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In vitro antioxidant and anti-inflammatory activities of fluidized bed ethanol extracts of Pleurotus ostreatus PQMZ91109, PBS281009, M2191 (obtained from a laboratory greenhouse), Agaricus bisporus, Marasmius oreades, Craterellus cornucopioides, and Tuber melanosporum were investigated using free radical scavenging activities, metal chelating effects, inhibition of lipid peroxidation (inhibition of peroxyl radicals), xanthine oxidase, and lipoxygenase, and identification of antioxidant compounds. Levels of different compounds with antioxidant properties demonstrated the presence of more tocopherols than polyphenol carboxylic acids and flavones. HPLC showed that T. melanosporum extracts had high correlation values between levels of different active compounds and antioxidant assay results. Tocopherol levels were highly correlated (R 2=0.867) with anti-inflammatory activities and inhibition of lipid peroxidation. Fluidized bed ethanol extracts from dried mushroom fruit bodies can be a source of products rich in tocopherols, and can constitute an important ingredient in food supplements.

A new mannose-recognizing lectin (MOL) was purified on an asialofetuin-column from fruiting bodies of Marasmius oreades grown in Japan. The lectin (MOA) from the fruiting bodies of the same fungi is well known to be a ribosome-inactivating type lectin that recognizes blood-group B sugar. However, in our preliminary investigation, MOA was not found in Japanese fruiting bodies of M. oreades, and instead, MOL was isolated. Gel filtration showed MOL is a homodimer noncovalently associated with two subunits of 13 kDa. The N-terminal sequence of MOL was blocked. The sequence of MOL was determined by cloning from cDNA and by protein sequencing of enzyme-digested peptides. The sequence shows mannose-binding motifs of bulb-type mannose-binding lectins from plants, and similarity to the sequences. Analyses of sugar-binding specificity by hemagglutination inhibition revealed the preference of MOL toward mannose and thyroglobulin, but asialofetuin was the strongest inhibitor of glycoproteins tested. Furthermore, glycan-array analysis showed that the specificity pattern of MOL was different from those of typical mannose-specific lectins. MOL preferred complex-type N-glycans rather than high-mannose N-glycans.

Nitric oxide (NO) is a gaseous, radical molecule that plays a role in various physiological processes. Previously, we reported that transduction of murine colon cancer cells (MC38) with herpes simplex virus thymidine kinase (HSV-tk) gene resulted in a significant over-expression of cyclooxygenase-2 (COX-2) and activation of NF-kB pathway. In this study we show that TNFα, but not LPS, was significantly able to stimulate the production of NO in HSV-tk transduced 9L glioblastoma cell lines, mediated by the up-regulation of iNOS transcript and iNOS protein. The TNFα-induced up-regulation of iNOS expression was mediated by MAPK and NF-κB signaling pathways as revealed by using selective pharmaceutical inhibitors. A culture liquid extract of the edible and medicinal mushroom Marasmius oreades that was previously shown to inhibit iNOS expression in MCF-7 was utilized to prepare fractions and evaluate their ability to affect TNFα-induced iNOS expression in HSV tk transduced 9L cell lines. While most of the tested fractions were shown to inhibit TNFα-induced iNOS expression, they targeted different signaling pathways in a selective fashion. Here, we report that fraction SiSiF1 interfered with IKBα phosphorylation and consequently interfered with NF-κB activation pathway. SiSiF1 showed minimal interference with the phosphorylation of p38 and JNK proteins. In contrast, fraction SiSiF3 selectively inhibited the phosphorylation of p38 and fractions SiSiF4 and SiSiF5 selectively inhibited the phosphorylation of JNK with no observed effect against IKBα and p38 phosphorylation. Our data illustrate the complexity of iNOS regulation in HSV tk transduced 9L cell lines and also the richness of natural products with bioactive substances that may act synergistically through different signaling pathways to affect iNOS gene expression.

MCF7 breast cancer cell line, carrying a luciferase reporter gene under the control of nuclear factor kappa B (NF-kappaB)-responsive promoter, was established and used for the screening of fungal organic extracts for their ability to interfere with the NF-kappaB activation pathway. Twenty-eight crude fungal extracts, out of 242, were found to inhibit NF-kappaB reporter activity by more than 40%. Furthermore, positive extracts were used to evaluate their antiproliferative activity as well as their ability to influence the phosphorylation and degradation levels of IkappaBa. Fungal extracts prepared from Marasmius oreades and Cyathus striatus showed significant inhibitory effects on the NF-kappaB activation pathway. Taken together, our results support the notion of the presence of novel activities that might be utilized as cancer therapeutics.

A novel lectin from the mushroom Marasmius oreades (MOA) has been shown to bind to human blood group B oligosaccharides [1]. In the present work we examine the binding of a series of analogues of the blood group B-trisaccharide, alphaGal(1-3)[alphaFuc(1-2)]betaGal-OR (1, R = (CH2)8COOMe). MOA was biotinylated and immobilized on a micro column (9.8 microL) for evaluation by Frontal Affinity Chromatography-Mass Spectrometry (FAC-MS) [2]. The trisaccharide 1 was found to be the epitope needed for maximum recognition (Kd = 3.6 microM). A series of synthetic deoxygenated and O-methylated analogues of the B-trisaccharide (R = OMe) were then screened against the lectin, and the key structural elements for binding were determined. OH-4 of the beta-Gal residue and OH-2 of the alpha-Gal residue were found to be critical for recognition. The FAC-MS technique also proved powerful in evaluating mixtures of compounds. Since the solution NMR structure and crystal structure of the B-trisaccharide are known [3], we propose the specific surface of the trisaccharide that is recognized by the lectin.

The impact on the environment of Cd, Pb and Zn emissions from the A1 motorway, one of the busiest in France, was based on a simultaneous analysis of lichens, fungi and soil.The study was conducted in 12 rest areas located along a 55 kmstretch of the motorway. Cd, Pb and Zn concentrations in the top soil horizon (0–10 cm) largely exceeded their concentrations at the control site.The Cd-, Pb- and Zn levels in fruiting bodies of fungi Armillaria mellea, Coprinus comatus, Marasmius oreades and Paxillus involutus were highly variable between sites. The contamination degree of transplanted thalliof Parmelia caperata was well correlated to the cumulated traffic on the western side. On the eastern side, heavy metals originated from the traffic but also from neighbouring sources (urbanization and agricultural practices). Although lead levels are decreasing, leaded petrol is still on offer and lead remains an indicator of vehicle pollution.

Except when cultures were well aerated by shaking,Marasmius oreades (Bolt.) Fr., the fairy ring mushroom, was able to utilize, in pure culture studies, fractions of humus from sources as divergent as the A horizon of a chernozemic soil and the Bh horizon of a Podzol. The importance of this ability to soil infestation was discussed and it was suggested that improved soil aeration be studied as a control measure.