• Title/Summary/Keyword: High-throughput analysis

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Caenorhabditis elegans: A Model System for Anti-Cancer Drug Discovery and Therapeutic Target Identification

  • Kobet, Robert A.;Pan, Xiaoping;Zhang, Baohong;Pak, Stephen C.;Asch, Adam S.;Lee, Myon-Hee
    • Biomolecules & Therapeutics
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    • 제22권5호
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    • pp.371-383
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    • 2014
  • The nematode Caenorhabditis elegans (C. elegans) offers a unique opportunity for biological and basic medical researches due to its genetic tractability and well-defined developmental lineage. It also provides an exceptional model for genetic, molecular, and cellular analysis of human disease-related genes. Recently, C. elegans has been used as an ideal model for the identification and functional analysis of drugs (or small-molecules) in vivo. In this review, we describe conserved oncogenic signaling pathways (Wnt, Notch, and Ras) and their potential roles in the development of cancer stem cells. During C. elegans germline development, these signaling pathways regulate multiple cellular processes such as germline stem cell niche specification, germline stem cell maintenance, and germ cell fate specification. Therefore, the aberrant regulations of these signaling pathways can cause either loss of germline stem cells or overproliferation of a specific cell type, resulting in sterility. This sterility phenotype allows us to identify drugs that can modulate the oncogenic signaling pathways directly or indirectly through a high-throughput screening. Current in vivo or in vitro screening methods are largely focused on the specific core signaling components. However, this phenotype-based screening will identify drugs that possibly target upstream or downstream of core signaling pathways as well as exclude toxic effects. Although phenotype-based drug screening is ideal, the identification of drug targets is a major challenge. We here introduce a new technique, called Drug Affinity Responsive Target Stability (DARTS). This innovative method is able to identify the target of the identified drug. Importantly, signaling pathways and their regulators in C. elegans are highly conserved in most vertebrates, including humans. Therefore, C. elegans will provide a great opportunity to identify therapeutic drugs and their targets, as well as to understand mechanisms underlying the formation of cancer.

플라스틱 광섬유를 사용한 통신망에서 OCDMA의 성능 분석 (Performance Analysis of OCDMA on Plastic Optical Fiber Access Network)

  • 장극;서희종
    • 한국전자통신학회논문지
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    • 제11권11호
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    • pp.1083-1092
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    • 2016
  • 본 논문은 플라스틱 광섬유(POF)를 연결한 네트워크에서 광 부호분할다중접근(OCDMA)기술의 성능을 분석한다. 최근에 POF는 무게가 가볍고, 중심선의 직경이 크며, 유연하고, 쉽게 설치할 수 있는 특성이 있기 때문에 POF를 이용한 광전송은 커다란 주목을 받고 있다. 특히 POF의 광대역(broad band) 특성은 OCDMA기술에 기초한 네트워크 연결 전송매체로서 각광을 받고 있다. 일반적인 OCDMA 시스템은 전송로의 수량이 BER특성에 의해서 단지 제한된 단위만을 동시에 연결하고 전송할 수 있다. 이러한 문제를 해결하기 위하여 본 논문에서는 새로운 다우선 예약프로토콜을 제안하였다. 이 프로토콜과 분산 조정 알고리듬을 이용하여, 통신로 내부와 목표지로 가는 정보의 충돌을 피할 수 있다. 이 프로토콜은 시간지연이 서로 다른 다매체정보의 전송을 효과적으로 지원할 수 있다. 데이터 분석과 모의실험을 실행하여, 여러 시스템 매개변수에 의존하는 네트워크 출력과 평균지연을 조사했다. 그 결과, OCDMA기술에 근거한 POF를 네트워크를 사용하는 다우선 예약 프로토콜이 효율적이라는 것을 확인할 수 있었다.

Prediction Model for the Cellular Immortalization and Transformation Potentials of Cell Substrates

  • Lee, Min-Su;Matthews Clayton A.;Chae Min-Ju;Choi, Jung-Yun;Sohn Yeo-Won;Kim, Min-Jung;Lee, Su-Jae;Park, Woong-Yang
    • Genomics & Informatics
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    • 제4권4호
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    • pp.161-166
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    • 2006
  • The establishment of DNA microarray technology has enabled high-throughput analysis and molecular profiling of various types of cancers. By using the gene expression data from microarray analysis we are able to investigate diagnostic applications at the molecular level. The most important step in the application of microarray technology to cancer diagnostics is the selection of specific markers from gene expression profiles. In order to select markers of Immortalization and transformation we used c-myc and $H-ras^{V12}$ oncogene-transfected NIH3T3 cells as our model system. We have identified 8751 differentially expressed genes in the immortalization/transformation model by multivariate permutation F-test (95% confidence, FDR<0.01). Using the support vector machine algorithm, we selected 13 discriminative genes which could be used to predict immortalization and transformation with perfect accuracy. We assayed $H-ras^{V12}$-transfected 'transformed' cells to validate our immortalization/transformation dassification system. The selected molecular markers generated valuable additional information for tumor diagnosis, prognosis and therapy development.

Analysis of MAPK Signaling Pathway Genes in the Intestinal Mucosal Layer of Necrotic Eenteritis-Afflicted Two Inbred Chicken Lines

  • Truong, Anh Duc;Hong, Yeojin;Lee, Janggeun;Lee, Kyungbaek;Lillehoj, Hyun S.;Hong, Yeong Ho
    • 한국가금학회지
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    • 제44권3호
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    • pp.199-209
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    • 2017
  • Mitogen-activated protein kinase (MAPK) signaling pathways play a key role in innate immunity, inflammation, cell proliferation, cell differentiation, and cell death. The main objective of this study was to investigate the expression level of candidate MAPK pathway genes in the intestinal mucosal layer of two genetically disparate chicken lines (Marek's disease-resistant line 6.3 and Marek's disease-susceptible line 7.2) induced with necrotic enteritis (NE). Using high-throughput RNA sequencing, we investigated 178 MAPK signaling pathway related genes that were significantly and differentially expressed between the intestinal mucosal layers of the NE-afflicted and control chickens. In total, 15 MAPK pathway genes were further measured by quantitative real-time PCR(qRT-PCR) and the results were consistent with the RNA-sequencing data. All 178 identified genes were annotated through Gene Ontology and mapped onto the KEGG chicken MAPK signaling pathway. Several key genes of the MAPK pathway, ERK1/2, JNK1-3, p38 MAPK, MAP2K1-4, $NF-{\kappa}B1/2$, c-Fos, AP-1, Jun-D, and Jun, were differentially expressed in the two chicken lines. Therefore, we believe that RNA sequencing and qRT-PCR analysis provide resourceful information for future studies on MAPK signaling of genetically disparate chicken lines in response to pathogens.

Mutational Analysis of Extranodal NK/T-Cell Lymphoma Using Targeted Sequencing with a Comprehensive Cancer Panel

  • Choi, Seungkyu;Go, Jai Hyang;Kim, Eun Kyung;Lee, Hojung;Lee, Won Mi;Cho, Chun-Sung;Han, Kyudong
    • Genomics & Informatics
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    • 제14권3호
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    • pp.78-84
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    • 2016
  • Extranodal natural killer (NK)/T-cell lymphoma, nasal type (NKTCL), is a malignant disorder of cytotoxic lymphocytes of NK or T cells. It is an aggressive neoplasm with a very poor prognosis. Although extranodal NKTCL reportedly has a strong association with Epstein-Barr virus, the molecular pathogenesis of NKTCL has been unexplored. The recent technological advancements in next-generation sequencing (NGS) have made DNA sequencing cost- and time-effective, with more reliable results. Using the Ion Proton Comprehensive Cancer Panel, we sequenced 409 cancer-related genes to identify somatic mutations in five NKTCL tissue samples. The sequencing analysis detected 25 mutations in 21 genes. Among them, KMT2D, a histone modification-related gene, was the most frequently mutated gene (four of the five cases). This result was consistent with recent NGS studies that have suggested KMT2D as a novel driver gene in NKTCL. Mutations were also found in ARID1A, a chromatin remodeling gene, and TP53, which also recurred in recent NGS studies. We also found mutations in 18 novel candidate genes, with molecular functions that were potentially implicated in cancer development. We suggest that these genes may result in multiple oncogenic events and may be used as potential bio-markers of NKTCL in the future.

Comparative Transcriptome Analysis of Queen, Worker, and Larva of Asian Honeybee, Apis cerana

  • Kim, Woo Jin;Lee, Seok Hee;An, Saes Byeol;Kim, Song Eun;Liu, Qin;Choi, Jae Young;Je, Yeon Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • 제27권2호
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    • pp.271-276
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    • 2013
  • The Asian honeybee, Apis cerana, is a native honeybee species in Korea which is important in agriculture for pollination and honey production. For better understanding of the physiology of A. cerana, high-throughput Illumina transcriptome sequencing was performed to analyze the gene expression profiles of queen, worker, and larva. A total of 219,799,682 clean reads corresponding to 22.2 Gb of nucleotide sequences was obtained from the whole body total RNA samples. The Apis mellifera reference mRNA sequence database was used to measure the gene expression level with Bowtie2 and eXpress software, and the Illumina short reads were then mapped to 11,459 out of 11,736 A. mellifera reference genes. Total of 9,221 genes with FPKM value greater than 5 of each sample group were subjected to eggNOG with BLASTX for gene ontology analysis. The differential gene expression between queen and worker, and worker and larva were analyzed to screen the overexpressed genes in each sample group. In the queen and worker sample group, total of 1,766 genes were differentially expressed with 887 and 879 genes overexpressed over two folds in queen and worker, respectively. In the worker and larva sample group, total of 1,410 genes were differentially expressed with 1,009 and 401 genes overexpressed over two folds in worker and larva, respectively.

차별화된 호 처리 기법을 지원하는 SIP6 (SIP6 supporting the Differentiated Call Processing Scheme)

  • 김진철;최병욱;장천현;김기천;한선영
    • 한국정보과학회논문지:정보통신
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    • 제30권5호
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    • pp.621-630
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    • 2003
  • 본 논문에서는 NGN(Next Generation Network)을 고려하여 IPv6와 차별화된 호 처리 기법을 지원하는 SIP(Session Initiation Protocol) 프로토콜을 설계하고 구현하였다. NGN에서 다양한 응용 서비스들은 SIP를 통하여 호를 설정한다. 따라서, 소프트스위치(Softswitch) 및 SIP 서버는 호를 처리 할 때 팩스(Fax), 망 관리(Network Management), 홈 네트워킹(Home Networking)과 같이 호 설정 시간에 아주 민감한 서비스에 우선순위를 부여할 수 있어야 한다. 또한, ALL-IP를 고려하여 IPv6를 지원하여야 한다. 이러한 문제들을 해결하기 위해, 본 논문에서는 차별화된 호 처리 기법을 제안하였다. 차별화된 호 처리 기법은 우선순위에 따라 세 단계의 서비스품질을 사용하여, 호 처리에 대한 차별화 정책을 지원한다. 또한, 본 논문에서 정의된 서비스품질은 IPv6 헤더의 Flow Label 필드를 통하여 값을 설정한다. 성능분석 결과, SIP 서버 내에서 호가 처리 될 때 높은 우선순위를 가지는 호에 대한 처리율이 현저히 빠르게 나타났다. 이러한 결과는 NGN에서 호 설정 시간에 민감한 응용 서비스에 적용 할 경우, 호 설정 시간을 줄임으로서 응용 서비스의 성능을 향상시킬 수 있음을 증명한다.

Comparative analysis of commonly used peak calling programs for ChIP-Seq analysis

  • Jeon, Hyeongrin;Lee, Hyunji;Kang, Byunghee;Jang, Insoon;Roh, Tae-Young
    • Genomics & Informatics
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    • 제18권4호
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    • pp.42.1-42.9
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    • 2020
  • Chromatin immunoprecipitation coupled with high-throughput DNA sequencing (ChIP-Seq) is a powerful technology to profile the location of proteins of interest on a whole-genome scale. To identify the enrichment location of proteins, many programs and algorithms have been proposed. However, none of the commonly used peak calling programs could accurately explain the binding features of target proteins detected by ChIP-Seq. Here, publicly available data on 12 histone modifications, including H3K4ac/me1/me2/me3, H3K9ac/me3, H3K27ac/me3, H3K36me3, H3K56ac, and H3K79me1/me2, generated from a human embryonic stem cell line (H1), were profiled with five peak callers (CisGenome, MACS1, MACS2, PeakSeq, and SISSRs). The performance of the peak calling programs was compared in terms of reproducibility between replicates, examination of enriched regions to variable sequencing depths, the specificity-to-noise signal, and sensitivity of peak prediction. There were no major differences among peak callers when analyzing point source histone modifications. The peak calling results from histone modifications with low fidelity, such as H3K4ac, H3K56ac, and H3K79me1/me2, showed low performance in all parameters, which indicates that their peak positions might not be located accurately. Our comparative results could provide a helpful guide to choose a suitable peak calling program for specific histone modifications.

Validation of exercise-response genes in skeletal muscle cells of Thoroughbred racing horses

  • Kim, Doh Hoon;Lee, Hyo Gun;Sp, Nipin;Kang, Dong Young;Jang, Kyoung-Jin;Lee, Hak Kyo;Cho, Byung-Wook;Yang, Young Mok
    • Animal Bioscience
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    • 제34권1호
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    • pp.134-142
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    • 2021
  • Objective: To understand the athletic characteristics of Thoroughbreds, high-throughput analysis has been conducted using horse muscle tissue. However, an in vitro system has been lacking for studying and validating genes from in silico data. The aim of this study is to validate genes from differentially expressed genes (DEGs) of our previous RNA-sequencing data in vitro. Also, we investigated the effects of exercise-induced stress including heat, oxidative, hypoxic and cortisol stress on horse skeletal muscle derived cells with the top six upregulated genes of DEGs. Methods: Enriched pathway analysis was conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool with upregulated genes in horse skeletal muscle tissue after exercise. Among the candidates, the top six genes were analysed through geneMANIA to investigate gene networks. Muscle cells derived from neonatal horse skeletal tissue were maintained and subjected to exercise-related stressors. Transcriptional changes in the top six genes followed by stressors were investigated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Results: The inflammation response pathway was the most commonly upregulated pathway after horse exercise. Under non-cytotoxic conditions of exercise-related stressors, the transcriptional response of the top six genes was different among types of stress. Oxidative stress yielded the most similar expression pattern to DEGs. Conclusion: Our results indicate that transcriptional change after horse exercise in skeletal muscle tissue strongly relates to stress response. The qRT-PCR results showed that stressors contribute differently to the transcriptional regulation. These results would be valuable information to understand horse exercise in the stress aspect.

Comparative Microbiome Analysis of Three Species of Laboratory-Reared Periplaneta Cockroaches

  • Lee, Seogwon;Kim, Ju Yeong;Yi, Myung-hee;Lee, In-Yong;Lee, Won-Ja;Moon, Hye Su;Yong, Dongeun;Yong, Tai-Soon
    • Parasites, Hosts and Diseases
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    • 제58권5호
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    • pp.537-542
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    • 2020
  • Cockroaches inhabit various habitats, which will influence their microbiome. Although the microbiome can be influenced by the diet and environmental factors, it can also differ between species. Therefore, we conducted 16S rDNA-targeted high-throughput sequencing to evaluate the overall bacterial composition of the microbiomes of 3 cockroach species, Periplaneta americana, P. japonica, and P. fuliginosa, raised in laboratory for several generations under the same conditions. The experiments were conducted using male adult cockroaches. The number of operational taxonomic units (OTUs) was not significantly different among the 3 species. With regard to the Shannon and Pielou indexes, higher microbiome values were noted in P. americana than in P. japonica and P. fuliginosa. Microbiome composition was also evaluated, with endosymbionts accounting for over half of all OTUs in P. japonica and P. fuliginosa. Beta diversity analysis further showed that P. japonica and P. fuliginosa had similar microbiome composition, which differed from that of P. americana. However, we also identified that P. japonica and P. fuliginosa host distinct OTUs. Thus, although microbiome compositions may vary based on multiple conditions, it is possible to identify distinct microbiome compositions among different Periplaneta cockroach species, even when the individuals are reared under the same conditions.