕЙ АКТИВНОСТИ МАЗЕЙ, ПОЛУЧЕННЫХ НА ОСНОВЕ НЕКОТОРЫХ БИОЛОГИЧЕСКИ АКТИВНЫХ КОМПЛЕКСОВ ИЗ ВЫСШИХ БАЗИДИАЛЬНЫХ ГРИБОВ

Introduction. In connection with the acquisition of antibiotic resistance of pyogenic microbiota, the problem of topical (drug) treatment of wounds has become aggravated. Aim. Comparative in vivo evaluation of the medicinal properties of ointments prepared on the basis of some biologically active complexes from higher basidiomycetes. Materials and methods. Experimental ointments were obtained on the basis of melanins from the natural raw materials of the сhaga Inonotus obliquus and from its deep-liquid culture, total polysaccharides from the oyster mushroom Pleurotus ostreatus, as well as the melanin polysaccharide complex from these mushrooms. Melanins were obtained by alkaline hydrolysis. Melanin-containing ointments were prepared using two formulations of ointment bases (the ointment base included saline, vaseline oil, emulsifiers T-2 and Tween-60, in the second case — saline, lanolin as an emulsifier and vaseline) in the first case. Total polysaccharides were obtained by 96% ethyl alcohol precipitation from an aqueous extract of a fungal homogenate heated in a boiling water bath for 12 h, followed by purification from fractions insoluble in distilled water. Melanin-polysaccharide-containing ointments were prepared on the basis of melanins from the natural raw materials of the сhaga Inonotus obliquus and from its deep culture, total polysaccharides from the oyster mushroom Pleurotus ostreatus, the ratios in the ointment formulations being (1:1 and 1:2). The ointment base of polysaccharide-containing and melanin-polysaccharide-containing ointments was similar to the specified second formulation. The wound healing efficacy of the ointments was evaluated in eight groups (20 mice each) of healthy non-inbred mice of both sexes, weighing 17–19 g, of the ICR colony of the State Scientific Center for Virology and Biotechnology Vector. The control group was a group of mice, in the treatment of which Levomicon-TFF reference ointment was used (20 mice in the group). The treatment of mice was continued until their incised skin wounds healed. The motor activity, appetite, and dynamics of wound healing were evaluated daily in mice of all groups, the change in their body weight was assessed every three days, and a histomorphological study of the incised skin wound was performed. The areas of wounds, the percentage of reduction in the area of wounds, the rate of wound contraction, and the rate of wound healing were determined. Results. It was shown that the components of the ointment bases of the two tested ointment formulations, melanin from the natural raw materials of the сhaga and from its deep-liquid culture, the total polysaccharides of the oyster mushroom, as well as the melanin polysaccharide complexes of these mushrooms, do not have toxicity for experimental animals in the concentrations of the tested ointment formulations. Ointments containing melanins from natural raw materials and deep-liquid culture of the сhaga Inonotus obliquus have similar regenerative properties, ointments with a fivefold increase in the content of these melanins, polysaccharide-containing and melanin-polysaccharide-containing ointments, have provided significantly accelerated wound healing in experimental animals. At the same time, the development of granulation tissue in the wound and the replacement of the wound defect in the skin with stratified squamous epithelium as early as on the 12th day in these groups of animals, were demonstrated in comparison with mice of the control group treated with the reference ointment Levomicon-TFF. The epidermal defect in animals of the control group was eliminated only on the 21st day. Conclusion. Ointments containing melanins from the natural raw materials of сhaga and from its deep-liquid culture, polysaccharides from the oyster mushroom, as well as a complex of these substances from these mushrooms, are superior in wound healing effectiveness, as compaired to the control ointment Levomicon-TFF, and can be considered as promising for topical treatment of wounds.

In vitro Digestion and Fermentation Characteristics of Polysaccharides from Camellia nitidissima Chi

Polysaccharides were isolated from Camellia nitidissima Chi by water extraction and alcohol precipitation, then the polysaccharide extract was separated and purified by using DEAE-52 cellulose chromatography column. The digestion characteristics of the polysaccharides were studied in vitro by employing a simulated upper gastrointestinal model and analyzing molecular weight, reducing sugar and free monosaccharide ontents of digested products at different digestion times. The fermentation characteristics of polysaccharides were studied in vitro by using human fecal fermentation model and analyzing pH, total sugar, reducing sugar and short chain fatty acid content ,as well as the Lactobacillus, Bifidobacterium and Escherichia coli number. The results showed that, TPS1, TPS2, and TPS3 were obtained after purification of the polysaccharide from Camellia nitidissima Chi. During in vitro digestion, the molecular weight of TPS1, TPS2, TPS3 and the reducing sugar content did not changed by simulated saliva and simulated intestinal juice. Simulated gastric juice caused a slight decrease in the molecular weight of TPS1, TPS2, and TPS3, and the content of reducing sugar increased from 0.129±0.016, 0.155±0.026, 0.147±0.017 mmol/L to 0.223±0.018, 0.319±0.013, 0.294±0.030 mmol/L. Free monosaccharides were not detected during the digestion. During the fermentation process, the pH of the group added with TPS1, TPS2, and TPS3 was significantly lower than that of the blank control group, the sugar consumption rate was 91.4%, 89.0%, 94.5%, and the reducing sugar content increased during 0~18 h, reduced during 18~48 h; Compared with the blank control group, the levels of acetic acid, propionic acid, and butyric acid were all significantly increased and the order of increasing multiple size was TPS3>TPS2>TPS1. Further, TPS1, TPS2 and TPS3 could promote the growth of Lactobacillus and Bifidobacterium, inhibit the growth of Escherichia coli, and the order of probiotic effects was TPS3>TPS2>TPS1. In summary, the polysaccharides were isolated from Camellia nitidissima Chi were not digested in vitro, and could be used by fecal microflora to produce a large amount of acetic acid, propionic acid and butyric acid, which had potential probiotic effects. And TPS3 had better probiotic effects than TPS2 and TPS1.

Structural Characterization and Immunomodulatory Activities of Polysaccharides from Suillus granulatus

The present study was aimed to investigate the structural characteristics and immunomodulatory activity of Suillus granulatus polysaccharides. Two polysaccharide fractions(SGP3-1 and SGP3-2) were purified from the fruiting bodies of Suillus granulatus by water extraction and alcohol precipitation, deproteinization, and column chromatography separation. The physicochemical properties of SGP3-1 and SGP3-2 were characterized by high-performance gel permeation chromatography(HPGPC), high-performance liquid chromatography(HPLC), Fourier transform infrared spectroscopy(FT-IR), nuclear magnetic resonance spectroscopy(NMR), X-ray diffraction(XRD), etc. The results showed that the weight-average molecular weight of SGP3-1 and SGP3-2 were 170.48 kDa and 14.52 kDa, respectively. Both SGP3-1 and SGP3-2 were low-crystallinity amorphous polymers, and had no triple-helix conformation. Additionally, SGP3-1 characterized by α-type glycosidic linkage was mainly composed glucose(55.47%), mannose(20.63%) and galactose(14.04%), while SGP3-2 characterized by β-type glycosidic linkage was mainly composed of glucose(82.43%). The results from RAW264.7 cells in vitro suggested that SGP3-1 and SGP3-2 showed no cytotoxicity in the concentration range from 5 to 40 μg/mL. Moreover, SGP3-1 and SGP3-2 could significantly enhance the phagocytic activity and promote the production of nitric oxide(NO), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) through activating mitogen-activated protein kinases(MAPKs), phosphatidylinositol-3-kinase(PI3K)/Akt, and nuclear factor-κB(NF-κB) signaling pathways. Furthermore, SGP3-2 exhibited better immunomodulatory activity than SGP3-1. It showed that SGP3-1 and SGP3-2 could be used to develop functional foods, as dietary supplements for people with low immunity, and provide a theoretical basis and research basis for the further development of Boletus edulis.

Research on structure and antioxidant activity of polysaccharides from Ginkgo biloba leaves

Ginkgo biloba extract is regarded as one of the most popular natural plant preparations, among which polysaccharide is considered to be an important functional factor. In the study, polysaccharide was extracted from Ginkgo biloba leaves powder by ultrasonic-enzyme assisted water-soluble alcohol precipitation. The content of crude polysaccharide was 82.4% by the phenol sulfuric acid method. Structure and chemical composition of Ginkgo biloba leaves polysaccharide were investigated by chromatographic and nuclear magnetic resonance spectoscocopy analyses. Results showed that the purified Ginkgo biloba leaves polysaccharide was an acidic polysaccharide with a molecular weight of 12.08 kDa, containing glycosidic linkages of β-D-Rhap-(1→, β-L-Xylp-(1→, →4)-β-D-Manp-(1→, →4)-β-D-Glcp-(1→, →6)-β-D-Manp-(1→, →3)-β-L-Araf-(1→, →4)-β-D-Galp-(1→, →3,4)-β-D-Glcp-(1→. The monosaccharides were composed of rhamnose, arabinose, xylose, mannose, glucose and galactose with a molar ratio of 5.56:3.84:2.34:7.54:8.34:1, respectively. Scanning electron microscopy analysis showed that the microscopic conformation of the purified Ginkgo biloba leaves polysaccharide was fluffy and irregular honeycomb with certain pores. Antioxidant activities in vitro, including scavenging 1,1-Diphenyl-2-picrylhydrazine, scavenging hydroxyl radical activity and scavenging superoxide anion were 78.8%, 79.2% and 43.7%, respectively, which suggested that the purified Ginkgo biloba leaves polysaccharide had strong antioxidant activities. These findings indicate that Ginkgo. biloba leaves polysaccharides have potential as natural antioxidants in food processing.

Protective role of ergothioneine isolated from Pleurotus ostreatus against dextran sulfate sodium-induced ulcerative colitis in rat model.

The well-acknowledged sulfur-containing amino acid ergothioneine (EGT), which is usually perceived to be produced from fungi, cyanobacteria, and mushrooms, has been increasingly demonstrated as a diet-derived antioxidant with therapeutic potential. However, little is known regarding the therapeutic benefit against ulcerative colitis of EGT. Meanwhile, few studies have reported whether EGT could be produced by Pleurotus ostreatus, which is the second most cultivated edible mushroom worldwide, with ideal economic and ecological values and medicinal properties. In the current study, EGT was primarily extracted and isolated from P. ostreatus by water extraction and alcohol precipitation. Then, the Sephadex G-10 gel permeation chromatography and preparative high-performance liquid chromatography were employed in order to purify and collect EGT. Based on nuclear magnetic resonance spectrum characterization, the content of EGT extracted from P. ostreatus is 1.916mg/g (the weight of EGT is the same as that of dry P. ostreatus). Finally, both the antioxidant activity in vitro and potential effects of EGT on dextran sulfate sodium-induced ulcerative colitis in rats were investigated. The results showed that the collected EGT exhibited strong antioxidant activity with a concentration-dependent manner in 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis (3-ethylbenzoline-6-sulphonic acid) assays. The oral administration of EGT can significantly alleviate colon length shortening and colonic pathological damage, mediated by downregulating the expression of pro-inflammatory factors and inhibiting TLR4/MyD88/NF-κB signaling pathway. Therefore, this study demonstrated that EGT from P. ostreatus would be an excellent dietary compound to ameliorate ulcerative colitis. PRACTICAL APPLICATION: In order to develop Pleurotus ostreatus as a functional food and related products for the prevention and treatment of ulcerative colitis, the separation and purification technology of ergothioneine (EGT) from P. ostreatus was studied, which not only provided a new and broader source for the acquisition and development of EGT, but also provided a basis for the development of EGT as a drug against UC.

Pharmacological Investigations in Traditional Utilization of Alhagi maurorum Medik. in Saharan Algeria: In Vitro Study of Anti-Inflammatory and Antihyperglycemic Activities of Water-Soluble Polysaccharides Extracted from the Seeds.

The anti-inflammatory and antihyperglycemic effects of polysaccharides extracted from Alhagi maurorum Medik. seeds, spontaneous shrub collected in Southern of Algerian Sahara were investigated. Their water extraction followed by alcoholic precipitation was conducted to obtain two water-soluble polysaccharides extracts (WSPAM1 and WSPAM2). They were characterized using Fourier transform infrared, 1H/13C Nuclear Magnetic Resonance, Gas Chromatography-Mass Spectrometry and Size Exclusion Chromatography coupled with Multi-Angle Light Scattering. The capacity of those fractions to inhibit α-amylase activity and thermally induced Bovine Serum Albumin denaturation were also investigated. WSPAM1 and WSPAM2 were galactomannans with a mannose/galactose ratio of 2.2 and 2.4, respectively. The SEC-MALLS analysis revealed that WSPAM1 had a molecular weight of 1.4 × 106 Da. The investigations highlighted antinflammatory and antihyperglycemic effects in a dose-dependant manner of WSPAM1 and WSPAM2.

Genetic markers associated with insecticide resistance and resting behaviour in Anopheles gambiae mosquitoes in selected sites in Kenya

BackgroundMolecular diagnostic tools have been incorporated in insecticide resistance monitoring programmes to identify underlying genetic basis of resistance and develop early warning systems of vector control failure. Identifying genetic markers of insecticide resistance is crucial in enhancing the ability to mitigate potential effects of resistance. The knockdown resistance (kdr) mutation associated with resistance to DDT and pyrethroids, the acetylcholinesterase-1 (ace-1R) mutation associated with resistance to organophosphates and carbamates and 2La chromosomal inversion associated with indoor resting behaviour, were investigated in the present study.MethodsAnopheles mosquitoes sampled from different sites in Kenya and collected within the context of malaria vector surveillance were analysed. Mosquitoes were collected indoors using light traps, pyrethrum spray and hand catches between August 2016 and November 2017. Mosquitoes were identified using morphological keys and Anopheles gambiae sensu lato (s.l.) mosquitoes further identified into sibling species by the polymerase chain reaction method following DNA extraction by alcohol precipitation. Anopheles gambiae and Anopheles arabiensis were analysed for the presence of the kdr and ace-1R mutations, while 2La inversion was only screened for in An. gambiae where it is polymorphic. Chi-square statistics were used to determine correlation between the 2La inversion karyotype and kdr-east mutation.ResultsThe kdr-east mutation occurred at frequencies ranging from 0.5 to 65.6% between sites. The kdr-west mutation was only found in Migori at a total frequency of 5.3% (n = 124). No kdr mutants were detected in Tana River. The ace-1R mutation was absent in all populations. The 2La chromosomal inversion screened in An. gambiae occurred at frequencies of 87% (n = 30), 80% (n = 10) and 52% (n = 50) in Baringo, Tana River and Migori, respectively. A significant association between the 2La chromosomal inversion and the kdr-east mutation was found.ConclusionThe significant association between the 2La inversion karyotype and kdr-east mutation suggests that pyrethroid resistant An. gambiae continue to rest indoors regardless of the presence of treated bed nets and residual sprays, a persistence further substantiated by studies documenting continued mosquito abundance indoors. Behavioural resistance by which Anopheles vectors prefer not to rest indoors may, therefore, not be a factor of concern in this study’s malaria vector populations.