• Title/Summary/Keyword: Additional substrate

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Chemical Modification of Brain Glutamate Dehydrogenase Isoproteins with Phenylglyoxal

  • Ahn, Jee-Yin;Cho, Eun-Hee;Lee, Kil-Soo;Choi, Soo-Young;Cho, Sung-Woo
    • BMB Reports
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    • v.32 no.5
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    • pp.515-520
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    • 1999
  • Incubation of two types of glutamate dehydrogenase isoproteins from bovine brain with the arginine-specific dicarbonyl reagent phenylglyoxal resulted in a biphasic loss of enzyme activity. Reaction of the glutamate dehydrogenase isoproteins with phenylglyoxal caused a rapid loss of 53~62% of the enzyme activities and modification of two residues of arginine per enzyme subunit. Prolonged incubation of the glutamate dehydrogenase isoproteins with phenylglyoxal resulted in the modification of an additional four residues of arginine per enzyme subunit without further loss of the residual activities. Partial protection against inactivation was provided by the coenzyme NADH or substrate 2-oxoglutarate. The most marked decrease in the rate of inactivation was observed by the combined addition of NADH and 2-oxoglutarate, suggesting that the first two modified arginine residues are in the vicinity of the catalytic site. However, inactivation of the glutamate dehydrogenase isoproteins by phenylglyoxal appears to be partial with approximately 40% activity remained after an extended reaction time with excess reagent, suggesting that the modified arginine residues may not be directly involved in catalysis. The lack of complete protection by substrates also suggest the possibility that the modified arginine residues are not directly involved at the active site, and the partial loss of activity by the modification of arginine residues may be due to a conformational change. There were no significant differences between the two glutamate dehydrogenase isoproteins in sensitivities to inactivation by phenylglyoxal, indicating that the microenvironmental structures of the glutamate dehydrogenase isoproteins are very similar to each other.

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Fabrication of a Bottom Electrode for a Nano-scale Beam Resonator Using Backside Exposure with a Self-aligned Metal Mask

  • Lee, Yong-Seok;Jang, Yun-Ho;Bang, Yong-Seung;Kim, Jung-Mu;Kim, Jong-Man;Kim, Yong-Kweon
    • Journal of Electrical Engineering and Technology
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    • v.4 no.4
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    • pp.546-551
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    • 2009
  • In this paper, we describe a self-aligned fabrication method for a nano-patterned bottom electrode using flood exposure from the backside. Misalignments between layers could cause the final devices to fail after the fabrication of the nano-scale bottom electrodes. A self-alignment was exploited to embed the bottom electrode inside the glass substrate. Aluminum patterns act as a dry etching mask to fabricate glass trenches as well as a self-aligned photomask during the flood exposure from the backside. The patterned photoresist (PR) has a negative sidewall slope using the flood exposure. The sidewall slopes of the glass trench and the patterned PR were $54.00^{\circ}$ and $63.47^{\circ}$, respectively. The negative sidewall enables an embedment of a gold layer inside $0.7{\mu}m$ wide glass trenches. Gold residues on the trench edges were removed by the additional flood exposure with wet etching. The sidewall slopes of the patterned PR are related to the slopes of the glass trenches. Nano-scale bottom electrodes inside the glass trenches will be used in beam resonators operating at high resonant frequencies.

Investigations on Possible Roles of C-Terminal Propeptide of a Ca-Independent ${\alpha}$-Amylase from Bacillus

  • Salimi, Ali;Yousefi, Fatemeh;Ghollasi, Marzieh;Daneshjou, Sara;Tavoli, Hesam;Ghobadi, Sirous;Khajeh, Khosro
    • Journal of Microbiology and Biotechnology
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    • v.22 no.8
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    • pp.1077-1083
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    • 2012
  • Previously, an extracellular ${\alpha}$-amylase (BKA) had been purified from the culture of Bacillus sp. KR8104. Subsequently, the crystal structure of the active enzyme revealed a 422 amino acids polypeptide. In this study, the bka was cloned into E. coli, which encoded a polypeptide of 659 amino acids including two additional fragments: one 44 residues N-terminal fragment and another 193 residues C-terminal fragment. In order to investigate the role of the C-terminal fragment, two constructs with and without this region [$BKA{\Delta}$(N44) and $BKA{\Delta}$(N44C193)] were designed and expressed in E. coli BL21. The optimum pH, thermal stability, and the end-products of starch hydrolysis were found to be similar in both constructs. The $K_m$ and $V_{max}$ values for $BKA{\Delta}$(N44) were lower than $BKA{\Delta}$(N44C193), using either starch or ethylidene-blocked 4-nitrophenylmaltoheptaoside as a substrate.

Interaction of Bovine Growth Hormone with Buffalo Adipose Tissue and Identification of Signaling Molecules in Its Action

  • Sodhi, R.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1030-1038
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    • 2007
  • Results on localization of growth hormone receptor (GHR), interaction of growth hormone (GH) with receptor in buffalo adipose tissue and identification of activated signaling molecules in the action of GH are presented. Bovine GH (bGH) was labeled with fluorescein or biotin. Fluorescein-labelled bGH was used for localization of GHRs in buffalo adipocytes. The receptors were present on the cell surface. The affinity of binding of GH to its receptor was determined by designing an experiment in which buffalo adipose tissue explants, biotinylated GH and streptavidin-peroxidase conjugate were employed. The affinity constant was calculated to be $2{\times}10^8M^{-1}$. The receptor density on adipose tissue was found to be 1 femto mole per mg of tissue. Signalling molecules generated in the action of GH were tentatively identified by employing Western blot and enhanced chemiluminescence techniques using anti-phosphotyrosine antibody. Based on molecular weights of proteins reactive to anti-phosphotyrosine antibody, three signaling molecules viz. insulin receptor substrate, Janus activated kinase (Jak) and mitogen activated protein were tentatively identified. These signaling molecules appeared in a time (incubation time of explants with growth hormone) dependent way. The activation of Jak2 was confirmed by employing anti-Jak2 antibody in a Western blot. The activation of Jak2 occurred during 5 min incubation of buffalo adipose tissue explants with GH and incubation for an additional period, viz. 30 min. or 60 min., resulted in a drastic reduction in activation. The results suggest that Jak2 activation is an early event in the action of GH in buffalo adipose tissue.

Electrical and Optical Characteristics of Plasma Display Panel Fabricated by Vacuum In-line Sealing (진공 인라인 실장에 의해 제작된 플라즈마 디스플레이 패널의 전기적ㆍ광학적 특성)

  • Park, Sung-Hyun;Lee, Neung-Hun
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.4
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    • pp.344-349
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    • 2005
  • The optical and electrical characteristics of plasma display panel(PDP) using the vacuum in-line sealing technology compared with the conventional sealing process in this research. This PDP consisted of MgO protecting layer by e-beam evaporation and battier rib, transparent dielectric layer, dielectric layer, and electrodes by screen printer and then sealed off on Ne-Xe(4 %) 400 Torr and 430。C. The brightness and luminous efficiency were good as the base vacuum level was higher, and it was to check the advantage of high vacuum level sealing, one of the strong points of the vacuum in-line sealing process. However, the brightness and luminous efficiency was dropped sharply because of a crack on MgO protecting layer by the difference of the expansion and contraction stress on high temperature in the vacuum states between MgO and substrate. Fortunately, the crack was prevented by MgO was deposited on higher temperature than 300。C. Finally, the PDP, was fabricated by the vacuum in-line sealing process, resulted the lower brightness than processing only the thermal annealing treatment in the vacuum chamber, but the luminous efficiency was increased by the reducing power consumption with the decreasing luminous current. The vacuum in-line sealing technology was not to need the additional thermal annealing process and could reduce the fabrication process and bring the excellent optical and electrical properties without the crack of MgO protecting layer than the conventional sealing process.

The Improvement of Electrical Characteristics of Inkjet-printed Cu films with Stress Relaxation during Thermal Treatment (잉크젯 프린팅된 Cu 박막의 응력해소를 통한 전기적 특성 개선)

  • Yi, Seol-Min;Joo, Young-Chang
    • Journal of the Microelectronics and Packaging Society
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    • v.21 no.4
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    • pp.57-62
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    • 2014
  • Using flexible bismaleimide-triazine co-polymer as a substrate, inkjet-printed Cu films were also investigated for low-cost and process feasibility of flexible electronics. After annealing at $200^{\circ}C$ for 1 h under various reducing ambient, surface color was changed to red and electrical resistivity was decreased to the level of conductor under formic acid ambient. However, its resistivity was much higher than conventional copper films due to surface crack. In order to reduce the residual film stress after annealing, additional isothermal treatment was inserted before anneal hiring the stress relaxation applied in processes of amorphous materials. As a result, no surface crack was observed and electrical resistivity of $3.4{\mu}{\Omega}cm$ was measured after annealing at $230^{\circ}C$ with stress relaxation while electrical resistivity of $7.4{\mu}{\Omega}cm$ was observed after normal annealing without relaxation. The effect of stress relaxation was also confirmed by observing surface crack after decreasing the relaxation time to 0 min.

Development and testing of multicomponent fuel cladding with enhanced accidental performance

  • Krejci, Jakub;Kabatova, Jitka;Manoch, Frantisek;Koci, Jan;Cvrcek, Ladislav;Malek, Jaroslav;Krum, Stanislav;Sutta, Pavel;Bublikova, Petra;Halodova, Patricie;Namburi, Hygreeva Kiran;Sevecek, Martin
    • Nuclear Engineering and Technology
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    • v.52 no.3
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    • pp.597-609
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    • 2020
  • Accident Tolerant Fuels have been widely studied since the Fukushima-Daiichi accident in 2011 as one of the options on how to further enhance the safety of nuclear power plants. Deposition of protective coatings on nuclear fuel claddings has been considered as a near-term concept that will reduce the high-temperature oxidation rate and enhance accidental tolerance of the cladding while providing additional benefits during normal operation and transients. This study focuses on experimental testing of Zr-based alloys coated with Cr-based coatings using Physical Vapour Deposition. The results of long-term corrosion tests, as well as tests simulating postulated accidents, are presented. Zr-1%Nb alloy used as nuclear fuel cladding serves as a substrate and Cr, CrN, CrxNy layers are deposited by unbalanced magnetron sputtering and reactive magnetron sputtering. The deposition procedures are optimized in order to improve coating properties. Coated as well as reference uncoated samples were experimentally tested. The presented results include standard long-term corrosion tests at 360℃ in WWER water chemistry, burst (creep) tests and mainly single and double-sided high-temperature steam oxidation tests between 1000 and 1400℃ related to postulated Loss-of-coolant accident and Design extension conditions. Coated and reference samples were characterized pre- and post-testing using mechanical testing (microhardness, ring compression test), Thermal Evolved Gas Analysis analysis (hydrogen, oxygen concentration), optical microscopy, scanning electron microscopy (EDS, WDS, EBSD) and X-ray diffraction.

Kinetic Properties of the Dye-Coupled Cytoplasmic Polyol Dehydrogenase from Gluconobacter melanogenus (Gluconobacter melanogenus 로부터의 폴리올 탈수소효소에 대한 반응속도론적 특성에 관한 연구)

  • Kang-Wha Kim;Hyun-Jae Lee
    • Journal of the Korean Chemical Society
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    • v.24 no.4
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    • pp.315-321
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    • 1980
  • A steady-state kinetic study on a dye-coupled cytoplasmic polyol dehydrogenase from G. melanogenus was carried by the initial velocity measurements in the direction of the polyol oxidation and the product inhibition by D-fructose. For the initial rate experiments, D-mannitol and D-sorbitol were employed as the specific polyol substrates and 2,6-dichlorophenolin-dophenol (DPIP) as the specific cofactor substrate for the enzyme. When the polyol and DPIP were examined by varying one of substrates and by fixing the second, the corresponding reciprocal plots showed the typical parallel pattern. This suggests that the enzyme from G. melanogenus proceeds by a Ping Pong Bi-Bi mechanism in which the polyol may account as the first reactant-in, and the ketose formed as the first product-out, respectively. The product inhibition patterns obtained by D-fructose (one no-inhibition, one non-competitive, and two competitive) may also provide an additional conformatory evidence for the above mechanism. Based on the kinetic parameters obtained, it was also suggested that the rate-limiting step in the direction of polyol oxidation is associated with the release of the ketose from the Enzyme${\cdot}$Polyol complex.

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Magnetic Properties of FePt:C Nanocomposite Film

  • Ko, Hyun-Seok;A. Perumal;Shin, Sung-Chul
    • Proceedings of the Korean Magnestics Society Conference
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    • 2003.06a
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    • pp.220-221
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    • 2003
  • Equiatomic FePt and CoPt alloy thin films have received considerable attention as possible magnetic and magneto-optic recording because of their high magnetic anisotropy energy and high coercivity. The high coercivity in these thin films is due to the presence of finely dispersed ordered FePt phase mixed with disordered FePt phase. However, a high temperature treatment, either substrate heating during deposition or post annealing, is needed to obtain the ordered L1$\_$0/ phase with high value of magneto crystalline anisotropy. Recent microstructural studies on these films suggest that the average grain size ranges from 10-50 nm and the grains are magnetically coupled between each other. On the other hand, the ultrahigh-density magnetic recording media with low media noise imposes the need of a material, which consists of magnetically isolated grains with size below 10 nm. The magnetic grain isolation can be controlled by the amount of additional non-magnetic element in the system which determines the interparticle separation and therefore the interparticle interactions. Recently, much research work has been done on various non-magnetic matrices. Preliminary studies showed that the samples prepared in B$_2$O$_3$ and Carbon matrices have shown strong perpendicular anisotropy and fine grain size down to 4nm, which suggest these nanocomposite films are very promising and may lead to the realization of a magnetic medium capable of recording densities beyond 1 Tb/in$^2$. So, in this work, the effect of Carbon doping on the magnetic properties of FePt nanoparticles were investigated.

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In-situ Annealing of $MgB_2$ Thin Films Prepared By rf Magnetron Co-Sputtering (Rf co-sputtering으로 제작한 MgB$_2$ 박막의 in-situ 열처리 효과)

  • 김윤원;안종록;이순걸;이규원;김인선;박용기
    • Progress in Superconductivity
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    • v.5 no.2
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    • pp.105-108
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    • 2004
  • We have studied effects of in-situ annealing on the fabrication of superconducting MgB$_2$ thin films prepared by rf magnetron co-sputtering. The Films were deposited on A1$_2$O$_3$ (1102) substrates at room temperature by using Mg and B targets. To trap remnant $O_2$ gas in the chamber, we used 20 mtorr Af sputter-gas balanced with 5 mol % of H$_2$ gas. To enhance adhesion to the substrate a thin layer of B was deposited prior to the codeposition of Mg and B. After completion of the film deposition, an additional Mg layer was deposited on top to compensate for Mg loss during the subsequent in-situ annealing. We have investigated the effects of two most important annealing parameters that are the Mg-to-B composition ratio and the annealing temperature. The range of the Mg-to-B composition ratio was from 0.42 to 0.85, and that of the annealing temperature was 500 $^{\circ}C$∼750 $^{\circ}C$. The Best result was obtained for the composition ratio of about 10% Mg excess from the stoichiometry and the annealing temperature of 700 $^{\circ}C$. Based on these results, we obtained films with T$_{c}$ : 36.5 K by further refining the fabrication process.s.

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