• Title/Summary/Keyword: Activation method

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Characteristics of Glucose-6-phosphate Dehydrogenase from Leuconostoc mesenteroides (Leuconostoc mesenteroides에서 分離한 Glucose-6-phosphate Dehydrogenase의 特性)

  • Byun Si Myung;Yang Do Choi;Moon H. Han
    • Journal of the Korean Chemical Society
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    • v.23 no.4
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    • pp.248-258
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    • 1979
  • Glucose 6-phosphate dehydrogenase of Leuconostoc mesenteroides which was purifid by an affinity chromatography was studied on the characterization, kinetics and chemical modification. The apparent molecular weight of the enzyme was 112,000 by the gel filtration method of Sephadex G-200 column. The optimum temperature of $NAD^+$-linked reation was 50$^{circ}C$ and the activation energy and the heat of inactivation were 8.36 kcal/mole and -58.2kcal/mole, respectively. The steady state kinetic study showed KG6P, Kemp, and CX KNADP to be 76.9 PM, 7.46${\mu}M$ and 7.14 ${\mu}M$, respectively, and KGGP, KNAD,and aKNm to be 53.7${\mu}M$, 115.2${\mu}M$ and 702.2${\mu}M$ for the $NAD^+$-linked reaction at pH 7.8, optimum pH. The pH dependent kinetic constants suggested that the two ionizing groups whose pKa is 7.2 .and pKb is 9.0-9.6 were involved in the enzyme-substrate interaction. Evidence by photooxidation and carboxymethylation of the enzyme suggested that the imidazole group of histidine with pKa group may participate in the catalytic site.

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Growth and optical characterization of $CuInSe_2$ single crystal thin film for solar cell application (태양전지용 $CuInSe_2$단결정 박막 성장과 광학적 특성)

  • 백승남;홍광준
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.12 no.4
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    • pp.202-209
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    • 2002
  • The stochiometric mix of evaporating materials for the $CuInSe_2$single crystal thin films was prepared from horizontal furnace. To obtain the single crystal thin films, $CuInSe_2$compound crystal was deposited on thoroughly etched semi-insulating GaAs(100) substrate by the Hot Wall Epitaxy (HWE) system. The source and substrate temperature were $620^{\circ}C$ and $410^{\circ}C$, respectively. The crystalline structure of single crystal thin films was investigated by the photoluminescence and double crystal X-ray diffraction (DCXD). The carrier density and mobility of $CuInSe_2$single crystal thin films measured from Hall effect by van der Pauw method. From the photocurrent spectrum by illumination of perpendicular light on the c-axis of the $CuInSe_2$single crystal thin film, we have found that the values of spin orbit splitting $\Delta$So and the crystal field splitting $\Delta$Cr. From the photoluminescence measurement on $CuInSe_2$single crystal thin film, we observed free exciton ($E_x$) existing only high quality crystal and neutral bound exciton ($A^{\circ}$, X) having very strong peak intensity. Then, the full-width-at-half-maximum (FWHM) and binding energy of neutral donor bound exciton were 7 meV and 5.9 meV, respectivity. By haynes rule, an activation energy of impurity was 59 meV.

Inhibition of Nitric Oxide Production by ladybug extracts(Harmonia axyridis) in LPS-activated BV-2 cells (무당벌레(Harmonia axyridis) 추출물에 의한 BV-2 세포주의 Nitric Oxide 생성 저해 활성)

  • Han Sang-Mi;Lee Sang-Han;Yun Chi-Young;Kang Seok-Woo;Lee Kyung-Gill;Kim Ik-Soo;Yun Eun-Young;Lee Pyeong-Jae;Kim Sun-Yeou;Hwang Jae-Sam
    • Korean journal of applied entomology
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    • v.45 no.1 s.142
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    • pp.31-36
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    • 2006
  • Inflammation in the brain has known to be associated with the development of a various neurologiacal diseases. The hallmark of neuro-inflammation is the activation of microglia, brain macrophage. Pro-inflammatory compounds including nitric oxide(NO) are the main cause of neuro-degenerative disease such as Alzheimer's disease. In the study, we examined whether Harmonia axyridis extracts inhibit the NO production by a direct method using Griess reagent, western blotting and by RT-PCR(Reverse Transcription-Polymerase Chain Reactionin) the gene expression of inducible nitric oxide synthase(iNOS). Distilled water$(H_2O)$ and methanol(MeOH) extracts of H. axyridis inhibited the protein expression of TNF-a(Tumor Necrosis Factor) and IL-6(Interleukin) in LPS (Lipopolysaccharide) stimulated BV-2 cells at the concentration of 100 ng/ml. Incubation of BV-2 cells with the extracts of $H_2O$ of MeOH inhibited the LPS induced NO and iNOS protein. And this inhibition of iNOS protein is concordant with the inhibition of iNOS mRNA expression. These data suggested that H. axyridis extracts may play a crucial role in inhibiting the NO production.

Characteristics of Isotherm, Kinetic and Thermodynamic Parameters for the Adsorption of Acid Red 66 by Activated Carbon (활성탄에 의한 Acid Red 66의 흡착에 대한 등온선, 동력학 및 열역학적 특성)

  • Lee, Jong-Jib
    • Clean Technology
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    • v.26 no.1
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    • pp.30-38
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    • 2020
  • The kinetic and thermodynamic parameters of Acid Red 66, adsorbed by granular activated carbon, were investigated on areas of initial concentration, contact time, and temperature. The adsorption equilibrium data were applied to Langmuir, Freundlich, Temkin, Redlich-Peterson, and Temkin isotherms. The agreement was found to be the highest in the Freundlich model. From the determined Freundlich separation factor (1/n = 0.125 ~ 0.232), the adsorption of Acid Red 66 by granular activated carbon could be employed as an effective treatment method. Temkin's constant related to adsorption heat (BT = 2.147 ~ 2.562 J mol-1) showed that this process was physical adsorption. From kinetic experiments, the adsorption process followed the pseudo-second order model with good agreement. The results of the intraparticle diffusion equation showed that the inclination of the second straight line representing the intraparticle diffusion was smaller than that of the first straight line representing the boundary layer diffusion. Therefore, it was confirmed that intraparticle diffusion was the rate-controlling step. From thermodynamic experiments, the activation energy was determined as 35.23 kJ mol-1, indicating that the adsorption of Acid Red 66 was physical adsorption. The negative Gibbs free energy change (ΔG = -0.548 ~ -7.802 kJ mol-1) and the positive enthalpy change (ΔH = +109.112 kJ mol-1) indicated the spontaneous and endothermic nature of the adsorption process, respectively. The isosteric heat of adsorption increased with the increase of surface loading, indicating lateral interactions between the adsorbed dye molecules.

Propofol protects human keratinocytes from oxidative stress via autophagy expression

  • Yoon, Ji-Young;Jeon, Hyun-Ook;Kim, Eun-Jung;Kim, Cheul-Hong;Yoon, Ji-Uk;Park, Bong-Soo;Yu, Su-Bin;Kwak, Jin-Won
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.17 no.1
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    • pp.21-28
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    • 2017
  • Background: The skin consists of tightly connected keratinocytes, and prevents extensive water loss while simultaneously protecting against the entry of microbial pathogens. Excessive cellular levels of reactive oxygen species can induce cell apoptosis and also damage skin integrity. Propofol (2,6-diisopropylphenol) has antioxidant properties. In this study, we investigated how propofol influences intracellular autophagy and apoptotic cell death induced by oxidative stress in human keratinocytes. Method: The following groups were used for experimentation: control, cells were incubated under normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) without propofol; hydrogen peroxide ($H_2O_2$), cells were exposed to $H_2O_2$ ($300{\mu}M$) for 2 h; propofol preconditioning (PPC)/$H_2O_2$, cells pretreated with propofol ($100{\mu}M$) for 2 h were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)/PPC/H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability, apoptosis, and migration capability were evaluated. Relation to autophagy was detected by western blot analysis. Results: Cell viability decreased significantly in the $H_2O_2$ group compared to that in the control group and was improved by propofol preconditioning. Propofol preconditioning effectively decreased $H_2O_2$-induced cell apoptosis and increased cell migration. However, pretreatment with 3-MA inhibited the protective effect of propofol on cell apoptosis. Autophagy was activated in the $PPC/H_2O_2$ group compared to that in the $H_2O_2$ group as demonstrated by western blot analysis and autophagosome staining. Conclusion: The results suggest that propofol preconditioning induces an endogenous cellular protective effect in human keratinocytes against oxidative stress through the activation of signaling pathways related to autophagy.

Propofol protects against oxidative-stress-induced COS-7 cell apoptosis by inducing autophagy

  • Yoon, Ji-Young;Baek, Chul-Woo;Kim, Eun-Jung;Park, Bong-Soo;Yu, Su-Bin;Yoon, Ji-Uk;Kim, Eok-Nyun
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.17 no.1
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    • pp.37-46
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    • 2017
  • Background: In oxidative stress, reactive oxygen species (ROS) production contributes to cellular dysfunction and initiates the apoptotic cascade. Autophagy is considered the mechanism that decreases ROS concentration and oxidative damage. Propofol shows antioxidant properties, but the mechanisms underlying the effect of propofol preconditioning (PPC) on oxidative injury remain unclear. Therefore, we investigated whether PPC protects against cell damage from hydrogen peroxide ($H_2O_2$)-induced oxidative stress and influences cellular autophagy. Method: COS-7 cells were randomly divided into the following groups: control, cells were incubated in normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) for 24 h without propofol; $H_2O_2$, cells were exposed to $H_2O_2$ ($400{\mu}M$) for 2 h; $PPC+H_2O_2$, cells pretreated with propofol were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)+PPC+H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide thiazolyl blue (MTT) reduction. Apoptosis was determined using Hoechst 33342 staining and fluorescence microscopy. The relationship between PPC and autophagy was detected using western blot analysis. Results: Cell viability decreased more significantly in the $H_2O_2$ group than in the control group, but it was improved by PPC ($100{\mu}M$). Pretreatment with propofol effectively decreased $H_2O_2$-induced COS-7 cell apoptosis. However, pretreatment with 3-MA inhibited the protective effect of propofol during apoptosis. Western blot analysis showed that the level of autophagy-related proteins was higher in the $PPC+H_2O_2$ group than that in the $H_2O_2$ group. Conclusion: PPC has a protective effect on $H_2O_2$-induced COS-7 cell apoptosis, which is mediated by autophagy activation.

Purification of Recombinant CTP-Conjugated Human prostatic acid phosphatase for activation of Dendritic Cell (수지상세포 활성화를 위한 세포투과 펩타이드가 결합된 재조합 전립성 산성 인산분해효소의 정제)

  • Yi, Ki-Wan;Ryu, Kang
    • KSBB Journal
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    • v.24 no.1
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    • pp.80-88
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    • 2009
  • Human prostatic acid phosphatase (PAP), with comprehensive homology to glandular kallikrein, are representative serum biomarkers of prostate cancer. Dendritic cell (DC), which is the potent antigen-presenting cells(APC) in the immune system, can induce strong T cell responses against viruses, microbial pathogens, and tumors. Therefore, the immunization using DC loaded with tumor-associated antigens is a powerful method for inducing anti-tumor immunity. The CTP (Cytoplasmic Transduction Peptide) technology developed by Creagene which can transport attached bio-polymers like nucleic acids or proteins into the cell with high permeation efficiency. As the active forms of PAP can mediate apoptotic processing, we used multimer forms of PAP as an inactive form for antigen pulsing of DCs. In this study, multimeric forms of CTP-rhPAP was obtained according to the advanced purification process and subsequently confirmed by gel filtration chromatography, western blot and Dynamic Light Scattering. Therefore, CTP-conjugated PA multimers transduced into the cytoplasm were efficiently presented on the cell surface without any harm effect on cells via MHC class I molecules and result in induction of a large number of effector cell.

Engineering Performance and Applicability of Environmental Friendly Porous Concrete for a Marine Ranch Using Steel Industry By-products (철강산업 부산물을 활용한 해양목장 조성용 친환경 다공질 콘크리트의 공학적 성능 및 적용성)

  • Lee, Byung-Jae;Jang, Young-Il;Kim, Yun-Yong
    • Journal of the Korea Concrete Institute
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    • v.25 no.1
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    • pp.115-123
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    • 2013
  • The steel industry, a representative industry that significantly consumes raw materials and energy, produces steel as well as a large amount of by-product steel slag through the production process. The vast habitat foundation of marine life has been destroyed due to recent reckless marine development and environment pollution, resulting in intensification of the decline of marine resources, and a solution to this issue is imperative. In order to propose a method to recycle large amounts of by-product slag into a material that can serve as an alternative to natural aggregate, the engineering properties and applicability for each mixing factor of environment friendly porous concrete as a material for the composition of marine ranches were evaluated in this study. The test results for percentage of voids per mixing ratio revealed that the margin of error for all conditions was within 2.5%. The compressive strength test results showed that the most outstanding environmental friendly porous concrete can be manufactured when mixing 30% slag aggregate and 10% specially treated granular fertilizer for the optimum volume fraction. As concrete for marine applications, the best seawater resistance was obtained with mixing conditions for high compression strength. An assessment of the ability to provide a marine life habitat foundation of environmentally friendly porous concrete showed that a greater percentage of voids facilitated implantation and inhabitation of marine life, and the mixing of specially treated granular fertilizer led to active initial implantation and activation of inhabitation. The evaluation of harmfulness to marine life depending on the mixture of slag aggregate and specially treated granular fertilizer revealed that the stability of fish is secured.

A Study on the Switching Characteristcs of PLT(10) Thin Films (PLT(10) 박막의 Switching 특성에 관한 연구)

  • Kang, Seong-Jun;Chang, Dong-Hoon;Yoon, Yung-Sup
    • Journal of the Korean Institute of Telematics and Electronics D
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    • v.36D no.11
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    • pp.63-70
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    • 1999
  • A PLT(10) thin film has been deposited on $Pt/TiO_2/SiO_2/Si$ substrate by sol-gel method, and its switching characteristics have been investigated with various top electrode areas, input pulse voltages and loan resistances. As the external input pulse voltage increases from 2V to 5V, the switching time decreases from $0.49{\mu}s$ to $0.12{\mu}s$. The activation energy ($E_a$) obtained from the relations between the switching time and the applied pulse voltage is evaluated as 209kV/cm. The switched charge densities at 5V obtained from the hysteresis loop and the polarization switching are $11.69{\mu}C/cm^2$ and $13.02{\mu}C/cm^2$, respectively, which agree relatively well with each other and show the difference of 10%. When the top electrode area increases from TEX>$3.14{\times}10^{-4}cm^2$ to $5.03{\times}10^{-3}cm^2$ and the load resistance increases from 50${\Omega}$ to 3.3$k{\Omega}$, the switching time increases from $0.12{\mu}s$ to $1.88{\mu}s$ and from $0.12{\mu}s$ to $9.7{\mu}s$, respectively. These switching characteristics indicate that PLT(10) thin film can be well applied in nonvolatile memory devices.

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Imyosan induces caspases-mediated apoptosis in human colorectal cancer HCT116 cells (이묘산(二妙散)에 의한 대장암 세포주 HCT116의 Caspases 활성화를 매개로 한 세포사멸)

  • Kim, Sun-Mo;Yun, Hyun-Jeung;Lee, Hyun-Woo;Kim, Pan-Jun;Lee, Chang-Hyun;Park, Won-Hwan;Park, Sun-Dong
    • Herbal Formula Science
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    • v.14 no.2
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    • pp.21-32
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    • 2006
  • The purpose of this study was to investigate the effect of Imyosan on apoptosis in human colorectal cancer HCT116 cells. Phellodendron amurense Rupr. and Atratylodes lancea D.C. compose Imyosan. First of all, to study the cytotoxic effect of methanol extract of Imyosan (IMS-MeOH) on HCT116 cells, the cells were treated with various concentrations of IMS-MeOH and then cell viability was determined by XTT reduction method. IMS-MeOH reduced viability of HCT116 cells in a dose and time-dependent manner. To confirm the induction of apoptosis, the c1eavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of caspase-3, procaspase-8 and procaspase-9 were examined by western blot analysis. IMS-MeOH decreased procaspase-3, procaspase-8 and procaspase-9 levels in a dose-dependent manner and induced the clevage of PARP. IMS-MeOH triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome c from mitochondria to cytosol. Furthermore, IMS-MeOH also downregulated the anti-apoptotic Bcl-2 and upregulated the pro-apoptotic-Bax. Therefore, these results suggest that IMS-MeOH induced HCT1l6 cell death through the mitochondrial pathway. To explore whether the activities of caspases was required for induction of apoptosis by IMS-MeOH, caspase-3, -8, -9 activity measured by using substrates, respectively. IMS-MeOH increased caspase-3, -8, -9 activity. Co-treatment with inhibitors of caspase-3, -8, -9 and IMS-MeOH significantly blocked IMS-MeOH-triggered apoptosis in HCT1l6 cells. These results suggest that IMS-MeOH induces caspases-mediated apoptosis.

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