• Title/Summary/Keyword: linear assay

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Cobra (Ophiphagus hannah) 독으로부터 세포 독성 단백질의 분리

  • 김영식;이병무;안미영;박재갑;현진원;임경화;박호군
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.205-205
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    • 1994
  • 동물, 곤충, 해조류 등으로 부터 얻어진 독소는 해로운 것으로 알려져 있지만 의학적으로 유용한 화합물을 내포하고 있다. 본 연구는 이러한 점에 착안하여 독사, 또는 곤충의 독 30여종을 대상으로 위암 세포인 SNU-1에 대해서 MTT assay에 의한 세포 독성 시험을 실시하였다. 이 중 세포 독성 활성이 가장 놓은 코브라 계통인 Ophiphagus hannah의 독으로 부터 세포 독성 물질의 정제를 시도하였다. HPLC-GPC와 HPLC-lEX 또는 RP-HPLC를 이용하여 정제하였고 각 분획들을 암세포주에 대해서 시험하였다. TSK-2000SW로 부터 2개의 분획을 얻어 분획 I에서 $IC_{50}$/ 의 값이 0.5 - 3 $\mu\textrm{g}$/ml의 범위 내에서 확인되었다. 분자량이 작은 분획 II에서는 세포독성이 작게 나타났지만 농축하여 성분 분석을 시도하였다. 분획 I을 RP-HPLC를 이용하여 TFA와 acetonitrile의 linear gradient에 의해 더욱 분리를 하였고, phopholipase $A_2$ 활성의 가능성도 측정하였다. 분획 II는 Mono S 칼람을 이용하여 ammonium acetate농도에 따른 pH gradient에 의한 분리를 시도하여 순도를 SDS-PAGE에 의해서 확인하였다. 각각의 크로마토그리피로 부터 얻은 분획에 대해서 세포 독성을 실시중에 있으며 성질도 동정중에 있다.

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Development of a One-step Two-site Enzyme Immunoassay for Measuring Human Alpha-fetoprotein by Eliminating Hook-effect

  • Kim, Se-Ho
    • BMB Reports
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    • v.34 no.1
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    • pp.47-50
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    • 2001
  • A one-step, two-site enzyme immunoassay was developed for measuring human alpha-fetoprotein (AFP) in serum and amniotic fluid using monoclonal antibodies (McAb) by eliminating the high-dose hook effect. Three McAbs that recognize different epitopes were selected among 16 different clones on the basis of epitope mapping, two for immobilization and one for horseradish peroxidase conjugation. This one-step immunoassay system is more convenient and rapid compared to a conventional two-step sandwich immunoassay system. It did not exhibit the hook effect to around 2.7 mg/ml of AFP, which is probably one of the highest concentrations of AFP in the serum. The dose-response curve of the system was linear to 500 mg/ml of AFP and the system could differentiate as low as 1 mg/ml of AFP The intra- and inter-assay variations were in an acceptable range; 95~104% and 97~105% respectively Its correlation with other commercial systems was around 95%.

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Development of a Rapid Spectrophotometric Method for Detecting Bacterial Mucinase Complex

  • Kim, Yoon-Hee;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.345-348
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    • 2002
  • A rapid spectrophotometric method for detecting the mucinase complex was developed. Bovine submaxillary mucin is cleaved by commercial mucinase between the oligosaccharide chain and the side chain of peptide linkage, thereby liberating the N-acetyl neuraminic acid (NANA). The release of NANA resulted in an increase of absorbance at 280 nm. The susceptibility to NANA by the new method was found to be at least 10-fold more sensitive than the thiobarbituric acid method. Moreover, the quantification of NANA released from mucin by commercial neuraminidase and partially purified Vibrio parahaemolyticus mucinase showed a good linear correlation in proportion to the concentration of the enzyme used. These results demonstrate that the rapid identification of mucin degradation can be determined by a spectrophotometric assay, thereby providing a new, fast, and sensitive method for assaying the bacterial mucinase complex.

Monitoring of Leuconostoc Population During Sauerkraut Fermentation by Quantitative Real-Time Polymerase Chain Reaction

  • Kim, So-Young;Yoo, Ki-Seon;Kim, Yu-Jin;Seo, Eun-Young;Kim, Beom-Soo;Han, Nam-Soo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1069-1072
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    • 2011
  • A real-time PCR assay method was established to monitor Leuconostoc spp. populations via specific amplification of the dextransucrase gene. Quantification of L. mesenteroides B-512F using both genomic DNA and cell suspensions yielded a log-linear correlation spanning approximately 5 log units. By using this method, monitoring changes of Leuconostoc spp. during sauerkraut fermentation was successfully accomplished with accuracy after inoculation of starter and sugars (sucrose and maltose).

Radiobiological Evaluation in Pig Bred in the Vicinity of Yeonggwang Nuclear Power Station Using Micronuclei in Cytokinesis-blocked Lymphocyte (림프구의 미소핵을 지표로 영광 원자력발전소 주변 사육 돼지의 방사선 생물학적 평가)

  • 김세라;강창모;김성호
    • Journal of Veterinary Clinics
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    • v.21 no.3
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    • pp.286-290
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    • 2004
  • Cytogenetic and hematological analysis was performed in peripheral blood of pig in the vicinity of Yeonggwang nuclear power station and control area. The frequency of micronuclei (MN) in peripheral blood lymphocytes from pig was used as a biomarker of radiobiological effects resulting from exposure to environmental radiation. An estimated dose of radiation was calculated by a best fitting linear-quadratic model based on the radiation-induced MN formation from the swine lymphocytes exposed in vitro to radiation over the range from 0 Gy to 4 Gy. MN rates in lymphocytes of pig from Yeonggwang nuclear power station and control area were 10.60/1,000 and 11.10/1,000, respectively. There were no significant differences in MN frequencies and hematological values in pig between Yeonggwang and control area. The study indicates that the MN assay in lymphocyte of pig is a rapid, sensitive and accurate method that can be used to monitor a large population exposed to radiation.

Voltammetric Investigation of Vitamin B_6 at a Glassy Carbon Electrode and Its Application in Determination

  • Wu, Yun-Hua;Song, Fa-Jun
    • Bulletin of the Korean Chemical Society
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    • v.29 no.1
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    • pp.38-42
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    • 2008
  • The voltammetic behavior of Vitamin B6 (VB6) was studied at a glassy carbon electrode in phosphate buffers using cyclic, linear sweep and differential pulse voltammetry. The oxidation process was shown to be irreversible over the entire pH range studied (4.0-10.0) and was adsorption controlled. The adsorption amount of VB6 on the glassy carbon electrode was examined by chronocoulometry and the value of n, product of transfer coefficient and number of electrons transferred in the rate determining step, was determined from Tafel plot. VB6 was determined by differential pulse voltammetry and the peak current was found linearly with its concentration in the range of 3 10-7-2 10-4 mol L-1. The detection limit was 1 10-7 mol L-1. The procedure was successfully applied for the assay of VB6 in tablets.

Pure Culture Assay with Streptomyces viridosporus T7A for Biodegradability Determination of Oxidized Potato Starch/Polyethylene Films

  • Kim, Meera;Kim, Sung-Hong
    • Preventive Nutrition and Food Science
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    • v.6 no.2
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    • pp.112-116
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    • 2001
  • Oxidized potato starch/polyethylene (PE) cast films were prepared with different percentages of linear low density PE (LLDPE), oxidized potato starch and prooxidant. For the determination of biodegradability of the films, lignocellulose-degrading Streptomyces viridosporus T7A (ATCC 39115) was used. Films were chemically disinfected and incubated with S. viridosporus by shaking at 100 rpm at 37$^{\circ}C$ for eight weeks. Hydroxyl indices of the films by Fourier-Transform Infrared Spectroscopy, mechanical Properties of the films by Instron and film morphology by scanning electron microscope (SEM) were measured. The hydroxyl index of the film containing the oxidized potato starch incubated with S. viridosporus T7A was higher than that of the corresponding control. All the films containing 5% and 10% oxidized starch showed a decrease of tensile strength on the films after incubation when the corresponding uninoculated film was compared. In the oxidized starch/PE film incubated with S. viridosporus T7A, partial destruction of starch and PE was examined by SEM.

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A Dipstick-Type Electrochemical Immunosensor for The Detection of The Organophosphorus Insecticide Fenthion

  • Cho, Young-Ae;Cha, Geun-Sig;Lee, Yong-Tae;Lee, Hye-Sung
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.743-746
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    • 2005
  • A dipstick-type immunochemical biosensor for the detection of the organophosphorus insecticide fenthion was developed using a screen-printed electrode system as an amperometric transducer with polyclonal antibodies against fenthion as a bioreceptor. The assay of the biosensor involved competition between the pesticide in the sample and pesticide-glucose oxidase conjugate for binding to the antibody immobilized on the membrane. This was followed by measurement of the activity of the bound enzyme by the supply of the enzyme substrate (glucose) and amperometric determination of the enzyme reaction product ($H_2O_2$). The activity of the bound enzyme was inversely proportional to the concentration of pesticide. The optimized sensor system showed a linear response against the logarithm of the pesticide concentration ranging from $10^{-2}$ to $10^3\;{\mu}g/L$.

Sound Quality Index Development of Electrically Powered Vehicle Roller Blind (차량용 전동 롤러 블라인드의 음질지수 개발)

  • Sung, Weonchan;Jo, Hyeonho;Kim, Seonghyeon;Park, Dongchul;Kang, Yeonjune
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2014.10a
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    • pp.785-790
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    • 2014
  • The purpose of this study is to identify the significant sound quality metric and compose the sound quality index of motor driven roller blind which is part of vehicle sunroof. Before subjective evaluation, sound characteristics of roller blind was analyzed and set the target operating sound for subjective evaluation. Thus, transfer sound of roller blind which has the characteristics of sound modulation was used for subjective evaluation. Linear regression was carried out by chosen Zwicker's metrics which are pointed by comments of jurors. Loudness and sharpness related metrics are prime metrics in sound quality index we composed. Effect of roller blind assay when it is attached to real vehicle was identified to evaluate the validity of index.

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Pharmacokinetic-Pharmacodynamic Modeling of a Direct Thrombin Inhibitor, Argatroban, in Rats

  • Park, Eun-Hye;Shin, Beom-Soo;Yun, Chi-Ho;Lee, Mann-Hyung;Yoo, Sun-Dong
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.373-379
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    • 2009
  • This study was conducted to develop a pharmacokinetic-pharmacodynamic (PK/PD) model of a direct thrombin inhibitor, argatroban to predict the concentration-effect profiles in rats. Argatroban was i.v. injected to rats at 0. 2, 0.8 and 3.2 mg/kg doses (n = 4-5 per dose), and plasma drug levels were determined by a validated LC/MS/MS assay. The pharmacokinetics of argatroban was linear over the i.v. dose range studied. The thrombin time (TT) and the activated partial thromboplastin time (aPTT) were measured in rat plasma and they were found to linearly increase with increasing the dose. A 2-compartment pharmacokinetic model linked with an indirect response pharmacodynamic model was successfully utilized to evaluate the drug concentration-response relationship.