• Title/Summary/Keyword: microarray experiment

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The Genome-Wide Expression Profile of Saussurea lappa Extract on House Dust Mite-Induced Atopic Dermatitis in Nc/Nga Mice

  • Lim, Hye-Sun;Ha, Hyekyung;Shin, Hyeun-Kyoo;Jeong, Soo-Jin
    • Molecules and Cells
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    • v.38 no.9
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    • pp.765-772
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    • 2015
  • Saussurea lappa has been reported to possess anti-atopic properties. In this study, we have confirmed the S. lappa's anti-atopic properties in Nc/Nga mice and investigated the candidate gene related with its properties using microarray. We determined the target gene using real time PCR in in vitro experiment. S. lappa showed the significant reduction in atoptic dermatitis (AD) score and immunoglobulin E compared with the AD induced Nc/Nga mice. In the results of microarray using back skin obtained from animals, we found that S. lappa's properties are closely associated with cytokine-cytokine receptor interaction and the JAK-STAT signaling pathway. Consistent with the microarray data, real-time RT-PCR confirmed these modulation at the mRNA level in skin tissues from S. lappa-treated mice. Among these genes, PI3Kca and $IL20R{\beta}$ were significantly downregulated by S. lappa treatment in Nc/Nga mouse model. In in vitro experiment using HaCaT cells, we found that the S. lappa components, including alantolactone, caryophyllene, costic acid, costunolide and dehydrocostus lactone significantly decreased the expression of PI3Kca but not $IL20R{\beta}$ in vitro. Therefore, our study suggests that PI3Kca-related signaling is closely related with the protective effects of S. lappa against the development of atopic-dermatitis.

CLUSTERING DNA MICROARRAY DATA BY STOCHASTIC ALGORITHM

  • Shon, Ho-Sun;Kim, Sun-Shin;Wang, Ling;Ryu, Keun-Ho
    • Proceedings of the KSRS Conference
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    • 2007.10a
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    • pp.438-441
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    • 2007
  • Recently, due to molecular biology and engineering technology, DNA microarray makes people watch thousands of genes and the state of variation from the tissue samples of living body. With DNA Microarray, it is possible to construct a genetic group that has similar expression patterns and grasp the progress and variation of gene. This paper practices Cluster Analysis which purposes the discovery of biological subgroup or class by using gene expression information. Hence, the purpose of this paper is to predict a new class which is unknown, open leukaemia data are used for the experiment, and MCL (Markov CLustering) algorithm is applied as an analysis method. The MCL algorithm is based on probability and graph flow theory. MCL simulates random walks on a graph using Markov matrices to determine the transition probabilities among nodes of the graph. If you look at closely to the method, first, MCL algorithm should be applied after getting the distance by using Euclidean distance, then inflation and diagonal factors which are tuning modulus should be tuned, and finally the threshold using the average of each column should be gotten to distinguish one class from another class. Our method has improved the accuracy through using the threshold, namely the average of each column. Our experimental result shows about 70% of accuracy in average compared to the class that is known before. Also, for the comparison evaluation to other algorithm, the proposed method compared to and analyzed SOM (Self-Organizing Map) clustering algorithm which is divided into neural network and hierarchical clustering. The method shows the better result when compared to hierarchical clustering. In further study, it should be studied whether there will be a similar result when the parameter of inflation gotten from our experiment is applied to other gene expression data. We are also trying to make a systematic method to improve the accuracy by regulating the factors mentioned above.

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The Study of X Chromosome Inactivation Mechanism in Klinefelter's Syndrome by cDNA Microarray Experiment

  • Jeong, Yu-Mi;Chung, In-Hyuk;Park, Jung Hoon;Lee, Sook-Hwan;Chung, Tae-Gyu;Kim, Yong Sung;Kim, Nam-Soon;Yoo, Hyang-Sook;Lee, Suman
    • Genomics & Informatics
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    • v.2 no.1
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    • pp.30-35
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    • 2004
  • To investigate the XIST gene expression and its effect in a Klinefelter's patient, we used Klinefelter's syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter's syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.

Cross Platform Data Analysis in Microarray Experiment (서로 다른 플랫폼의 마이크로어레이 연구 통합 분석)

  • Lee, Jangmee;Lee, Sunho
    • The Korean Journal of Applied Statistics
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    • v.26 no.2
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    • pp.307-319
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    • 2013
  • With the rapid accumulation of microarray data, it is a significant challenge to integrate available data sets addressing the same biological questions that can provide more samples and better experimental results. Sometimes, different microarray platforms make it difficult to effectively integrate data from several studies and there is no consensus on which method is the best to produce a single and unified data set. Methods using median rank score, quantile discretization and standardization (which directly combine rescaled gene expression values) and meta-analysis (which combine the results of individual studies at the interpretative level) are reviewed. Real data examples downloaded from GEO are used to compare the performance of these methods and to evaluate if the combined data set detects more reliable information from the separated data sets or not.

DNA Microarray Analysis of Immediate Response to EGF Treatment in Rat Schwannoma Cells

  • OH, Min-Kyu;Scoles, Daniel R.;Pulst, Stefan-M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.5
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    • pp.444-450
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    • 2005
  • Epidermal growth factor (EGF) activates many intracellular effector molecules, which subsequently influence the expression levels of many genes involved in cell growth, apoptosis and signal transduction, etc. In this study, the early response of gene expressions due to EGF treatment was monitored using oligonucleotide DNA microarrays in rat schwannoma cell lines. An immunoblotting experiment showed the successful activation of EGF receptors and an effector protein, STAT5, due to EGF treatment. The microarray study showed that 35 genes were significantly induced and 2 were repressed within 60 min after the treatment. The list of induced genes included early growth response 1, suppressor of cytokine signaling 3, c-fos, interferon regulatory factor 1 and early growth response 2, etc. According to the microarray data, six of these were induced by more than 10-fold, and showed at least two different induction patterns, indicating complicated regulatory mechanisms in the EGF signal transduction.

Finding associations between genes by time-series microarray sequential patterns analysis

  • Nam, Ho-Jung;Lee, Do-Heon
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.161-164
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    • 2005
  • Data mining techniques can be applied to identify patterns of interest in the gene expression data. One goal in mining gene expression data is to determine how the expression of any particular gene might affect the expression of other genes. To find relationships between different genes, association rules have been applied to gene expression data set [1]. A notable limitation of association rule mining method is that only the association in a single profile experiment can be detected. It cannot be used to find rules across different condition profiles or different time point profile experiments. However, with the appearance of time-series microarray data, it became possible to analyze the temporal relationship between genes. In this paper, we analyze the time-series microarray gene expression data to extract the sequential patterns which are similar to the association rules between genes among different time points in the yeast cell cycle. The sequential patterns found in our work can catch the associations between different genes which express or repress at diverse time points. We have applied sequential pattern mining method to time-series microarray gene expression data and discovered a number of sequential patterns from two groups of genes (test, control) and more sequential patterns have been discovered from test group (same CO term group) than from the control group (different GO term group). This result can be a support for the potential of sequential patterns which is capable of catching the biologically meaningful association between genes.

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Developing a Molecular Prognostic Predictor of a Cancer based on a Small Sample

  • Kim Inyoung;Lee Sunho;Rha Sun Young;Kim Byungsoo
    • Proceedings of the Korean Statistical Society Conference
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    • 2004.11a
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    • pp.195-198
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    • 2004
  • One Important problem in a cancer microarray study is to identify a set of genes from which a molecular prognostic indicator can be developed. In parallel with this problem is to validate the chosen set of genes. We develop in this note a K-fold cross validation procedure by combining a 'pre-validation' technique and a bootstrap resampling procedure in the Cox regression . The pre-validation technique predicts the microarray predictor of a case without having seen the true class level of the case. It was suggested by Tibshirani and Efron (2002) to avoid the possible over-fitting in the regression in which a microarray based predictor is employed. The bootstrap resampling procedure for the Cox regression was proposed by Sauerbrei and Schumacher (1992) as a means of overcoming the instability of a stepwise selection procedure. We apply this K-fold cross validation to the microarray data of 92 gastric cancers of which the experiment was conducted at Cancer Metastasis Research Center, Yonsei University. We also share some of our experience on the 'false positive' result due to the information leak.

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Improved Statistical Testing of Two-class Microarrays with a Robust Statistical Approach

  • Oh, Hee-Seok;Jang, Dong-Ik;Oh, Seung-Yoon;Kim, Hee-Bal
    • Interdisciplinary Bio Central
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    • v.2 no.2
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    • pp.4.1-4.6
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    • 2010
  • The most common type of microarray experiment has a simple design using microarray data obtained from two different groups or conditions. A typical method to identify differentially expressed genes (DEGs) between two conditions is the conventional Student's t-test. The t-test is based on the simple estimation of the population variance for a gene using the sample variance of its expression levels. Although empirical Bayes approach improves on the t-statistic by not giving a high rank to genes only because they have a small sample variance, the basic assumption for this is same as the ordinary t-test which is the equality of variances across experimental groups. The t-test and empirical Bayes approach suffer from low statistical power because of the assumption of normal and unimodal distributions for the microarray data analysis. We propose a method to address these problems that is robust to outliers or skewed data, while maintaining the advantages of the classical t-test or modified t-statistics. The resulting data transformation to fit the normality assumption increases the statistical power for identifying DEGs using these statistics.

An Iterative Normalization Algorithm for cDNA Microarray Medical Data Analysis

  • Kim, Yoonhee;Park, Woong-Yang;Kim, Ho
    • Genomics & Informatics
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    • v.2 no.2
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    • pp.92-98
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    • 2004
  • A cDNA microarray experiment is one of the most useful high-throughput experiments in medical informatics for monitoring gene expression levels. Statistical analysis with a cDNA microarray medical data requires a normalization procedure to reduce the systematic errors that are impossible to control by the experimental conditions. Despite the variety of normalization methods, this. paper suggests a more general and synthetic normalization algorithm with a control gene set based on previous studies of normalization. Iterative normalization method was used to select and include a new control gene set among the whole genes iteratively at every step of the normalization calculation initiated with the housekeeping genes. The objective of this iterative normalization was to maintain the pattern of the original data and to keep the gene expression levels stable. Spatial plots, M&A (ratio and average values of the intensity) plots and box plots showed a convergence to zero of the mean across all genes graphically after applying our iterative normalization. The practicability of the algorithm was demonstrated by applying our method to the data for the human photo aging study.

Surface Modification of Glass Chip for Peptide Microarray (펩타이드 Microarray를 위한 유리 칩의 표면 개질)

  • Cho, Hyung-Min;Lim, Chang-Hwan;Neff, Silke;Jungbauer, Alois;Lee, Eun-Kyu
    • KSBB Journal
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    • v.22 no.4
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    • pp.260-264
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    • 2007
  • Peptides are frequently studied as candidates for new drug development. Recently, synthesized peptide library is screened for a certain functionality on a microarray biochip format. In this study, in order to replace the conventional cellulose membrane with glass for a microarray chip substrate for peptide library screening, we modified the glass surface from amines to thiols and covalently immobilized the peptides. Using trypsin-FITC (fluorescein isothiocyanate) conjugate that could specifically bind to a trypsin binding domain consisting of a 7-amino acid peptide, we checked the degree of surface modification. Because of the relatively lower hydrophilicity and reduced surface roughness, the conjugation reaction to the glass required a longer reaction time and a higher temperature. It took approximately 12 hr for the reaction to be completed. From the fluorescence signal intensity, we could differentiate between the target and the control peptides. This difference was confirmed by a separate experiment using QCM. Furthermore, a smaller volume and higher concentration of a spot showed a higher fluorescence intensity. These data would provide the basic conditions for the development of microarray peptide biochips.