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Effect of Heat, Pressure, and Acid Treatments on DNA and Protein Stability in GM Soybean (GM 콩 DNA와 단백질의 안정성에 대한 열, 압력 및 산 처리의 영향)

  • Pack, In-Soon;Jeong, Soon-Chun;Yoon, Won-Kee;Park, Sang-Kyu;Youk, Eun-Soo;Kim, Hwan-Mook
    • Korean Journal of Food Science and Technology
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    • v.36 no.4
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    • pp.677-682
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    • 2004
  • Debates on safety of genetically modified (GM) crops have led to mandatory-labeling legislation of GM foods in many countries including Korea. Effects of heat, pressure, and acid treatments on degradation of DNAs or proteins in GM soybean at levels below detection limits of qualitative PCR and lateral flow strip test (LFST) methods were examined. Results showed that genomic DNAs and proteins were degraded into fragment sizes no longer possible for detection of inserted gene depending on thermal, or thermal and pressure treatment period. Detectaability of LFST for toasted meal increased in weakly treated soybean. DNA and protein detection methods were barely effective for detection of GM ingredient after $121^{\circ}C$ and 1.5 atmospheric treatment for 20 min. These results will be useful in determining GM labeling requirements of processed foods.

Rapid Agrobacterium-mediated genetic rice transformation method using liquid media (액체배양을 이용한 단기 벼 형질전환 방법)

  • Yang, Dae-Hwa;Chang, Ahn-Cheol;Ahn, Il-Pyung;Kim, Hae-Jung;Kim, Dong-Hern;Lee, Hyo-Yeon;Suh, Seok Cheol
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.37-42
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    • 2013
  • Rice is one of the most important cereal crops as a model plant for functional genomics of monocotyledons and usually transformed using Agrobacterium tumefaciens. However, the transformation's process using previous method is still time consuming and uneconomical, low efficiency. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the infection and co-cultivation, Agrobacterium elimination, infected calli's selection steps using liquid media. We directly inoculated Agrobacterium containing a ZjLsL gene under the control of constitutive promoter into the 1- to 3-week-old rice calli derived from mature seeds. After 3 days of co-cultivation, the infected calli were transferred onto liquid media of Agrobacterium elimination and calli's selection for 3 days. The calli were transferred to calli's growth solid media for 14 days and then the calli transferred to shoot induction and root induction media. Putative transformants were initially selected on the medium containing phosphinothricin, and the PAT protein verified by PAT strip test. This method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

The Studies on the Removal of Harmful Microorganisms in Water by Using Antimicrobial Copper (친환경 항균성 구리의 수질 내 생물학적 오염원 제거 가능성 연구)

  • Ji, Keunho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.9
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    • pp.384-390
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    • 2018
  • Air and soil are being contaminated by the environmental pollution as a result of climate change and urbanization, resulting in water pollution reaching serious levels. In this studies, we investigated the use of antimicrobial copper for the removal of biological pollutants from water system. Specifically, we tested its effects against E. coli, B. subtilis, S. aureus, K. pneumoniae and P. aeruginosa. Made a sphere shape having a diameter of 2cm using a strip-shaped copper wire of 0.5g, 1g and 2g. And then, to confirm the antimicrobial activities, each copper ball was equipped in the broth which inoculated each pathogens. The results showed that bacterial growth of the five test bacteria was inhibited by more than 99% after reaction with a 0.5 g copper ball for at least 20 minutes. Based on the these results, if perform the further experiment such cytotoxicity, it is expected that will be enough to be used as a filter for water quality purification. The developed technique is expected to be widely applied in various industries.

Carbonization Patten and Operation Characteristics of a 1Φ 2 W MCCB Damaged by PCITS (PCITS에 의해 소손된 1Φ 2 W MCCB의 탄화 패턴 및 작동 특성)

  • Lee, Jae-Hyuk;Choi, Chung-Seog
    • Fire Science and Engineering
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    • v.28 no.5
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    • pp.8-13
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    • 2014
  • The purpose of this study is to analyze the damage pattern when overcurrent is applied to a thermal magnetic type molded case circuit breaker (MCCB) using a Primary Current Injection Test System (PCITS). When an overcurrent of 150 A was applied to the PCITS for 5 seconds with the trip bar of an MCCB being damaged, it was found that the surface of the temperature control device (bimetallic strip) positioned at the right was significantly carbonized. When an overcurrent of 300 A was applied to the PCITS for 5 s under the same conditions, the entire temperature control device was deteriorated, becoming flattened and in close contact with the MCCB. When an overcurrent of 450 A was applied to the PCITS for 5 s, the coil of the temperature control device was melted and disconnected. In addition, it was observed that the contacts, the enclosure and upper cover were deformed and there was a trace of carbonization on them. When approximately 3 s had elapsed after an overcurrent of 600 A was applied, white smoke occurred inside the MCCB and a flame was radiated out, after which the overcurrent supply stopped with "phutt" (whomp) sound. It was observed that when the same type of MCCB is damaged by a general flame, the surfaces of its handle, terminal, arc divider (extinguisher) and temperature control device were carbonized uniformly. In addition, it was found that the trip bar of the operating mechanism was melted down and the metal operation pin was moved while being tripped.

Analysis of Human Serum Amyloid A-1 Concentrations Using a Lateral Flow Immunoassay with CdSe/ZnS Quantum Dots (Human Serum Amyloid A-1 단백질 농도 분석을 위한 CdSe/ZnS 양자점 기반의 Lateral Flow Immunoassay 방법 개발)

  • Fajri, Aidil;Goh, Eunseo;Lee, Sanghyuk;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.30 no.4
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    • pp.429-434
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    • 2019
  • A lateral flow immunoassay platform utilizing antibody functionalized water soluble CdSe/ZnS semiconductor quantum dots (QDs) was developed for the analysis of human serum amyloid A-1 (hSAA1) in a buffer solution. hSAA1 was chosen as a target protein because it is regarded as a potential biomarker associated with early diagnosis and prognosis in patients of lung cancer. The immunoassay strip on a nitrocellulose membrane was fabricated by spraying two lines composed of a test line with a monoclonal antibody against hSAA1 (10G1) (anti hSAA1) and a control line of anti-chicken IgY. While the CdSe/ZnS QDs synthesized in an organic phase were transferred to a water phase by ligand exchange using carboxylic acid modified alkane thiol. The QDs was then conjugated to monoclonal antibody against hSAA1 (14F8) [anti hSAA1 (14F8)] and used as a fluorescent detection probe. The sequential lateral flow of hSAA1 in different concentration and QDs-anti hSAA1 (14F8) complex allowed to form the surface sandwich complex of anti hSAA1 (10G1)/hSAA1/QD-anti hSAA1 (14F8), which was then analyzed using fluorescence microscope. A 100 nM concentration of hSAA1 protein can be detected by naked eyes under an optimized lateral flow buffer condition with a sensing time of 5 mins.

Screening Methods for Plant-Coating Materials and Transpiration Inhibitory Effect of Soybean Oil to Crops (식물 코팅 소재 선발법과 작물들에 대한 콩 오일의 증산 억제 효과)

  • Jung, In Hong;Park, No Bong;Kim, Sang-Yeol;Na, Young-Eun;Kim, Soon-Il
    • Korean Journal of Plant Resources
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    • v.27 no.4
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    • pp.380-391
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    • 2014
  • Plants as well as crops are damaged by a combination of the hot and dry winds that has been a major factor in the reduction of crop production. A means to protect them from damaging conditions is to consider a coating material. In this study, we established laboratory screening methods to find a coating material to protect a crop from rapid transpiration caused by various factors. In a test measuring the weight loss of kidney bean seedlings for 6 days, Avion treatments decreased its weight loss (P=0.05). Owing to long-time spend in completing this assay, we performed a more simple method using a cobalt chloride paper strip, which changes from blue to red colors under water condition. Beewax, guagum, paraffin liquid, soybean oil, and PE-635 gave a waterproofing effect above 37 and 43% at 0.5 and 1 h after treatment, respectively. However, these tested materials did not show significant waterproofing results at 2 h. Although the methods produced reasonable results, a screening method to obtain more objective data is needed. An alternative is to use an instrument that can detect the transpiration of crop leaves. In a preliminary test using barley leaves, a portable photosynthesis system showed transpiration inhibition of 2% soybean oil and 10 times-diluted Avion under field conditions. In another test using the leaves of maize seedlings and apricot tree, 2% liquid paraffin and plant oils such as apricot oil, linseed oil, olive oil, and soybean oil showed significant transpiration inhibition (P=0.05). Especially, paraffin liquid and soybean oil selected from above tests gave good transpiration inhibitory effects against rice at 2%. In addition, the mixture of 2% soybean oil and a spreader showed more elevated inhibition results comparing with soybean oil or the spreader alone indicating that the spreader may be attributed to more uniform diffusion of the hydrophobic material onto the leaf surface of maize seedlings. The hydrophobic material coated physically the stomata and cuticle layers on leaf surfaces of rice. These hydrophobic materials screened in this study are expected to be used as plant coating materials.

Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors (Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換))

  • Son, Suk Gyu;Hyu, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.164-172
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    • 1998
  • This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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Effects of Potassium-Channel Opener on Thallium-201 Kinetics: In-vitro Study in Rat Myocyte Preparations and In-vivo Mice Biodistribution Study (K-통로개방제가 배양심근세포와 생쥐 체내의 Thallium-201역동학에 미치는 영향)

  • Lee, Jae-Tae;Kim, Eun-Ji;Ahn, Byeong-Cheol;Sohn, Kang-Kyun;Lee, Kyu-Bo;Ha, Jeoung-Hee;Kim, Chun-K.
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.4
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    • pp.507-515
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    • 1996
  • Background : Potassium channel opener (K-opener) opens ATP-sensitive K'-channel located at cell membrane and induces potassium efflux from cytosol, resulting in intracellular hyperpolarization. Newly synthesized K-opener is currently examined for pharmacologic potency by means of rubidium release test from smooth muscle strip pre-incubated with Rb-86. Since in-vivo behavior of thallium is similar to that of rubidium, we hypothesized that K-opener can alter T1-201 kinetics in vivo. Purpose : This study was prepared to investigate the effects of pinacidil (one of potent K-openers) on the T1-201 uptake and clearance in cultured myocyte, and in-vivo biodistribution in mice. Methods : Spontaneous contracting myocytes were prepared to imitate in-vivo condition from 20 hearts of 3-5 days old Sprague-Dawley rat and cultured for 3-5 days before use ($5{\times}10^5$ cells/ml). Pinacidil was dissolved in 10% DMSO solution at a final concentration of 100nM or l0uM and was co-incubated with T1-201 in HBSS buffer for 20-min to evaluate its effect on cellular T1-uptake, or challenged to cell preparation pre-incubated with T1-201 for washout study. Two, 40 or $100{\mu}g$ of pinacidil was injected intravenously into ICR mice at 10 min after $5{\mu}Ci$ T1-201 injection, and organ uptake and whole body retention rate were measured at different time points. Results : Co-incubation of pinacidil with T1-201 resulted in a decrease in T1-201 uptake into cultured myocyte by 1.6 to 2.5 times, depending on pinacidil concentration and activity of T1-201 used. Pinacidil enhanced T1-201 washout by 1.6-3.1 times from myocyte preparations pre-incubated with T1-201. Pinacidil treatment appears to be resulted in mild decreases in blood and liver activity in normal mice, in contrast, renal and cardiac uptake were mildly decreased in a dose dependent manner. Whole body retention ratios of T1-201 were lower at 24 hour after injection with $100{\mu}g$ of pinacidil than control. Conclusion : These results suggest that treatment with K-opener may affect the interpretation of T1-201 myocardial images, due to decreasing thallium accumulation and enhancing washout from myocardium.

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Effect of working time on the film thickness of dental resin cements (레진 시멘트의 혼합 후 시간에 따른 피막도의 변화)

  • Yi, Yu-Seung;Kim, Sung-Hun;Lee, Jai-Bong;Han, Jung-Suk;Yeo, In-Sung;Ha, Seung-Ryong;Kim, Hee-Kyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.53 no.4
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    • pp.325-329
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    • 2015
  • Purpose: The aim of this study was to compare the film thicknesses of several resin cements as a function of time after mixing and to examine the effect of working time on the film thicknesses. Materials and methods: The film thickness (${\mu}m$) of 4 resin cements (n=10), 1 composite resin (Panavia F 2.0), 3 self-adhesive resin (Clearfil SA luting, Zirconite, RelyX U200) cements was measured at 20-second intervals after mixing of the cements up to 200 seconds under a load of 50 N. Linear regression was fitted to verify the effect of working time on the film thickness of each cement. Data were compared to the working time recommended by manufacturers using Wilcoxon test ($\alpha$=.05). Results: All of the materials showed a positive linear correlation between the film thickness and working time. There was no statistically significant difference between the working time based on our results and the values recommended by the manufacturers even though there was a discrepancy between those two values. Conclusion: The film thickness of resin cements could increase with the increase of working time. Working time to meet the ISO standard of $50-{\mu}m$ maximum film thickness could be different from the manufacturer's recommended value.

Analysis of rpoB Gene in Rifampin-Resistant M. Tuberculosis by Direct Sequencing and Line Probe Assay (염기서열결정과 Line Probe 분석법에 의한 Rifampin내성 결핵균의 rpoB 유전자 분석)

  • Lee, Min-Ki;Kim, Yun-Seong;Lee, Hyo-Jin;Cheon, Du-Su;Yun, Sang-Myung;Park, Sam-Seok;Kim, Cheol-Min;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.251-263
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    • 1997
  • Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

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