• Title/Summary/Keyword: screening conditions

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Development of Efficient Screening Methods for Resistant Cucumber Plants to Meloidogyne incognita (오이 뿌리혹선충병에 대한 효율적인 저항성 검정법 확립)

  • Hwang, Sung Min;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.20 no.2
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    • pp.119-125
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    • 2014
  • Root-knot nematodes represent a significant problem in cucumber, causing reduction in yield and quality. To develop screening methods for resistance of cucumber to root-knot nematode Meloidogyne incognita, development of root-knot nematode of four cucumber cultivars ('Dragonsamchuk', 'Asiastrike', 'Nebakja' and 'Hanelbakdadaki') according to several conditions such as inoculum concentration, plant growth stage and transplanting period was investigated by the number of galls and egg masses produced in each seedling 45 days after inoculation. There was no difference in galls and egg masses according to the tested condition except for inoculum concentration. Reproduction of the nematode on all the tested cultivars according to inoculum concentration increased in a dose-dependent manner. On the basis of the result, the optimum conditions for root-knot development on the cultivars is to transplant period of 1 week, inoculum concentration of 5,000 eggs/plant and plant growth stage of 3-week-old in a greenhouse ($25{\pm}5^{\circ}C$). In addition, under optimum conditions, resistance of 45 commercial cucumber cultivars was evaluated. One rootstock cultivar, Union was moderately resistant to the root-knot nematode. However, no significant difference was in the resistance of the others cultivar. According to the result, we suggest an efficient screening method for new resistant cucumber to the root-knot nematode, M. incognita.

Mithramycin Inhibits Etoposide Resistance in Glucose-deprived HT-29 Human Colon Carcinoma Cells

  • Lee, Eun-Mi;Park, Hae-Ryong;Hwang, Ji-Hwan;Park, Dong-Jin;Chang, Kyu-Seob;Kim, Chang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1856-1861
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    • 2007
  • Physiological cell conditions such as glucose deprivation and hypoxia play roles in the development of drug resistance in solid tumors. These tumor-specific conditions cause decreased expression of DNA topoisomerase $II{\alpha}$, rendering cells resistant to topo II target drugs such as etoposide. Thus, targeting tumor-specific conditions such as a low glucose environment may be a novel strategy in the development of anticancer drugs. On this basis, we established a novel screening program for anticancer agents with preferential cytotoxic activity in cancer cells under glucose-deprived conditions. We recently isolated an active compound, AA-98, from Streptomyces sp. AA030098 that can prevent stress-induced etoposide resistance in vitro. Furthermore, LC-MS and various NMR spectroscopic methods identified AA-98 as mithramycin, which belongs to the aureolic acid group of antitumor compounds. We found that mithramycin prevents the etoposide resistance that is induced by glucose deprivation. The etoposide-chemosensitive action of mithramycin was just dependent on strict low glucose conditions, and resulted in the selective cell death of etoposide-resistant HT-29 human colon cancer cells.

Development of an Efficient In Vitro Screening Method for Selection of Resistant Lily Cultivars Against Fusarium oxysporum f. sp. lilii (백합 구근부패병 저항성 검정을 위한 기내 간편 검정법 개발)

  • Jang, Ji-Young;Moon, Ki-Beom;Ha, Jang-Ho;Park, Ji-Sun;Kim, Mi-Jin;Jeon, Jae-Heung;Lee, Geung-Joo;Kim, Hyun-Soon
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.883-890
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    • 2015
  • The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.

A Method and Application on Reliability Test (신뢰성 시험의 방법과 응용에 관한 연구)

  • 김재중;김원중
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.20 no.44
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    • pp.393-399
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    • 1997
  • This study is concerned with reliability technology. In order to achieve the quality level of items for consumer's satisfaction, tests for the item's reliability are essential. This article deals with a method and real field application to plan reliability testing. Especially the environmental conditions and methods such as screening test for electronic components will be shown. As well, we will explore methods and field applications with respect to mechanic destructive tests and non destructive tests.

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Establishment of Artificial Screening Methods and Evaluation of Barley Germplasms for Resistance to Fusarium Head Blight (보리 붉은곰팡이병 검정법과 저항성 품종 선발)

  • Han Ouk-Kyu;Kim Jung-Gon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.3
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    • pp.191-196
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    • 2005
  • Fusarium head blight (FHB) is a severe disease problem that affects the quality and yield of barley grain. The evaluation of FHB resistance is difficult because environmental conditions greatly influence FHB infection and development. The objectives of this study were to: 1) establish an efficient screening method for selecting resistant barley to FHB, 2) compare FHB severity between the cut-spike method and pot-plant method for development of mass screening, and 3) estimate FHB resistance for barley germplasms. Barley cultivars and lines were evaluated for reaction to FHB in controlled-greenhouse condition. Spikes were spray-inoculated with a suspension $(5.0\times10^5\;macroconidia\;mL^{-1})$ of Fusarium graminearum SCK-O4 strain, and then kept in a greenhouse at $18-25^{\circ}C$ with $80-100\%$ relative humidity. Inoculation were employed at 3 different heading growth stages (heading date, three days after heading, and five days after heading). The inoculation was performed in 2 consecutive days in order to avoid escapes. The inoculated plants were maintained in the greenhouse at 4 different free moisture periods (1, 3, 5, and 7 days). The percentage of FHB severity was scored from 0 to 9 according to the rate of infected kernels per spike, and three spikes were evaluated per replication with 3 replicates. There were significant differences of FHB severity depending on the different free moisture periods, but not by the inoculation at different heading stages. The optimum evaluation point of FHB severity in the greenhouse condition was on the 7th day under free moisture condition after inoculation at the heading date. Infection level in cut-spike method highly correlated with that in pot-plant method. This suggested that cut-spike method is useful in evaluating of FHB resistance in barley. Six cultivars, such as Jinkwang, Buheung, Atahualpha 92, Chevron-b, Gobernadora-d, and MNBrite-c, were selected as resistant varieties to FHB. Correlation coefficient for the FHB severity evaluated by the pot-plant method between two seasons was 0.794, indicating the stability and accuracy of the screening method.

Pepper Blight Disease Inhibition Metagenome Clone Screening Using Soil Metagenome Library (토양 Metagenome Library로부터 고추역병 저해 클론 탐색)

  • Park, Hae-Chul;Sung, So-Ra;Kim, Dong-Gwan;Koo, Bon-Sung;Jeong, Byeong-Moon;Kim, Jin-Heung;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.228-231
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    • 2009
  • We have purified Phytophthora capsici alpha and beta tubulin from Escherchia coli BL21(DE3). The recombinant alpha and beta tubulins were assembled into microtubule in vitro with specific conditions. The metagenome library was isolated from soil in the Mt. Yeo-Ki, Suwon, Korea and manufactured with the method mentioned in experiment contents for in vitro screening of microtubule assembly screening. FRET effect was used for microtubule assembly inhibitor screening with metagenome library. We got 2 metagenome clones from in vitro screening, and these 2 hit clones showed P. capsici growth inhibition activity on the growing pepper plants. These results suggest that new development of potent inhibitor for pepper blight disease and new approach to prevention of pepper blight disease.

Structural Characteristics of Expression Module of Unidentified Genes from Metagenome (메타게놈 유래 미규명 유전자의 발현에 관련된 특성분석)

  • Park, Seung-Hye;Jeong, Young-Su;Kim, Won-Ho;Kim, Geun-Joong;Hur, Byung-Ki
    • KSBB Journal
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    • v.21 no.2
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    • pp.144-150
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    • 2006
  • The exploitation of metagenome, the access to the natural extant of enormous potential resources, is the way for elucidating the functions of organism in environmental communities, for genomic analyses of uncultured microorganism, and also for the recovery of entirely novel natural products from microbial communities. The major breakthrough in metagenomics is opened by the construction of libraries with total DNAs directly isolated from environmental samples and screening of these libraries by activity and sequence-based approaches. Screening with activity-based approach is presumed as a plausible route for finding new catabolic genes under designed conditions without any prior sequence information. The main limitation of these approaches, however, is the very low positive hits in a single round of screening because transcription, translation and appropriate folding are not always possible in E. coli, a typical surrogate host. Thus, to obtain information about these obstacles, we studied the genetic organization of individual URF's(unidentified open reading frame from metagenome sequenced and deposited in GenBank), especially on the expression factors such as codon usage, promoter region and ribosome binding site(rbs), based on DNA sequence analyses using bioinformatics tools. And then we also investigated the above-mentioned properties for 4100 ORFs(Open Reading Frames) of E. coli K-12 generally used as a host cell for the screening of noble genes from metagenome. Finally, we analyzed the differences between the properties of URFs of metagenome and ORFs of E. coli. Information derived from these comparative metagenomic analyses can provide some specific features or environmental blueprint available to screen a novel biocatalyst efficiently.

Development of an aequorin-based assay for the screening of corticotropin-releasing factor receptor antagonists (CRF1 길항제 스크리닝을 위한 에쿼린 기반 세포실험 개발연구)

  • Noh, Hyojin;Lee, Sunghou
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.11
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    • pp.7575-7581
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    • 2015
  • Corticotropin-releasing factor(CRF), one of the stress driven neuropeptides, was widely proposed to influence hair loss and re-growth. For the development of receptor antagonists, the screening system based on intracellular calcium signal process was developed and optimized. The aequorin parental cells were transfected with CRF1 receptor and alpha 16 promiscuous G protein cDNA to establish HEK293a16/hCRF1, a stable cell line for the human CRF1 receptor. In HEK293a16/hCRF1 cells, the range of sauvagine dose response was 12-fold higher($EC_{50}:15.21{\pm}1.83nM$) than in the transiently expressed cells, hence essential conditions for the antagonist screening experiments such as the robust signals and high solvent tolerance were secured. The standard antagonists for the CRF1 receptor, antalarmin and CP154526, resulted $IC_{50}$ values of $414.1{\pm}5.5$ and $290.7{\pm}1.9nM$, respectively. Similar results were presented with frozen HEK293a16/hCRF1 cells. Finally, our HEK293a16/hCRF1 cells with the aequorin based cellular functional assay can be a model system for the development of functional cosmetics and modulators that can have a clinical efficacy on hair re-growth.

Effectiveness of stages-matched educational program for cervical cancer screening among adult women in a community (일개 지역사회 여성 주민의 자궁경부암 조기검진 수검에 관한 행동변화단계별 교육 프로그램의 효과)

  • Kim, Young-Bok
    • Korean Journal of Health Education and Promotion
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    • v.24 no.5
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    • pp.23-37
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    • 2007
  • Background: Even though cervical cancer poses a significant global cancer burden, successful implementations for early detection offer an opportunity to prevent deaths and reduce the cancer burden. In Korea, cervical cancer is the most prevalent type of cancer among adult women, but it is one of the few cancers in which a consensus-approved screening test exists for early diagnosis, Pap test, that can be combined with highly efficacious treatment regimens for early-stage disease. Purpose: This study was carried out to identify the cognitive-behavioral factors associated with cervical cancer screening behavior among adult women, aged 40 to 59, and to develop tailored messages and to evaluate the effectiveness of stage-matched educational program. Method: A total of 283 women who aged 40 years or older was recruited in Seoul, from September, 1st to November, 14th, 2003. The intervention group (N=162) and the control group (N=121) were selected from five sub-districts in Seocho-gu, Seoul. Building on the TTM, a quasi-experimental study was conducted to test the effectiveness of stages-matched intervention addressed at the five stages of cervical cancer screening behavior. Women in the intervention group were randomly assigned to one of two conditions, internet or postal services. Results: In our results, 88.9% of participants had received a Pap test at least once in their life-time, and 65.4% had got it in the past two years. With regard to cognitive-behavioral factors, the stages-matched educational program increased attitude and process of change for cervical cancer screening. The percentage changed was the largest in maintenance stage. With regard to delivery methods for tailored messages, the print materials were more effective at increasing screening adherence than the e-mail. Whereas the postal service group showed remarkable the change of behavior stage, the internet service group did not. Also it was not shown any difference of the satisfaction with stages-matched educational program between internet and postal service groups. Conclusion: This study suggested that cervical cancer screening behavior could be changed by tailored messages which had developed with cognitive-behavioral factors. The stages-matched educational program was effective to promote the screening adherence for cervical cancer.