• Title/Summary/Keyword: Screening Method

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Intersubunit Communication of Escherichia coli Tryptophan Synthase (대장균 트립토판 생성효소의 소단위체간 상호조절)

  • Cho, Won Jin;Lim, Woon Ki
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1410-1414
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    • 2017
  • Escherichia coli tryptophan synthase (TS) contains ${\alpha}_2{\beta}_2$, which catalyzes the final two steps in Trp biosynthesis. A molecular tunnel exists between the two active sites of ${\alpha}$ and ${\beta}$ subunits in TS. Via intersubunit communication, TS increases catalytic efficiency, including substrate channeling. The ${\beta}$ subunit of TS is composed of two domains, one of which, the COMM (communication) domain, plays an important role in intersubunit communication. The ${\alpha}$ subunit has a TIM barrel structure. This protein has functional regions at the C terminal of ${\beta}$ pleated sheets and in its loop regions. Three regions of the ${\alpha}$ subunit (${\alpha}L6$ [${\alpha}-loop$ L6], ${\alpha}L2$, and ${\alpha}L3$) are implicated in intersubunit communication. In the present study, conformational changes in ${\alpha}L6$ were monitored by measuring the sensitivity of mutant proteins in these regions to trypsin. The addition of a ${\alpha}$ subunit-specific ligand, D,L-${\alpha}$-glycerophosphate (GP), partially restored the sensitivity of mutant proteins to trypsin. In contrast, the addition of the ${\beta}$ subunit-specific ligand L-serine (Ser) resulted in varied sensitivity to trypsin, with an increase in PT53 (substitution of Pro with Thr at residue 53) and DG56, decrease in NS104 and wild type, and no change in GD51 and PH53. This finding may be related to several reaction intermediates formed under this condition. The addition of both GP and Ser led to a highly stable state of the complex. The present results are consistent with the current model. The method used herein may be useful for screening residues involved in intersubunit communication.

The Relationship Between Serum Prostate Specific Antigen and Prostate Volume of Measured by Transrectal Ultrasonography (혈청 전립선특이항원과 경직장초음파로 측정된 전립선 용적과의 상관관계)

  • Kim, Mi-Young
    • Journal of radiological science and technology
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    • v.33 no.3
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    • pp.239-243
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    • 2010
  • Even though clinical use DRE, PSA and TRUS for screening while diagnosing prostate cancer early, however, we cannot achieve to diagnose it accurately by one method. Thus, mutual supplements are strongly required in diagnosis. It means that we should identify relationship between prostate volume and serum PSA under different ages for better diagnosis. The subjects(405 men) visited the department of diagnostic radiology in a general university hospital from January 2008 to December 2008, who was going to take the prostatic evaluation by TRUS. 13 men, who did definite diagnosis of prostate cancer before through biopsy, were excluded in this study. All data were expressed as mean and standard deviations by using SPSS 14.0 package programs and were analyzed by using one-way ANOVA to evaluate the effect of different ages. And Scheffe Post hoc comparison is accomplished in order to make significant difference in prostate volume, PSA and PSAD according to ages. Pearson correlation was used for ages, prostate volume, PSA and PSAD. According to examination, the subjects' prostate volume and serum PSA level increase as person's age. We can see significant difference between group over 70 years and group from 40 and 50 years. Moreover, age, prostate volume and serum PSA have significant positive correlation. Even though we can get variations in serum PSA level by changing prostate volume according to ages, it is worthwhile to mention that more strong association for prostate volume is observed than ages in elevation of serum PSA. Therefore, it is strongly recommended to consider prostate volume weightly when it comes to evaluate for men with elevated PSA level. Finally, TRUS is an excellent extra equipment to diagnose prostate cancer because it can contribute to early diagnose and pertinent treatment of prostate cancer.

Study on the Characteristics of DNA Comet Assay for Irradiated Vegetables and Grains (방사선조사된 채소류 및 곡물류의 DNA Comet Assay 특성 연구)

  • Seo, Jung-Eun;Oh, Se-Wook;Kim, Yun-Ji;Lee, Nam-Hyouck;Hong, Sang-Pill;Kim, Young-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.4
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    • pp.472-476
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    • 2008
  • The possibility of DNA comet assay as a rapid method for screening the irradiated vegetables and grains was evaluated. Vegetables (spring onion, garlic, and tomato) irradiated at $0{\sim}3$ kGy and grains (rice flour and black soybean) irradiated at $0{\sim}9$ kGy were used as samples. Optimum DNA comet assay conditions, such as elution, sedimentation of suspension, and lysis time of cell, were established. The optimum conditions for vegetables were 10 min for the elution time, 0 min for the sedimentation time, and 5 min for the lysis time. The optimum conditions for grains were 15 min for the elution time, 60 min for the sedimentation time, and 30 min for the lysis time. For the food application of DNA comet assay, it was possible to screen various food samples irradiated at the following doses: spring onion at 2 kGy, garlic at 3 kGy, tomato at 1 kGy, rice flour at 9 kGy, and black soybean at 3 kGy. Each sample showed different forms and sizes in DNA comet. Therefore, further studies on various methods using comet shape, concentration, or area in DNA comet assay are necessary.

Sorption and Leaching Characteristics of Diesel-Contaminated Soils Treated by Cold Mix Asphalt (Cold Mix Asphalt로 처리한 디젤 오염 토양의 흡착 및 용출특성)

  • Seo Jin-Kwon;Hwang Inseong;Park Joo-Yang
    • Journal of Soil and Groundwater Environment
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    • v.9 no.4
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    • pp.24-31
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    • 2004
  • A cold mix asphalt (CMA) treatment process was proposed as a tool to recycle soils contaminated with petroleum hydrocarbons. Experimental studies were conducted to characterize performances of the CMA process in treating soils contaminated with diesel or diesel compounds. From the screening experiments, it was found that performances of five types of asphalt emulsions that contained a cationic or an anionic or a nonionic surfactant were not substantially different. In consideration of higher affinity for soils and higher sorption coefficients obtained, an emulsion containing Lauryl Dimethyl Benzyl Ammonium Chloride (LDBAC) was selected as a promising asphalt emulsion for treating diesel-contaminated soils. When the asphalt emulsion LDBAC was applied to treat three compounds that originated from diesel, the removal efficiencies obtained in the order of decreasing efficiencies were as follows: docosane > pentadecane > undecane. Leaching experiments on the specimen formulated by the emulsion LDBAC found that the selected treatment method could treat soils with diesel concentrations as high as 10,000 mg/kg. Leaching of the diesel from the specimen was controlled by diffusion for the first four days and then leaching rate diminished substantially. The latter behavior was characterized as depletion, which represents that the contaminant released amounts to more than $50\%$ of the total amount of the contaminant that can be leached. The amounts of three diesel compounds leached from the specimen in the order of decreasing amount were undecane, pentadecane, and docosane. The curing of the soil contaminated with pentadecane was relatively slow.

Direct Imaging of Polarization-induced Charge Distribution and Domain Switching using TEM

  • O, Sang-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.99-99
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    • 2013
  • In this talk, I will present two research works in progress, which are: i) mapping of piezoelectric polarization and associated charge density distribution in the heteroepitaxial InGaN/GaN multi-quantum well (MQW) structure of a light emitting diode (LED) by using inline electron holography and ii) in-situ observation of the polarization switching process of an ferroelectric Pb(Zr1-x,Tix)O3 (PZT) thin film capacitor under an applied electric field in transmission electron microscope (TEM). In the first part, I will show that strain as well as total charge density distributions can be mapped quantitatively across all the functional layers constituting a LED, including n-type GaN, InGaN/GaN MQWs, and p-type GaN with sub-nm spatial resolution (~0.8 nm) by using inline electron holography. The experimentally obtained strain maps were verified by comparison with finite element method simulations and confirmed that not only InGaN QWs (2.5 nm in thickness) but also GaN QBs (10 nm in thickness) in the MQW structure are strained complementary to accommodate the lattice misfit strain. Because of this complementary strain of GaN QBs, the strain gradient and also (piezoelectric) polarization gradient across the MQW changes more steeply than expected, resulting in more polarization charge density at the MQW interfaces than the typically expected value from the spontaneous polarization mismatch alone. By quantitative and comparative analysis of the total charge density map with the polarization charge map, we can clarify what extent of the polarization charges are compensated by the electrons supplied from the n-doped GaN QBs. Comparison with the simulated energy band diagrams with various screening parameters show that only 60% of the net polarization charges are compensated by the electrons from the GaN QBs, which results in the internal field of ~2.0 MV cm-1 across each pair of GaN/InGaN of the MQW structure. In the second part of my talk, I will present in-situ observations of the polarization switching process of a planar Ni/PZT/SrRuO3 capacitor using TEM. We observed the preferential, but asymmetric, nucleation and forward growth of switched c-domains at the PZT/electrode interfaces arising from the built-in electric field beneath each interface. The subsequent sideways growth was inhibited by the depolarization field due to the imperfect charge compensation at the counter electrode and preexisting a-domain walls, leading to asymmetric switching. It was found that the preexisting a-domains split into fine a- and c-domains constituting a $90^{\circ}$ stripe domain pattern during the $180^{\circ}$ polarization switching process, revealing that these domains also actively participated in the out-of-plane polarization switching. The real-time observations uncovered the origin of the switching asymmetry and further clarified the importance of charged domain walls and the interfaces with electrodes in the ferroelectric switching processes.

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Evaluation of Diagnostic Performance of a Polymerase Chain Reaction for Detection of Canine Dirofilaria immitis (개 심장사상충을 진단하기 위한 중합연쇄반응검사 (PCR)의 진단적 특성 평가)

  • Pak, Son-Il;Kim, Doo
    • Journal of Veterinary Clinics
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    • v.24 no.2
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    • pp.77-81
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    • 2007
  • Diagnostic performance of polymerase chain reaction (PCR) for detecting Dirofilaria immitis in dogs was evaluated when no gold standard test was employed. An enzyme-linked immunosorbent assay test kit (SnapTM, IDEXX, USA) with unknown parameters was also employed. The sensitivity and specificity of the PCR from two-population model were estimated by using both maximum likelihood using expectation-maximization (EM) algorithm and Bayesian method, assuming conditional independence between the two tests. A total of 266 samples, 133 samples in each trial, were randomly retrieved from the heartworm database records during the year 2002-2004 in a university animal hospital. These data originated from the test results of military dogs which were brought for routine medical check-up or testing for heartworm infection. When combined 2 trials, sensitivity and specificity of the PCR was 96.4-96.7% and 97.6-98.8% in EM and 94.4-94.8% and 97.1-98% in Bayesian. There were no statistical differences between estimates. This finding indicates that the PCR assay could be useful screening tool for detecting heartworm antigen in dogs. This study was provided further evidences that Bayesian approach is an alternative approach to draw better inference about the performance of a new diagnostic test in case when either gold test is not available.

Screening for Growth Inhibitory Effects of Medicinal Plants Used in Traditional Korean Medicine in NCI-H1229 Human Lung Cancer Cells (국산 한약재로 이용되는 약용식물의 NCI-H1229 인간 폐암 세포주에 대한 성장 억제효과 분석)

  • Nho, Jong Hyun;Kim, A Hyeon;Jung, Ho Kyung;Lee, Mu Jin;Jang, Ji Hun;Lee, Ki Ho;Lee, Hyun Joo;Park, Ho;Cho, Hyun Woo
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.4
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    • pp.281-290
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    • 2018
  • Background: Lung cancer, the most common malignant disease worldwide, is the predominant cause of cancer deaths, particularly amongst men. Therefore, various researchers have focused on the growth inhibitory effects of medicinal plants used in traditional Korean medicine. This study aimed to investigate the growth inhibitory effects of ethanol extracts of Rubiae radix, Inulae flos, Nelumbinis receptaculum, Astilbe radix, and Lagerstroemia flos on NCI-H1229 cells. Method and Results: The viability of NCI-H1229 cells was evaluated in vitro using an MTS assay. Treatment with the ethanol extracts of the selected medicinal plants at $500{\mu}g/m{\ell}$ reduced NCI-H1229 cell viability and increased apoptotic cell death and caspase-3 activation. In addition, treatment with ethanol extracts of Inulae flos and Astilbe radix increases DNA fragmentation, as measured by the TUNEL assay. Conclusions: These results indicated that ethanol extracts of Rubiae radix, Inulae flos, Nelumbinis receptaculum, Astilbe radix, and Lagerstroemia flos exhibited growth inhibitory effects, inducing apoptotic cell death, DNA fragmentation and caspase-3 activation in NCI-H1229 cells. Therefore, these medicinal plant extracts may be used in the development of natural medicines to inhibit the growth of lung cancers. However, further study is needed to determine the active ingredients of the ethanol extracts from medicinal plants that are reposible for the inhibitory effect on lung cancer cell grwoth.

Expression of Anthrax Lethal Factor, a Major Virulence Factor of Anthrax, in Saccharomyces cerevisiae (Yeast내에서 탄저병 원인균인 Bacillus anthracis의 치사독소인 Lethal Factor 단백질 발현)

  • Hwang Hyehyun;Kim Joungmok;Choi Kyoung-Jae;Chung Hoeil;Han Sung-Hwan;Koo Bon-Sung;Yoon Moon-Young
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.275-280
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    • 2005
  • Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.

Biofilm Forming Ability and Production of Curli and Cellulose in Clinical Isolates of Enterobacteriaceae (생물막형성 장내세균의 Curli 및 Cellulose 세포외 바탕질 분석)

  • Choi, Yeh-Wan;Lee, Hee-Woo;Kim, Sung-Min;Lee, Je-Chul;Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek;Kim, Jung-Min
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.335-341
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    • 2011
  • In this study, 22 clinical isolates of Enterobacteriaceae including Citrobacterfreundii (6 strains), Enterobacter cloacae (5 strains), Enterobacter aerogenes (3 strains), Serratia marcescens (7 strains) and Pantoea spp. (1 strain) were investigated for the biofilm forming ability and biosynthesis of curli and cellulose. Biofilm forming ability was the highest among the isolates of E. cloacae and the lowest among the isolates of E. aerogenes. The expression of the biofilm-forming extracellular matrix components, cellulose and curli fimbriae, was examined by Congo-red (CR) staining and calcofluor staining methods. PCR screening for the presence of curli gene (csgA) revealed that 4 strains of E. cloacae and 1 strain of C. freundii carried the csgA, showing a good correlation between the phenotypic detection of curli fimbriae by CR staining method and the genotypic detection of curli gene by PCR in E. cloacae.

Anti-Androgenic Activity of Phthalate Esters (Di(2-ethylhexyl) Phthalate, Di(n-butyl) Phthalate, and Butylbenzyl Phthalate) in the Rodent 10-day Hershberger Assay using Immature Castrated Male Rats

  • Kang, Il-Hyun;Kim, Hyung-Sik;Kim, Tae-Sung;Moon, Hyun-Ju;Kim, In-Young;Kang, Tae-Seok;Park, Kui-Lea;Choi, Kwang-Sik;Han, Soon-Young
    • Toxicological Research
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    • v.21 no.3
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    • pp.187-193
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    • 2005
  • The rodent Hershberger assay is considered as a potential short term in vivo screening method for the detection of androgenic or anti-androgenic compounds. The objective of this study was to evaluate the anti-androgenic activities of di(2-ethylhexyl) phthalate (DEHP), di(n-butyl) phthalate (DBP), and butylbenzyl phthalate (BBP). A 10-day Hershberger assay was performed using immature Sprague-Dawley male rats castrated at 6 weeks of age. Tastosterone propionate (TP, 0.4 mg/kg/day) was administered s.c. to castrated male rats and followed by flutamide (1, 5, 10, or 20 mg/kg/day) treatment for 10 days by oral gavage. Similarly, DEHP, DBP, or BBP were also administered by oral gavage at 250, 500, or 1000 mg/kg/day after TP (0.4 mg/kg/day) administration. As expected, flutamide significantly inhibited the TP-induced re-growth of seminal vesicles, ventral prostate, and Levator ani plus bulbocavernosus muscles (LABC) at 1 mg/kg/day and above, and Cowper's glands and glans penis at 5 mg/kg/day and above. DEHP significantly (p<0.05) decreased the seminal vesicles, ventral prostate, LABC and Cowper's glands weights at 1000 mg/kg/day. BBP at 1000 mg/kg/day significantly inhibited TP-induced re-growth of the LABC in the immature castrated male rats, whereas ventral prostate, seminal vesicles, and Cowper's glands weights were unaffected. In contrast to DEHP, DBP did not affect accessory sex organ weights at any concentration. Body weights, combined adrenal glands, and kidney weights were not affected, but liver weights were significantly increased at high dosages in the DEHP, DBP, and BBP treatment groups. Our observations strongly suggest that DEHP acts as an androgen antagonist at the high dose (i.e., 1000 mg/kg/day).