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Detection of Lamivudine-Resistant Mutations of HBV DNA Polymerase Gene Using PCR-Direct Sequencing

  • Lee, Kyung-Ok;Lee, Hye-Jung;Byun, Ji-Young;Lee, Sung-Yeun;Kim, Jeong-Sook;Jung, Na-Young;Chung, Soo-Jin;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.196-202
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    • 2006
  • Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

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Contrasting Styles of Gold and Silver Mineralization in the Central and Southeastern Korea (한국 중부와 동남부지역 금·은광화작용의 성인적 특성)

  • Choi, Seon-Gyu;Choi, Sang-Hoon
    • Economic and Environmental Geology
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    • v.28 no.6
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    • pp.587-597
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    • 1995
  • Two distinct precious-metal mineralizations actively occur at central and southeastern Korea which display consistent relationships among geologic, geochemical and genetic environments. A large number of preciousmetal vein deposits in the central Korea occur in or near Mesozoic granite batholiths elongated in a NE-SW direction. Whereas, gold and/or silver deposits in the southeastern Korea occur within Cretaceous volcanic and sedimentary rocks. However, most of the precious-metal deposits in the southeastern Korea show characteristics of the silver-rich deposits than the gold-rich deposits in the central Korea. Two epochs of main igneous activities are recognized: a) Jurassic Daebo igneous activity between 121 and 183 Ma, and b) Cretaceous Bulgugsa igneous activity between 60 and 110 Ma. Precious-metal mineralization took place between 158 and 71 Ma, coinciding with portions of the two magmatic activities. Contrasts in the style of mineralization, together with radiometric age data and differences in geologic settings reflect the genetically variable natures of hydrothermal activities from middle Jurassic to late Cretaceous time. The compilation and re-evaluation of these data suggest that the genetic types of hydrothermal precious-metal vein deposits in the central and southeastern Korea varied with time. The Jurassic and early Cretaceous mineralizations are characterized by the Au-dominant type, but tend to change to the Au-Ag and/or Ag-dominant types at late Cretaceous. The Jurassic Au-dominant deposits commonly show several characteristics; prominent associations with pegmatites, simple massive vein morphologies, high fmeness values in ore-concentrating parts, and a distinctively simple ore mineralogy such as Fe-rich sphalerite, galena, chalcopyrite, Au-rich electrum, pyrrhotite and/or pyrite. The Cretaceous precious-metal deposits are generally characterized by some- features such as complex vein morphologies, low to medium fmeness values in the ore concentrates, and abundance of ore minerals including Ag sulfosalts, Ag sulfides, Ag tellurides and native silver. Mineralogical and fluid inclusion studies indicate that the Jurassic Au-dominant deposits in the central area were formed at the high temperature (about $300^{\circ}$ to $500^{\circ}C$) and pressure (about 4 to 5 kbars), whereas mineralizations of the Cretaceous Au-Ag and Ag-dominant deposits were occurred at the low temperature (about $200^{\circ}$ to $350^{\circ}C$) and pressure (<0.5 kbars) from the ore fluids containing more amounts of less-evolved meteoric waters.

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Isolation of Glucoamylase Producing Yeasts and its Enzymatic Characteristics (Glucoamylase 생성효모의 분리와 효소학적 특성)

  • Park, So-Young;Choi, Soon-Young;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.386-393
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    • 1999
  • Ethanol is considered as one of the most suitable substitutes for the petroleum, since it offers attractive functional features at an economical cost. Glucoamylase producing yeasts were isolated and characterized. Based on the morphological character, carbon fermentations, assimilation of carbon and nitrate, growth on vitamine-free medicine, and urease activity, five isolates of Saccharomyces diastaticus, two isolates of Saccharomycopsis fibuligera, and two of Schwanniomyces occidentalis, and each isolate of Ambrosiozyma monospora and Lipomyces kononenkoae were identified. Among 12 isolates, one of the S. diastaticus, E3 showed the highest activity of glucoamylase and identified as Saccharomyces diastaticus. The hydrolysis of starch by the E3 strain showed the release of considerable amount of reducing sugar, along with the reduction in iodine staining capacity. The product of action of glucoamylase, glucose was determined by thin-layer chromatography. The enzyme activity was found to be stable in broad pH range of $5.0{\sim}7.0$ with optimal activity at pH $5.0{\sim}6.0$. The enzyme showed optimal antivity at $50^{\circ}C{\sim}60^{\circ}C$. Soluble starch and glucose were better carbon sources for the enzyme production than xylose and glycerol. $Na^+\;and\;Mg^{2+}$ increased the glucoamylase activity, however $Hg^{2+}\;and\;Ag^{2+}$ inhibited the activity. Soluble starch was the best substrate for the enzyme activity.

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High Efficiency Solar Cell(I)-Fabrication and Characteristics of $N^+PP^+$ Cells (고효율 태양전지(I)-$N^+PP^+$ 전지의 제조 및 특성)

  • 강진영;안병태
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.18 no.3
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    • pp.42-51
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    • 1981
  • Boron was predeposited into p (100) Si wafer at 94$0^{\circ}C$ for 60minutes to make the back surface field. High tempreature diffusion process at 1145$^{\circ}C$ for 3 hours was immediately followed without removing boron glass to obtain high surface concentration Back boron was annealed at 110$0^{\circ}C$ for 40minutes after boron glass was removed. N+ layer was formed by predepositing with POCI3 source at 90$0^{\circ}C$ for 7~15 minutes and annealed at 80$0^{\circ}C$ for 60min1es under dry Of ambient. The triple metal layers were made by evaporating Ti, Pd, Ag in that order onto front and back of diffused wafer to form the front grid and back electrode respectively. Silver was electroplated on front and back to increase the metal thickness form 1~2$\mu$m to 3~4$\mu$m and the metal electrodes are alloyed in N2 /H2 ambient at 55$0^{\circ}C$ and followed by silicon nitride antireflection film deposition process. Under artificial illumination of 100mW/$\textrm{cm}^2$ fabricated N+PP+ cells showed typically the open circuit voltage of 0.59V and short circuit current of 103 mA with fill factor of 0.80 from the whole cell area of 3.36$\textrm{cm}^2$. These numbers can be used to get the actual total area(active area) conversion efficiency of 14.4%(16.2%) which has been improved from the provious N+P cell with 11% total area efficiency by adding P+ back.

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A Comparison Analysis on the Efficiency of Solar Cells of Shingled Structure with Various ECA Materials (다양한 ECA 소재를 활용한 shingled 구조의 태양전지 효율 비교 분석)

  • Jang, Jae Joon;Park, Jeong Eun;Kim, Dong Sik;Choi, Won Seok;Lim, Donggun
    • Journal of the Korean Solar Energy Society
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    • v.39 no.4
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    • pp.1-9
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    • 2019
  • Modules using 6 inch cells have problems with loss due to empty space between cells. To solve this problem made by shingled structure which can generate more power by utilizing empty space by increasing the voltage level than modules made in 6inch cell. Thus, in this paper, the c-Si cutting cells were produced using nanosecond green laser, and then the ECA was sprayed and cured to perform cutting cell bonding. Three types of ECA materials (B1, B2, B3) with Ag as the main component were used, and experimental conditions varied from 5 to 120 seconds of curing time, 130 to $210^{\circ}C$ of curing temperature, and 1 to 3 of curing numbers. As a results of experiments varying curing time, B1 showed efficiency 19.88% in condition of 60 seconds, B2 showed efficiency 20.15% in 90 seconds, and B3 showed efficiency 20.27% in 60 seconds. In addition, experiments with varying curing temperature, It was confirmed highest efficiency that 20.04% in condition of $170^{\circ}C$ with B1, 20.15% in condition of $150^{\circ}C$ with B2, 20.27% in condition of $150^{\circ}C$ with B3. These are because the Ag particles are densely formed on the surface to make the conduction path. After optimizing the conditions of temperature and curing time, the secondary-tertiary curing experiments were carried out. as the structural analysis, conditions of secondary-tertiary curing showed cracks that due to damp heat aging. As a result, it was found that the ECA B3 had the highest efficiency of 20.27% in condition of 60 seconds of curing time, $150^{\circ}C$ of curing temperature, and single number of curing, and that it was suitable for the manufacture of Solar cell of shingled structure rather than ECA B1 and B2 materials.

A Study on the Design and Fabrication of High Performance Large Current Mica Capacitor for Energy Storage Facility Applications (에너지 저장설비 응용을 위한 고 성능 대 전류 마이카 커패시터 설계 및 제작에 관한 연구)

  • Jung, Myung-Hee;Yun, Eui-Jung
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.60 no.10
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    • pp.1888-1894
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    • 2011
  • In this study, large-current (75 - 400 A), high-voltage (500 - 1000 $V_{rms}$), reliable capacitors with capacitances (C) of 100 - 1000 nF were developed for energy storage facility applications. Mica was used as the dielectric of the capacitors. In order to form a parallel stack of a capacitor element, 50 ${\mu}m$ thick mica sheets with a size of $30mm{\times}35mm$ were used with lead foils for the plate lead type of mica capacitors (HCM-L), while the same sizes of mica sheets coated by Ag paste were employed with lead foils for the parallel plate terminal type (HCM-C). The developed capacitors exhibited well behaviored device characteristics which meet the requirements of the capacitors. The developed capacitors also showed excellent characteristics for thermal shock test. The stability characteristics of developed capacitors for large current stress was superior to those measured for the capacitors prepared recently by CDETm.

Purification and Charaterization of Antifungal Chitinase from Indigenous Antagonistic Microorganism Serratia sp. 3095

  • Lee, Eun-Tag;Kim, Sang-Dal
    • Journal of Applied Biological Chemistry
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    • v.42 no.1
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    • pp.7-11
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    • 1999
  • An extracellular chitinase of the selected strong antifungal microorganism, Serratia sp. 3095, was purified by salting out, affinity adsorption, Sepadex G-100 gel fitration, Sepadex G-75 gel fitration and DEAE Sepadex A-50 chromatography. The molecular weight of the purified chitinase was estimated to be 62,000 dalton by SDS-PAGE. Optimal pH and temperature of the chitinase were pH 7.5 and 45, respectively. The enzyme retained more than 80% of the activity between pH 5.5 and pH 10.5, and below $50^{\circ}C$ but was unstable above $60^{\circ}C$, below pH 5.0. The activity of the chitinase was inhibited about 60% by $Sn^{2+}$, 40% by $Hg^{2+}$ and $Ag^+$, 70% by AHA, 40% by iodoacetate, 35% by thiourea and p-CMB, but stabilized by SDS. $K_m$ value of the purified chitinase was 3.68 mg/ml for colloidal chitin. The chitinase from Serratia sp. 3095 showed antifungal activity to Fusariurm solani.

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Analysis of Void Effects on Mechanical Property of BGA Solder Joint (솔더 접합부에 생성된 Void의 JEDEC 규격과 기계적 특성에 미치는 영향)

  • Lee, Jong-Gun;Kim, Kwang-Seok;Yoon, Jeong-Won;Jung, Seung-Boo
    • Journal of the Microelectronics and Packaging Society
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    • v.18 no.4
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    • pp.1-9
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    • 2011
  • Understanding the void characterization in the solder joints has become more important because of the application of lead free solder materials and its reliability in electronic packaging technology. According to the JEDEC 217 standard, it describes void types formed in the solder joints, and divides into some categories depending on the void position and formation cause. Based on the previous papers and the standards related to the void, reliability of the BGA solder joints is determined by the size of void, as well as the location of void inside the BGA solder ball. Prior to reflow soldering process, OSP(organic surface preservative) finished Cu electrode was exposed under $85^{\circ}C$/60%RH(relative humidity) for 168 h. Voids induced by the exposure of $85^{\circ}C$/60%RH became larger and bigger with increasing aging times. The void position has more influence on mechanical strength property than the amount of void growth does.

Purification and Characterization of a Chitinase from Cytophaga sp. HJ Isolated from Sea Sand

  • Lee, Dong-Mi;Noh, Hee-Jung;Lee, Kang-Man
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.839-846
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    • 1999
  • An extracellular chitinase-producing bacterial strain induced by colloidal chitin was isolated from sea sand and was identified to be a member of the genus Cytophaga. The chitinase was purified successively by 30-60% ammonium sulfate fractionation, and DEAE-Bio gel A column, Octyl-Sepharose CL-4B column, and DEAE-Bio gel A column chromatographies. The enzyme had a molecular mass of 59.75 kDa, and the amino terminal amino acid sequence was ATPNAPVISW MPTDXXLQNXS. The enzyme acted better on colloidal chitin as a substrate than on chitosan. For colloidal chitin and chitosan (Degree of Acetylation, 15-25%), $K_{cat}$ values were 0.60U/mg and 0.08U/mg, respectively. HPLC analysis of the enzymatic reaction products showed that the chitinase produced mostly N-acetyl-D-glucosarnine and di-N-acetylchitobiose. The optimum temperature and pH for the enzyme were $50^{\circ}C$ and 4.0, respectively. N-Bromosuccinimide and $Hg^{2+}$ inhibited the chitinase activity as much as 90%, and $Sb^{3+}$, diethylpyrocarbonate, and $Ag^{+}$ inhibited it by 50-70%.

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Combined Therapy of Alfa-Interferon and Thymodulin on Children with Chronic Active Hepatitis B (소아의 B형 만성 활동성 간염에서 저용량 ${\alpha}$-Interferon과 Thymodulin의 병용 치료 효과)

  • Choe, Byung-Ho;Ko, Cheol-Woo
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.1 no.1
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    • pp.79-89
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    • 1998
  • Purpose: Though many antiviral or immunomodulatory agents have been used in patients with chronic HBV hepatitis, interferon is considered to be the only effective therapeutic agent so far. Among immunomodulatory agents, thymodulin, the oral form of thymosin, is currently in clinical trial. We compared the efficacy of alfa-interferon therapy alone with a combined therapy of alfa-interferon and thymodulin in children with chronic active hepatitis B. Method: Twenty three children aged 4.4~13.7 years who were known to be positive for HBsAg and HBeAg in serum for at least 6 months and who had biopsy-proven chronic active hepatitis were given either combined therapy of alfa-interferon and thymodulin or alfa-interferon alone, and all children were HBV DNA positive in their serum at the beginning. Follow-ups have been done for at least 1 year after a 6 month course of therapy and clearance of viral replication markers has been evaluated. Results: 1) During follow up period, 11 (48%) children were seroconverted to anti-HBe and were cleared of HBV DNA from their serum. However, 2 of them relapsed after discontinuance of interferon therapy. 2) Seroconversion occurred more frequently among those who had not been vertically transmitted, had elevated serum ALT levels and low HBV DNA levels before interferon therapy. 3) There was no significant advantage of the combined therapy with thymodulin compared to interferon therapy alone. Conclusion: Combined therapy of alfa-interferon and thymodulin failed to demonstrate synergistic effect. We think that combination therapies of alfa-interferon with other antiviral or immunomodulatory agents need to be studied in order to achieve better therapeutic responses.

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