• Title/Summary/Keyword: subtractive

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Molecular Biological Analysis of Fish Behavior as a Biomonitoring System for Detecting Diazinon

  • Shin, Sung-Woo;Chon, Tae-Soo;Kim, Jong-Sang;Lee, Sung-Kyu;Koh, Sung-Cheol
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2002.10a
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    • pp.156-156
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    • 2002
  • The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.

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Localization of Weel and Other Cell Cycle Machinery in the Mouse Primordial and Growing Follicles

  • Park, Chang-Eun;Kim, Young-Hoon;Jeon, Eun-Hyun;Lee, Suman;Lee, Sook-Hwan;Lee, Kyung-Ah
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.02a
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    • pp.21-23
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    • 2003
  • Mechanisms regulate the arrest and growth of the resting primordial follicles are very poorly understood. To elucidate genes involved in the early folliculogenesis, we conducted suppression subtractive hybridization using mRNA from day1 and day5 ovaries and selected weel for further analysis, since it was most frequent gene in the day1-subtracted cDNA library (1). Expression of weel and correlated components of the cell cycle machinery, such as cdc2, cyclin B1, cdc25C, and phosphorylated cdc2 was evaluated by immunohistochemistry. In primordial follicles, expression of weel, cdcw, and cyclin B1 was cytoplasmic in oocytes, but phosphorylated cdc2 was weakly expressed in oocytes. While cdc25C expression was in ovarian somatic and in some theca cells. None of components was expressed in the pre-granulosa cells of the primordial follicles, while weel weakly, and cdc2 and cyclin B1 was strongly expressed in the granulosa cells of the growing follicles. Results from the present study suggest that 1) the mejotic arrest of the oocytes may not due to of cell cycle machinery, and 2) the weel may arrest meiosis by sequestering cdc2 and cyclin B1 in the cytoplasm by protein-protein interactions and/or by inhibitory phosphorylation.

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Comparative proteomic analysis of plant responses to sound waves in Arabidopsis

  • Kwon, Young Sang;Jeong, Mi-Jeong;Cha, Jaeyul;Jeong, Sung Woo;Park, Soo-Chul;Shin, Sung Chul;Chung, Woo Sik;Bae, Hanhong;Bae, Dong-Won
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.261-272
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    • 2012
  • Environmental factors greatly influence the growth, development, and even genetic characteristics of plants. The mechanisms by which sound influences plant growth, however, remain obscure. Previously, our group reported that several genes were differentially regulated by specific frequenciesof sound treatmentusing a sound-treated subtractive library. In this study, we used a proteomic approach to investigate plant responses to sound waves in Arabidopsis. The plants were exposed to 250-Hz or 500-Hz sound waves, and total proteins were extracted from leaves 8 h and 24 h after treatment. Proteins extracted from leaves were subjected to 2-DE analysis. Thirty-eight spots were found to be differentially regulated in response to sound waves and were identified using MALDI-TOF MS and MALDI-TOF/TOF MS. The functions of the identified proteins were classified into photosynthesis, stress and defense, nitrogen metabolism, and carbohydrate metabolism. To the best of our knowledge, this is the first report on the analysis of protein changes in response to sound waves in Arabidopsis leaves. These findings provide a better understanding of the molecular basis of responses to sound waves in Arabidopsis.

An Acidic PATHOGENESIS-RELATED1 Gene of Oryza grandiglumis is Involved in Disease Resistance Response Against Bacterial Infection

  • Shin, Sang Hyun;Pak, Jung-Hun;Kim, Mi Jin;Kim, Hye Jeong;Oh, Ju Sung;Choi, Hong Kyu;Jung, Ho Won;Chung, Young Soo
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.208-214
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    • 2014
  • Wild rice, Oryza grandiglumis shows hyper-resistance response to pathogen infection. In order to identify genes necessary for defense response in plants, we have carried out a subtractive hybridization coupled with a cDNA macroarray. An acidic PATHOGENESIS-RELATED1 (PR1) gene of the wild rice is highly identical to the acidic PR1 genes of different plant species. The OgPR1a cDNA has an apparent single open reading frame with a predicted molecular mass 40,621 Da and an isoelectic point of 5.14. Both in silico analysis and a transient expression assay in onion epidermal cells revealed that the OgPR1a protein could be localized in intercellular space in plants. The OgPR1a mRNA was strongly transcribed by the exogenous treatment with ethylene and jasmonic acid as well as protein phosphatase inhibitors. Additionally, ectopic expression of the OgPR1a conferred disease resistance on Arabidopsis to the bacterial and fungal infections.

Simplification of Boundary Representation Models Based on Stepwise Volume Decomposition (단계적 볼륨분해에 기반한 경계표현 모델의 단순화)

  • Kim, Byung Chul;Mun, Duhwan
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.37 no.10
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    • pp.1305-1313
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    • 2013
  • In this study, a method to apply feature-based simplification to boundary representation models is proposed. For feature-based simplification, a volume decomposition tree is created from a boundary representation model. The volume decomposition tree is represented by regularized Boolean operations of additive volumes, subtractive volumes, and fillet/round/chamfer volumes, and it is generated by stepwise volume decomposition, which consists of fillet/round/chamfer decomposition, wrap-around decomposition, volume split decomposition, and cell-based decomposition. After the volume decomposition tree is transformed to an infix expression, the CAD model can be simplified by reordering the volumes. To verify the proposed method, a prototype system was implemented, and experiments on test cases were conducted. From the results of the experiments, it is verified that the proposed method is useful for simplifying CAD models based on boundary representation.

Genes Expressed During Fruiting Body Formation of Agrocybe cylindracea

  • Shim, Sung-Mi;Kim, Sang-Beom;Kim, Hey-Young;Rho, Hyun-Su;Lee, Hyun-Sook;Lee, Min-Woong;Lee, U-Youn;Im, Kyung-Hoan;Lee, Tae-Soo
    • Mycobiology
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    • v.34 no.4
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    • pp.209-213
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    • 2006
  • Agrocybe cylindracea, an edible mushroom belonging to Bolbitiaceae, Agaricales, is widely used as invaluable medicinal material in the oriental countries. This study was initiated to find the genes expressed during the fruiting body formation of A. cylindracea. The cDNAs expressed differentially during fruiting body morphogenesis of A. cylindracea were isolated through subtractive hybridization between vegetative mycelia and fruiting bodies. The cDNAs expressed in the fruiting body morphogenesis of A. cylindracea were cloned and twenty genes were identified. Eleven were homologous to genes of known functions, three were homologous to genes in other organism without any function known. Six were completely novel genes specific to A. cylindracea so far examined. Some genes with known functions were a pleurotolysin, a self-assembling poreforming cytolysins; Aa-Pril and Pir2p, specifically induced genes during fruiting initiation of other mushroom, Agrocybe aegerita; an amino acid permease; a cytochrome P450; a MADS-box gene; a peptidylprolyl isomerase; and a serine proteinase. For other clones, no clear function was annotated so far. We believe the first report of the differentially expressed genes in fruiting process of A. cylindracea will be great helps for further research.

Down-regulation of Tcf-1 Expression by Activation-induced Apoptosis of T Cell Hybridoma

  • Jeong, Sun-Joo;Jeon, Sung-Ho;Yim, Jeong-Bin;Park, Sang-Dai;Rho, Hyun-Seung
    • Animal cells and systems
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    • v.2 no.3
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    • pp.403-410
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    • 1998
  • The Tcf-1 (T cell specific factor-1) is a transcription factor uniquely expressed in T-lineage cells. Its expression is developmentally regulated, which is high in the specific stage of immature thymocytes, but is much lower in mature T cells. We cloned the Tcf-1 gene by subtractive hybridization and found it to be highly expressed in the thymus compared to the mRNA level in the spleen as expected. Since apoptosis occurs enormously in the thymus, we were interested in whether Tcf-1 gene expression could be regulated by such a high level of apoptotic assault. By using T cell hybridoma 70.7 cells, we induced apoptosis by incubating cells with anti-CD3 antibody in vitro. After apoptosis induction, Tcf-1 mRNA level was found to be significantly reduced compared to normal cells. Since Tcf-1 is a transcription factor for the CD3-e gene, we tested how CD3-e expression is regulated in apoptotic cells. The surface level of CD3-e protein is also down-regulated after apoptosis induction. Such a down-modulation of CD3-e protein would reduce the TCR/CD3 complex on the cell surface, which would be an important regulator for T cell apoptosis.

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The Hypernodulating nts Mutation Induces Jasmonate Synthetic Pathway in Soybean Leaves

  • Seo, Hak Soo;Li, Jinjie;Lee, Sun-Young;Yu, Jae-Woong;Kim, Kil-Hyun;Lee, Suk-Ha;Lee, In-Jung;Paek, Nam-Chon
    • Molecules and Cells
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    • v.24 no.2
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    • pp.185-193
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    • 2007
  • Symbiotic nitrogen fixation with nitrogen-fixing bacteria in the root nodules is a distinctly beneficial metabolic process in legume plants. Legumes control the nodule number and nodulation zone through a systemic negative regulatory system between shoot and root. Mutation in the soybean NTS gene encoding GmNARK, a CLAVATA1-like serine/threonine receptor-like kinase, causes excessive nodule development called hypernodulation. To examine the effect of nts mutation on the gene expression profile in the leaves, suppression subtractive hybridization was performed with the trifoliate leaves of nts mutant 'SS2-2' and the wild-type (WT) parent 'Sinpaldalkong2', and 75 EST clones that were highly expressed in the leaves of the SS2-2 mutant were identified. Interestingly, the expression of jasmonate (JA)-responsive genes such as vspA, vspB, and Lox2 were upregulated, whereas that of a salicylate-responsive gene PR1a was suppressed in the SS2-2 mutant. In addition, the level of JA was about two-fold higher in the leaves of the SS2-2 mutant than in those of the WT under natural growth conditions. Moreover, the JA-responsive gene expression persists in the leaves of SS2-2 mutant without rhizobia infection in the roots. Taken together, our results suggest that the nts mutation increases JA synthesis in mature leaves and consequently leads to constitutive expression of JA-responsive genes which is irrelevant to hypernodulation in the root.

A Study on Introduction of Division Algorithm in Mathematics Textbooks : Focussing on Elementary Math Textbooks and Manuals Applied 2009 Revised Curriculum (자연수 세로 나눗셈 알고리즘 도입 방법 고찰: 2009 개정 교육과정의 초등학교 수학 교과서와 지도서를 중심으로)

  • Kang, Ho-Jin;Kim, Ju-Chang;Lee, Kwang-Ho;Lee, Jae-Hak
    • Education of Primary School Mathematics
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    • v.20 no.1
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    • pp.69-84
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    • 2017
  • The purpose of this study is to review how to introduce a division algorithm in mathematics textbooks which were applied 2009 revised curriculum. As a result, the textbooks do not introduce the algorithm in the context of division by equal part. The standardized division algorithm was introduced apart from the stepwise division algorithms and there is no explanation in between them. And there is a lack connectivity between activities and algorithms. This study is expected to help new curriculum and textbook to introduce division algorithm in proper way.

A Study on Feature-Based Multi-Resolution Modelling - Part II: System Implementation and Criteria for Level of Detail (특징형상기반 다중해상도 모델링에 관한 연구 - Part II: 시스템 구현 및 상세수준 판단기준)

  • Lee K.Y.;Lee S.H.
    • Korean Journal of Computational Design and Engineering
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    • v.10 no.6
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    • pp.444-454
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    • 2005
  • Recently, the requirements of multi-resolution models of a solid model, which represent an object at multiple levels of feature detail, are increasing for engineering tasks such as analysis, network-based collaborative design, and virtual prototyping and manufacturing. The research on this area has focused on several topics: topological frameworks for representing multi-resolution solid models, criteria for the level of detail (LOD), and generation of valid models after rearrangement of features. As a solution to the feature rearrangement problem, the new concept of the effective zone of a feature is introduced in the former part of the paper. In this paper, we propose a feature-based non-manifold modeling system to provide multi-resolution models of a feature-based solid or non-manifold model on the basis of the effective feature zones. To facilitate the implementation, we introduce the class of the multi-resolution feature whose attributes contain all necessary information to build a multi-resolution solid model and extract LOD models from it. In addition, two methods are introduced to accelerate the extraction of LOD models from the multi-resolution modeling database: the one is using an NMT model, known as a merged set, to represent multi-resolution models, and the other is storing differences between adjacent LOD models to accelerate the transition to the other LOD. We also suggest the volume of the feature, regardless of feature type, as a criterion for the LOD. This criterion can be used in a wide range of applications, since there is no distinction between additive and subtractive features unlike the previous method.