No abstract

The study was designed to investigate the effects of extract of Clitocybe nuda (CNE) on type 1 diabetes mellitus and dyslipidemia in streptozotocin- (STZ-) induced diabetic mice. Diabetes was induced by injection of STZ. Diabetic mice were randomly divided into five groups and given orally CNE (C1: 0.2, C2: 0.5, and C3: 1.0 g/kg body weight) or metformin (Metf) or vehicle for 4 weeks. STZ induction decreased in the levels of insulin, body weight, and the weight of skeletal muscle, whereas the levels of blood glucose, hemoglobin nonenzymatically (percent HbA1c), and circulating triglyceride (P < 0.001, P < 0.001, and P < 0.01, resp.) were increased. CNE decreased the levels of blood glucose, HbA1c, and triglyceride levels, whereas it increased the levels of insulin and leptin compared with the vehicle-treated STZ group. STZ induction caused a decrease in the protein contents of skeletal muscular and hepatic phosphorylation of AMP-activated protein kinase (phospho-AMPK) and muscular glucose transporter 4 (GLUT4). Muscular phospho-AMPK contents were increased in C2-, C3-, and Metf-treated groups. CNE and Metf significantly increased the muscular proteins of GLUT4. Liver phospho-AMPK showed an increase in all CNE- and Metf-treated groups combined with the decreased hepatic glucose production by decreasing phosphenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), and 11beta hydroxysteroid dehydroxygenase (11β-HSD1) gene, which contributed to attenuating diabetic state. The study indicated that the hypoglycemic properties of CNE were related to both the increased muscular glucose uptake and the reduction in hepatic gluconeogenesis. CNE exerts hypolipidemic effect by increasing gene expressions of peroxisome proliferator-activated receptor α (PPARα) and decreasing expressions of fatty acid synthesis, including acyl-coenzyme A: diacylglycerol acyltransferase (DGAT) 2. Therefore, amelioration of diabetic and dyslipidemic state by CNE in STZ-induced diabetic mice occurred by regulation of GLUT4, PEPCK, DGAT2, and AMPK phosphorylation.

This study was designed to characterize seed-colonizing microbial communities that were previously shown to be involved in the suppression of seedling disease caused by Pythium ultimum in a municipal biosolids compost. Selective microbial inhibitors were employed to inactivate portions of the microbial community associated with seed germinated in a compost medium to evaluate their impact on disease suppression. After initial screenings for toxicity to both cucumber and P. ultimum, six selective inhibitors were eventually used to assess the impact of seed treatment on the reduction of bacterial and fungal populations and on disease suppression. Rifampicin was the most effective inhibitor for inactivating disease suppression. Bacterial communities that colonized cucumber seed sown in compost medium for 8 h and seed sown in compost medium for 8 h followed by a 3-h treatment of either rifampicin at 500 ppm or water were dislodged from seed surfaces and subjected to RNA extraction and reverse transcription to cDNA. Differences in the composition of seed-colonizing bacterial communities were assessed using terminal restriction fragment length polymorphisms (T-RFLP) of polymerase chain reaction-amplified 16S rDNA genes. T-RFLP profiles revealed a diversity of distinct bacterial taxa, a number of which dominate seed surfaces within 8 h of sowing. Analysis of similarity (ANOSIM) using terminal restriction fragment (T-RF) presence or absence showed that community profiles of nontreated and water-treated seed were quite similar whereas community profiles from rifampicin-treated seed were distinct. Differences in community profiles based on T-RF abundance (peak height and peak area) indicated that all treatments were unique (ANOSIM, all pairwise comparisons P < 0.05) Peaks heights and areas of relatively few T-RFs were reduced to zero following rifampicin treatment and 34 T-RFs explained 85% of the observed difference between treatments. Tentative taxon assignments for each of the T-RFs that contributed to the treatment differences revealed a preponderance of sequences with affinities to the α-, β-, and γ-Proteobacteria and Firmicutes. Limited sequencing of clones associated with water-treated and rifampicin-treated seed revealed the presence of similar taxa dominated by members of the γ-Proteobacteria. Many species within these taxa (such as Pseudomonas spp., Enterobacter spp., and Bacillus spp.) are known to be suppressive to Pythium diseases. Results of our study have confirmed that Pythium disease suppression in a municipal biosolids compost is mediated by compost-associated bacteria that colonize seed within hours after sowing. By focusing on actively growing microbes in the infection court during important stages of pathogen infection, we believe we can more efficiently determine the mechanisms of disease suppression and the microbes involved. Although specific to this pathosystem and compost, our results have a much broader scope of inference and illustrate the utility of such a targeted approach in identifying a rela...

This work represents the first evaluation of the effects of water extract of C. nuda (WE-CN), an edible mushroom, on murine bone marrow-derived dendritic cells (BMDCs) and the potential pathway through which the effects are mediated. Our experimental results show that WE-CN could induce phenotypic maturation of DCs, as shown by the increased expression of MHC and costimulatory molecules. In addition, it also induced the proinflammatory cytokines expression on DCs and enhanced both the proliferation and IFN-γ secretion of allogenic T cells. Therefore, since WE-CN did not induce maturation of DCs generated from mice with mutated TLR-4 or TLR-2, suggesting that TLR4 and TLR2 might function as membrane receptors for WE-CN. Moreover, the mechanism of action of WE-CN may be mediated by increased phosphorylation of ERK, p38, and JNK mitogen-activated protein kinase (MAPK) and increased NF-κB p65 activity, which are important signaling molecules downstream of TLR-4 and TLR-2. Finally, coimmunization of mice with WE-CN and a HER-2/neu DNA vaccine induced a HER-2/neu-specific Th1 response that resulted in significant inhibition of HER-2/neu overexpressing mouse bladder tumor (MBT-2) growth. These data suggest that WE-CN induces DC maturation through TLR-4 and/or TLR-2 and that WE-CN can be used as an adjuvant in cancer vaccine immunotherapy.

The objective of this study was to evaluate the antihyperlipidemic and antihyperglycemic effects and mechanism of the extract of Clitocybe nuda (CNE), in high-fat- (HF-) fed mice. C57BL/6J was randomly divided into two groups: the control (CON) group was fed with a low-fat diet, whereas the experimental group was fed with a HF diet for 8 weeks. Then, the HF group was subdivided into five groups and was given orally CNE (including C1: 0.2, C2: 0.5, and C3: 1.0 g/kg/day extracts) or rosiglitazone (Rosi) or vehicle for 4 weeks. CNE effectively prevented HF-diet-induced increases in the levels of blood glucose, triglyceride, insulin (P < 0.001, P < 0.01, P < 0.05, resp.) and attenuated insulin resistance. By treatment with CNE, body weight gain, weights of white adipose tissue (WAT) and hepatic triacylglycerol content were reduced; moreover, adipocytes in the visceral depots showed a reduction in size. By treatment with CNE, the protein contents of glucose transporter 4 (GLUT4) were significantly increased in C3-treated group in the skeletal muscle. Furthermore, CNE reduces the hepatic expression of glucose-6-phosphatase (G6Pase) and glucose production. CNE significantly increases protein contents of phospho-AMP-activated protein kinase (AMPK) in the skeletal muscle and adipose and liver tissues. Therefore, it is possible that the activation of AMPK by CNE leads to diminished gluconeogenesis in the liver and enhanced glucose uptake in skeletal muscle. It is shown that CNE exhibits hypolipidemic effect in HF-fed mice by increasing ATGL expression, which is known to help triglyceride to hydrolyze. Moreover, antidiabetic properties of CNE occurred as a result of decreased hepatic glucose production via G6Pase downregulation and improved insulin sensitization. Thus, amelioration of diabetic and dyslipidemic states by CNE in HF-fed mice occurred by regulation of GLUT4, G6Pase, ATGL, and AMPK phosphorylation.