• Title/Summary/Keyword: High-throughput analysis

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Quantitative Proteomics Towards Understanding Life and Environment

  • Choi, Jong-Soon;Chung, Keun-Yook;Woo, Sun-Hee
    • Korean Journal of Environmental Agriculture
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    • v.25 no.4
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    • pp.371-381
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    • 2006
  • New proteomic techniques have been pioneered extensively in recent years, enabling the high-throughput and systematic analyses of cellular proteins in combination with bioinformatic tools. Furthermore, the development of such novel proteomic techniques facilitates the elucidation of the functions of proteins under stress or disease conditions, resulting in the discovery of biomarkers for responses to environmental stimuli. The ultimate objective of proteomics is targeted toward the entire proteome of life, subcellular localization biochemical activities, and the regulation thereof. Comprehensive analysis strategies of proteomics can be classified into three categories: (i) protein separation via 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification via either Edman sequencing or mass spectrometry (MS), and (iii) proteome quantitation. Currently, MS-based proteomics techniques have shifted from qualitative proteome analysis via 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, toward quantitative proteome analysis. In vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes. protein-labeling tagging with isotope-coded affinity tags, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope-labeled amino acids can be in vivo labeled into live culture cells via metabolic incorporation. MS-based proteomics techniques extend to the detection of the phosphopeptide mapping of biologically crucial proteins, which ale associated with post-translational modification. These complementary proteomic techniques contribute to our current understanding of the manner in which life responds to differing environment.

The Data Envelopment Analysis of Container Terminals to Transshipment Cargo (환적화물의 컨테이너 터미널 효율성 분석)

  • Park, Hong-Gyun
    • Journal of Korea Port Economic Association
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    • v.26 no.1
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    • pp.1-19
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    • 2010
  • This paper focuses measuring the efficiency of container yards on container terminals in Busan (Gasungdae, Shinsundae, Gamman, New Gamman, Uam, Gamchon, PNC) and Gwangyang(GICT, KEC, Dongbu, KIT) using Data Envelopment Analysis(DEA) approach. Container terminals in Busan and Gwangyang play an important role in the region's economic development. The results show that Shinsundae was an efficient DMU during the period of 2007 to 2009, while Gamman, New Gamman and PNC were efficient terminals in 2009. The very inefficient terminals were shown to be GICT, KEC, Dongbu and KIT. GICT(2009), KEC(2009), Dongbu(2008-2009), KIT(2009) on Gwangyang Port were found to be relatively the inefficient terminals in terms of the returns to scale. This study also finds that the efficiency of Shinsundae terminal was so high as to be abel to keep its efficiency in spite of the additional increase of the inputs from 2007 to 2009. Gamman terminal was in the decreasing returns to scale in 2009, while the other terminals were in the increasing returns to scale. It means that we are able to improve the efficiency of the Gamman terminal with increasing returns to scale through enlarging the scale.

Effect of Soil Properties and Soil Bacterial Community on Early Growth Characteristics of Wild-simulated Ginseng (Panax ginseng C. A. Meyer) in Coniferous and Mixed Forest (침엽수림과 혼효림에서 토양특성과 토양세균 군집이 산양삼 초기 생육특성에 미치는 영향)

  • Kim, Ki Yoon;Kim, Hyun Jun;Um, Yurry;Jeon, Kwon Seok
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.3
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    • pp.183-194
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    • 2020
  • Background: This study investigated the effect of soil properties and soil bacterial community on early growth characteristics of wild-simulated ginseng (Panax ginseng C. A. Meyer) in coniferous and mixed forest experimental fields. Methods and Results: The soil bacterial community was analyzed using a high throughput sequencing technique (Illumina MiSeq sequencing). The relationship between the soil bacterial community, soil properties, and growth characteristics of wild-simulated ginseng were analyzed using principal coordinate analysis (PCoA) and the Pearson's correlation analysis. Soil properties and soil bacterial community showed significant difference with forest physiognomy. Results of Pearson's correlation analysis and PCoA showed that the soil properties (soil pH, organic matter, total nitrogen, and cation exchange capacity) and soil bacterial community had significant correlation with tree species ratio and early growth characteristics of wild-simulated ginseng. Conclusions: This study clearly demonstrated the effect of soil properties and soil bacterial community on early growth characteristics of wild-simulated ginseng in coniferous and mixed forest. Moreover, these results will help in the selection of suitable cultivation sites for wild-simulated ginseng.

Quantitative and Rapid Analysis of Transglutaminase Activity Using Protein Arrays in Mammalian Cells

  • Kwon, Mi-Hye;Jung, Jae-Wan;Jung, Se-Hui;Park, Jin-Young;Kim, Young-Myeong;Ha, Kwon-Soo
    • Molecules and Cells
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    • v.27 no.3
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    • pp.337-343
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    • 2009
  • We developed a novel on-chip activity assay using protein arrays for quantitative and rapid analysis of transglutaminase activity in mammalian cells. Transglutaminases are a family of $Ca^{2+}$-dependent enzymes involved in cell regulation as well as human diseases such as neurodegenerative disorders, inflammatory diseases and tumor progression. We fabricated the protein arrays by immobilizing N,N'-dimethylcasein (a substrate) on the amine surface of the arrays. We initiated transamidating reaction on the protein arrays and determined the transglutaminase activity by analyzing the fluorescence intensity of biotinylated casein. The on-chip transglutaminase activity assay was proved to be much more sensitive than the $[^3H]putrescine$-incorporation assay. We successfully applied the on-chip assay to a rapid and quantitative analysis of the transglutaminase activity in all-trans retinoic acid-treated NIH 3T3 and SH-SY5Y cells. In addition, the on-chip transglutaminase activity assay was sufficiently sensitive to determine the transglutaminase activity in eleven mammalian cell lines. Thus, this novel on-chip transglutaminase activity assay was confirmed to be a sensitive and high-throughput approach to investigating the roles of transglutaminase in cellular signaling, and, moreover, it is likely to have a strong potential for monitoring human diseases.

Early Diagnosis of ABCB11 Spectrum Liver Disorders by Next Generation Sequencing

  • Lee, Su Jeong;Kim, Jung Eun;Choe, Byung-Ho;Seo, An Na;Bae, Han-Ik;Hwang, Su-Kyeong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.20 no.2
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    • pp.114-123
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    • 2017
  • Purpose: The goal of this study was the early diagnosis of ABCB11 spectrum liver disorders, especially those focused on benign recurrent intrahepatic cholestasis and progressive familial intrahepatic cholestasis. Methods: Fifty patients presenting neonatal cholestasis were evaluated to identify underlying etiologies. Genetic analysis was performed on patients suspected to have syndromic diseases or ABCB11 spectrum liver disorders. Two families with proven ABCB11 spectrum liver disorders were subjected to genetic analyses to confirm the diagnosis and were provided genetic counseling. Whole exome sequencing and Sanger sequencing were performed on the patients and the family members. Results: Idiopathic or viral hepatitis was diagnosed in 34%, metabolic disease in 20%, total parenteral nutrition induced cholestasis in 16%, extrahepatic biliary atresia in 14%, genetic disease in 10%, neonatal lupus in 2%, congenital syphilis in 2%, and choledochal cyst in 2% of the patients. The patient with progressive familial intrahepatic cholestasis had novel heterozygous mutations of ABCB11 c.11C>G (p.Ser4*) and c.1543A>G (p.Asn515Asp). The patient with benign recurrent intrahepatic cholestasis had homozygous mutations of ABCB11 c.1331T>C (p.Val444Ala) and heterozygous, c.3084A>G (p.Ala1028Ala). Genetic confirmation of ABCB11 spectrum liver disorder led to early liver transplantation in the progressive familial intrahepatic cholestasis patient. In addition, the atypically severe benign recurrent intrahepatic cholestasis patient was able to avoid unnecessary liver transplantation after genetic analysis. Conclusion: ABCB11 spectrum liver disorders can be clinically indistinguishable as they share similar characteristics related to acute episodes. A comprehensive genetic analysis will facilitate optimal diagnosis and treatment.

Comparative Analysis of Gene Expression in the Female Reproductive Organs

  • Kim, Min-Goo;Seo, Hee-Won;Choi, Yo-Han;Lee, Chang-Kyu;Ka, Hak-Hyun
    • Journal of Embryo Transfer
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    • v.24 no.2
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    • pp.77-87
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    • 2009
  • To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

Transcriptome Profiling and In Silico Analysis of the Antimicrobial Peptides of the Grasshopper Oxya chinensis sinuosa

  • Kim, In-Woo;Markkandan, Kesavan;Lee, Joon Ha;Subramaniyam, Sathiyamoorthy;Yoo, Seungil;Park, Junhyung;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1863-1870
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    • 2016
  • Antimicrobial peptides/proteins (AMPs) are present in all types of organisms, from microbes and plants to vertebrates and invertebrates such as insects. The grasshopper Oxya chinensis sinuosa is an insect species that is widely consumed around the world for its broad medicinal value. However, the lack of available genetic information for this species is an obstacle to understanding the full potential of its AMPs. Analysis of the O. chinensis sinuosa transcriptome and expression profile is essential for extending the available genetic information resources. In this study, we determined the whole-body transcriptome of O. chinensis sinuosa and analyzed the potential AMPs induced by bacterial immunization. A high-throughput RNA-Seq approach generated 94,348 contigs and 66,555 unigenes. Of these unigenes, 36,032 (54.14%) matched known proteins in the NCBI database in a BLAST search. Functional analysis demonstrated that 38,219 unigenes were clustered into 5,499 gene ontology terms. In addition, 26 cDNAs encoding novel AMPs were identified by an in silico approach using public databases. Our transcriptome dataset and AMP profile greatly improve our understanding of O. chinensis sinuosa genetics and provide a huge number of gene sequences for further study, including genes of known importance and genes of unknown function.

Application of Competitive ELISA Method for Estimation of Urinary Aflatoxin M1 Level (ELISA 방법을 이용한 요중 아플라톡신 M1 측정)

  • Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.23 no.2
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    • pp.306-310
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    • 2013
  • We compared the efficacy of the competitive ELISA method for measuring the level of urinary aflatoxin M1 (AFM1) with that of the HPLC-fluorescence detector (HPLC-FLD) method. The recovery rate of AFM1 with the ELISA method was 105% (73-124%), and the coefficient of variation of the analysis was 6.85%. The ELISA method showed a 0.20 pg/ml and 0.62 pg/ml limit of detection and limit of quantitation, respectively. In correlation analysis, the two methods showed a very strong and statistically significant correlation (R=0.96, p<0.01). However, in spite of the strong correlation, the ELISA method tended to overestimate the urinary AFM1 concentration compared to the HPLC-FLD method. These results suggest that the competitive ELISA method may be a useful technique for measuring the AFM1 level in high-throughput urine samples, but it needs to be corrected with a regression equation from regression analysis with the HPLC-FLD method.

Expression profiles of microRNAs in skeletal muscle of sheep by deep sequencing

  • Liu, Zhijin;Li, Cunyuan;Li, Xiaoyue;Yao, Yang;Ni, Wei;Zhang, Xiangyu;Cao, Yang;Hazi, Wureli;Wang, Dawei;Quan, Renzhe;Yu, Shuting;Wu, Yuyu;Niu, Songmin;Cui, Yulong;Khan, Yaseen;Hu, Shengwei
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.757-766
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    • 2019
  • Objective: MicroRNAs are a class of endogenous small regulatory RNAs that regulate cell proliferation, differentiation and apoptosis. Recent studies on miRNAs are mainly focused on mice, human and pig. However, the studies on miRNAs in skeletal muscle of sheep are not comprehensive. Methods: RNA-seq technology was used to perform genomic analysis of miRNAs in prenatal and postnatal skeletal muscle of sheep. Targeted genes were predicted using miRanda software and miRNA-mRNA interactions were verified by quantitative real-time polymerase chain reaction. To further investigate the function of miRNAs, candidate targeted genes were enriched for analysis using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment. Results: The results showed total of 1,086 known miRNAs and 40 new candidate miRNAs were detected in prenatal and postnatal skeletal muscle of sheep. In addition, 345 miRNAs (151 up-regulated, 94 down-regulated) were differentially expressed. Moreover, miRanda software was performed to predict targeted genes of miRNAs, resulting in a total of 2,833 predicted targets, especially miR-381 which targeted multiple muscle-related mRNAs. Furthermore, GO and KEGG pathway analysis confirmed that targeted genes of miRNAs were involved in development of skeletal muscles. Conclusion: This study supplements the miRNA database of sheep, which provides valuable information for further study of the biological function of miRNAs in sheep skeletal muscle.

Effect of Interference Mitigation Technique and Performance Analysis for Small Cell in Homogeneous Networks (동종네트워크 상에서 셀 소형화 간섭 완화 기법 및 성능 분석)

  • Jang, Ye-Ok;Cho, Eun-Hyung;Hong, Een-Kee
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.39C no.10
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    • pp.937-945
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    • 2014
  • As various services requiring high data rate are supported by introducing LTE/LTE-adv., mobile traffic increases rapidly. To cope with the continuous growth of traffic demand, small cell technology is considered as one of the most promising one. Small cell can increase system capacity by increasing the number of base stations with reduced cell radius. In this paper, we analyze the effect of cell densification with small cells in terms of SINR and average UE throughput considering cell split and the number of UE per unit area. As the cell becomes smaller, SINR degradation arises from high ICI(Inter Cell Interference) and we evaluate the effect of interference mitigation scheme in small cell environment where the proper interference mitigation technique is applied.