• Title/Summary/Keyword: EGL

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Ethanol Extract of Ganoderma lucidum Augments Cellular Anti-oxidant Defense through Activation of Nrf2/HO-1

  • Lee, Yoo-hwan;Kim, Jung-hee;Song, Choon-ho;Jang, Kyung-jeon;kim, Cheol-hong;Kang, Ji-Sook;Choi, Yung-hyun;Yoon, Hyun-Min
    • Journal of Pharmacopuncture
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    • v.19 no.1
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    • pp.59-69
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    • 2016
  • Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

The Endo-β-1,4-Glucanase of Bacillus amyloliquefaciens Is Required for Optimum Endophytic Colonization of Plants

  • Fan, Xiaojing;Yang, Ruixian;Qiu, Sixin;Cai, Xueqing;Zou, Huasong;Hu, Fangping
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.946-952
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    • 2016
  • The eglS gene in Bacillus amyloliquefaciens encodes an endo-β-1,4-glucanase that belongs to glycosyl hydrolase family 5. In this study, a disruption mutant of gene eglS was constructed to examine its role in bacterial adaptation in plants. The mutant TB2k, eglS gene inactivated bacterial strain, was remarkably impaired in extracellular cellulase activity. When inoculated on Brassica campestris, the TB2k population was reduced by more than 60% compared with the wild-type strain in the root, stem, and leaf tissues. Overexpression of eglS in the wild-type strain increased the bacteria population in the plant tissues. Further studies revealed that the transcription level of eglS was correlated with bacterial population. These data demonstrate that endo-β-1,4-glucanase of B. amyloliquefaciens is required for its optimal endophytic colonization.

Cloning, Expression, Purification, and Properties of an Endoglucanase Gene (Glycosyl Hydrolase Family 12) from Aspergillus niger VTCC-F021 in Pichia pastoris

  • Pham, Thi Hoa;Quyen, Dinh Thi;Nghiem, Ngoc Minh;Vu, Thu Doan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1012-1020
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    • 2011
  • A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.

A Study on the Dielectric Properties of Glass Fiber-Reinforced Plastic Composites (유리 섬유 강화 복합재료의 유전 특성에 관한 연구)

  • Lee, B.S.;Whang, M.W.;Kim, J.S.;Cho, G.S.;Yuk, J.H.;Lee, D.C.
    • Proceedings of the KIEE Conference
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    • 1996.07c
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    • pp.1615-1617
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    • 1996
  • In this study, epoxidized bisphenolic resins laminated with glass fiber mat(GFRP) are ivestigated on surface, bulk aspect and dielectric constant(${\varepsilon}'$ and ${\varepsilon}''$) vs. frequency characteristics with temperature. The investigation shows the different characteristics accordig to the attachments of fiber surface, filler content, matrix properties, and the others. Especially, dielectric properties of this sample are highly increased above $100^{\circ}C$ and decreased with the rise of frequency. There is a resonance at the high frequency region ($1MHz{\sim}10MHz$). So, dielectric properties show the shift with frequency and temperature. Dielectric properties of EGL 10 are higher than those of EGL 40 with the frequency. Generally, dielectric properties of EGL 10 are more unstable than those of EGL 40 on the shift of frequency and temperature.

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Recycling Technology of Waste Product in Electro Galvanizing Line of Steel Company

  • Lee, Jae-Young;Lee, H. H.;Kim, D. Y.;J. G. Sohn
    • Proceedings of the IEEK Conference
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    • 2001.10a
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    • pp.281-285
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    • 2001
  • This technology Provides an economical Production of high value added goods applicable to electro chemicals by recycling of waste products in EGL(Electro Galvanizing Line). The waste products produced in EGL contain potassium chloride (KCI), nickel and zinc. Highly pure KCI and Zinc Chloride which are raw material of electro plating, can be produced by the development of the recycling process. The scope of this study ranges from laboratory experiments to pilot test in plant. We have developed the whole process of recycling technology such as purification method of waste products, fabrication methods of electro chemicals, basic design of plant, pilot scale production and evaluation of pilot goods, Developed electro chemicals were pure enough to satisfy the specification of steel company.

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Impact of Genetically Modified Enterobacter cloacae on Indigenous Endophytic Community of Citrus sinensis Seedlings

  • Fernando Dini;Mortatti, Marcelo-Jose;Souza, Andre-Oliveira de;Walter Maccheroni;Joao Lucio;Welington Luiz
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.169-173
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    • 2004
  • Enterobacter cloacae (strain PR2/7), a genetically modified endophyte(GME) in citrus plants, carrying different plasmids (pEC3.0/18, pCelE, pEglA and pGFP), was inoculated into Citrus sinensis seedlings under greenhouse conditions. The impact of this on the indigenous bacterial endophytic community was studied by analyses of 2 different morphologic groups. The germination rates of inoculated seeds were evaluated in greenhouse, and plasmid stability under in vitro conditions. Results demonstrated a great and diverse endophytic community inside plants, and specialization in tissue colonization by some bacterial groups, in different treatments. Shifts in seed germination rate were observed among treatments: in general, the PR2/7 harboring pEglA bacterial- clone significantly reduced seed germination, compared to the PR2/7 harboring pEC3.0/18 clone. This suggests that the presence of the pEglA plasmid changes bacteria-seed interactions. The endophytic community of citrus seedlings changed according to treatment. In seedlings treated with the PR2/7 with pEglA clone, the population of group II decreased significantly, within the context of the total endophytic community. These results indicate that the application of GMEs induces shifts in the endophytic bacterial community of citrus seedlings.

Immunoreactivity of PCNA in the Cerebellum of Developing Guinea Pig

  • Kim, Dong-joon;Jun, Yonghyun
    • International Journal of Oral Biology
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    • v.43 no.2
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    • pp.93-100
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    • 2018
  • The investigation of the embryonic development of the cerebellum has a long history. The postnatal normal development of the cerebellum in rodents and other animals became a popular topic for morphological investigations nearly a century ago. However, surprisingly, only a few studies are available regarding the prenatal normal development of the rodent cerebellum, especially in guinea pigs. Cell proliferation is essential for the development of the nervous system. The assessment of cell proliferation can be achieved by using various methods. In this study, we investigated the cell proliferation of the cerebellar cortex in guinea pigs at different stages of pregnancy and in postnatal life. Fetuses were obtained by cesarean section at 50 or 60 days of gestation (dg). Immunohistochemistry was performed with proliferating cell nuclear antigen (PCNA) antibody in the cerebellum. Strong PCNA immunoreactivity was observed in the external granular layer (EGL), which is a neurogenic zone in the cerebellum. The proportion of PCNA-IR cells was greater at 1 week than at 60 dg in lobule I, but not lobule VIII. After 50 dg, the width of the EGL continued to decline until 1 week, due to the maturation of the EGL cells. These results demonstrate the pattern of PCNA immunoreactivity in the developing cerebellum of guinea pigs. This serves as a guideline to study abnormal cerebellum development.

Wear for Polisher Brush of EGL Plating Cell using Finite Element Analysis (유한요소해석을 이용한 EGL 도금조 Polisher Brush의 마모예측)

  • Ku, J.K.;Noh, H.G.;Heo, S.C.;Song, W.J.;Ku, T.W.;Kim, J.;Kang, B.S.
    • Proceedings of the Korean Society for Technology of Plasticity Conference
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    • 2009.10a
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    • pp.342-345
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    • 2009
  • Electro galvanized steel is electroplated cold roller steel for improving corrosion resistance and paintability, and is widely used in automobiles and home appliances. In the electroplating line for manufacturing electro galvanized steel if plating process is carried out with impurity on conductor roll surface, the defects in manufacturing process occurs because of steel fault. For quality, polishing is always required to separate impurity on surface of conductor roll. In this study, finite element analysis of wear for polisher brush is carried out for replaced time of it.

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The evaluation on the biological safety of diagnostic ultrasound using radiation-induced apoptosis in the external granular layer of mouse cerebellum (마우스 소뇌과립층의 apoptosis를 지표로 한 진단용 초음파의 안전성 검증)

  • Oh, Heon;Lee, Song-eun;Yang, Jung-ah;Jo, Sung-kee;Chung, Chi-young;Son, Chang-ho;Kim, Sung-ho
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.628-634
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    • 1999
  • We have studied, by a nonisotopic in situ end-labeling(ISEL) technique, frequency of apoptosis in the external granular layer(EGL) of the cerebellum of immature mice by ${\gamma}$-rays irradiation from $^{60}Co$ or diagnostic ultrasound exposure. The total number of normal cells and cells showing morphological features of apoptosis were counted. The frequency of apoptotic cells was expressed as a percentage of the total number of cells in EGL. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 6~8 hours after exposure. The immature mice that received 18, 36, 54, 108, 198, 396 cGy of ${\gamma}$-rays or diagnostic ultrasound(7.5MHz, 4.2mW, $I_{SPTA}=7.9mW/cm^2$, $I_{SPTA}=114.3W/cm^2$) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after ${\gamma}$-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed by a linear-quadratic model ; frequency of apoptotic cell in the EGL was y = $(0.1349{\pm}0.01175)D$+$(-0.0001522{\pm}0.0000334)D^2$+0.048($r^2$ = 0.981, D = dose in cGy). In the experiment of ultrasound exposure, the frequency of apoptotic cell was $0.106{\pm}0.130$(10 minutes exposure) and $0.167{\pm}0.220$(30 minutes exposure). We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curve from the ${\gamma}$-irradiation. The relative dose of ultrasound exposure compared with ${\gamma}$-irradiation were 0.432 cGy(10 minutes exposure) and 0.885 cGy(30 minutes exposure). We have found that there is no evidence to indicate that diagnostic ultrasound involves a significant risk.

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Cloning of a Paenibacillus sp. Endo-${\circ}$-1,4-Glucanase Gene and Its Coexpression with the Endomyces fibuliger ${\circ}$-Glucosidase Gene in Saccharomyces cerevisiae

  • KIM, HYUNJIN;JI-YOUNG YANG;HYEON-GYU LEE;JAEHO, CHA
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.685-692
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    • 2001
  • A gene, Egl, from Paenibacillus sp. KCTC 8848P encoding endo-${\circ}$-1,4-glucanase was cloned and expressed in Escherichia coli. It consisted of an open reading frame of 1,191 bp for a protein that consisted of 397 amino acids with a molecular weight of 44,539 Da. The deduced amino acid sequence of the endo-${\circ}$-1,4-glucanase gene had a 94% similarity to the endo-$\beta$-1,4-glucanase of Bacillus polymyxa. The Egl gene was also expressed in Saccharomyces cerevisiae secreting Endomyces fibuliger $\beta$-glucosidase (BGL1) under the control of the alcohol dehydrogenase (ADC1) gene promoter, S. cerevisiae transformant producing both endo-${\circ}$-1,4-glucanase and ${\circ}$-glucosidase grew on carboxymethyl cellulose as the sole carbon source.

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