본 연구에서는 버섯추출물 (42개)의 in vitro 항알레르 기 효능탐색을 위해 랫드비만세포 (RBL-2H3 cell)에서 면역글로불린 (IgE)가 매개한 탈과립에 대한 저해 효과를 실험하였다. 이를 위해 anti-DNP IgE 및 DNP-HSA에 의 해 알레르기반응이 유발된 랫드비만세포에서 버섯 추출물 의 IL-4과 β-hexosaminidase 분비량에 대한 저해활성과 세포생존에 대한 영향이 분석되었다. 실험결과, IL-4 분비 에 대해서는 팽이버섯 물추출물 등 5개의 버섯 추출물이 20% 이상의 우수한 저해효과를 나타내었으며, β- hexosaminidase 분비에 대해서는 영지버섯의 물추출물 등 8개의 버섯 추출물이 20% 이상의 비교적 우수한 저해활성 을 나타내었다. 세포증식에 대해서는 잎새버섯의 물추출물 등 대부분의 버섯 추출물이 우수한 세포증식효과를 나타 내었다. 팽이버섯의 물추출물과 상황버섯의 물추출물은 β- hexosaminidase 및 IL-4 분비에 대해 모두 비교적 우수한 저해효과를 나타내었다. 추가로 2, 10, 50 ug/ml에서 실험 된 영지버섯, 편각영지버섯, 눈꽃동충하초, 상황버섯 그리 고 느타리버섯의 물추출물들은 β-hexosaminidase의 분비 량을 농도-의존적으로 감소시켰다. 이상의 결과를 살펴볼 때, 이들 in vitro 항알레르기 효과를 나타낸 버섯 추출물 들은 추가실험을 통해 항알레르기 소재로의 활용성 검토 가 필요하다고 사료되었다.

In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phos-phodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, 67.57±4.11% aging, 44.61±6.4%) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of con-trol group intensity rate (51.53.±3.80), aging group (68.10±5.54) and treatment of caffeine (45.04±2.98). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging (90.44±10.18 VS 67.88±7.72). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro

This study was carried out to investigate comparision anaylsis of metabolites in mandarin leave inoculated by Elsinoe fawcetii. For the analysis of metabolites, we performed using by UPLC-MS spectrometry in gradient eluent condition by acetonitrile and water. Flow rate was 0.4ml/min. Instruments conditions were followed by detection ion mode(positive ion), scan range(MS:m/z100-1000), spray voltage (4kV) ,capillary voltage(35v), capillary temp(300℃). Several flavones were detected in inoculated pathogen compared to control.

Three edible mushrooms; "Gumbit"(Pleurotus cornucopiae var. citrinopileatus) and Noeul"(Pleurotus salmoneostramineus) and Tricholoma spectabilis which are widely distributed in China, Japan and Korea. For the study of anti-aging activities, we were performed on adriamycin-induced cellular senescence model. Human dermal fibroblasts(HDFs) and human umbilical endothelial cells(HUVECs) treated with adriamycin for inducing aging were used for cytotoxicity and senescence-associated beta-galactosidase(SA-β-gal) activity assay.

Cordyceps militar is (Clavicipitales) is an edible mushroom which is widely distributed in China, Japan and Korea. Various phytochemical constituents, cordycepin, homocitrully laminoadenosine and sterols have been reported from this source and a wide range of biological activities, including antimicrobial, macrophage activation, anticancer, immune modulatory effects were studied. In a continuing search for bioactive constituents from Korean mushrooms, we performed a phytochemical investigation of the MeOH extract from the fruiting bodies of C. militaris. By repeated column chromatographic separation of the extract, fourteen compounds, were isolated. The identification and structural elucidation of the compound was based on NMR spectral data. anti-aging activities were performed on adriamycin-induced cellular senescence model. Human dermal fibroblasts(HDFs) and human umbilical endothelial cells(HUVECs) treated with adriamycin for inducing aging were used for cytotoxicity and senescence-associated beta-galactosidase(SA-β-gal) activity assay.

Autophagy is conserved response to starvation by which cells catabolize their components to create an internal supply of essential nutrients. Ceramide is known to induce autophagy in many cells through down-regulation of amino acid and glucose transporters. The mechanism of starvation induced-autophagy in mouse embryo remains unclear. In order to understand the mechanism by which starvation regulates autophagy, in this study, we investigated nutrient transporters expression and the effect of c2-ceramide on the in vitro development, apoptosis and autophagy via starvation in mouse embryo. Glucose transporters (Glut1 and Glut 3), high levels of transcript were expressed from 1 to 2 cells and gradually decreased through the morula and blastocyst (BL) stages. Amino acid transporters (LAT-1 and 4F2hc) gradually decreased from the zygote to the BL stage. Furthermore, the expression of nutrient transporters (Glut1, 3, LAT-1 and 4F2hc) were significantly reduced at the BL stage after ceramide treatment. Especially, mTOR expression after ceramide treatment of embryos was significantly higher than controls. Ceramide treated embryos exhibited significantly reduced developmental rates and total cell numbers, and increased apoptotic cell death at the BL stage. Consequently, we next evaluated the effect of ceramide treatment on mitochondrial number and morphology. There was a significant decrease in the average mtDNA copy number and the mitochondrial area in ceramide treated BL stage embryos. Both the expression of autophagy-related genes, Lc3, Gabarap, Atg4A and Atg4B, and the synthesis of LC3 were significantly induced at the BL stage. These results suggest that autophagy under starvation condition influences the in vitro development and apoptosis and autophagy, and may play a role in early mouse embryogenesis.

This study was carried out to investigate anticancer activities fruiting body extracts and fractions of Cordyceps militaris. Fruiting body of this mushroom was extracted using by 80% MeOH. Fractionations of these extracts were performed by n-hexane, methylene chloride, ethyl acetate, n-BuOH. AGS(human gastric cancer line) was cultured in media conditions (10% FBS, 1% Penicillin-Streptomycin in RPMI). Anticancer activities of each fractions of Cordyceps militaris were examined by using MTT, Cell titer Glo.

This study was carried out to investigate neuronal protective activity of fruiting body of Hericium erinaceum. In order to search the effective active compound against amyloid beta peptide-induced oxidative stress on neuronal cells, rat pheochromocytoma cells (PC12), Extracts of Hericium erinaceum were screened and evaluated using both the 2’,7’-dichlorofluorescin diacetate assay (DCF-DA), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to above assays, Solvent partitions of extracts were selected for further purification and isolation of anti-Alzheimer’s disease compound as it exerted the highest protective effects against hydrogen peroxide (H2O2)-induced oxidative stress.

Several bacteria have been known as the causal agents of certain diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). It is well known as bacterial diseases of the cultivated mushroom such as brown blotch, mummy disease, bacterial pit, bacterial rot and weeping disease, ginger blotch, and drippy gill. Brown blotch is the most critical cause of crop loss in the commercial mushroom industry. The classical bacterial blotch disease of mushrooms is caused by a fluorescent pseudomonad, Pseudomonas tolaasii. Affected mushrooms show lesions which become dark chocolate-brown, are wet, and deeply pit the caps and stalks. Although Pseudomonas tolaasii has been known as the casual agent of bacterial blotch, much controversy exists regarding the identification of this bacterium and whether blotch may be caused by more than one organism. This study was carried out to investigate characterization and biological control of Pseudomonas tolaasi and other possible browning pathogens isolated from cultivated mushrooms. One hundred seventy four bacteria were isolated from the cultivated mushroom and collected from main producing districts throughout the country. The isolates were classified into Pseudomonas tolaasii(20 strains), Pseudomonas gingeri(1 strains), Pseudomonas agarici(4 strains), Pseudomonas putida(11 strains), Pseudomonas sp.(46 strains), Ewingella americana(14 strains), Stenotrophomonas sp.(4 strains), and others(74 strains) on the basis of 16 rDNA analysis. The most dominants of these species were Pseudomonas tolaasii and Ewingella americana. Pseudomonad isolates were mainly divided into two groups in white line test and a sharply defined white line of precipitate forms in Pseudomonas agar F(Difco) between the opaque white colonies of P. tolaasii and translucent colonies of certain unidentified pseudomonads. The white line test was positive when 20 isolates of P. tolaasi from different countries were examined, whereas 62 isolates of pseudomonads did not give the white line reaction with a reacting translucent colony Pseudomonas. All the isolates tested for white line forming bacteria including P. tolaasi were highly pathogenic to mushroom tissue. Although browning of mushrooms in host tests does not perfectly help in the identification of P. tolaasi, a conspicuous pitting produced at the cut surface of mushroom tissue is as specific as the white line test in detecting P. tolaasii in suspension in distilled water. URP2F primers of 20-mer were used to assess the genetic diversity of white line forming bacteria. The phylogenetic tree was constructed by using the neighbor-joining method. In the analysis of RAPD pattern, all isolates of white line precipitate have some of the different genetic traits as collected districts. Phylogenetic analysis of 16S rDNA revealed that twenty isolates including white line forming bacteria were closely related to P. tolaasii and showed high similarity. To biological control on bacterial browning disease of cultivated mushrooms, six hundreds plant extracts (332 EtOH extracts, 268 water extracts) was used for control of mushroom disease. Thirty plant extracts in bacterial disease(Pseudomonas tolaasii, P. agarici, B. gladioli, E. americana) and thirty three in fungus disease(T. harzianum, C. mycophilum, V. fungicola) showed strong anti-microbes activity. They showed stronger anti-microbes activity at ethanol extracts than water extracts. MIC of extract BCW128 on Pseudomonas tolaasii was 700ppm and HDE17 was 330ppm. MIC of extract YCE107 on P. agarici was 330ppm, JGE96 was 330ppm and BCW128 was 700ppm. The bacteria inhibit tolaasin secreted by Pseudomonas tolaasii was selected three genus(Bacillus sp. etc). Now we are carrying out more research on these bacteria.

This study was carried out to investigate characteristic pattern of fruiting body of Ganoderma lucidum and their antioxidant activity. Mycelia of all strains were firstly inoculated into potato dextrose agar(PDA) and then transfered to a media of saw dust which contained 20% rice bran. These mycelia of saw dust were then inoculated into oak tree in polyethylene bags which has been sterilized for 8h at 120℃. The polyethylene bags were sent to a growth room for growth of fruit bodies. Antioxidant activities of each fruiting body were investigated by DPPH method.

Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor p27KIP1. Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.