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Characterization of proteases of Toxoplasma gondii (Toxoplasma gondii에서 단백질 분해 효소의 특징)

  • Choe, Won-Yeong;Nam, Ho-U;Yun, Ji-Hye
    • Parasites, Hosts and Diseases
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    • v.27 no.3
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    • pp.161-170
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    • 1989
  • The proteases of Toxoplasma gcndii were purified partially and characterisrd for some biochemical properties including various chromatographic patterns, major catalytic classes, and conditions to promote the activity of these enzymes. When Toxoplasma extract was incubated with 3H-casein at various pH, peak hydrolysis of casein was observed at pH 6.0 and at pH 8.5. Proteasfs working at pH 6.0 and at pH 8.5 were purified partially by conventional methods of chromatographies of DE52 anion rxchange, Sephadex G-200 gel permeation, and hydroxylapatite chromatography. Partially purified enzymes were tested by site-specific inhibitors and promotorf. The protease working at pH 6.0 was inactivated by iodoacetamide with LDso of 10-5 M and promoted by dithiothreitol, while the protease working at pH 8.5 was inhibited by phenylmethylsulfonyl fluoride with LD50 of 10-5 M and was Promoted by ATP (excess ATP beyond 2 mM inhibited the activity reversely). The protease of pH 8.5 had the activity of ATPase which might exert the energy to its action. Therefore the former was referred to as a cysteinyl acid protease and the latter, ATP-dependent neutral serine protease.

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Demonstration of species-specific and cross reactive components of Paragonimus tvestermani crude worm antigen by EITB (Immunoblot를 이용한 폐흡충 비항원의 특이 항원대의 증명)

  • Joo, Kyoung-Hwan;Ahn, Hyuck;Chung, Myung-Sook;Lim, Han-Jong
    • Parasites, Hosts and Diseases
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    • v.27 no.1
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    • pp.9-14
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    • 1989
  • Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

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Changes of Chemical Bond in Woody Charcoal from Different Carbonization Temperatures (목질탄화물 내의 화학 결합 변화)

  • Jo, Tae-Su;Lee, Oh-Kyu;Choi, Joon-Weon;Cho, Sung-Taig;Kim, Suk-Kuwon
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.1
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    • pp.87-93
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    • 2009
  • Properties and chemical bonding of wood charcoal were investigated to understand the chemistry occurring in wood carbonization. From the pH changes of wood charcoal, it is revealed that it becomes acidic to weakly basic for charcoal carbonized at about $300^{\circ}C$, whereas it turns to basic at higher carbonization temperature higher than $600^{\circ}C$. Also, the ratio of carbon atoms in the charcoal was increased with increasing the carbonization temperature, while those of oxygen and hydrogen atoms. This tendency was significant when the carbonization temperature was increased up to $600^{\circ}C$ and the ratio changes of the atoms became stable at above $600^{\circ}C$. In the changes of chemical bonding, the ratio of C-C bonding was increased and those of C-O-H and C-O-R bonding was decreased significantly. It is considered that bondings connected to oxygen atoms tends to be broken, and the ratio of C-C bonding increased. Consequently, it is expected that this change may causes occurrence of new functional groups. In addition to that, it seems to be that the chemical bondings undergo the partial decomposition, formation, and recombination steps, Because ratio of C=O bonding tended to be increased or decreased by increasing the carbonization temperature. This understanding of chemical bond changes in charcoal can be a compensative consideration on the knowledges made only by physical parameters in the properties of micro-pore which has limited to explain the phenomenon. Also, it is considered that this can be treated as a basic knowledge for upgrading and development of use of wood charcoal.

Synthesis of L-3-[$^{123}I$]iodo-${\alpha}$-methyltyrosine and Biodistribution in 9L Glioma Bearing Rats (L-3-[$^{123}I$iodo-${\alpha}$-methyltyrosine 합성과 9L Glioma 이식 백서 분포조사)

  • Yang, Seung-Dae;Lim, Sang-Moo;Woo, Kwang-Sun;Chung, Wee-Sup;Chun, Kwon-Soo;Suh, Yong-Sup;Lim, Jong-Seok;Park, Hyon;Yun, Yong-Ki;Lee, Jong-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.1
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    • pp.105-109
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    • 1995
  • L-3-[$^{123}I$]iodo-${\alpha}$-methyltyrosine([$^{123}I$] IMT) was synthesized by electrophilic radio-iodination using chloramine-T and Iodobead in phosphate buffered solution. And the biodistribution was examined in 9L glioma bearing rats. The radiosynthesis of [$^{123}I$]IMT with iodobead was simpler and higher in radiochemical yield(88%) than the method using chloramine-T(83%) as radioiodinating reagent. The highest yield was obtained from the reaction using 1 piece of Iodobead, $200{\mu}g$ ${\alpha}$-methyltyrosine in $100{\mu}l$ phosphate-buffered solution(pH 5.5) and the reaction was completed in 7min. 24 hours after the injection, the biodistribution in 9L glioma transplanted rats revealed the in vivo deiodination, the excretion via kidney, and 3 times higher uptake in the tumor than normal brain. These results suggest the promising clinical use of [$^{123}I$] IMT in the various malignancies.

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Prevention of P-i Interface Contamination Using In-situ Plasma Process in Single-chamber VHF-PECVD Process for a-Si:H Solar Cells

  • Han, Seung-Hee;Jeon, Jun-Hong;Choi, Jin-Young;Park, Won-Woong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.204-205
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    • 2011
  • In thin film silicon solar cells, p-i-n structure is adopted instead of p/n junction structure as in wafer-based Si solar cells. PECVD is a most widely used thin film deposition process for a-Si:H or ${\mu}c$-Si:H solar cells. For best performance of thin film silicon solar cell, the dopant profiles at p/i and i/n interfaces need to be as sharp as possible. The sharpness of dopant profiles can easily achieved when using multi-chamber PECVD equipment, in which each layer is deposited in separate chamber. However, in a single-chamber PECVD system, doped and intrinsic layers are deposited in one plasma chamber, which inevitably impedes sharp dopant profiles at the interfaces due to the contamination from previous deposition process. The cross-contamination between layers is a serious drawback of a single-chamber PECVD system in spite of the advantage of lower initial investment cost for the equipment. In order to resolve the cross-contamination problem in single-chamber PECVD systems, flushing method of the chamber with NH3 gas or water vapor after doped layer deposition process has been used. In this study, a new plasma process to solve the cross-contamination problem in a single-chamber PECVD system was suggested. A single-chamber VHF-PECVD system was used for superstrate type p-i-n a-Si:H solar cell manufacturing on Asahi-type U FTO glass. A 80 MHz and 20 watts of pulsed RF power was applied to the parallel plate RF cathode at the frequency of 10 kHz and 80% duty ratio. A mixture gas of Ar, H2 and SiH4 was used for i-layer deposition and the deposition pressure was 0.4 Torr. For p and n layer deposition, B2H6 and PH3 was used as doping gas, respectively. The deposition temperature was $250^{\circ}C$ and the total p-i-n layer thickness was about $3500{\AA}$. In order to remove the deposited B inside of the vacuum chamber during p-layer deposition, a high pulsed RF power of about 80 W was applied right after p-layer deposition without SiH4 gas, which is followed by i-layer and n-layer deposition. Finally, Ag was deposited as top electrode. The best initial solar cell efficiency of 9.5 % for test cell area of 0.2 $cm^2$ could be achieved by applying the in-situ plasma cleaning method. The dependence on RF power and treatment time was investigated along with the SIMS analysis of the p-i interface for boron profiles.

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MOCVD를 이용한 자발성장 InAs 양자점의 적층 성장 시 발생하는 파장변화량 제어

  • Choe, Jang-Hui;An, Seong-Su;Yu, Su-Gyeong;Lee, Jong-Min;Park, Jae-Gyu;Lee, Dong-Han;Jo, Byeong-Gu;Han, Won-Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.150-151
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    • 2011
  • 양자점 Laser Diode(LD)는 낮은 문턱전류, 높은 미분 이득을 갖으며 또한 온도변화에도 안정적이기 때문에 광통신분야에서 광원으로 양자점 LD를 사용하기 위한 연구가 계속되고 있다. 양자점은 fill factor가 낮기 때문에 양자점의 밀도를 높이거나 양자점을 적층 성장하여 fill factor를 높인다. 그러나 양자점을 적층 성장하면 각 층간의 응력, 수직적 결합, 전기적인 결합이 생기며 이는 양자점의 전기적, 광학적 특성에 영향을 미친다. 본 연구에서는 metal organic chemical vapor deposition (MOCVD)을 이용하여 InP기판 위에 자발성장 법으로 InAs 양자점을 다주기 성장하였으며 photoluminescence (PL)을 이용하여 광학적 특성을 분석하였다. precursor는 trimethylindium (TMI), trimethylgalium (TMGa), $PH_3$, $AsH_3$를 사용하였으며 carrier gas는 $H_2$를 사용하였다. InAs 양자점은 1100 nm의 파장을 갖는 InGaAsP barrier 위에 성장하였고, InAs와 InGaAsP의 성장온도는 $520^{\circ}C$이며 InAs 양자점 성장시 V/III 비는 3.66으로 일정하게 유지하였다. 그림 1은 양자점 성장시간을 0.11분으로 고정하여 3주기(A), 5주기(B), 8주기(C) 성장한 구조이며 그림 2는 양자점 성장시간을 3주기마다 0.01분씩 줄여가며 3주기는 0.11분${\times}$3(D), 6주기는 0.11분${\times}$3+0.10분${\times}$3(E), 9주기는 0.11분${\times}$3+0.10분${\times}$3+0.09분${\times}$3(F) 으로 성장한 성장구조이다. 각 성장한 시료는 PL을 이용하여 파장과 반치폭을 측정하였다. 그림 3은 양자점 성장시간을 고정한 시료 A, B, C의 PL파장과 PL반치폭 데이터이다. PL파장은 A, B, C 시료 각각 1504 nm, 1571 nm, 1702 nm이며 반치폭은 각각 140 meV, 140 meV, 150 meV이다. PL파장과 반치폭은 각각 3주기에서 6주기로 증가할 때 67 nm, 0 meV 6주기에서 9주기로 증가할 때는 131 nm, 10 meV 증가하였다. 다음 그림4는 양자점 성장시간을 조절하여 성장한 양자점 시료 D, E, F의 PL파장과 PL반치폭 데이터이다. PL파장은 D, E, F 시료 각각 1509 nm, 1556 nm, 1535 nm이며 반치폭은 각각 137 meV, 138 meV, 144 meV이다. PL파장과 반치폭은 각각 3주기에서 6주기로 증가할 때 47 nm, 1 meV 증가하였고, 6주기에서 9주기로 증가할 때는 21 nm 감소, 6 meV 증가하였다. 양자점 성장시간을 고정하여 다주기를 성장하였고 또 3주기마다 양자점 성장시간을 달리하여 다주기를 성장하였으며 PL을 이용해 광학적 특성을 연구하였다. 성장된 양자점의 PL 파장과 PL 반치폭 변화를 통해 적층구조에서 성장 주기가 늘어날수록 양자점의 크기가 증가하는 것을 확인하였고 또한 적층성장을 할 때 양자점 성장시간을 줄임으로써 양자점의 크기 변화를 제어 할 수 있었다.

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Purification and Characterization of Lipase from Acinetobacter sp. B2 Isolated from Oil­contaminated Soil (유류오염지역에서 분리한 Acinetobacter sp. B2로부터의 Lipase 정제 및 특성)

  • Son Seung Hwa;Park Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.320-327
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    • 2004
  • Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil sam­ples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of p­nitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.

Characteristics of Parathion Hydrolase by Pseudomonas rhodesiae H5 (Pseudomonas rhodesiae H5가 생산하는 Parathion Hydrolase의 특성)

  • Yun Nam Kyung;Park Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.199-204
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    • 2004
  • The parathion hydrolase (OPH) produced by Pseudomonas rhodesiae H5 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Parathion hydrolase from crude extracts of P. rhodesiae H5 has two components designated as OPH $I_1$ and OPH $I_2$, Optimum pH and temperature of OPH $I_1$and OPH $I_2$ were pH 7.2 and $30^{\circ}C$, and pH 7.6 and $37^{\circ}C$, respectively. The activation energy of OPH $I_1$ for the hydrolysis of parathion was 3.01 ㎉/I, II, III in the temperature range of $4^{\circ}C$ to $30^{\circ}C$, and Michaelis constant ($K_m$) for parathion was 69.2 ${\mu}M$. The activation energy of OPH $I_2$ for the hydrolysis of parathion was 4.07㎉/㏖ in the temperature range of $4^{\circ}C$ to $37^{\circ}C$, and Michaelis constant for parathion was 150.9${\mu}M$. Furthermore OPH $I_1$ was completely inhibited by 1 mM $Ca^2+$, $Cu^2+$, $Mg^2+$, $Ni^2+$, but OPH $I_2$ was less inhibited than OPH $I_1$ by the metals used in this study.

Studies on the Spore Formation of Aspergillus niger in Potassium Acetate Medium (아세트산염 배지에서의 검정곰팡이(Aspergillus niger)의 포자형성에 관한 연구)

  • Lee, Ho-Young;Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.15 no.3
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    • pp.149-157
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    • 1987
  • This study was undertaken to investigate the differentiation, from spore germination to hyphae growth and phialide formation, of Aspergillus niger through the method of synchronous and submerged culture. Through continuous experiments by shake culture with potassium acetate medium, we observed the formation of spores at appropriate concentration and pH. Potassium acetate medium was set pH 5.5, 6.0, 6.5 and 7.0 on each scale, and control, 20 mM, and 40 mM, 80 mM and 160 mM concentrations on the other scale. Aspergillus niger was cultured in the defined media at $28^{\circ}C$, and mycelial dry weight, changes of pH and the onset of sporulation were checked. The mycelial dry weight, increased in potassium acetate medium, and pH increased during mycelial growth and gradually decreased after the spore formation. When pH increased excessively in Potassium acetate medium with pH 7.0, the mycelia could not adapt and mycelial dry weight decreased gradually. At pH 5.5, the onset of sporulation was done within one day at 20 mM it took, at 80 mM three days and at 160 mM concentration. in two days, at 40 mM one to four days were taken, 80 mM concentration respectively. At pH 6.5, the onset of sporulation was done in three days and four days at 80 mM concentrations respectively. Spore formation was not shown at pH 7.0. In controlled medium with all levels of pH, spore formation was not shown.

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Differentiation Trend of Rare Earth Elements of the Skaergaard Intrusion (Skaergaard 암체의 희토류의 분화경향)

  • Yun D. Jang
    • Economic and Environmental Geology
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    • v.34 no.6
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    • pp.617-625
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    • 2001
  • The Skaergaard intrusion is widely considered a type example of a strongly fractionated, layered intrusion that has undergone extensive in situ igneous differentiation. The Intrusion, therefore, should be a good locality for modeling trace element vriation in a closed system. Previous studios (Haskin and Haskin, 1968; Faster et al., 1974), however, have suggested thats the rare earth elements in whole rocks and mineeral separates from the Intrusion did not fellow the expected trend for closed system crystatllization. Trace element modeling using published distribution coefficients, modal abundances of the coexisting minerals, and the concentration of trace elements In whole rocks and mineral separates from the Skaergaard Intrusion, reveals that the rare earth elements were significantly Influenced by the crystallization of abundant apatite in the Layered Series suring the final stages of crystallization. The results of trace element modeling also suggcsts that apatite, which appears sporadically in the UBS, is not a primary liquidus phase in these samples as previously suggested (Naslund, 1984) but an interstitial phase that (lid not directly effect trace element abundances In the evolving magma As the Skaergaard magma coaled convection, or convected as small Isolated cells during the final stages of differentiation, an elebated $P_{H2O}$ Induced by accumulation of volatile elements near the roof of the magma chamber ingibited or delayed the precipitation of primary apatite in the UBS If the Skaergaard differentiation Is modeler assuming primary apatite crystallization In the upper par of the LS where abundant modal apatite is present, and only late stage crystallization of apatite In the UBS where apatite Is less abundant, rare earth elements abundances follow a closed system variation trend. These results rule but any differentiation model for the Skaergaard Intrusion that Includesvolumetrically significant injections or discharges of magma Into or out of the chamber during the final 20% of the crystallization history.

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