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Effects of Fermented Rice Wine by Using Mycelium of Phellinus linteus on the Expression of Inflammation-Related Proteins in Human Hepatoma Cells and Rat Liver (상황버섯 균사체를 이용한 발효주가 인체간암세포와 흰쥐 간의 염증관련 단백질 발현에 미치는 영향)

  • Ahn Seung-Min;Lee Jun-Hyuk;Choi Yung-Hyun;Lee Yong-Tae;Chung Kyung-Tae;Jeong Young-Kee;Jo Un-Bock;Choi Byung-Tae
    • Journal of Life Science
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    • v.16 no.1
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    • pp.101-107
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    • 2006
  • We have recently discovered that mycelium of Phellinus linteus, popular medical mushrooms in Korea, possess alcohol dehydrogenase and produce alcohol. In the present study, it was examined that the effect of fermented rice wine made by using mycelium of P. linteus (FLMP) on the expression of in-flammation-related proteins in both $HepG_2$ cells and rats. To examine the effect of FLMP on the morphology and expression of inflammatory proteins in $HepG_2$ cells, the cells were incubated with ethanol, and FLMP for 24 hours, and then analyzed by microscopic observation and Western blot and reverse transcription polymerase chain reaction (RT-PCR). While ethanol induced the morphological change accompanied with cell debris formation and scattering on $HepG_2$ cells, FLMP had no effect. The results of Western blot and RT-PCR analyses showed that the level of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-1 and COX-2 was induced by ethanol, however, FLMP inhibited the expression of these proteins and its mRNAs. In the animal model, the value of flutamate oxaloacetate transaminase and glutamate pyruvate transaminase was significantly increased by administration with ethanol. But the group administrated with FLMP showed lower levels on the changes of these markers compared with ethanol-administrated group. Besides, the results of Western blot and RT-PCR analyses showed that the expression of inflammatory proteins such as iNOS, COX-1 and COX-2 was not affected by FLMP administration in rat liver. About histopathological and immunohistochemical observations, inflammatory loci were markedly decreased in the FLMP-administrated rat compared to ethanol-administrated rats and showed weaker COX-2 and iNOS jmmunoreactions. These results suggested that FLMP showed slight changes on the inflammatory proteins expression compared to ethanol and FLMP may be used as a functional alcoholic beverage.

Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells (택란 메탄올 추출물에 의한 인체 폐암 세포주 A549의 G1 arrest 유발)

  • Park, Hyun-Jin;Jin, Soojung;Oh, You Na;Yun, Seung-Geun;Lee, Ji-Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1109-1117
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    • 2013
  • Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells Lycopus lucidus, a herbaceous perennial, is used as a traditional remedy in East Asia, including China and Korea. It has been reported that L. lucidus has anti-allergic effects, inhibitory effects on cholesterol acyltransferase in high glucose-induced vascular inflammation, and anti-proliferative effects in human breast cancer cells. However, the molecular mechanisms of the anti-cancer effects of L. lucidus have not yet been fully determined. In this study, we evaluated the anti-cancer effect and the mechanism of action of L. lucidus in human lung adenocarcinoma A549 cells using methanol extracts of L. lucidus (MELL). MELL treatment showed cytotoxic activity in a dose-dependent manner and induced G1 arrest in A549 cells. The induction of G1 arrest by MELL was associated with the up-regulation of phospho-CHK2 and the down-regulation of Cdc25A phosphatase. In addition, MELL treatment induced decreased expression of G1/S transition-related proteins, including CDK2, CDK4, CDK6, cyclin D1 and cyclin E. MELL also regulated the mRNA expression of CDK2 and cyclin E. On the other hand, the expression of p53 and the cyclin-dependent kinase inhibitor p21 was not induced by MELL. Collectively, these results suggest that MELL may exert an anti-cancer effect by cell cycle arrest at G1 phase through the ATM/CHK2/Cdc25A/CDK2 pathway in A549 cells.

Expression Analysis of Oryza sativa Ascorbate Peroxidase 1 (OsAPx1) in Response to Different Phytohormones and Pathogens (벼 ascobate peroxidase 단백질의 병원균 및 식물호르몬에 대한 발현 분석)

  • Wang, Yiming;Wu, Jingni;Choi, Young Whan;Jun, Tae Hwan;Kwon, Soon Wook;Choi, In Soo;Kim, Yong Chul;Gupta, Ravi;Kim, Sun Tae
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1091-1097
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    • 2015
  • We have isolated and characterized an ascorbate peroxidase (APx) gene, OsAPx1 from rice. Northern and Western blot analyses indicated that at young seedling stage, OsAPx1 mRNA was expressed highly in root, shoot apical meristem (SAM) and leaf sheath than leaf. In mature plant, OsAPx1 gene expressed highly in root, stem and flower but weakly in leaf. OsAPx1 gene and protein expression level was induced in leaves inoculated with Magnaporthe oryzae (M. oryzae) and Xanthomonas oryzae pv. oryzae (Xoo). Phytohormones treatment showed that OsAPx1 was up-regulated by jasmonic acid (JA), but was down regulated by ABA and SA co-treatments with JA, resulting that they have antagonistic effect on pathogen responsive OsAPx1 expression. Phylogenetic analysis illustrated that Arabidopsis AtAPx1 has a close relationship with OsAPx1. In AtAPx1 knock out lines, the accumulation of O2- and H2O2 are all highly detected than wild type, revealing that the high concentration of exogenous H2O2 cause the intercellular superoxide anion and hydrogen peroxide accumulation in AtAPx1 knockout plant. These results suggested that OsAPx1 gene may be associated with the pathogen defense cascades as the mediator for balancing redox state by acting ROS scavenger and is associated with response to the pathogen defense via Jasmonic acid signaling pathway.

The Characteristics of Microbial Community for Biological Activated Carbon in Water Treatment Plant (생물활성탄 공정에서 활성탄 재질에 따른 부착미생물 군집특성)

  • Son, Hee-Jong;Park, Hong-Ki;Lee, Soo-Ae;Jung, Eun-Young;Jung, Chul-Woo
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.12
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    • pp.1311-1320
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    • 2005
  • The purpose of this research is to survey characteristics of microbial community and the removal efficiency of organic materials for biological activated carbon in water treatment plant. Coal based activated carbon retained more attached bacterial biomass on the surface of the activated carbon than the other activated carbon with operating time and materials. The heterotrophic plate count(HPC), eubacteria(EUB) and 4,6-diamidino-2-phenylindole(DAPI) counts were ranged from $0.95{\times}10^7$ to $52.4{\times}10^7$ CFU/g, from $3.8{\times}10^8$ to $134.2{\times}10^8$ cells/g and from $7.0{\times}10^8$ to $250.2{\times}10^8$ cells/g, respectively. The biomass of EUB and DAPI appeared to be much more $10^2$ than HPC, which were increasing in bed volume of 20,000 at the stage of steady-state. The change of microbial community by analyzing fluorescent in situ hybridization(FISH) method with rRNA-targeted oligonucleotide probes, the dominant group was $\alpha$-proteobacteria($\alpha$ group) and high G+C content bacteria(HGC) the lowest distributing rate before reaching the bed volume of 20,000. After reaching the bed volume of 20,000, $\alpha$ group and other groups of bacteria became decreased, on the other hand, the proportion of both $\beta$-proteobacteria($\beta$ group) and $\gamma$-proteobacteri($\gamma$ group) were increasing. Coconut and wood based activated carbons had similar trend with coal based activated carbon, but the rate of $\alpha$ group on coal based activated carbon had gradually increased. Bacterial production with the operating period appeared highest in coal based activated carbon at the range of $1.2{\sim}3.4\;mg-C/m^3{\cdot}h$ while the coconut and wood based activated carbon were ranged from 1.1 to 2.6 $mg-C/m^3{\cdot}h$ and from 0.7 to 3.5 $mg-C/m^3{\cdot}h$ respectively. The removal efficiency of assimilable organic carbon(AOC) showed to be highly correlated with bacterial production. The correlation coefficient between removal efficiency of AOC and bacterial production were 0.679 at wood based activated carbon, 0.291 at coconut based activated carbon and 0.762 at coal based activated carbon, respectively.

Anti-inflammatory Effect of Oxya chinensis sinuosa Ethanol Extract in LPS-induced RAW 264.7 Cells (LPS로 유도된 RAW 264.7세포에 대한 벼메뚜기(Oxya chinensis sinuosa) 에탄올 추출물의 항염증 효과)

  • Yoon, Young-Il;Chung, Mi Yeon;Hwang, Jae-Sam;Goo, Tae-Won;Ahn, Mi-Young;Lee, Young-Bo;Han, Myung-Sea;Yun, Eun-Young
    • Journal of Life Science
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    • v.24 no.4
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    • pp.370-376
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    • 2014
  • Although the grasshopper Oxya chinensis sinuosa has long been used as food in Korea, there is little data on its functional effects. In this study, we investigated the anti-inflammatory effect of O. c. sinuosa ethanol extract (OCE) in RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for induction of inflammation. First, we determined that there is no cytotoxicity at $2,000{\mu}g/ml$ or less of OCE in RAW 264.7 cells. To evaluate the anti-inflammatory effects of OCE, we investigated expression levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, and pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether OCE could inhibit translocation of NF-${\kappa}B$ p65 into the nucleus in LPS induced RAW 264.7 cells. As a result, we found that the mRNA and protein levels of TNF-${\alpha}$ and IL-6 decreased in LPS-induced RAW 264.7 cells after treatment with OCE in a dose-dependent manner. In addition, we confirmed a $2,000{\mu}g/ml$ concentration of OCE inhibited translocation of NF-${\kappa}B$ p65 by immunnostaining and Western blot analysis, and a decrease in the protein expression levels of iNOS and COX-2. Accordingly, we suppose that OCE has an anti-inflammatory effect through down-regulation of TNF-${\alpha}$, IL-6, iNOS, and COX-2 related to ${\kappa}B$ p65 inflammatory signaling pathways.

Combined Treatment of Nonsteroidal Anti-inflammatory Drugs and Genistein Synergistically Induces Apoptosis via Induction of NAG-1 in Human Lung Adenocarcinoma A549 Cells (인간 A549 폐암세포에서 비스테로이드성 항염증제와 genistein의 복합처리에 의한 NAG-1 의존적 세포사멸 증진 효과)

  • Kim, Cho-Hee;Kim, Min-Young;Lee, Su-Yeon;Moon, Ji-Young;Han, Song-Iy;Park, Hye-Gyeong;Kang, Ho-Sung
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1073-1080
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    • 2009
  • A number of studies have demonstrated that the regular use of nonsteroidal anti-inflammatory drugs (NSAIDs) can reduce the risks of colorectal, oesophageal and lung cancers. NSAIDs have been shown to exert their anti-cancer effects through inducing apoptosis in cancer cells. The susceptibility of tumor cells to anti-tumor drug-induced apoptosis appears to depend on the balance between pro-apoptotic and anti-apoptotic programs such as nuclear factor kB (NF-kB), phosphatidylinositol 3-kinase (PI3K)-Akt/protein kinase B (PKB) and MEK1/2-ERK1/2 pathways. We examined the effects of pro-survival PI3K and ERK1/2 signal pathways on cell cycle arrest and apoptosis in response to NSAIDs including sulindac sulfide and NS398. We show that simultaneous inhibition of the Akt/PKB and ERK1/2 signal cascades could synergistically enhance the potential pro-apoptotic activities of sulindac sulfide and NS398. Similar enhancement was observed in cells treated with sulindac sulfide or NS398 and 100 ${\mu}$M genistein, an inhibitor of receptor tyrosine kinases (RTKs) that are upstream of PI3K and MEK1/2 signaling. We further demonstrate that NAG-1 is induced and plays a critical role(s) in apoptosis by NSAIDs-based combined treatment. In sum, our results show that combinatorialtreatment of sulindac sulfide or NS398 and genistein results in a highlysynergistic induction of apoptotic cell death to increase the chemopreventive effects of the NSAIDs, sulindac sulfide and NS398.

Cloning of Low-molecular-weight Glutenin Subunit Genes and Identification of their Protein Products in Common Wheat (Triticum aestivum L.) (보통 밀에서 저분자글루테닌 유전자 클로닝 및 단백질 동정)

  • Lee, Jong-Yeol;Kim, Yeong-Tae;Kim, Bo-Mi;Lee, Jung-Hye;Lim, Sun-Hyung;Ha, Sun-Hwa;Ahn, Sang-Nag;Nam, Myung-Hee;Kim, Young-Mi
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.547-554
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    • 2010
  • Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.

Development of Anti-Aging Products (Anti-Wrinkle) like Epidermal Growth Factor(EGF) Materials using Supercritical Heat-Treated Extract Radish (초임계 열처리된 무 성분을 이용한 상피세포성장인자(EGF) 유사소재 개발 및 광노화(주름개선) 효과)

  • Kim, Hyun Kyoung
    • The Journal of the Convergence on Culture Technology
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    • v.4 no.3
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    • pp.197-207
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    • 2018
  • The radish skin and radish greens (mucheong) are an edible part of the radish. But they are removed before eating the radish and used as a byproduct or an animal feed material because of their tough and rough texture. This study was conducted to investigate the effect of supercritical heat-treated radish-extract on UV-induced Hos: HRM-2 wrinkled mouse animal model on anti-aging wrinkles. Supercritical heat-treated radish-extract was applied on the back of seven-weeks old HRM-2 mice. The effect of HRE on skin thickness, elasticity and wrinkle formation of the mice was observed by using UVB lamp to induce melanogenesis and wrinkle formation. As the result, increased depth of wrinkles was observed in the negative control group in comparison to the normal group. In contrast, decreased depth of wrinkles was observed in the radish-extract-free group compared to the negative control group. In the study of the effect of radish-extract on wrinkle-formation related gene expression and protein what protein expression, MMP-2 and MMP-2 gene expression significantly increased in the negative control group compared to the normal group. The gene expression reduced independence to the mass of radish-extract treated. Similar to quantitative results of mRNA expression, the expression of MMP-2 protein increased as a result of UVB-irradiation. The MMP-2 expression was inhibited in dependence to the mass of radish-extract treated. In conclusion, the supercritical heat-treated radish-extract has an effect on improving skin wrinkles not only when it is applied to the skin but also when orally ingested. Thus, it can be effectively used as a composition to health functional products. Thereafter, we can also conclude that radish, a food that does not show any side-effects even upon long-term intake, can reduce wrinkle formation as well as improve skin elasticity when taken regularly for a long period.

(Image Analysis of Electrophoresis Gels by using Region Growing with Multiple Peaks) (다중 피크의 영역 성장 기법에 의한 전기영동 젤의 영상 분석)

  • 김영원;전병환
    • Journal of KIISE:Software and Applications
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    • v.30 no.5_6
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    • pp.444-453
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    • 2003
  • Recently, a great interest of bio-technology(BT) is concentrated and the image analysis technique for electrophoresis gels is highly requested to analyze genetic information or to look for some new bio-activation materials. For this purpose, the location and quantity of each band in a lane should be measured. In most of existing techniques, the approach of peak searching in a profile of a lane is used. But this peak is improper as the representative of a band, because its location does not correspond to that of the brightest pixel or the center of gravity. Also, it is improper to measure band quantity in most of these approaches because various enhancement processes are commonly applied to original images to extract peaks easily. In this paper, we adopt an approach to measure accumulated brightness as a band quantity in each band region, which Is extracted by not using any process of changing relative brightness, and the gravity center of the region is calculated as a band location. Actually, we first extract lanes with an entropy-based threshold calculated on a gel-image histogram. And then, three other methods are proposed and applied to extract bands. In the MER method, peaks and valleys are searched on a vertical search line by which each lane is bisected. And the minimum enclosing rectangle of each band is set between successive two valleys. On the other hand, in the RG-1 method, each band is extracted by using region growing with a peak as a seed, separating overlapped neighbor bands. In the RG-2 method, peaks and valleys are searched on two vertical lines by which each lane is trisected, and the left and right peaks nay be paired up if they seem to belong to the same band, and then each band region is grown up with a peak or both peaks if exist. To compare above three methods, we have measured the location and amount of bands. As a result, the average errors in band location of MER, RG-1, and RG-2 were 6%, 3%, and 1%, respectively, when the lane length is normalized to a unit value. And the average errors in band amount were 8%, 5%, and 2%, respectively, when the sum of band amount is normalized to a unit value. In conclusion, RG-2 was shown to be more reliable in the accuracy of measuring the location and amount of bands.

Inhibitory Effects of Ginger and Beopje Ginger on DSS-induced Colitis in Mice (생강과 법제생강의 DSS(Dextran Sulfate Sodium)로 유도된 마우스의 대장염 억제 효과)

  • Kim, Sin-Jeong;Kim, So-Hee;Lim, Yaung-Iee;Kim, Yong-Gyu;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.477-484
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    • 2014
  • The purpose of this study was to examine the anti-inflammatory effects of ginger and processed (Beopje) ginger on colitis induced by 2.5% dextran sulfate sodium (DSS) in Balb/c mice. Beopje means a process that herbal medicines are treated by a specific Korean traditional method in order to obtain better pharmacological effects. Mice were fed saline or two different doses of ethanol extracts (ginger and processed (Beopje) ginger) once a day for 14 days. Colitis was induced from day 7 to 14 via administration of 2.5% DSS in drinking water. Experimental animals were divided into four groups: Nor (Normal, 200 ${\mu}L$ of saline without 2.5% DSS-treated group), Con (Control, 200 ${\mu}L$ of saline and 2.5% DSS treated group), G (500 mg/kg of ginger and 2.5% DSS treated group), and BG (500 mg/kg of Beopje ginger and 2.5% DSS treated group). Body weights of both ginger-administered groups increased compared to the control. Colon length increased to 7.6, and 8.0 cm in the G and BG groups, respectively, whereas that of control was 5.7 cm. Histological colon injury induced by DSS-induced colitis was reduced (P<0.05). In serum and DSS-treated colon tissues, mRNA expression levels of IFN-${\gamma}$, IL-6, TNF-${\alpha}$, and IL-12 of the Beopje ginger-treated group were significantly suppressed compared to those of the ginger-treated groups. Expression levels of iNOS and COX-2 of the Beopje ginger-treated group were significantly reduced compared to those of the ginger-treated groups (P<0.05), and BG showed stronger anti-inflammatory effects on colitis. These results indicated that ginger exerted anti-inflammatory effects on DSS-induced colitis in mice, and its effects could be increased through Beopje.