• Korean Journal of Agricultural Science
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• v.16 no.2
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• pp.225-238
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• 1989

These studies were conducted to investigate the extraction, purification and characterization of oriental pear (Niitaka. Pyrus pyrifolia Nak.) protease, and the results obtained were as follows: 1. Oriental pear protease was effectively extracted by the method of homogenizing pear pulp with 0.7 volume of 0.1M-sodium phosphate buffer, pH 6.5 containing 5mM-cysteine, 40mM-2-mercaptoethanol and 2mM-EDTA at 10,000 rpm for 5 min. 2. The protease was purified by ammonium sulfate fractionation, Sephadex G-100 filtration and DEAE-Sephadex A-50 column chromatography, and the purified enzyme gave a single protein band on polyacrylamide gel electrophoresis. 3. The specific activity of purified enzyme was 29.65 unit/mg protein and the yield was 7.22%. 4. The moecular weight of the protease was estimated to be about 51,000 by SDS-polyacrylamide gel electrophoresis, and the enzyme had Km value of 54.5 mg/ml for casein. 5. The purified enzyme had a maximum activity at pH 6.0 and $50^{\circ}C$, and was stable from pH 5.5-6.5 and at temperatures below $50^{\circ}C$ 6. Casein was a better substrate for this protease compared to hemoglobin. 7. The enzyme activity was markedly inhibited by p-chloromercuribenzoic acid and heavy metal salts such as $HgCl_2$ and $MnSO_4$ also considerably inhibited the enzyme activity.

• KSCE Journal of Civil and Environmental Engineering Research
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• v.41 no.4
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• pp.347-355
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• 2021

During the earthquake, for multi-story structure, if the first floor is soft, the deformation will concentrate on that floor causing a serious damage to the column members which might leads to the collapse of the whole structure like Piloti structure during the Pohang earthquake in Korea. According to the 2016 National Disaster Management Research Institute's "Investigation of Seismic Reinforcement and Cost Analysis of Domestic Non-seismic Buildings", the rate of seismic resistance of private reinforced concrete buildings was 38.3 %. Among them, it was reported that the seismic-resistance ratio of the two to five-story structures was less than 50 %. Accordingly, the government is trying to improve the seismic rate through support projects, but the conventional seismic reinforcement methods are still expensive, and emergency construction is difficult. Therefore, in this study, the field applicability was evaluated by improving the reinforcement method using Velcro, which was developed through the research project of the Ministry of Land, Transport and Maritime Affairs in 2014. In order to improve the performance of the Velcro reinforcement method, introducing the initial tension of Velcro using high foaming rigid urethane filling between the Velcro and concrete of the columns was applied. Additionally, an experiment was conducted to evaluate the ductility of Velcro specimen from the concrete confinement effect. As a result, the ductility of the Velcro specimen was improved compare to Normal specimen. However, the energy dissipation capacity of VELCRO2 is better than VELCRO1, yet the maximum ductility of those two specimens did not show a significant difference. Therefore, the improvement of the internal filler material is still needed to have a better maximum ductility.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.16-22
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• 2004

Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.323-330
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• 1995

Protease activity was identified in crude extracts of Pnrqgonimw westermnni eggs which were purified from infected dog lungs, isolated on 14 weeks after metacercarial challenge. The eggs were used after removing possibly contaminated host or worm tissues on their shell surfaces. In the crude egg extracts, high proteolytic activities against carboBfrb enzoyl - ph enylalanyl - arginyl-4- methoxy- β- naphthylamide (Cbz - phe - arg- MNA) and Azocoll were detected whereas those against succinyl-alanyl-propyl-phenylalanyl-p- nitroanilide (Suc-ala-pro-phe-pNA) were not revealed. The eVe eBdlibited the maximal activity at pH 6. Its activity was inhibited by specific cysteine protease inhibitors, 105 M I- trans-epoxysuccinylleucylamido (4-guanidino) butane (E-64) and 1 mM iodoacetamide (LAA) while potentiated by 6.5-fold in the presence of 2.5 mM dithiothreitol (DTT) . When the enzyme was purified partially by Sephacryl S-300 High Resolution gel filtration, it migrated as a single homogeneous band at 35 kDa. The 35 kDa cysteine protease has been recognized neither in the metacercariae nor in the adult. These findings indicated the presence of at least one protease of cathepsin family in immature eggs of f westernani.

• Korean Chemical Engineering Research
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• v.49 no.5
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• pp.571-576
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• 2011

Characteristics of bubble driven wakes were investigated in a simulated GTL process(0.102 m ${\times}$ 1.5 m in height) with viscous liquid medium. Effects of gas velocity(0.04 ~ 0.12 m/s) and liquid viscosity(0.001 ~ 0.050 $Pa{\cdot}s$) on the wake characteristics such as rising velocity, frequency, size and holdup were determined by employing a resistivity probe method. The wake phase formed behind the rising multi-bubbles as well as single bubbles were detected effectively from the conductivity fluctuations measured by the probe. Compressed, filtered and regulated air and aqueous solutions of Carboxy Methyl Cellulose(CMC) were used as a dispersed gas phase and a continuous liquid medium, respectively. It was found that the rising velocity and size of wake phase increased with an increase in gas velocity or liquid viscosity. The holdup and frequency of wake phase increased with increasing gas velocity due to the increase of gas input into the process with increasing gas velocity. However, the values of holdup and frequency of wake phase decreased with increasing liquid viscosity, since the size of bubbles and thus that of wakes increased with increasing liquid viscosity. The ratio of wake holdup to that of gas phase, which was in the range of 0.25 ~ 0.48, increased with an increase in liquid viscosity but decreased with gas velocity. The wake characteristics were well correlated in terms of operating variables within this experimental conditions.

• Journal of Korean Association for Spatial Structures
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• v.11 no.1
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• pp.121-130
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• 2011

Space frame structures have the advantage of constructing a large space structures without column and it may be considered as a shell structure. Nevertheless, with the characteristics of thin and long term of spacing, the unstable problem of space structure could not be set up clearly, and there is a huge difference between theory and experiment. Therefore, in this work, the tangential stiffness matrix of space frame structures is studied to solve the instability problem, and the nonlinear incremental analysis of the structures considering rise-span ratio(${\mu}$) and the ratio of load($R_L$) is performed for searching unstable points. Basing on the results of the example, global buckling can be happened by low rise-span ratio(${\mu}$), nodal buckling can be occurred by high rise-span ratio(${\mu}$). And in case of multi node space structure applying the ratio of load($R_L$), the nodal buckling phenomenon occur at low the ratio of load($R_L$), the global buckling occur a1 high the ratio of load($R_L$). In case of the global buckling, the load of bifurcation is about from 50% to 70% of perfect one's snap-through load.

• Korean Journal of Microbiology
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• v.43 no.1
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• pp.19-22
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• 2007

Acetohydroxyacid synthase (E.C.2.2.1.6., AHAS) is the enzyme that catalyses the first step in the synthesis of the branched-chain amino acids valine, leucine and isoleucine. The AHAS gene (TIGR access code HI2585) from Heamophilus influenzae was cloned into the bacterial expression vector pET-28a and expressed in the Escherichia coli strain BL21(DE3). The expressed enzyme was purified by $Ni^{2+}-charged$ HiTrap chelating HP column. The purified enzyme appears as a single band on SDS-PAGE with a molecular mass of about 63.9 kDa. The enzyme exhibits absolute dependence on the three cofactors FAD, $MgCl_{2}$ and thiamine diphosphate for activity. Specific activity of purified enzyme has 3.22 unit/mg and optimum activity in the pH 7.5 at $37^{\circ}C$. This enzyme activity has an effect on the buffer. When comparing the enzyme activity against the organic solvent, it followed in type and the difference it is but even from the aqueous solution where the organic solvent is included with the fact that the enzyme activity is maintained.

• The Korean Journal of Pesticide Science
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• v.14 no.4
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• pp.371-380
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• 2010

This study was performed to develop a precise single residue analytical method of fungicide cyprodinil in representative crops for using as general residue analytical methods which could be applied to most of crops. Apple, mandarin, Korean cabbage and green pepper were selected as representative crops, and they were macerated, extracted with acetonitrile, concentrated and partitioned with n-hexane. Then the extracts were concentrated and cleaned-up through silica gel column with ethyl acetate:n-hexane (15:85, v/v) before concentration and analysis with HPLC. LOQ (limit of quantitation) of cyprodinil was 5 ng (S/N>10) and MQL (method qnantitation limit) was 0.05 mg/kg. Recoveries were measured at three fortification levels (MQL, 10MQL and 100MQL) on crop samples and ranged from 82.0 to 108.2% and coefficients of variation were less than 10% regardless of sample type.

• Journal of Pharmaceutical Investigation
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• v.41 no.4
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• pp.255-262
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• 2011

Method using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed and validated for the determination of pregabalin in plasma samples. Acquisition was performed by monitoring the transitions: m/z 160.1${\rightarrow}$142.2 for pregabalin and m/z 423.2${\rightarrow}$207.1 for losartan (as an internal standard). After cold acetonitrileinduced protein precipitation of the plasma samples, separation was performed with C18 column by isocratic mobile phase consisted of 10 mM ammonium acetate and acetonitrile (15:85, v/v). Results were linear over the concentration ranged from 0.1 to $10{\mu}g$/mL and the correlation coefficients (r) were $\geq0.99$. Intra- and inter-day precisions were $\leq6.02$ and $\leq11.04%$, respectively, and intra- and inter-day accuracies were 96.60-101.09 and 98.10-102.60%, respectively. This validated method was successfully applied to a bioequivalence study of two formulations of pregabalin, Daewoong pregabalin capsule (Daewoong Pharm. Co., Ltd.) and Lyrica$^{(R)}$ capsule (Pfizer Korea Ltd.) in twenty eight healthy Korean volunteers. The subjects received a single oral dose of each formulation (150 mg as pregabalin) in a randomized $2{\times}2$ crossover study and plasma samples were obtained from each subject at predetermined time intervals. Then, the pharmacokinetic parameters ($AUC_{0-t}$, $C_{max}$ and $T_{max}$) were calculated and statistically analyzed to assess the differences between two formulations. The 90% confidence intervals for the log-transformed data were acceptable range of log 0.8-log 1.25 (e.g., log 1.0048-log 1.0692 for AUC0-t, log 0.9142-log 1.0421 for $C_{max}$). Thus, $AUC_{0-t}$ and $C_{max}$ met the criteria of the Korea Food and Drug Administration (KFDA) for bioequivalence test indicating that Daewoong pregabalin capsule was bioequivalent to Lyrica$^{(R)}$ capsule.

• The Korean Journal of Nuclear Medicine
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• v.14 no.2
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• pp.19-27
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• 1980

Although primary application of radioisotope scanning technics to the liver has been of use in the detection of the intra-hepatic space occupying lesion from the normal functioning liver parenchyme, there has been on increasing awareness of its use in evalution of Liver function. In this study, the diseases of the liver were classified into group A,B,C and D by the liver scanning findings, conventional liver function tests and clinical findings. Following were the results: 1. The colloidal radiogold liver scan appeared normal in the group A, also the albumin in serum, alkaline phosphatase activity and prothrombin time were within normal levels in this group. 2. In the group B, there were acute hepatitis 24(48%), chronic hepatitis 5(10%), toxic hepatitis 3(6%), subacute hepatic necrosis 3(6%), typhoid liver 4(8%), hepatic tuberculosis 2(4%), diabetes mellitus 3(6%) and others 3(6%). In this group, SGOT and SGPT were increased predominantly as compared with group A, and the liver scan showed small amount of mottling of activity and faintly visualized spleen. 3. In the group C, there were postnecrotic liver cirrhosis 30(60%), Laennec cirrhosis 10(20%), cardiac cirrhosis 1(2%), cholangiocarcinoma 1(2%), chronic active hepatitis 6(12%), hepatic milliary tuberculosis 1(2%) and gall bladder cancer 1(2%). In this group, the albumin in serum and prothrombin time were lowered significantly and the liver scan showed severe mottling of activity with extra-hepatic uptake in the spleen and bone marrow along the vertebral column. 4. In the group D, there were primary hepatoma 26(52%), hepatoma with liver cirrhosis 7(14%), metastatic liver cancer 5(10%), liver abscess 10(20%), multiple liver cyst 1(2%) and cystic duct adenoma 1(2%), In this group, the alkaline phosphatase activity was elevated with single or multiple intrahepatic space occupying lesion in the radiogold colloid liver scan.