• Title/Summary/Keyword: Microarrays

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Quality Control Usage in High-Density Microarrays Reveals Differential Gene Expression Profiles in Ovarian Cancer

  • Villegas-Ruiz, Vanessa;Moreno, Jose;Jacome-Lopez, Karina;Zentella-Dehesa, Alejandro;Juarez-Mendez, Sergio
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.5
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    • pp.2519-2525
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    • 2016
  • There are several existing reports of microarray chip use for assessment of altered gene expression in different diseases. In fact, there have been over 1.5 million assays of this kind performed over the last twenty years, which have influenced clinical and translational research studies. The most commonly used DNA microarray platforms are Affymetrix GeneChip and Quality Control Software along with their GeneChip Probe Arrays. These chips are created using several quality controls to confirm the success of each assay, but their actual impact on gene expression profiles had not been previously analyzed until the appearance of several bioinformatics tools for this purpose. We here performed a data mining analysis, in this case specifically focused on ovarian cancer, as well as healthy ovarian tissue and ovarian cell lines, in order to confirm quality control results and associated variation in gene expression profiles. The microarray data used in our research were downloaded from ArrayExpress and Gene Expression Omnibus (GEO) and analyzed with Expression Console Software using RMA, MAS5 and Plier algorithms. The gene expression profiles were obtained using Partek Genomics Suite v6.6 and data were visualized using principal component analysis, heat map, and Venn diagrams. Microarray quality control analysis showed that roughly 40% of the microarray files were false negative, demonstrating over- and under-estimation of expressed genes. Additionally, we confirmed the results performing second analysis using independent samples. About 70% of the significant expressed genes were correlated in both analyses. These results demonstrate the importance of appropriate microarray processing to obtain a reliable gene expression profile.

Genome-wide Expression Profiling of Piperine and Piper nigrum Linne (호초(胡椒)와 Piperine에 의한 총체적 유전자 발현 비교)

  • Jo, Eun-Young;Jeong, Ji-Cheon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.5
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    • pp.831-836
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    • 2010
  • In addition to spice, black pepper (Piper nigrum Linne : PnL) has been used as herbal medicine because of its function in anti-oxidation, anti-inflammation, and anti-carcinogenesis. Recently, it has been reported that piperine, a component of PnL, inhibits adipocyte differentiation by repressing various adipogenic gene expressions. In this study, we determined whether piperine is a major constituent of PnL that confers the anti-adipogenic activity at whole genome level. Differentiation of 3T3-L1 pre-adipocytes was induced in presence of PnL extract or piperine. To compare genes that are regulated by PnL extract or piperine, we performed expression profiling using microarrays (Agilent Mouse 44k 4plex). RNA samples were labeled with Cy3 and Cy5, respectively. Labeled samples were hybridized to the microarrays. Results were filtered and cut off set p<0.05. Genes exhibiting significant differences in expression level were classified into Gene Ontology (GO)-based functional categories (http://www.geneontology.org) and KEGG (http://www.genome.jp/kegg/). Extract of PnL and its component piperine reduced lipid accumulation in 3T3-L1 cells during adipogenesis. Such anti-adipogenic activity appears to result from down-regulation of transcription factor genes involved in adipogenesis, and other genes involved in fatty acid synthesis, transport, triglyceride synthesis, and carbohydrate metabolism. These genome-wide studies lead to conclude that piperine, as a critical component of PnL, plays common role with PnL in anti-adipogenesis.

Design and Implementation of Integrated System for Microarray Data (마이크로어레이 실험 및 분석 데이터 처리를 위한 통합 관리 시스템의 설계와 구현)

  • 이미경;최정현;조환규
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.182-190
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    • 2003
  • As DNA microarrays are widely used recently, the amount of microarray data is exponentially increasing. Until now, however, no domestic system is available for the efficient management of such data. Because the number of experimental data in a specific laboratory is limited, it is necessary to avoid redundant experiments and to accumulate the results using a shared data management system for microarrays. In this paper, a system named WEMA (WEb management of Micro Arrays) was designed and implemented to manage and process the microarray data. WEMA system was designed to include the basic feature of MIAME (Minimal Information About a Microarray Experiment), and general data units were also defined in the system in order to systematically manage the data. The WEMA system has three main features: efficient management of microarray data, integration of input/ouput data, and metafile processing. The system was tested with actual microarray data produced by a molecular biology laboratory, and we found that the biologists could systematically manage and easily analyze the microarray data. As a consequence, the researchers could reduce the cost of data exchange and communication.

Photolithographic Fabrication of Poly(Ethylene Glycol) Microstructures for Hydrogel-based Microreactors and Spatially Addressed Microarrays

  • Baek, Taek-Jin;Kim, Nam-Hyun;Choo, Jae-Bum;Lee, Eun-Kyu;Seong, Gi-Hun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1826-1832
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    • 2007
  • We describe the fabrication of poly(ethylene glycol) diacrylate (PEG-DA) hydrogel microstructures with a high aspect ratio and the use of hydrogel microstructures containing the enzyme ${\beta}$-galactosidase (${\beta}$-Gal) or glucose oxidase (GOx)/horseradish peroxidase (HRP) as biosensing components for the simultaneous detection of multiple analytes. The diameters of the hydrogel microstructures were almost the same at the top and at the bottom, indicating that no differential curing occurred through the thickness of the hydrogel microstructure. Using the hydrogel microstructures as microreactors, ${\beta}$-Gal or GOx/HRP was trapped in the hydrogel array, and the time-dependent fluorescence intensities of the hydrogel array were investigated to determine the dynamic uptake of substrates into the PEG-DA hydrogel. The time required to reach steady-state fluorescence by glucose diffusing into the hydrogel and its enzymatic reactions with GOx and HRP was half the time required for resorufin ${\beta}$-D-galactopyranoside (RGB) when used as the substrate for ${\beta}$-Gal. Spatially addressed hydrogel microarrays containing different enzymes were micropatterned for the simultaneous detection of multiple analytes, and glucose and RGB solutions were incubated as substrates. These results indicate that there was no cross-talk between the ${\beta}$-Gal-immobilizing hydrogel micropatches and the GOx/HRP-immobilizing micropatches.

Large Scale Gene Expression Analysis in Rat Models of 4-Vessel Occlusion Ischemia (4-Vessel Occlusion 허혈동물모델에서의 대규모 유전자 발현 연구)

  • Kang, Bong-Joo;Hong, Seong-Gil;Kim, Yun-Taik;Kim, Young-Ok;Cho, Dong-Wuk
    • Korean Journal of Oriental Medicine
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    • v.6 no.1
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    • pp.89-98
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    • 2000
  • Cerebral ischemia, the most prevalent form of clinical stroke, is a medical problem of the first magnitude. Substantial efforts are being made to develop drugs which will protect the brain from the neurodegeneration followed by an ischemic stroke. A key factor in this process is the development of animal models that mimic the neuropathological consequences of stroke. Recently, there is increasing an evidence that free radical is involved in the mechanisms of ischemic brain damage. We investigated the macro scale gene expression analysis on the global ischemia induced by 4-vessel occlusion in Wister rats. The recent availability of microarrays provides an attractive strategy for elaborating an unbiased molecular profile of large number of genes during ischemic injury. This experimental approach offers the potential to identify molecules or cellular pathways not previously associated with ischemia. Ischemia was induced by 4-vessel occlusion for 10 minutes and reperfused again. RNA from sham control brain and time-dependent ischemed brain were hybridized to microarrays containing 4,000 rat genes. 589 genes were found to be at least 2 fold regulated at one or more time points. These survey data provide the foundation studies that should provide convincing proof for ischemia and oxidative stress on gene expression.

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Studies on Gene Expression of Yukmijihwang-tang using High-throughput Gene Expression Analysis Techniques (대규모 유전자 분석 기법을 이용한 육미지황원의 유전자 발현 연구)

  • Kang, Bong-Joo;Kim, Yun-Taik;Cho, Dong-Wuk
    • Korean Journal of Oriental Medicine
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    • v.8 no.2 s.9
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    • pp.95-107
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    • 2002
  • Yukmijihwang-tang(YM) is a noted herbal prescription in Chinese and Korean traditional medicines, and it has been known to reinforce the vital essence and has been widely used for a variety of disease such as stroke, osteoporosis, anti-tumor, and hypothyrodism. Regarding its traditional use, YM has been known to reinforce the Yin (vital essence) of liver and kidney. Also it has been known to reinforce nutrition and biological function in brain. Recently, studies suggested that YM increase antioxidant activities and exert the protective effect against oxidant-induced liver cell injury. We investigated the high-throughput gene expression analysis on the Yukmijihwang-tang administrated in SD rats. Microarray data were validated on a limited number of genes by semiquantitative RT-PCR and Western blot analyses. The recent availability of microarrays provides an attractive strategy for elaborating an unbiased molecular profile of large number of genes in drug discovery This experimental approach offers the potential to identify molecules or cellular pathways not previously associated with herbal medicine. Total RNA from normal control brain and Yukmijihwang-tang administrated brain were hybridized to microarrays containing 10,000 rat genes. The 52 genes were found to be up-regulated(twice or more) excluding EST gene. The nine genes were found to be down-regulated(twice or more) excluding EST gene. Gene array technology was used to identify for the first time many genes expression pathway analysis that arecell cycle pathway, apoptosis pathway, electron transport chain pathway, cytoplasmic ribosomal protein pathway, fatty acid degradation pathway, and TGF-beta signaling pathway. These differentially expressed genes pathway analysis have not previously been iavestigated in the context of herbal medicine efficacy and represent novel factors for further study of the mechanism of herbal medicine efficacy.

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Current status on expression profiling using rice microarray (벼 microarray를 이용한 유전자발현 profiling 연구동향)

  • Yoon, Ung-Han;Kim, Yeon-Ki;Kim, Chang-Kug;Hahn, Jang-Ho;Kim, Dong-Hern;Lee, Tae-Ho;Lee, Gang-Seob;Park, Soo-Chul;Nahm, Baek-Hie
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.144-152
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    • 2010
  • As the International Rice Genome Sequencing Project (IRGSP) was completed in 2005 and opened to the public, many countries are making a lot of investments in researches on the utilization of sequence information along with system development. Also, the necessity of the functional genomics researches using microarray is increased currently to secure unique genes related with major agricultural traits and analyze metabolic pathways. Microrarray enables efficient analysis of large scale gene expression and related transcription regulation. This review aims to introduce available microarrays made based on rice genome information and current status of gene expression analysis using these microarrays integrated with the databases available to the public. Also, we introduce the researches on the large scale functional analysis of genes related with useful traits and genetic networks. Understanding of the mechanism related with mutual interaction between proteins with co-expression among rice genes can be utilized in the researches for improving major agricultural traits. The direct and indirect interactions of various genes would provide new functionality of rice. The recent results of the various expression profiling analysis in rice will promote functional genomic researches in plants including rice and provide the scientists involved in applications researches with wide variety of expression informations.

Polychlorobiphenyl (PCB) 토양오염복원: PCB 제거 토양미생물들의 군집과 기능을 효과적으로 분석하는 신 genomics 방법개발에 관한 연구

  • Park Jun-Hong
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2005.04a
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    • pp.28-30
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    • 2005
  • Because of high population diversity in soil microbial communities, it is difficult to accurately assess the capability of biodegradation of toxicant by microbes in soil and sediment. Identifying biodegradative microorganisms is an important step in designing and analyzing soil bioremediation. To remove non-important noise information, it is necessary to selectively enrich genomes of biodegradative microorganisms fromnon-biodegradative populations. For this purpose, a stable isotope probing (SIP) technique was applied in selectively harvesting the genomes of biphenyl-utilizing bacteria from soil microbial communities. Since many biphenyl-using microorganisms are responsible for aerobic PCB degradation In soil and sediments, biphenyl-utilizing bacteria were chosen as the target organisms. In soil microcosms, 13C-biphenyl was added as a selective carbon source for biphenyl users, According to $13C-CO_2$ analysis by GC-MS, 13C-biphenyl mineralization was detected after a 7-day of incubation. The heavy portion of DNA(13C-DNA) was separated from the light portion of DNA (12C-DNA) using equilibrium density gradient ultracentrifuge. Bacterial community structure in the 13C-DNAsample was analyzed by t-RFLP (terminal restriction fragment length polymorphism) method. The t-RFLP result demonstates that the use of SIP efficiently and selectively enriched the genomes of biphenyl degrading bacteria from non-degradative microbes. Furthermore, the bacterial diversity of biphenyl degrading populations was small enough for environmental genomes tools (metagenomics and DNA microarrays) to be used to detect functional (biphenyl degradation) genes from soil microbial communities, which may provide a significant progress in assessing microbial capability of PCB bioremediation in soil and groundwater.

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Combining Support Vector Machine Recursive Feature Elimination and Intensity-dependent Normalization for Gene Selection in RNAseq (RNAseq 빅데이터에서 유전자 선택을 위한 밀집도-의존 정규화 기반의 서포트-벡터 머신 병합법)

  • Kim, Chayoung
    • Journal of Internet Computing and Services
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    • v.18 no.5
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    • pp.47-53
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    • 2017
  • In past few years, high-throughput sequencing, big-data generation, cloud computing, and computational biology are revolutionary. RNA sequencing is emerging as an attractive alternative to DNA microarrays. And the methods for constructing Gene Regulatory Network (GRN) from RNA-Seq are extremely lacking and urgently required. Because GRN has obtained substantial observation from genomics and bioinformatics, an elementary requirement of the GRN has been to maximize distinguishable genes. Despite of RNA sequencing techniques to generate a big amount of data, there are few computational methods to exploit the huge amount of the big data. Therefore, we have suggested a novel gene selection algorithm combining Support Vector Machines and Intensity-dependent normalization, which uses log differential expression ratio in RNAseq. It is an extended variation of support vector machine recursive feature elimination (SVM-RFE) algorithm. This algorithm accomplishes minimum relevancy with subsets of Big-Data, such as NCBI-GEO. The proposed algorithm was compared to the existing one which uses gene expression profiling DNA microarrays. It finds that the proposed algorithm have provided as convenient and quick method than previous because it uses all functions in R package and have more improvement with regard to the classification accuracy based on gene ontology and time consuming in terms of Big-Data. The comparison was performed based on the number of genes selected in RNAseq Big-Data.

Salt-Responsive Genes in Salt Tolerant Rice Mutants Revealed through Microarray Analysis

  • Song, Jae Young;Kim, Dong Sub;Lee, Myung-Chul;Kang, Si-Yong;Kim, Jin-Baek;Lee, Kyung Jun;Yun, Song Joong
    • Journal of Radiation Industry
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    • v.4 no.4
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    • pp.325-334
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    • 2010
  • Transcriptional regulation in response to salt in mutant lines was investigated using oligonucleotide microarrays. In order to characterize the salt-responsive genes in rice, the expression profiles of transcripts that responded to salt-treatment were monitored using the microarrays. In the microarray analysis, among 37,299 reliable genes, 5,101, 2,758 and 2,277 genes were up-regulated by more than 2-fold using the salt treatment, while the numbers of down-regulated genes were 4,619, 3,234, and 1,878 in the WT, ST-495, and ST-532, respectively. From genotype changes induced by gamma ray mutagenesis, 3,345 and 2,397 genes were up-regulated, while 2,745 and 2,075 genes were down-regulated more than 2-fold in the two untreated mutants lines compared with untreated WT, respectively. A total of 3,108 and 2,731 genes were up-regulated more than 2-fold, while 3,987 and 3,660 genes were down-regulated by more than 2-fold in the salt treatment of the two mutants lines compared with the salt treated WT, respectively. The expressions of membrane transporter genes such as OsAKT1, OsKUP, and OsNAC increased more severely in ST-495 and ST-532 than in the WT. The expressions of the proline accumulation related genes such as OsP5CS and OsP5CR were also markedly increased in the salt tolerant mutants when compared to the WT plant.