• Title/Summary/Keyword: Half-cell method

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The Effects of Co-cultivation Medium and Culture Conditions on Rice Transformation Efficiency (공동배양과정의 배지조성과 배양조건이 벼 형질전환효율에 미치는 영향)

  • Kim, Yul-Ho;Park, Hyang-Mi;Choi, Man-Soo;Yun, Hong-Tai;Choi, Im-Soo;Shin, Dong-Bum;Kim, Chung-Kon;Lee, Jang-Yong
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.252-260
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    • 2009
  • Rice is the most important cereal crop not only in supplying the basic staple food for more than half of the world's population but also as a model plant for functional genomic studies of monocotyledons. Although rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5 mg/L L-cysteine, 1 mM sodium thiosulfate, 1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5 mg/L silver nitrate). Co-cultivation temperature ($23.5^{\circ}C$ for 1 day, $26.5^{\circ}C$ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.

Three-Dimensional Magnetotelluric Modeling Using Integral Equations (적분방정식을 이용한 3차원 지자기 지전류 모델링)

  • Kim, Hee Joon;Lee, Dong Sung
    • Economic and Environmental Geology
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    • v.27 no.2
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    • pp.191-199
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    • 1994
  • We have developed an algorithm based on the method of integral equations to simulate the magnetotelluric (MT) responses of three-dimensional (3-D) bodies in a layered half-space. The inhomogeneities are divided into a number of cells and are replaced by an equivalent current distribution which is approximated by pulse basis functions. A matrix equation is constructed using the electric Green's tensor function appropriate to a layered earth, and is solved for the vector current in each cell. Subsequently, scattered fields are found by integrating electric and magnetic Green's tensor functions over the scattering current About a 3-D conductive body near the earth's surface, interpretation using 2-D transverse electric modeling schemes can imply highly erratic low resistivities at depth. This is why these routines do not account for the effect of boundary charges. However, centrally located profiles across elongate 3-D prisms may be modeled accurately with a 2-D transverse magnetic algorithm, which implicitly includes boundary charges in its formulation. Multifrequency calculations show that apparent resistivity and impedance phase are really two complementary parameters. Hence, they should be treated simultaneously in broadband MT interpretation.

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Electronic Structures of ANb2PS10 (A=Ag, Na) and AuNb4P2S20

  • Jung, Dong-Woon;Kim, Sung-Jin
    • Bulletin of the Korean Chemical Society
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    • v.24 no.6
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    • pp.739-743
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    • 2003
  • New quaternary compounds $ANb_2PS_{10}$ (A = Na, Ag) and $AuNb_4P_2S_{20}$ were synthesized and characterized. The structures of three compounds consist of one-dimensional infinite chains built by [$Nb_2S_{12}$] and [$PS_4$] units. Cation atoms are occupied within the van der Waals gap of sulfur atoms between infinite chains to make -S…$M^+$…S- contacts. There is only one Au atom site and so crystallographically a unit cell contains four equivalent Au atoms in $AuNb_4P_2S_{20}$. This is only the half of the numbers of Na or Ag atoms in $NaNb_2PS_{10}$ or $AgNb_2PS_{10}$. The ratio between $Nb_2PS_{10}$ matrix vs the cation is, therefore, 1 : 1 for Ag and Na, but it is 2 : 1 for Au. Mixed valency in Au or Nb was expected to balance the charge in the latter compound. The electronic structures calculated based on the extended Huckel tight-binding method show that $ANb_2PS_{10}$ (A = Ag, Na) are semiconducting, while $AuNb_4P_2S_{20}$ is metallic, which is not consistent with the experimental results of these three compounds that all exhibit semiconducting property. The result of calculation suggests that $AuNb_4P_2S_{20}$ might be a magnetic insulator. Magnetic measurement experiment exactly proved that the compound is a Slater antiferromagnetic material with the Neels' temperature of 45 K. It is recognized, therefore, that electronic structure analysis is very useful to understand the properties of compounds.

Enhanced In Vitro Protein Synthesis Through Optimal Design of PCR Primers

  • Ahn Jin-Ho;Son Jeong-Mi;Hwang Mi-Yeon;Kim Tae-Wan;Park Chang-Kil;Choi Cha-Yong;Kim Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.355-359
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    • 2006
  • The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.

Development of Competitive Direct Enzyme-linked Immunosorbent Assay for the Detection of Gentamicin Residues in the Plasma of Live Animals

  • Jin, Yong;Jang, Jin-Wook;Lee, Mun-Han;Han, Chang-Hoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1498-1504
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    • 2005
  • Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.

Dietary Intakes and Status of Folate in Koean Women of Child-bearing Potential (가임기 여성의 엽산 섭취량 및 엽산영양상태)

  • 임현숙
    • Journal of Nutrition and Health
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    • v.33 no.3
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    • pp.296-303
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    • 2000
  • We examined the folate intakes and assessed folate nutritional status of Korean women with childbearing potential. A total of 91 healthy women aged between 15 and 49 participated. They were divided into three groups by their age : A(15-24 yrs), B(25-34 yrs) and C(35-49 yrs). Folate intakes were determined by direct analysis. The foods consumed for 24 hours were collected proportionally and assessed folate. Their blood drawn in fasting state were analyzed folate levels. Folate contents of food homogenate, plasma and erythrocyte were determined a microbiological method using Lactobacillus. casei (ATCC 7469). Prior to the micro-assay, the food homogenate were treated with alpha-amylase, protease and folate conjugase. Mean daily folate intake of the total subjects was 145.8$\mu\textrm{g}$/d and in each group of A, B, and C was 114.0$\mu\textrm{g}$/d, 141.6$\mu\textrm{g}$/d, and 164.6$\mu\textrm{g}$/d, respectively. That of group C was significantly higher than that of group A(p<0.05). However, those of all the groups were lower than compared to the Korean Recommened Dietary Allowances(RDA) for folate. Especially the subjects in the group A consumed folate least that was below the half of the Korean RDA. The mean energy intake of all subjects was 1638㎉/d and those in each group of A, B, and C did not meet the Korean RDA for energy. The energy intake were significantly correlated with folate intakes(r=0.5050, p<0.001). Mean plasma and erythrocyte folate concentrations of total subjects were 6.9ng/mL and 266.3ng/mL, respectively. None were found to be deficient both in plasma(<3ng/mL)and erythrocyte (<140ng/mL) folate levels. There was only one subject who had red blood cell folate level below 157ng/mL concentration. These results show that folate status of the Korean women of reproductive age is not much bad. But it should be better that letting them improve their folate status by increasing energy intake, choosing high folate foods.

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A Review of the TCM Articles about Cauterization Treatment (낙법(烙法)에 관한 중의(中醫) 논문 고찰)

  • Jang, Ho-Jeong;Lee, Hyeon-Joo;Seo, Hyung-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.31 no.3
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    • pp.60-66
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    • 2018
  • Objectives : The purpose of this study is to review the chinese published papers on cauterization of traditional chinese medicine. Methods : We searched chinese published papers from 1958 until May 2017 via CNKI(China National Knowledge Infrastructure) with the keyword "cauterization". We analyzed the studies covered cauterization medical treatment, and classified them by 5 categories including periods, type of study, treatment site, experiment target site, and cauterizing method. Results : We reviewed 112 chinese papers which include 10 Original articles, 43 Review articles, and 59 Case reports. Examining yearly distributions, we can see that research on cauterization is becoming more active than in the past. In classification of 93 clinical studies by treatment site, the number of research on Tonsillitis accounts for almost half(43 studies), followed by Sore throat(14 studies). Among 10 experimental studies, Eye is the most frequently targeted organ which was used to study high intra-ocular pressure(5 studies), and retinal ganglion cell(2 studies). Lastly, there are various methods of cauterization used in papers: Branding iron(54 studies), and Red-hot needle(24 studies) are two major heating methods. Conclusions : This analysis shows that studies on the application of cauterization have been actively conducted in China these days. Furthermore, up-to-date cauterizing methods have been developed such as electric type and microwave type beyond traditional ways. We expect this article will encourage further research on cauterization in order to apply it to a variety of diseases. Then, it could become one of new effective medical treatments in Korean medicine.

Evaluate the Concrete mix by Type Accelerated Corrosion Test and Chloride Penetration Analysis with Artificial Seawater Cyclic Wet and Dry Condition (인공해수 건습반복조건에 따른 콘크리트배합별 부식촉진시험법과 염화물 침투해석평가)

  • Park, Sang-Soon;Kim, Min-Wook
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.1 no.3
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    • pp.211-218
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    • 2013
  • Cyclic wet and dry conditions in the marine environment structures corrosion is known to be the fastest rising. For that reason, accelerated corrosion test methods for the reproduction of tidal environment has been actively conducted. However, many studies have estimated threshold value for steel corrosion or concentrated in chloride penetration analysis. In this study, cyclic wet and dry conditions to reproduce the structure of the environment in accelerated corrosion and chloride penetration test analysis was performed. Corrosion was determined by the result of reinforcement corrosion monitoring based on galvanic potential measurement and half-cell potential method. Accelerated corrosion test results for each formulation was different corrosion periods, the order OPC> FA> BS> High-strength concrete. FEM durability interpretation program DuCOM was conducted under the same conditions as in accelerated corrosion test. The experimental RCPT tests demonstrated the validity of the result.

Characteristics for Reinforcement Corrosion and Chloride Ion Diffusion of High Volume Fly Ash Concrete (하이볼륨 플라이애시 콘크리트의 철근부식 및 염소이온 확산 특성)

  • Kwon, Soon-Oh;Bae, Su-Ho;Lee, Hyun-Jin;Jung, Sang-Hwa
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.2 no.1
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    • pp.34-39
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    • 2014
  • The purpose of this experimental research is to evaluate the resistance of reinforcement corrosion and chloride ion penetration of high volume fly ash (HVFA) concrete. For this purpose, concrete test specimens were made for various strength level and replacement ratio of fly ash, and then compressive strength and diffusion coefficient for chloride ion of them were measured for 28, 91 and 182 days, respectively. Also, corrosion monitoring by half cell potential method was carried out for the made lollypop concrete test specimens to detect the time of corrosion initiation for reinforcement in concrete. As a result, it was observed from the test results that compressive strength of HVFA concrete was decreased with increasing replacement ratio of fly ash but long-term resistance against reinforcement corrosion and chloride ion penetration of that was increased.

Presenilin Modulates Calcium-permeant, Magnesium-Nucleotide regulated channel, I(MgNUM)

  • Shin, Sun-Young;Jeong, Soon-Youn;Uhm, Dae-Yong;Sungkwon Chung
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.47-47
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    • 2003
  • The presenilin 1 (PS1) or PS2 is an essential component of the ${\gamma}$-secretase complex, which mediates the intramembrane proteolysis of selected type-I membrane, including the ${\beta}$-amyloid precursor protein (APP) to yield A${\beta}$. Familial Alzheimer's disease (FAD)-associated mutations in presenilins give rise to an increased production of a highly amyloidogenic A${\beta}$42. In addition to their well-documented proteolytic function, the presenilins play a role in calcium signaling. We have previously reported that presenilin FAD mutations cause highly consistent alterations in intracellular calcium signaling pathways, which include deficits in capacitative calcium entry (CCE), the refilling mechanism for depleted internal calcium stores. However, molecular basis for the presenilin-mediated modulation of CCE remains to be elucidated. In the present study, whole-cell patch clamp method was used to identify a specific calcium-permeable ion channel current(s) that is responsible for the CCE deficits associated with FAD-linked PS1 mutants. Unexpectedly, both voltage-activated and conventional store depletion-activated calcium currents I(CRAC), were absent in HEK293 cells, which were stably transfected either with wild-type or FAD mutant (L286V, M146L, and delta E9) forms of PS1. Recently, magnesium-nucleotide-regulated metal cation current, or I(MagNum), has been described and appears to share many common properties with I(CRAC) including calcium permeability and inhibitor sensitivity (e.g. 2-APB). We have detected I(MagNum) in all 293 cells tested. Interestingly, FAD mutant 293 cells developed only about half of currents compared to PS1 wild type cells.

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