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Kanno T.,Hokkaido University | Kawamura S.,Hokkaido University of Education | Harada E.,Iwade Research Institute of Mycology | Kameya H.,Japan National Food Research Institute | And 3 more authors.
Nippon Shokuhin Kagaku Kogaku Kaishi | Year: 2013

We evaluated a variety of radical scavenging abilities of hot extracts of mushrooms, by employing the electron spin resonance (ESR) spin trapping method and oxygen radical absorbance capacity (ORAC) assay. The ESR spin trapping method using 5-(2,2-dimethyl-1,3-propoxycyclophosphoryl)-5-methyl-1-pyrroline-N- oxide (CYPMPO), as the spin trapping reagent, identified the hydroxyl radical (OH), superoxide radical (O2 -), alkoxyl radical (RO) and singlet oxygen (1O2). The ESR adduct signals were sensitive and stable. The trapping activity indicated L-ascorbic acid (from OH), α-lipoic acid (from O2 -), glutathione (from RO) and 1O2 equivalents. The radical scavenging abilities of hot water extracts from 13 mushroom species did not correlate with the corresponding ORAC values. However, our results indicate that an important feature of methods for evaluating antioxidant activities should be the determination of the radical scavenging abilities by ESR spin trapping. Source


Endo M.,Health Science University | Beppu H.,Mie University | Akiyama H.,Health Science University | Wakamatsu K.,Health Science University | And 6 more authors.
Biochimica et Biophysica Acta - General Subjects | Year: 2010

Background: Mushrooms of the genus Agaricus are a common folk remedy against carcinoma. The active ingredients, polysaccharides and protein-polysaccharide complexes containing β-glucan, have been isolated and shown to have indirect tumor-suppressing activity via an immunological activation. Methods: The diffusible fraction of a hot-water extract of Agaricus blazei Murrill (ABM) powder was fractionated by HPLC based on the anti-tumor activity against leukemic cells in vitro. The structure of the anti-tumor substance was determined by NMR and MS analyses. Results: We purified a tumorcidal substance from the diffusible fraction of ABM and identified it as agaritine, β-N-(γ-l(+)-glutamyl)-4-(hydroxymethyl) phenylhydrazine, having a molecular mass of 267Da. This compound inhibited the proliferation of leukemic cell lines such as U937, MOLT4, HL60 and K562 with IC 50 values of 2.7, 9.4, 13.0, and 16.0μg/mL, respectively, but showed no significant effect on normal lymphatic cells at concentrations up to 40μg/mL. Although agaritine has been suspected of having genotoxic or carcinogenic properties, agaritine did not activate the umu gene of Salmonella, which reacts to carcinogens. General significance: The results indicate that agaritine from ABM has direct anti-tumor activity against leukemic tumor cells in vitro. This is in contrast to the carcinogenic activity previously ascribed to this compound. Our results also show that this activity is distinct from that of β-glucan, which indirectly suppresses proliferation of tumor cells. © 2010 Elsevier B.V. Source


Hosoki K.,Institute for Clinical Research | Hosoki K.,Mie University | Kainuma K.,Institute for Clinical Research | Toda M.,Mie University | And 9 more authors.
Biochemical and Biophysical Research Communications | Year: 2014

Epithelial to mesenchymal transition (EMT) is a mechanism by which eosinophils can induce airway remodeling. Montelukast, an antagonist of the cysteinyl leukotriene receptor, can suppress airway remodeling in asthma. The purpose of this study was to evaluate whether montelukast can ameliorate airway remodeling by blocking EMT induced by eosinophils. EMT induced was assessed using a co-culture system of human bronchial epithelial cells and human eosinophils or the eosinophilic leukemia cell lines, Eol-1. Montelukast inhibited co-culture associated morphological changes of BEAS-2b cells, decreased the expression of vimentin and collagen I, and increased the expression of E-cadherin. Montelukast mitigated the rise of TGF-β1 production and Smad3 phosphorylation. Co-culture of human eosinophils with BEAS-2B cells significantly enhanced the production of CysLTs compared with BEAS-2B cells or eosinophils alone. The increase of CysLTs was abolished by montelukast pre-treatment. Montelukast had similar effects when co-culture system of Eol-1 and BEAS-2B was used. This study showed that montelukast suppresses eosinophils-induced EMT of airway epithelial cells. This finding may explain the mechanism of montelukast-mediated amelioration of airway remodeling in bronchial asthma. © 2014 Elsevier Inc. All rights reserved. Source


Ito T.,Hokkaido University of Education | Kato M.,Nagoya University | Tsuchida H.,Aichi Mizuho College | Harada E.,Iwade Research Institute of Mycology | And 2 more authors.
Food Science and Technology Research | Year: 2011

We measured the anti-oxidative and anti-inflammatory activities of hot water extracts prepared from 11 species of mushrooms. Anti-oxidative activity was evaluated using the oxygen radical absorbance capacity method, and anti-inflammatory activity was examined by measuring the inhibition of lysine chloramine formation by hypochlorous acid. Hot water extracts of Grifola gargal (G. gargal) showed the strongest anti-oxidative activity and inhibitory effects on lysine chloramines. Hot water extracts of G. gargal were evaluated by HPLC and divided into 8 fractions. The most active fraction among these 8 fractions was further purified by preparative HPLC. The active component isolated by HPLC was identified as ergothioneine (EGT) using spectral analysis. On HPLC analysis, the EGT content in G. gargal was the highest among the 11 species of mushrooms. We also examined the protective role of EGT by examining the inflammatory response of adipocyte cells induced by tumor necrosis factor-α. Source


Harada E.,Mie University | Harada E.,Iwade Research Institute of Mycology | D'Alessandro-Gabazza C.N.,Mie University | Toda M.,Mie University | And 8 more authors.
Journal of Medicinal Food | Year: 2015

The beneficial effects of edible mushrooms for improving chronic intractable diseases have been documented. However, the antiatherogenic activity of the new medicinal mushroom Grifola gargal is unknown. Therefore, we evaluated whether Grifola gargal can prevent or delay the progression of atherosclerosis. Atherosclerosis was induced in ApoE lipoprotein-deficient mice by subcutaneous infusion of angiotensin II. Grifola gargal extract (GGE) was prepared and intraperitoneally injected. The weight of heart and vessels, dilatation/atheroma formation of thoracic and abdominal aorta, the percentage of peripheral granulocytes, and the blood concentration of MCP-1/CCL2 were significantly reduced in mice treated with GGE compared to untreated mice. By contrast, the percentage of regulatory T cells and the plasma concentration of SDF-1/CXCL12 were significantly increased in mice treated with the mushroom extract compared to untreated mice. In vitro, GGE significantly increased the secretion of SDF-1/CXCL12, VEGF, and TGF-β1 from fibroblasts compared to control. This study demonstrated for the first time that Grifola gargal therapy can enhance regulatory T cells and ameliorate atherosclerosis in mice. © Copyright 2015, Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition 2015. Source

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