• Title/Summary/Keyword: High-throughput Screening

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Applying a modified AUC to gene ranking

  • Yu, Wenbao;Chang, Yuan-Chin Ivan;Park, Eunsik
    • Communications for Statistical Applications and Methods
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    • v.25 no.3
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    • pp.307-319
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    • 2018
  • High-throughput technologies enable the simultaneous evaluation of thousands of genes that could discriminate different subclasses of complex diseases. Ranking genes according to differential expression is an important screening step for follow-up analysis. Many statistical measures have been proposed for this purpose. A good ranked list should provide a stable rank (at least for top-ranked gene), and the top ranked genes should have a high power in differentiating different disease status. However, there is a lack of emphasis in the literature on ranking genes based on these two criteria simultaneously. To achieve the above two criteria simultaneously, we proposed to apply a previously reported metric, the modified area under the receiver operating characteristic cure, to gene ranking. The proposed ranking method is found to be promising in leading to a stable ranking list and good prediction performances of top ranked genes. The findings are illustrated through studies on both synthesized data and real microarray gene expression data. The proposed method is recommended for ranking genes or other biomarkers for high-dimensional omics studies.

Magnetic beads separation using a multi-layered microfluidic channel (다층구조의 미세유체채널을 이용한 자성입자 분리)

  • Lee, Hye-Lyn;Song, Suk-Heung;Jung, Hyo-Il
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1685-1686
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    • 2008
  • This paper presents the design and experiment results of a multi-layered microsystem for magnetic bead applications. The magneto-microfluidic device is designed for capable of separating magnetic beads. In the presence of the magnetic field, magnetic beads are attracted and moved to high gradient magnetic fields. A multi-layered microfluidic channel consists of top and bottom layers in order to separate magnetic beads in the vertical direction. Our channel is easily integrated magnetic cell sorter, especially on-chip microelectromagnet or permanent magnet device. Fast separation of magnetic beads in top and bottom channels can be used in high throughput screening to monitor the efficiency of blood and drug compounds.

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Genotoxicity on Structural Derivatives of Sophoricoside, a Component of Sophora Japonica, in Bacterial and Mammalian Cells

  • Ryu, Jae-Chun;Kim, Youn-Jung;Kim, Mi-Soon;Kim, Min-Ji;Sarma, Sailendra Nath;Jung, Sang-Hun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.179-188
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    • 2005
  • To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.

Development of a Method for High throughput Screening of Antagonistic Substances against Rice Pathogens using Rice Leaf Explants (벼의 생엽절편을 이용한 병원균 억제물질의 대량 스크리닝 방법 개발)

  • Park, Sait-Byul;Lee, Choong-Hwan;Kim, Tae-Jong;Kang, Lin-Woo;Lee, Byoung-Moo;Kim, Jeong-Gu
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.39-42
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    • 2012
  • A new method for the high throughput screening of antagonistic substances against rice pathogens using rice leaf explants was developed. This method can be used to confirm the activities of any compound or mixture suppressing rice bacterial blight (BB) before field tests. Xanthomonas oryzae pv. oryzae (Xoo) culture medium was distributed in 96 well plates with equally sized explants and the active compounds were added to the wells. The strength suppressing BB was converted into an area percent of the lesion on the rice explants. The explants under BB suppressing activity remained uninfected maintaining their actual green color, while infected explants exhibited pale yellow-colored lesions. Based on the results, this method seems to be faster and easier, dose-dependent, and can be performed all-at-once with a small amount of unspecified compounds. This method also has the potential to be applied to inspection activities for the suppression of other waterborne crop diseases.

Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application

  • Kim, Jennifer Jooyoun;Kwon, Young-Kyung;Kim, Ji Hyung;Heo, Soo-Jin;Lee, Youngdeuk;Lee, Su-Jin;Shim, Won-Bo;Jung, Won-Kyo;Hyun, Jung-Ho;Kwon, Kae Kyoung;Kang, Do-Hyung;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1559-1565
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    • 2014
  • Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.

Metagenome, the Untapped Microbial Genome, toward Discovery of Novel Microbial Resources and Application into the Plant Pathology

  • Lee, Seon-Woo
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.93-98
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    • 2005
  • Molecular ecological studies of microbial communities revealed that only tiny fraction of total microorganisms in nature have been identified and characterized, because the majority of them have not been cultivated. A concept, metagenome, represents the total microbial genome in natural ecosystem consisting of genomes from both culturable microorganisms and viable but non-culturable bacteria. The construction and screening of metagenomic libraries in culturable bacteria constitute a valuable resource for obtaining novel microbial genes and products. Several novel enzymes and antibiotics have been identified from the metagenomic approaches in many different microbial communities. Phenotypic analysis of the introduced unknown genes in culturable bacteria could be an important way for functional genomics of unculturable bacteria. However, estimation of the number of clones required to uncover the microbial diversity from various environments has been almost impossible due to the enormous microbial diversity and various microbial population structure. Massive construction of metagenomic libraries and development of high throughput screening technology should be necessary to obtain valuable microbial resources. This paper presents the recent progress in metagenomic studies including our results and potential of metagenomics in plant pathology and agriculture.

Property-based Design of Ion-Channel-Targeted Library

  • Ahn, Ji-Young;Nam, Ky-Youb;Chang, Byung-Ha;Yoon, Jeong-Hyeok;Cho, Seung-Joo;Koh, Hun-Yeong;No, Kyoung-Tai
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.134-138
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    • 2005
  • The design of ion channel targeted library is a valuable methodology that can aid in the selection and prioritization of potential ion channel-likeness for ion-channel-targeted bio-screening from large commercial available chemical pool. The differences of property profiling between the 93 ion-channel active compounds from MDDR and CMC database and the ACDSC compounds were classified by suitable descriptors calculated with preADME software. Through the PCA, clustering, and similarity analysis, the compounds capable of ion channel activity were defined in ACDSC compounds pool. The designed library showed a tendency to follow the property profile of ion-channel active compounds and can be implemented with great time and economical efficiencies of ligand-based drug design or virtual high throughput screening from an enormous small molecule space.

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Combinatorial Solid Phase Peptide Synthesis and Bioassays

  • Shin, Dong-Sik;Kim, Do-Hyun;Chung, Woo-Jae;Lee, Yoon-Sik
    • BMB Reports
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    • v.38 no.5
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    • pp.517-525
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    • 2005
  • Solid phase peptide synthesis method, which was introduced by Merrifield in 1963, has spawned the concept of combinatorial chemistry. In this review, we summarize the present technologies of solid phase peptide synthesis (SPPS) that are related to combinatorial chemistry. The conventional methods of peptide library synthesis on polymer support are parallel synthesis, split and mix synthesis and reagent mixture synthesis. Combining surface chemistry with the recent technology of microelectronic semiconductor fabrication system, the peptide microarray synthesis methods on a planar solid support are developed, which leads to spatially addressable peptide library. There are two kinds of peptide microarray synthesis methodologies: pre-synthesized peptide immobilization onto a glass or membrane substrate and in situ peptide synthesis by a photolithography or the SPOT method. This review also discusses the application of peptide libraries for high-throughput bioassays, for example, peptide ligand screening for antibody or cell signaling, enzyme substrate and inhibitor screening as well as other applications.

Innate Color Preference of Zebrafish and Its Use in Behavioral Analyses

  • Park, Jong-Su;Ryu, Jae-Ho;Choi, Tae-Ik;Bae, Young-Ki;Lee, Suman;Kang, Hae Jin;Kim, Cheol-Hee
    • Molecules and Cells
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    • v.39 no.10
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    • pp.750-755
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    • 2016
  • Although innate color preference of motile organisms may provide clues to behavioral biases, it has remained a longstanding question. In this study, we investigated innate color preference of zebrafish larvae. A cross maze with different color sleeves around each arm was used for the color preference test (R; red, G; green, B; blue, Y; yellow). The findings showed that 5 dpf zebrafish larvae preferred blue over other colors (B > R > G > Y). To study innate color recognition further, tyrosinase mutants were generated using CRISPR/Cas9 system. As a model for oculocutaneous albinism (OCA) and color vision impairment, tyrosinase mutants demonstrated diminished color sensation, indicated mainly by hypopigmentation of the retinal pigment epithelium (RPE). Due to its relative simplicity and ease, color preference screening using zebrafish larvae is suitable for high-throughput screening applications. This system may potentially be applied to the analysis of drug effects on larval behavior or the detection of sensory deficits in neurological disorder models, such as autism-related disorders, using mutant larvae generated by the CRISPR/Cas9 technique.

Developing a Protein-chip for Depigmenting Agents Screening (미백제 스크리닝용 단백질칩의 개발)

  • Kim, Eun-Ki;Kwak, Eun-Young;Han, Jung-Sun;Lee, Hyang-Bok;Shin, Jung-Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.13-16
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    • 2005
  • For the high-throughput-screening system (HTS) of depigmenting agents using a protein chip, effects of oligonucleotide-inhibitor sequence on the binding of Mitf protein to E box of MC1R was investigated. The sequence of oligonucletide-inhibitor affected the binding of the target DNA to Mitf, depending on the location of the sequence variation in the inhibitor nucleotide. The oligonucletide-inhibitor that changed the CATGTG sequence didn't show enough inhibition of the target DNA to Mitf, whereas significant inhibition was observed when the sequence outside the CATGTG was changed. This result indicated that CATCTG is crucial sequence for the binding of Mitf to I-box which initiates the transcription of pigmenting genes.