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Sample records for based phosphoproteomic analysis

  1. Phosphoproteomics-based systems analysis of signal transduction networks

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    Hiroko eKozuka-Hata

    2012-01-01

    Full Text Available Signal transduction systems coordinate complex cellular information to regulate biological events such as cell proliferation and differentiation. Although the accumulating evidence on widespread association of signaling molecules has revealed essential contribution of phosphorylation-dependent interaction networks to cellular regulation, their dynamic behavior is mostly yet to be analyzed. Recent technological advances regarding mass spectrometry-based quantitative proteomics have enabled us to describe the comprehensive status of phosphorylated molecules in a time-resolved manner. Computational analyses based on the phosphoproteome dynamics accelerate generation of novel methodologies for mathematical analysis of cellular signaling. Phosphoproteomics-based numerical modeling can be used to evaluate regulatory network elements from a statistical point of view. Integration with transcriptome dynamics also uncovers regulatory hubs at the transcriptional level. These omics-based computational methodologies, which have firstly been applied to representative signaling systems such as the epidermal growth factor receptor pathway, have now opened up a gate for systems analysis of signaling networks involved in immune response and cancer.

  2. Selecting Sample Preparation Workflows for Mass Spectrometry-Based Proteomic and Phosphoproteomic Analysis of Patient Samples with Acute Myeloid Leukemia

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    Maria Hernandez-Valladares

    2016-08-01

    Full Text Available Global mass spectrometry (MS-based proteomic and phosphoproteomic studies of acute myeloid leukemia (AML biomarkers represent a powerful strategy to identify and confirm proteins and their phosphorylated modifications that could be applied in diagnosis and prognosis, as a support for individual treatment regimens and selection of patients for bone marrow transplant. MS-based studies require optimal and reproducible workflows that allow a satisfactory coverage of the proteome and its modifications. Preparation of samples for global MS analysis is a crucial step and it usually requires method testing, tuning and optimization. Different proteomic workflows that have been used to prepare AML patient samples for global MS analysis usually include a standard protein in-solution digestion procedure with a urea-based lysis buffer. The enrichment of phosphopeptides from AML patient samples has previously been carried out either with immobilized metal affinity chromatography (IMAC or metal oxide affinity chromatography (MOAC. We have recently tested several methods of sample preparation for MS analysis of the AML proteome and phosphoproteome and introduced filter-aided sample preparation (FASP as a superior methodology for the sensitive and reproducible generation of peptides from patient samples. FASP-prepared peptides can be further fractionated or IMAC-enriched for proteome or phosphoproteome analyses. Herein, we will review both in-solution and FASP-based sample preparation workflows and encourage the use of the latter for the highest protein and phosphorylation coverage and reproducibility.

  3. SILAC-Based Temporal Phosphoproteomics

    DEFF Research Database (Denmark)

    Francavilla, Chiara; Hekmat, Omid; Blagoev, Blagoy;

    2014-01-01

    signaling events. Here we provide an optimized SILAC-based proteomic workflow to analyze temporal changes in phosphoproteomes, which involve a generic three step enrichment protocol for phosphopeptides. SILAC-labeled peptides from digested whole cell lysates are as a first step enriched for phosphorylated...... pathways. Quantitative proteomics that combines stable isotope labeling by amino acid in cell culture (SILAC) with enrichment strategies for post-translational modification-bearing peptides and high-performance tandem mass spectrometry represents a powerful and unbiased approach to monitor dynamic...

  4. Quantitative Phosphoproteomic Analysis of T-Cell Receptor Signaling.

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    Ahsan, Nagib; Salomon, Arthur R

    2017-01-01

    TCR signaling critically depends on protein phosphorylation across many proteins. Localization of each phosphorylation event relative to the T-cell receptor (TCR) and canonical T-cell signaling proteins will provide clues about the structure of TCR signaling networks. Quantitative phosphoproteomic analysis by mass spectrometry provides a wide-scale view of cellular phosphorylation networks. However, analysis of phosphorylation by mass spectrometry is still challenging due to the relative low abundance of phosphorylated proteins relative to all proteins and the extraordinary diversity of phosphorylation sites across the proteome. Highly selective enrichment of phosphorylated peptides is essential to provide the most comprehensive view of the phosphoproteome. Optimization of phosphopeptide enrichment methods coupled with highly sensitive mass spectrometry workflows significantly improves the sequencing depth of the phosphoproteome to over 10,000 unique phosphorylation sites from complex cell lysates. Here we describe a step-by-step method for phosphoproteomic analysis that has achieved widespread success for identification of serine, threonine, and tyrosine phosphorylation. Reproducible quantification of relative phosphopeptide abundance is provided by intensity-based label-free quantitation. An ideal set of mass spectrometry analysis parameters is also provided that optimize the yield of identified sites. We also provide guidelines for the bioinformatic analysis of this type of data to assess the quality of the data and to comply with proteomic data reporting requirements.

  5. SILAC for global phosphoproteomic analysis.

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    Pimienta, Genaro; Chaerkady, Raghothama; Pandey, Akhilesh

    2009-01-01

    Establishing the phosphorylation pattern of proteins in a comprehensive fashion is an important goal of a majority of cell signaling projects. Phosphoproteomic strategies should be designed in such a manner as to identify sites of phosphorylation as well as to provide quantitative information about the extent of phosphorylation at the sites. In this chapter, we describe an experimental strategy that outlines such an approach using stable isotope labeling with amino acids in cell culture (SILAC) coupled to LC-MS/MS. We highlight the importance of quantitative strategies in signal transduction as a platform for a systematic and global elucidation of biological processes.

  6. TiO2-Based Phosphoproteomic Analysis of the Plasma Membrane and the Effects of Phosphatase Inhibitor Treatment

    DEFF Research Database (Denmark)

    Thingholm, Tine; Larsen, Martin Røssel; Ingrell, Christian

    2008-01-01

    Phosphorylation of plasma membrane proteins frequently initiates signal transduction pathways or attenuate plasma membrane transport processes. Because of the low abundance and hydrophobic features of many plasma membrane proteins and the low stoichiometry of protein phosphorylation, studies...... of the plasma membrane phosphoproteome are challenging. We present an optimized analytical strategy for plasma membrane phosphoproteomics that combines efficient plasma membrane protein preparation with TiO 2-based phosphopeptide enrichment and high-performance mass spectrometry for phosphopeptide sequencing....... We used sucrose centrifugation in combination with sodium carbonate extraction to achieve efficient and reproducible purification of low microgram levels of plasma membrane proteins from human mesenchymal stem cells (hMSCs, 10 (7) cells), achieving more than 70% yield of membrane proteins...

  7. Systems Analysis for Interpretation of Phosphoproteomics Data

    DEFF Research Database (Denmark)

    Munk, Stephanie; Refsgaard, Jan C; Olsen, Jesper V

    2016-01-01

    Global phosphoproteomics investigations yield overwhelming datasets with up to tens of thousands of quantified phosphosites. The main challenge after acquiring such large-scale data is to extract the biological meaning and relate this to the experimental question at hand. Systems level analysis...... provides the best means for extracting functional insights from such types of datasets, and this has primed a rapid development of bioinformatics tools and resources over the last decade. Many of these tools are specialized databases that can be mined for annotation and pathway enrichment, whereas others...... provide a platform to generate functional protein networks and explore the relations between proteins of interest. The use of these tools requires careful consideration with regard to the input data, and the interpretation demands a critical approach. This chapter provides a summary of the most...

  8. Phosphoproteomic analysis of chromoplasts from sweet orange during fruit ripening.

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    Zeng, Yunliu; Pan, Zhiyong; Wang, Lun; Ding, Yuduan; Xu, Qiang; Xiao, Shunyuan; Deng, Xiuxin

    2014-02-01

    Like other types of plastids, chromoplasts have essential biosynthetic and metabolic activities which may be regulated via post-translational modifications, such as phosphorylation, of their resident proteins. We here report a proteome-wide mapping of in vivo phosphorylation sites in chromoplast-enriched samples prepared from sweet orange [Citrus sinensis (L.) Osbeck] at different ripening stages by titanium dioxide-based affinity chromatography for phosphoprotein enrichment with LC-MS/MS. A total of 109 plastid-localized phosphoprotein candidates were identified that correspond to 179 unique phosphorylation sites in 135 phosphopeptides. On the basis of Motif-X analysis, two distinct types of phosphorylation sites, one as proline-directed phosphorylation motif and the other as casein kinase II motif, can be generalized from these identified phosphopeptides. While most identified phosphoproteins show high homology to those already identified in plastids, approximately 22% of them are novel based on BLAST search using the public databases PhosPhAt and P(3) DB. A close comparative analysis showed that approximately 50% of the phosphoproteins identified in citrus chromoplasts find obvious counterparts in the chloroplast phosphoproteome, suggesting a rather high-level of conservation in basic metabolic activities in these two types of plastids. Not surprisingly, the phosphoproteome of citrus chromoplasts is also characterized by the lack of phosphoproteins involved in photosynthesis and by the presence of more phosphoproteins implicated in stress/redox responses. This study presents the first comprehensive phosphoproteomic analysis of chromoplasts and may help to understand how phosphorylation regulates differentiation of citrus chromoplasts during fruit ripening.

  9. Gel-based phosphoproteomics analysis of sarcoplasmic proteins in postmortem porcine muscle with pH decline rate and time differences

    DEFF Research Database (Denmark)

    Huang, Honggang; Larsen, Martin Røssel; Karlsson, Anders H;

    2011-01-01

    Meat quality development is highly influenced by the pH decline caused by the postmortem (PM) glycolysis. Protein phosphorylation is an important mechanism in regulating the activity of glycometabolic enzymes. Here, a gel-based phosphoproteomic study was performed to analyze the protein phosphory...

  10. Analytical strategies in mass spectrometry-based phosphoproteomics

    DEFF Research Database (Denmark)

    Rosenqvist, Heidi; Ye, Juanying; Jensen, Ole N

    2011-01-01

    to reveal key regulatory events and phosphorylation-mediated processes in the cell and in whole organisms. We present an overview of sensitive and robust analytical methods for phosphopeptide analysis, including calcium phosphate precipitation and affinity enrichment methods such as IMAC and TiO(2). We......Phosphoproteomics, the systematic study of protein phosphorylation events and cell signaling networks in cells and tissues, is a rapidly evolving branch of functional proteomics. Current phosphoproteomics research provides a large toolbox of strategies and protocols that may assist researchers...

  11. Phosphoproteomics Analysis of Endometrium in Women with or without Endometriosis

    Institute of Scientific and Technical Information of China (English)

    Hong-Mei Xu; Hai-Teng Deng; Chong-Dong Liu; Yu-Ling Chen; Zhen-Yu Zhang

    2015-01-01

    Background:The molecular mechanisms underlying the endometriosis are still not completely understood.In order to test the hypothesis that the approaches in phosphoproteomics might contribute to the identification of key biomarkers to assess disease pathogenesis and drug targets,we carried out a phosphoproteomics analysis of human endometrium.Methods:A large-scale differential phosphoproteome analysis,using peptide enrichment of titanium dioxide purify and sequential elution from immobilized metal affinity chromatography with linear trap quadrupole-tandem mass spectrometry,was performed in endometrium tissues from 8 women with or without endometriosis.Results:The phosphorylation profiling of endometrium from endometriosis patients had been obtained,and found that identified 516 proteins were modified at phosphorylation level during endometriosis.Gene ontology annotation analysis showed that these proteins were enriched in cellular processes of binding and catalytic activity.Further pathway analysis showed that ribosome pathway and focal adhesion pathway were the top two pathways,which might be deregulated during the development of endometriosis.Conclusions:That large-scale phosphoproteome quantification has been successfully identified in endometrium tissues of women with or without endometriosis will provide new insights to understand the molecular mechanisms of the development of endometriosis.

  12. Dataset of the Botrytis cinerea phosphoproteome induced by different plant-based elicitors

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    Eva Liñeiro

    2016-06-01

    Full Text Available Phosphorylation is one of the main post-translational modification (PTM involved in signaling network in the ascomycete Botrytis cinerea, one of the most relevant phytopathogenic fungus. The data presented in this article provided a differential mass spectrometry-based analysis of the phosphoproteome of B. cinerea under two different phenotypical conditions induced by the use of two different elicitors: glucose and deproteinized Tomate Cell Walls (TCW. A total 1138 and 733 phosphoproteins were identified for glucose and TCW culture conditions respectively. Raw data are deposited at the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier (PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD003099. Further interpretation and discussion of these data are provided in our research article entitled “Phosphoproteome analysis of B.cinerea in response to different plant-based elicitors” (Liñeiro et al., 2016 [1].

  13. In Vivo SILAC-Based Proteomics Reveals Phosphoproteome Changes during Mouse Skin Carcinogenesis

    DEFF Research Database (Denmark)

    Zanivan, Sara; Meves, Alexander; Behrendt, Kristina;

    2013-01-01

    SILAC technology in combination with high-resolution mass spectrometry (MS) can be successfully used to measure phosphoproteomes in vivo. Here, Zanivan, Mann, and colleagues have applied SILAC-based MS to investigate phosphoproteomic changes during skin carcinogenesis, using the DMBA/TPA two...

  14. Phosphoproteomics analysis of a clinical Mycobacterium tuberculosis Beijing isolate: expanding the mycobacterial phosphoproteome catalog.

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    Fortuin, Suereta; Tomazella, Gisele G; Nagaraj, Nagarjuna; Sampson, Samantha L; Gey van Pittius, Nicolaas C; Soares, Nelson C; Wiker, Harald G; de Souza, Gustavo A; Warren, Robin M

    2015-01-01

    Reversible protein phosphorylation, regulated by protein kinases and phosphatases, mediates a switch between protein activity and cellular pathways that contribute to a large number of cellular processes. The Mycobacterium tuberculosis genome encodes 11 Serine/Threonine kinases (STPKs) which show close homology to eukaryotic kinases. This study aimed to elucidate the phosphoproteomic landscape of a clinical isolate of M. tuberculosis. We performed a high throughput mass spectrometric analysis of proteins extracted from an early-logarithmic phase culture. Whole cell lysate proteins were processed using the filter-aided sample preparation method, followed by phosphopeptide enrichment of tryptic peptides by strong cation exchange (SCX) and Titanium dioxide (TiO2) chromatography. The MaxQuant quantitative proteomics software package was used for protein identification. Our analysis identified 414 serine/threonine/tyrosine phosphorylated sites, with a distribution of S/T/Y sites; 38% on serine, 59% on threonine and 3% on tyrosine; present on 303 unique peptides mapping to 214 M. tuberculosis proteins. Only 45 of the S/T/Y phosphorylated proteins identified in our study had been previously described in the laboratory strain H37Rv, confirming previous reports. The remaining 169 phosphorylated proteins were newly identified in this clinical M. tuberculosis Beijing strain. We identified 5 novel tyrosine phosphorylated proteins. These findings not only expand upon our current understanding of the protein phosphorylation network in clinical M. tuberculosis but the data set also further extends and complements previous knowledge regarding phosphorylated peptides and phosphorylation sites in M. tuberculosis.

  15. Quantitative Phosphoproteomics Analysis of ERBB3/ERBB4 Signaling.

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    Wandinger, Sebastian K; Lahortiga, Idoya; Jacobs, Kris; Klammer, Martin; Jordan, Nicole; Elschenbroich, Sarah; Parade, Marc; Jacoby, Edgar; Linders, Joannes T M; Brehmer, Dirk; Cools, Jan; Daub, Henrik

    2016-01-01

    The four members of the epidermal growth factor receptor (EGFR/ERBB) family form homo- and heterodimers which mediate ligand-specific regulation of many key cellular processes in normal and cancer tissues. While signaling through the EGFR has been extensively studied on the molecular level, signal transduction through ERBB3/ERBB4 heterodimers is less well understood. Here, we generated isogenic mouse Ba/F3 cells that express full-length and functional membrane-integrated ERBB3 and ERBB4 or ERBB4 alone, to serve as a defined cellular model for biological and phosphoproteomics analysis of ERBB3/ERBB4 signaling. ERBB3 co-expression significantly enhanced Ba/F3 cell proliferation upon neuregulin-1 (NRG1) treatment. For comprehensive signaling studies we performed quantitative mass spectrometry (MS) experiments to compare the basal ERBB3/ERBB4 cell phosphoproteome to NRG1 treatment of ERBB3/ERBB4 and ERBB4 cells. We employed a workflow comprising differential isotope labeling with mTRAQ reagents followed by chromatographic peptide separation and final phosphopeptide enrichment prior to MS analysis. Overall, we identified 9686 phosphorylation sites which could be confidently localized to specific residues. Statistical analysis of three replicate experiments revealed 492 phosphorylation sites which were significantly changed in NRG1-treated ERBB3/ERBB4 cells. Bioinformatics data analysis recapitulated regulation of mitogen-activated protein kinase and Akt pathways, but also indicated signaling links to cytoskeletal functions and nuclear biology. Comparative assessment of NRG1-stimulated ERBB4 Ba/F3 cells revealed that ERBB3 did not trigger defined signaling pathways but more broadly enhanced phosphoproteome regulation in cells expressing both receptors. In conclusion, our data provide the first global picture of ERBB3/ERBB4 signaling and provide numerous potential starting points for further mechanistic studies.

  16. Quantitative Phosphoproteomics Analysis of ERBB3/ERBB4 Signaling.

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    Sebastian K Wandinger

    Full Text Available The four members of the epidermal growth factor receptor (EGFR/ERBB family form homo- and heterodimers which mediate ligand-specific regulation of many key cellular processes in normal and cancer tissues. While signaling through the EGFR has been extensively studied on the molecular level, signal transduction through ERBB3/ERBB4 heterodimers is less well understood. Here, we generated isogenic mouse Ba/F3 cells that express full-length and functional membrane-integrated ERBB3 and ERBB4 or ERBB4 alone, to serve as a defined cellular model for biological and phosphoproteomics analysis of ERBB3/ERBB4 signaling. ERBB3 co-expression significantly enhanced Ba/F3 cell proliferation upon neuregulin-1 (NRG1 treatment. For comprehensive signaling studies we performed quantitative mass spectrometry (MS experiments to compare the basal ERBB3/ERBB4 cell phosphoproteome to NRG1 treatment of ERBB3/ERBB4 and ERBB4 cells. We employed a workflow comprising differential isotope labeling with mTRAQ reagents followed by chromatographic peptide separation and final phosphopeptide enrichment prior to MS analysis. Overall, we identified 9686 phosphorylation sites which could be confidently localized to specific residues. Statistical analysis of three replicate experiments revealed 492 phosphorylation sites which were significantly changed in NRG1-treated ERBB3/ERBB4 cells. Bioinformatics data analysis recapitulated regulation of mitogen-activated protein kinase and Akt pathways, but also indicated signaling links to cytoskeletal functions and nuclear biology. Comparative assessment of NRG1-stimulated ERBB4 Ba/F3 cells revealed that ERBB3 did not trigger defined signaling pathways but more broadly enhanced phosphoproteome regulation in cells expressing both receptors. In conclusion, our data provide the first global picture of ERBB3/ERBB4 signaling and provide numerous potential starting points for further mechanistic studies.

  17. Quantitative phosphoproteomic analysis of prion-infected neuronal cells

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    Löwer Johannes

    2010-09-01

    Full Text Available Abstract Prion diseases or transmissible spongiform encephalopathies (TSEs are fatal diseases associated with the conversion of the cellular prion protein (PrPC to the abnormal prion protein (PrPSc. Since the molecular mechanisms in pathogenesis are widely unclear, we analyzed the global phospho-proteome and detected a differential pattern of tyrosine- and threonine phosphorylated proteins in PrPSc-replicating and pentosan polysulfate (PPS-rescued N2a cells in two-dimensional gel electrophoresis. To quantify phosphorylated proteins, we performed a SILAC (stable isotope labeling by amino acids in cell culture analysis and identified 105 proteins, which showed a regulated phosphorylation upon PrPSc infection. Among those proteins, we validated the dephosphorylation of stathmin and Cdc2 and the induced phosphorylation of cofilin in PrPSc-infected N2a cells in Western blot analyses. Our analysis showed for the first time a differentially regulated phospho-proteome in PrPSc infection, which could contribute to the establishment of novel protein markers and to the development of novel therapeutic intervention strategies in targeting prion-associated disease.

  18. Phosphoproteome analysis of Lotus japonicus seeds.

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    Ino, Yoko; Ishikawa, Akiyo; Nomura, Ayako; Kajiwara, Hideyuki; Harada, Kyuya; Hirano, Hisashi

    2014-01-01

    In this study, we report the first dataset of phosphoproteins of the seeds of a model plant, Lotus japonicus. This dataset might be useful in studying the regulatory mechanisms of seed germination in legume plants. By proteomic analysis of seeds following water absorption, we identified a total of 721 phosphopeptides derived from 343 phosphoproteins in cotyledons, and 931 phosphopeptides from 473 phosphoproteins in hypocotyls. Kinase-specific prediction analyses revealed that different kinases were activated in cotyledons and hypocotyls. In particular, many peptides containing ATM-kinase target motifs, X-X-pS/pT-Q-X-X, were detected in cotyledons. Moreover, by real-time RT-PCR analysis, we found that expression of a homolog of ATM kinase is upregulated specifically in cotyledons, suggesting that this ATM-kinase homolog plays a significant role in cell proliferation in the cotyledons of L. japonicus seeds. The data have been deposited to the ProteomeXchange with identifier PXD000053 (http://proteomecentral.proteomexchange.org/dataset/PXD000053).

  19. In Vivo SILAC-Based Proteomics Reveals Phosphoproteome Changes during Mouse Skin Carcinogenesis

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    Sara Zanivan

    2013-02-01

    Full Text Available Cancer progresses through distinct stages, and mouse models recapitulating traits of this progression are frequently used to explore genetic, morphological, and pharmacological aspects of tumor development. To complement genomic investigations of this process, we here quantify phosphoproteomic changes in skin cancer development using the SILAC mouse technology coupled to high-resolution mass spectrometry. We distill protein expression signatures from our data that distinguish between skin cancer stages. A distinct phosphoproteome of the two stages of cancer progression is identified that correlates with perturbed cell growth and implicates cell adhesion as a major driver of malignancy. Importantly, integrated analysis of phosphoproteomic data and prediction of kinase activity revealed PAK4-PKC/SRC network to be highly deregulated in SCC but not in papilloma. This detailed molecular picture, both at the proteome and phosphoproteome level, will prove useful for the study of mechanisms of tumor progression.

  20. Assessment of phosphopeptide enrichment/precipitation method for LC-MS/MS based phosphoproteomic analysis of plant tissue

    DEFF Research Database (Denmark)

    Ye, Juanying; Rudashevskaya, Elena; Hansen, Thomas Aarup

      Introduction Mass spectrometry (MS) is a powerful technology for study of PTMs, including protein phosphorylation. Due to the low abundance of many phosphoproteins and the relatively poor ionization efficiency of phosphopeptides, specific enrichment of phosphopeptides prior to MS analysis is ne...... (MSA) method was used for phosphopeptide fragmentation. The resulting fragment ion spectra were processed with Proteome Discoverer software (Thermo Electron, Bremen, Germany). Results We first investigated the global phosphorylation profile of plant plasma membrane proteins by enriching...

  1. Phosphoproteomic analysis of the response of maize leaves to drought, heat and their combination stress

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    Xiuli eHu

    2015-05-01

    Full Text Available Drought and heat stress, especially their combination, greatly affect crop production. Many studies have described transcriptome, proteome and phosphoproteome changes in response of plants to drought or heat stress. However, the study about the phosphoproteomic changes in response of crops to the combination stress is scare. To understand the mechanism of maize responses to the drought and heat combination stress, phosphoproteomic analysis was performed on maize leaves by using multiplex iTRAQ-based quantitative proteomic and LC-MS/MS methods. Five-leaf-stage maize was subjected to drought, heat or their combination, and the leaves were collected. Globally, heat, drought and the combined stress significantly changed the phosphorylation levels of 172, 149 and 144 phosphopeptides, respectively. These phosphopeptides corresponded to 282 proteins. Among them, 23 only responded to the combined stress and could not be predicted from their responses to single stressors; 30 and 75 only responded to drought and heat, respectively. Notably, 19 proteins were phosphorylated on different sites in response to the single and combination stresses. Of the seven significantly enriched phosphorylation motifs identified, two were common for all stresses, two were common for heat and the combined stress, and one was specific to the combined stress. The signaling pathways in which the phosphoproteins were involved clearly differed among the three stresses. Functional characterization of the phosphoproteins and the pathways identified here could lead to new targets for the enhancement of crop stress tolerance, which will be particularly important in the face of climate change and the increasing prevalence of abiotic stressors.

  2. Phosphoproteomic analysis reveals compensatory effects in the piriform cortex of VX nerve agent exposed rats.

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    Nirujogi, Raja Sekhar; Wright, James D; Manda, Srikanth S; Zhong, Jun; Na, Chan Hyun; Meyerhoff, James; Benton, Bernard; Jabbour, Rabih; Willis, Kristen; Kim, Min-Sik; Pandey, Akhilesh; Sekowski, Jennifer W

    2015-01-01

    To gain insights into the toxicity induced by the nerve agent VX, an MS-based phosphoproteomic analysis was carried out on the piriform cortex region of brains from VX-treated rats. Using isobaric tag based TMT labeling followed by titanium dioxide enrichment strategy, we identified 9975 unique phosphosites derived from 3287 phosphoproteins. Temporal changes in the phosphorylation status of peptides were observed over a time period of 24 h in rats exposed to a 1× LD50, intravenous (i.v.) dose with the most notable changes occurring at the 1 h postexposure time point. Five major functional classes of proteins exhibited changes in their phosphorylation status: (i) ion channels/transporters, including ATPases, (ii) kinases/phosphatases, (iii) GTPases, (iv) structural proteins, and (v) transcriptional regulatory proteins. This study is the first quantitative phosphoproteomic analysis of VX toxicity in the brain. Understanding the toxicity and compensatory signaling mechanisms will improve the understanding of the complex toxicity of VX in the brain and aid in the elucidation of novel molecular targets that would be important for development of improved countermeasures. All MS data have been deposited in the ProteomeXchange with identifier PXD001184 (http://proteomecentral.proteomexchange.org/dataset/PXD001184).

  3. Temporal proteomic analysis of HIV infection reveals remodelling of the host phosphoproteome by lentiviral Vif variants.

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    Greenwood, Edward Jd; Matheson, Nicholas J; Wals, Kim; van den Boomen, Dick Jh; Antrobus, Robin; Williamson, James C; Lehner, Paul J

    2016-09-30

    Viruses manipulate host factors to enhance their replication and evade cellular restriction. We used multiplex tandem mass tag (TMT)-based whole cell proteomics to perform a comprehensive time course analysis of >6500 viral and cellular proteins during HIV infection. To enable specific functional predictions, we categorized cellular proteins regulated by HIV according to their patterns of temporal expression. We focussed on proteins depleted with similar kinetics to APOBEC3C, and found the viral accessory protein Vif to be necessary and sufficient for CUL5-dependent proteasomal degradation of all members of the B56 family of regulatory subunits of the key cellular phosphatase PP2A (PPP2R5A-E). Quantitative phosphoproteomic analysis of HIV-infected cells confirmed Vif-dependent hyperphosphorylation of >200 cellular proteins, particularly substrates of the aurora kinases. The ability of Vif to target PPP2R5 subunits is found in primate and non-primate lentiviral lineages, and remodeling of the cellular phosphoproteome is therefore a second ancient and conserved Vif function.

  4. Temporal proteomic analysis of HIV infection reveals remodelling of the host phosphoproteome by lentiviral Vif variants

    Science.gov (United States)

    Greenwood, Edward JD; Matheson, Nicholas J; Wals, Kim; van den Boomen, Dick JH; Antrobus, Robin; Williamson, James C; Lehner, Paul J

    2016-01-01

    Viruses manipulate host factors to enhance their replication and evade cellular restriction. We used multiplex tandem mass tag (TMT)-based whole cell proteomics to perform a comprehensive time course analysis of >6500 viral and cellular proteins during HIV infection. To enable specific functional predictions, we categorized cellular proteins regulated by HIV according to their patterns of temporal expression. We focussed on proteins depleted with similar kinetics to APOBEC3C, and found the viral accessory protein Vif to be necessary and sufficient for CUL5-dependent proteasomal degradation of all members of the B56 family of regulatory subunits of the key cellular phosphatase PP2A (PPP2R5A-E). Quantitative phosphoproteomic analysis of HIV-infected cells confirmed Vif-dependent hyperphosphorylation of >200 cellular proteins, particularly substrates of the aurora kinases. The ability of Vif to target PPP2R5 subunits is found in primate and non-primate lentiviral lineages, and remodeling of the cellular phosphoproteome is therefore a second ancient and conserved Vif function. DOI: http://dx.doi.org/10.7554/eLife.18296.001 PMID:27690223

  5. Phosphoproteome analysis of E-coli reveals evolutionary conservation of bacterial Ser/Thr/Tyr phosphorylation

    DEFF Research Database (Denmark)

    Macek, B.; Gnad, F.; Soufi, Boumediene

    2008-01-01

    we use a recently developed proteomics approach based on phosphopeptide enrichment and high accuracy MS to analyze the phosphoproteome of the model Gram-negative bacterium Escherichia coli. We report 81 phosphorylation sites on 79 E. coli proteins, with distribution of Ser/Thr/Tyr phosphorylation...... sites 68%/23%/9%. Despite their phylogenetic distance, phosphoproteomes of E. coli and B. subtilis show striking similarity in size, classes of phosphorylated proteins, and distribution of Ser/Thr/Tyr phosphorylation sites. By combining the two datasets, we created the largest phosphorylation site...

  6. Quantitative phosphoproteomic analysis of porcine muscle within 24 h postmortem

    DEFF Research Database (Denmark)

    Huang, Honggang; Larsen, Martin Røssel; Palmisano, Giuseppe

    2014-01-01

    phosphorylation levels in muscle within 24 h PM. The high phosphorylation level of heat shock proteins (HSPs) in early PM may be an adaptive response to slaughter stress and protect muscle cell from apoptosis, as observed in the serine 84 of HSP27. This work indicated that PM muscle proteins underwent significant...... the dimethyl labeling combined with the TiSH phosphopeptide enrichment and LC-MS/MS strategy. This was the first high-throughput quantitative phosphoproteomic study in PM muscle of farm animals. In the work, both the proteome and phosphoproteome were analyzed, and the large number of identified peptides......, phosphopeptides and phosphorylation sites can greatly enrich the current farm animal protein database. The proteins involved in glycometabolism, muscle contraction and heat shock proteins (HSPs) showed significantly changed phosphorylation levels during PM meat development. This work indicated that PM muscle...

  7. Multidimensional electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) for quantitative analysis of the proteome and phosphoproteome in clinical and biomedical research.

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    Loroch, Stefan; Schommartz, Tim; Brune, Wolfram; Zahedi, René Peiman; Sickmann, Albert

    2015-05-01

    Quantitative proteomics and phosphoproteomics have become key disciplines in understanding cellular processes. Fundamental research can be done using cell culture providing researchers with virtually infinite sample amounts. In contrast, clinical, pre-clinical and biomedical research is often restricted to minute sample amounts and requires an efficient analysis with only micrograms of protein. To address this issue, we generated a highly sensitive workflow for combined LC-MS-based quantitative proteomics and phosphoproteomics by refining an ERLIC-based 2D phosphoproteomics workflow into an ERLIC-based 3D workflow covering the global proteome as well. The resulting 3D strategy was successfully used for an in-depth quantitative analysis of both, the proteome and the phosphoproteome of murine cytomegalovirus-infected mouse fibroblasts, a model system for host cell manipulation by a virus. In a 2-plex SILAC experiment with 150 μg of a tryptic digest per condition, the 3D strategy enabled the quantification of ~75% more proteins and even ~134% more peptides compared to the 2D strategy. Additionally, we could quantify ~50% more phosphoproteins by non-phosphorylated peptides, concurrently yielding insights into changes on the levels of protein expression and phosphorylation. Beside its sensitivity, our novel three-dimensional ERLIC-strategy has the potential for semi-automated sample processing rendering it a suitable future perspective for clinical, pre-clinical and biomedical research.

  8. Interleukin-2 signaling pathway analysis by quantitative phosphoproteomics

    DEFF Research Database (Denmark)

    Osinalde, Nerea; Moss, Helle; Arrizabalaga, Onetsine

    2011-01-01

    in modulation of the immune response. The complete characterization of the IL-2 pathway is essential to understand how aberrant IL-2 signaling results in several diseases such as cancer or autoimmunity and also how IL-2 treatments affect cancer patients. To gain insights into the downstream machinery activated...... by IL-2, we aimed to define the global tyrosine-phosphoproteome of IL-2 pathway in human T cell line Kit225 using high resolution mass spectrometry combined with phosphotyrosine immunoprecipitation and SILAC. The molecular snapshot at 5min of IL-2 stimulation resulted in identification of 172 proteins...... with increased abundance in the tyrosine-phosphorylated complexes, of which 34 were not previously described. In addition, chemical inhibition of the identified IL-2-mediated JAK, PI3K and MAPK signaling pathways, resulted in distinct alteration on the IL-2 dependent proliferation....

  9. Predicting Kinase Activity in Angiotensin Receptor Phosphoproteomes Based on Sequence-Motifs and Interactions

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    Bøgebo, Rikke; Horn, Heiko; Olsen, Jesper V;

    2014-01-01

    -arrestin dependent signalling. Two complimentary global phosphoproteomics studies have analyzed the complex signalling induced by the AT1aR. Here we integrate the data sets from these studies and perform a joint analysis using a novel method for prediction of differential kinase activity from phosphoproteomics data......Recent progress in the understanding of seven-transmembrane receptor (7TMR) signalling has promoted the development of a new generation of pathway selective ligands. The angiotensin II type I receptor (AT1aR) is one of the most studied 7TMRs with respect to selective activation of the β...... likely activated kinases. This suggested that AT1aR-dependent signalling activates 48 of the 285 kinases detected in HEK293 cells. Of these, Aurora B, CLK3 and PKG1 have not previously been described in the pathway whereas others, such as PKA, PKB and PKC, are well known. In summary, we have developed...

  10. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Tran H.; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese RW; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolic, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-11-11

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e., roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag 8-plex ITRAQ, enriched using Ni-NTA magnetic beads and subjected to nRPLC-MS/MS analysis using HCD and decision tree guided CID/ETD strategy. A total of 1,625 unique phosphopeptides, spanning 1,659 non-redundant phosphorylation sites, were detected from 1,126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5 fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  11. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum*

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese R.; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Paša-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-01-01

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e. roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5-fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation. PMID:22843990

  12. Quantitative phosphoproteomic analysis of soybean root hairs inoculated with Bradyrhizobium japonicum.

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T; Clauss, Therese R; Gritsenko, Marina A; Hixson, Kim K; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T; Stacey, Gary

    2012-11-01

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e. roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5-fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  13. Phosphoproteomic analysis of apoptotic hematopoietic stem cells from hemoglobin E/β-thalassemia

    Directory of Open Access Journals (Sweden)

    Roytrakul Sittiruk

    2011-06-01

    Full Text Available Abstract Background Hemoglobin E/β-thalassemia is particularly common in Southeast Asia and has variable symptoms ranging from mild to severe anemia. Previous investigations demonstrated the remarkable symptoms of β-thalassemia in terms of the acceleration of apoptotic cell death. Ineffective erythropoiesis has been studied in human hematopoietic stem cells, however the distinct apoptotic mechanism was unclear. Methods The phosphoproteome of bone marrow HSCs/CD34+ cells from HbE/β-thalassemic patients was analyzed using IMAC phosphoprotein isolation followed by LC-MS/MS detection. Decyder MS software was used to quantitate differentially expressed proteins in 3 patients and 2 normal donors. The differentially expressed proteins from HSCs/CD34+ cells were compared with HbE/β-thalassemia and normal HSCs. Results A significant change in abundance of 229 phosphoproteins was demonstrated. Importantly, the analysis of the candidate proteins revealed a high abundance of proteins that are commonly found in apoptotic cells including cytochrome C, caspase 6 and apoptosis inducing factors. Moreover, in the HSCs patients a significant increase was observed in a specific type of phosphoserine/threonine binding protein, which is known to act as an important signal mediator for the regulation of cell survival and apoptosis in HbE/β-thalassemia. Conclusions Our study used a novel method to investigate proteins that influence a particular pathway in a given disease or physiological condition. Ultimately, phosphoproteome profiling in HbE/β-thalassemic stem cells is an effective method to further investigate the cell death mechanism of ineffective erythropoiesis in β-thalassemia. Our report provides a comprehensive phosphoproteome, an important resource for the study of ineffective erythropoiesis and developing therapies for HbE/β-thalassemia.

  14. Phosphoproteomic analysis of the Chlamydia caviae elementary body and reticulate body forms.

    Science.gov (United States)

    Fisher, Derek J; Adams, Nancy E; Maurelli, Anthony T

    2015-08-01

    Chlamydia are Gram-negative, obligate intracellular bacteria responsible for significant diseases in humans and economically important domestic animals. These pathogens undergo a unique biphasic developmental cycle transitioning between the environmentally stable elementary body (EB) and the replicative intracellular reticulate body (RB), a conversion that appears to require extensive regulation of protein synthesis and function. However, Chlamydia possess a limited number of canonical mechanisms of transcriptional regulation. Ser/Thr/Tyr phosphorylation of proteins in bacteria has been increasingly recognized as an important mechanism of post-translational control of protein function. We utilized 2D gel electrophoresis coupled with phosphoprotein staining and MALDI-TOF/TOF analysis to map the phosphoproteome of the EB and RB forms of Chlamydia caviae. Forty-two non-redundant phosphorylated proteins were identified (some proteins were present in multiple locations within the gels). Thirty-four phosphorylated proteins were identified in EBs, including proteins found in central metabolism and protein synthesis, Chlamydia-specific hypothetical proteins and virulence-related proteins. Eleven phosphorylated proteins were identified in RBs, mostly involved in protein synthesis and folding and a single virulence-related protein. Only three phosphoproteins were found in both EB and RB phosphoproteomes. Collectively, 41 of 42 C. caviae phosphoproteins were present across Chlamydia species, consistent with the existence of a conserved chlamydial phosphoproteome. The abundance of stage-specific phosphoproteins suggests that protein phosphorylation may play a role in regulating the function of developmental-stage-specific proteins and/or may function in concert with other factors in directing EB-RB transitions.

  15. Analysis of EGFR signaling pathway in nasopharyngeal carcinoma cells by quantitative phosphoproteomics

    Directory of Open Access Journals (Sweden)

    He Qiu-Yan

    2011-06-01

    Full Text Available Abstract Background The epidermal growth factor receptor (EGFR is usually overexpressed in nasopharyngeal carcinoma (NPC and is associated with pathogenesis of NPC. However, the downstream signaling proteins of EGFR in NPC have not yet been completely understood at the system level. The aim of this study was identify novel downstream proteins of EGFR signaling pathway in NPC cells. Results We analyzed EGFR-regulated phosphoproteome in NPC CNE2 cells using 2D-DIGE and mass spectrometry analysis after phosphoprotein enrichment. As a result, 33 nonredundant phosphoproteins including five known EGFR-regulated proteins and twenty-eight novel EGFR-regulated proteins in CNE2 were identified, three differential phosphoproteins were selectively validated, and two differential phosphoproteins (GSTP1 and GRB2 were showed interacted with phospho-EGFR. Bioinformatics analysis showed that 32 of 33 identified proteins contain phosphorylation modification sites, and 17 identified proteins are signaling proteins. GSTP1, one of the EGFR-regulated proteins, associated with chemoresistance was analyzed. The results showed that GSTP1 could contribute to paclitaxel resistance in EGF-stimulated CNE2 cells. Furthermore, an EGFR signaling network based on the identified EGFR-regulated phosphoproteins were constructed using Pathway Studio 5.0 software, which includes canonical and novel EGFR-regulated proteins and implicates the possible biological roles for those proteins. Conclusion The data not only can extend our knowledge of canonical EGFR signaling, but also will be useful to understand the molecular mechanisms of EGFR in NPC pathogenesis and search therapeutic targets for NPC.

  16. Comparative gel-based phosphoproteomics in response to signaling molecules

    KAUST Repository

    Marondedze, Claudius

    2013-09-03

    The gel-based proteomics approach is a valuable technique for studying the characteristics of proteins. This technique has diverse applications ranging from analysis of a single protein to the study of the total cellular proteins. Further, protein quality and to some extent distribution can be first assessed by means of one-dimensional gel electrophoresis and then more informatively, for comparative analysis, using the two-dimensional gel electrophoresis technique. Here, we describe how to take advantage of the availability of fluorescent dyes to stain for a selective class of proteins on the same gel for the detection of both phospho- and total proteomes. This enables the co-detection of phosphoproteins as well as total proteins from the same gel and is accomplished by utilizing two different fluorescent stains, the ProQ-Diamond, which stains only phosphorylated proteins, and Sypro Ruby, which stains the entire subset of proteins. This workflow can be applied to gain insights into the regulatory mechanisms induced by signaling molecules such as cyclic nucleotides through the quantification and subsequent identification of responsive phospho- and total proteins. © Springer Science+Business Media New York 2013.

  17. Quantitative phosphoproteomic analysis reveals system-wide signaling pathways downstream of SDF-1/CXCR4 in breast cancer stem cells.

    Science.gov (United States)

    Yi, Tingfang; Zhai, Bo; Yu, Yonghao; Kiyotsugu, Yoshikawa; Raschle, Thomas; Etzkorn, Manuel; Seo, Hee-Chan; Nagiec, Michal; Luna, Rafael E; Reinherz, Ellis L; Blenis, John; Gygi, Steven P; Wagner, Gerhard

    2014-05-27

    Breast cancer is the leading cause of cancer-related mortality in women worldwide, with an estimated 1.7 million new cases and 522,000 deaths around the world in 2012 alone. Cancer stem cells (CSCs) are essential for tumor reoccurrence and metastasis which is the major source of cancer lethality. G protein-coupled receptor chemokine (C-X-C motif) receptor 4 (CXCR4) is critical for tumor metastasis. However, stromal cell-derived factor 1 (SDF-1)/CXCR4-mediated signaling pathways in breast CSCs are largely unknown. Using isotope reductive dimethylation and large-scale MS-based quantitative phosphoproteome analysis, we examined protein phosphorylation induced by SDF-1/CXCR4 signaling in breast CSCs. We quantified more than 11,000 phosphorylation sites in 2,500 phosphoproteins. Of these phosphosites, 87% were statistically unchanged in abundance in response to SDF-1/CXCR4 stimulation. In contrast, 545 phosphosites in 266 phosphoproteins were significantly increased, whereas 113 phosphosites in 74 phosphoproteins were significantly decreased. SDF-1/CXCR4 increases phosphorylation in 60 cell migration- and invasion-related proteins, of them 43 (>70%) phosphoproteins are unrecognized. In addition, SDF-1/CXCR4 upregulates the phosphorylation of 44 previously uncharacterized kinases, 8 phosphatases, and 1 endogenous phosphatase inhibitor. Using computational approaches, we performed system-based analyses examining SDF-1/CXCR4-mediated phosphoproteome, including construction of kinase-substrate network and feedback regulation loops downstream of SDF-1/CXCR4 signaling in breast CSCs. We identified a previously unidentified SDF-1/CXCR4-PKA-MAP2K2-ERK signaling pathway and demonstrated the feedback regulation on MEK, ERK1/2, δ-catenin, and PPP1Cα in SDF-1/CXCR4 signaling in breast CSCs. This study gives a system-wide view of phosphorylation events downstream of SDF-1/CXCR4 signaling in breast CSCs, providing a resource for the study of CSC-targeted cancer therapy.

  18. Quantitative Analysis of Human Pluripotency and Neural Specification by In-Depth (PhosphoProteomic Profiling

    Directory of Open Access Journals (Sweden)

    Ilyas Singec

    2016-09-01

    Full Text Available Controlled differentiation of human embryonic stem cells (hESCs can be utilized for precise analysis of cell type identities during early development. We established a highly efficient neural induction strategy and an improved analytical platform, and determined proteomic and phosphoproteomic profiles of hESCs and their specified multipotent neural stem cell derivatives (hNSCs. This quantitative dataset (nearly 13,000 proteins and 60,000 phosphorylation sites provides unique molecular insights into pluripotency and neural lineage entry. Systems-level comparative analysis of proteins (e.g., transcription factors, epigenetic regulators, kinase families, phosphorylation sites, and numerous biological pathways allowed the identification of distinct signatures in pluripotent and multipotent cells. Furthermore, as predicted by the dataset, we functionally validated an autocrine/paracrine mechanism by demonstrating that the secreted protein midkine is a regulator of neural specification. This resource is freely available to the scientific community, including a searchable website, PluriProt.

  19. Phosphoproteomic Analysis of the Highly-Metastatic Hepatocellular Carcinoma Cell Line, MHCC97-H

    Directory of Open Access Journals (Sweden)

    Miaomiao Tian

    2015-02-01

    Full Text Available Invasion and metastasis of hepatocellular carcinoma (HCC is a major cause for lethal liver cancer. Signaling pathways associated with cancer progression are frequently reconfigured by aberrant phosphorylation of key proteins. To capture the key phosphorylation events in HCC metastasis, we established a methodology by an off-line high-pH HPLC separation strategy combined with multi-step IMAC and LC–MS/MS to study the phosphoproteome of a metastatic HCC cell line, MHCC97-H (high metastasis. In total, 6593 phosphopeptides with 6420 phosphorylation sites (p-sites of 2930 phosphoproteins were identified. Statistical analysis of gene ontology (GO categories for the identified phosphoproteins showed that several of the biological processes, such as transcriptional regulation, mRNA processing and RNA splicing, were over-represented. Further analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG annotations demonstrated that phosphoproteins in multiple pathways, such as spliceosome, the insulin signaling pathway and the cell cycle, were significantly enriched. In particular, we compared our dataset with a previously published phosphoproteome in a normal liver sample, and the results revealed that a number of proteins in the spliceosome pathway, such as U2 small nuclear RNA Auxiliary Factor 2 (U2AF2, Eukaryotic Initiation Factor 4A-III (EIF4A3, Cell Division Cycle 5-Like (CDC5L and Survival Motor Neuron Domain Containing 1 (SMNDC1, were exclusively identified as phosphoproteins only in the MHCC97-H cell line. These results indicated that the phosphorylation of spliceosome proteins may participate in the metastasis of HCC by regulating mRNA processing and RNA splicing.

  20. The Arabidopsis thaliana Cyclic-Nucleotide-Dependent Response – a Quantitative Proteomic and Phosphoproteomic Analysis

    KAUST Repository

    Alqurashi, May M.

    2013-11-01

    Protein phosphorylation governs many regulatory pathways and an increasing number of kinases, proteins that transfer phosphate groups, are in turn activated by cyclic nucleotides. One of the cyclic nucleotides, cyclic adenosine monophosphate (cAMP), has been shown to be a second messenger in abiotic and biotic stress responses. However, little is known about the precise role of cAMP in plants and in the down-stream activation of kinases, and hence cAMP-dependent phosphorylation. To increase our understanding of the role of cAMP, proteomic and phosphoproteomic profiles of Arabidopsis thaliana suspension culture cells were analyzed before and after treatment of cells with two different concentrations of 8-Bromo-cAMP (1 µM and 100 nM) and over a time-course of one hour. A comparative quantitative analysis was undertaken using two- dimensional gel electrophoresis and the Delta 2D software (DECODON) followed by protein spot identification by tandem mass spectrometry combined with Mascot and Scaffold. Differentially expressed proteins and regulated phosphoproteins were categorized according to their biological function using bioinformatics tools. The results revealed that the treatment with 1 µM and 100 nM 8-Bromo-cAMP was sufficient to induce specific concentration- and time-dependent changes at the proteome and phosphoproteome levels. In particular, different phosphorylation patterns were observed overtime preferentially affecting proteins in a number of functional categories, notably phosphatases, proteins that remove phosphate groups. This suggests that cAMP both transiently activates and deactivates proteins through specific phosphorylation events and provides new insight into biological mechanisms and functions at the systems level.

  1. Nuclear phosphoproteome analysis of 3T3-L1 preadipocyte differentiation reveals system-wide phosphorylation of transcriptional regulators

    DEFF Research Database (Denmark)

    Rabiee, Atefeh; Schwämmle, Veit; Sidoli, Simone;

    2017-01-01

    HIGHLIGHTS: Mass spectrometry (MS) based quantitative proteomics and phosphoproteomics applied to monitor the alteration of nuclear proteins during the early stages (4 hours) of preadipocyte differentiation. A total of 4072 proteins including 2434 phosphorylated proteins identified, a majority......), in particular phosphorylation, play a major role in activating and propagating signals within TR networks upon induction of adipogenesis by extracellular stimulus. We applied mass spectrometry (MS) based quantitative proteomics and phosphoproteomics to monitor the alteration of nuclear proteins during the early....... New insights into phosphorylation-dependent signaling networks that impact on nuclear proteins and controls adipocyte differentiation and cell fate. Adipocytes (fat cells) are important endocrine and metabolic cells critical for systemic insulin sensitivity. Both adipose excess and insufficiency...

  2. Phosphoproteome analysis of functional mitochondria isolated from resting human muscle reveals extensive phosphorylation of inner membrane protein complexes and enzymes

    DEFF Research Database (Denmark)

    Zhao, Xiaolu; Leon, Ileana R; Bak, Steffen;

    2011-01-01

    for protein kinase A, protein kinase C, casein kinase II and DNA-dependent protein kinase. Our results demonstrate the feasibility of performing phosphoproteome analysis of organelles isolated from human tissue and provide novel targets for functional studies of reversible phosphorylation in mitochondria......Mitochondria play a central role in energy metabolism and cellular survival, and consequently mitochondrial dysfunction is associated with a number of human pathologies. Reversible protein phosphorylation emerges as a central mechanism in the regulation of several mitochondrial processes....... In skeletal muscle, mitochondrial dysfunction is linked to insulin resistance in humans with obesity and type 2 diabetes. We performed a phosphoproteomic study of functional mitochondria isolated from human muscle biopsies with the aim to obtain a comprehensive overview of mitochondrial phosphoproteins...

  3. Analytical strategies for phosphoproteomics

    DEFF Research Database (Denmark)

    Thingholm, Tine E; Jensen, Ole N; Larsen, Martin R

    2009-01-01

    sensitive and specific strategies. Today, most phosphoproteomic studies are conducted by mass spectrometric strategies in combination with phospho-specific enrichment methods. This review presents an overview of different analytical strategies for the characterization of phosphoproteins. Emphasis...... will be on the affinity methods utilized specifically for phosphoprotein and phosphopeptide enrichment prior to MS analysis, and on recent applications of these methods in cell biological applications....

  4. Phosphoproteomics in cereals.

    Science.gov (United States)

    Yang, Pingfang

    2015-01-01

    Cereals are the most important crop plant supplying staple food throughout the world. The economic importance and continued breeding of crop plants such as rice, maize, wheat, or barley require a detailed scientific understanding of adaptive and developmental processes. Protein phosphorylation is one of the most important regulatory posttranslational modifications and its analysis allows deriving functional and regulatory principles in plants. This minireview summarizes the current knowledge of phosphoproteomic studies in cereals.

  5. Motif-specific sampling of phosphoproteomes.

    Science.gov (United States)

    Ruse, Cristian I; McClatchy, Daniel B; Lu, Bingwen; Cociorva, Daniel; Motoyama, Akira; Park, Sung Kyu; Yates, John R

    2008-05-01

    Phosphoproteomics, the targeted study of a subfraction of the proteome which is modified by phosphorylation, has become an indispensable tool to study cell signaling dynamics. We described a methodology that linked phosphoproteome and proteome analysis based on Ba2+ binding properties of amino acids. This technology selected motif-specific phosphopeptides independent of the system under analysis. MudPIT (Multidimensional Identification Technology) identified 1037 precipitated phosphopeptides from as little as 250 microg of proteins. To extend coverage of the phosphoproteome, we sampled the nuclear extract of HeLa cells with three values of Ba2+ ions molarity. The presence of more than 70% of identified phosphoproteins was further substantiated by their nonmodified peptides. Upon isoproterenol stimulation of HEK cells, we identified an increasing number of phosphoproteins from MAPK cascades and AKAP signaling hubs. We quantified changes in both protein and phosphorylation levels of 197 phosphoproteins including a critical kinase, MAPK1. Integration of differential phosphorylation of MAPK1 with knowledge bases constructed modules that correlated well with its role as node in cross-talk of canonical pathways.

  6. Phosphoproteome Analysis Reveals the Molecular Mechanisms Underlying Deoxynivalenol-Induced Intestinal Toxicity in IPEC-J2 Cells

    Science.gov (United States)

    Zhang, Zhi-Qi; Wang, Song-Bo; Wang, Rui-Guo; Zhang, Wei; Wang, Pei-Long; Su, Xiao-Ou

    2016-01-01

    Deoxynivalenol (DON) is a widespread trichothecene mycotoxin that commonly contaminates cereal crops and has various toxic effects in animals and humans. DON primarily targets the gastrointestinal tract, the first barrier against ingested food contaminants. In this study, an isobaric tag for relative and absolute quantitation (iTRAQ)-based phosphoproteomic approach was employed to elucidate the molecular mechanisms underlying DON-mediated intestinal toxicity in porcine epithelial cells (IPEC-J2) exposed to 20 μM DON for 60 min. There were 4153 unique phosphopeptides, representing 389 phosphorylation sites, detected in 1821 phosphoproteins. We found that 289 phosphopeptides corresponding to 255 phosphoproteins were differentially phosphorylated in response to DON. Comprehensive Gene Ontology (GO) analysis combined with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment revealed that, in addition to previously well-characterized mitogen-activated protein kinase (MAPK) signaling, DON exposure altered phosphatidylinositol 3-kinase/Akt (PI3K/Akt) and Janus kinase/signal transducer, and activator of transcription (JAK/STAT) pathways. These pathways are involved in a wide range of biological processes, including apoptosis, the intestinal barrier, intestinal inflammation, and the intestinal absorption of glucose. DON-induced changes are likely to contribute to the intestinal dysfunction. Overall, identification of relevant signaling pathways yielded new insights into the molecular mechanisms underlying DON-induced intestinal toxicity, and might help in the development of improved mechanism-based risk assessments in animals and humans. PMID:27669298

  7. Quantitative Phosphoproteomic Analysis of Arabidopsis in Response to Salt and Hydrogen Peroxide Stresses

    Institute of Scientific and Technical Information of China (English)

    Yanmei Chen

    2012-01-01

    Salinity and oxidative stresses are major factors in affecting and limiting the productivity of agricultural crops.The study of biochemical and molecular responses of plants in response to those stresses is important for crop genetics and breeding.Extensive evidence shows that reversible protein phosphorylation plays a central role in mediating stress-regulated physiological responses,but little is known about its extent and function.Mass spectrometry provides a powerful tool for the in-depth analysis of systems biology.In this study,we performed a global quantitative analysis of the Arabidopsis phosphoproteomics in response to a time course of stress treatments using 15N-metabolic labeling and subcellular fractionation approaches.In total,we found 176 phosphoproteins showed to be regulated under stresses.Nine SnRK2 kinases identified to be differentially phosphorylated at multiple serine/threonine residues in their kinase domains following stress treatments,demonstrating different temporal phosphorylation induction of the various isoforms.K+ and Na+ transporters showed coordinated phosphorylation regulation under salt stress.In particular,nuclear proteins and protein kinases have high phosphorylation site occupancy in response to stress treatment.This suggests that the wide range of signaling and cellular processes that are modulated in this study.

  8. An automated platform for analysis of phosphoproteomic datasets: application to kidney collecting duct phosphoproteins.

    Science.gov (United States)

    Hoffert, Jason D; Wang, Guanghui; Pisitkun, Trairak; Shen, Rong-Fong; Knepper, Mark A

    2007-09-01

    Large-scale phosphoproteomic analysis employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) often requires a significant amount of manual manipulation of phosphopeptide datasets in the post-acquisition phase. To assist in this process, we have created software, PhosphoPIC (PhosphoPeptide Identification and Compilation), which can perform a variety of useful functions including automated selection and compilation of phosphopeptide identifications from multiple MS levels, estimation of dataset false discovery rate, and application of appropriate cross-correlation (XCorr) filters. In addition, the output files generated by this program are compatible with downstream phosphorylation site assignment using the Ascore algorithm, as well as phosphopeptide quantification via QUOIL. In this report, we utilized this software to analyze phosphoproteins from short-term vasopressin-treated rat kidney inner medullary collecting duct (IMCD). A total of 925 phosphopeptides representing 173 unique proteins were identified from membrane-enriched fractions of IMCD with a false discovery rate of 1.5%. Of these proteins, 106 were found only in the membrane-enriched fraction of IMCD cells and not in whole IMCD cell lysates. These identifications included a number of well-studied ion and solute transporters including ClC-1, LAT4, MCT2, NBC3, and NHE1, all of which contained novel phosphorylation sites. Using a label-free quantification approach, we identified phosphoproteins that changed in abundance with vasopressin exposure including aquaporin-2 (AQP2), Hnrpa3, IP3 receptor 3, and pur-beta.

  9. Ultradeep Human Phosphoproteome Reveals a Distinct Regulatory Nature of Tyr and Ser/Thr-Based Signaling

    Directory of Open Access Journals (Sweden)

    Kirti Sharma

    2014-09-01

    Full Text Available Regulatory protein phosphorylation controls normal and pathophysiological signaling in eukaryotic cells. Despite great advances in mass-spectrometry-based proteomics, the extent, localization, and site-specific stoichiometry of this posttranslational modification (PTM are unknown. Here, we develop a stringent experimental and computational workflow, capable of mapping more than 50,000 distinct phosphorylated peptides in a single human cancer cell line. We detected more than three-quarters of cellular proteins as phosphoproteins and determined very high stoichiometries in mitosis or growth factor signaling by label-free quantitation. The proportion of phospho-Tyr drastically decreases as coverage of the phosphoproteome increases, whereas Ser/Thr sites saturate only for technical reasons. Tyrosine phosphorylation is maintained at especially low stoichiometric levels in the absence of specific signaling events. Unexpectedly, it is enriched on higher-abundance proteins, and this correlates with the substrate KM values of tyrosine kinases. Our data suggest that P-Tyr should be considered a functionally separate PTM of eukaryotic proteomes.

  10. Phosphoproteomic analysis of protein kinase C signaling in Saccharomyces cerevisiae reveals Slt2 mitogen-activated protein kinase (MAPK)-dependent phosphorylation of eisosome core components.

    Science.gov (United States)

    Mascaraque, Victoria; Hernáez, María Luisa; Jiménez-Sánchez, María; Hansen, Rasmus; Gil, Concha; Martín, Humberto; Cid, Víctor J; Molina, María

    2013-03-01

    The cell wall integrity (CWI) pathway of the model organism Saccharomyces cerevisiae has been thoroughly studied as a paradigm of the mitogen-activated protein kinase (MAPK) pathway. It consists of a classic MAPK module comprising the Bck1 MAPK kinase kinase, two redundant MAPK kinases (Mkk1 and Mkk2), and the Slt2 MAPK. This module is activated under a variety of stimuli related to cell wall homeostasis by Pkc1, the only member of the protein kinase C family in budding yeast. Quantitative phosphoproteomics based on stable isotope labeling of amino acids in cell culture is a powerful tool for globally studying protein phosphorylation. Here we report an analysis of the yeast phosphoproteome upon overexpression of a PKC1 hyperactive allele that specifically activates CWI MAPK signaling in the absence of external stimuli. We found 82 phosphopeptides originating from 43 proteins that showed enhanced phosphorylation in these conditions. The MAPK S/T-P target motif was significantly overrepresented in these phosphopeptides. Hyperphosphorylated proteins provide putative novel targets of the Pkc1-cell wall integrity pathway involved in diverse functions such as the control of gene expression, protein synthesis, cytoskeleton maintenance, DNA repair, and metabolism. Remarkably, five components of the plasma-membrane-associated protein complex known as eisosomes were found among the up-regulated proteins. We show here that Pkc1-induced phosphorylation of the eisosome core components Pil1 and Lsp1 was not exerted directly by Pkc1, but involved signaling through the Slt2 MAPK module.

  11. Refined phosphopeptide enrichment by phosphate additive and the analysis of human brain phosphoproteome.

    Science.gov (United States)

    Tan, Haiyan; Wu, Zhiping; Wang, Hong; Bai, Bing; Li, Yuxin; Wang, Xusheng; Zhai, Bo; Beach, Thomas G; Peng, Junmin

    2015-01-01

    Alzheimer's disease (AD) is the most common form of dementia, characterized by progressive loss of cognitive function. One of the pathological hallmarks of AD is the formation of neurofibrillary tangles composed of abnormally hyperphosphorylated tau protein, but global deregulation of protein phosphorylation in AD is not well analyzed. Here, we report a pilot investigation of AD phosphoproteome by titanium dioxide enrichment coupled with high resolution LC-MS/MS. During the optimization of the enrichment method, we found that phosphate ion at a low concentration (e.g. 1 mM) worked efficiently as a nonphosphopeptide competitor to reduce background. The procedure was further tuned with respect to peptide-to-bead ratio, phosphopeptide recovery, and purity. Using this refined method and 9 h LC-MS/MS, we analyzed phosphoproteome in one milligram of digested AD brain lysate, identifying 5243 phosphopeptides containing 3715 nonredundant phosphosites on 1455 proteins, including 31 phosphosites on the tau protein. This modified enrichment method is simple and highly efficient. The AD case study demonstrates its feasibility of dissecting phosphoproteome in a limited amount of postmortem human brain. All MS data have been deposited in the ProteomeXchange with identifier PXD001180 (http://proteomecentral.proteomexchange.org/dataset/PXD001180).

  12. Quantitative phosphoproteomic analysis of early seed development in rice (Oryza sativa L.).

    Science.gov (United States)

    Qiu, Jiehua; Hou, Yuxuan; Tong, Xiaohong; Wang, Yifeng; Lin, Haiyan; Liu, Qing; Zhang, Wen; Li, Zhiyong; Nallamilli, Babi R; Zhang, Jian

    2016-02-01

    Rice (Oryza sativa L.) seed serves as a major food source for over half of the global population. Though it has been long recognized that phosphorylation plays an essential role in rice seed development, the phosphorylation events and dynamics in this process remain largely unknown so far. Here, we report the first large scale identification of rice seed phosphoproteins and phosphosites by using a quantitative phosphoproteomic approach. Thorough proteomic studies in pistils and seeds at 3, 7 days after pollination resulted in the successful identification of 3885, 4313 and 4135 phosphopeptides respectively. A total of 2487 proteins were differentially phosphorylated among the three stages, including Kip related protein 1, Rice basic leucine zipper factor 1, Rice prolamin box binding factor and numerous other master regulators of rice seed development. Moreover, differentially phosphorylated proteins may be extensively involved in the biosynthesis and signaling pathways of phytohormones such as auxin, gibberellin, abscisic acid and brassinosteroid. Our results strongly indicated that protein phosphorylation is a key mechanism regulating cell proliferation and enlargement, phytohormone biosynthesis and signaling, grain filling and grain quality during rice seed development. Overall, the current study enhanced our understanding of the rice phosphoproteome and shed novel insight into the regulatory mechanism of rice seed development.

  13. Reconstruction of Insulin Signal Flow from Phosphoproteome and Metabolome Data

    Directory of Open Access Journals (Sweden)

    Katsuyuki Yugi

    2014-08-01

    Full Text Available Cellular homeostasis is regulated by signals through multiple molecular networks that include protein phosphorylation and metabolites. However, where and when the signal flows through a network and regulates homeostasis has not been explored. We have developed a reconstruction method for the signal flow based on time-course phosphoproteome and metabolome data, using multiple databases, and have applied it to acute action of insulin, an important hormone for metabolic homeostasis. An insulin signal flows through a network, through signaling pathways that involve 13 protein kinases, 26 phosphorylated metabolic enzymes, and 35 allosteric effectors, resulting in quantitative changes in 44 metabolites. Analysis of the network reveals that insulin induces phosphorylation and activation of liver-type phosphofructokinase 1, thereby controlling a key reaction in glycolysis. We thus provide a versatile method of reconstruction of signal flow through the network using phosphoproteome and metabolome data.

  14. Elucidating the CXCL12/CXCR4 signaling network in chronic lymphocytic leukemia through phosphoproteomics analysis.

    Directory of Open Access Journals (Sweden)

    Morgan O'Hayre

    Full Text Available BACKGROUND: Chronic Lymphocytic Leukemia (CLL pathogenesis has been linked to the prolonged survival and/or apoptotic resistance of leukemic B cells in vivo, and is thought to be due to enhanced survival signaling responses to environmental factors that protect CLL cells from spontaneous and chemotherapy-induced death. Although normally associated with cell migration, the chemokine, CXCL12, is one of the factors known to support the survival of CLL cells. Thus, the signaling pathways activated by CXCL12 and its receptor, CXCR4, were investigated as components of these pathways and may represent targets that if inhibited, could render resistant CLL cells more susceptible to chemotherapy. METHODOLOGY/PRINCIPAL FINDINGS: To determine the downstream signaling targets that contribute to the survival effects of CXCL12 in CLL, we took a phosphoproteomics approach to identify and compare phosphopeptides in unstimulated and CXCL12-stimulated primary CLL cells. While some of the survival pathways activated by CXCL12 in CLL are known, including Akt and ERK1/2, this approach enabled the identification of additional signaling targets and novel phosphoproteins that could have implications in CLL disease and therapy. In addition to the phosphoproteomics results, we provide evidence from western blot validation that the tumor suppressor, programmed cell death factor 4 (PDCD4, is a previously unidentified phosphorylation target of CXCL12 signaling in all CLL cells probed. Additionally, heat shock protein 27 (HSP27, which mediates anti-apoptotic signaling and has previously been linked to chemotherapeutic resistance, was detected in a subset (approximately 25% of CLL patients cells examined. CONCLUSIONS/SIGNIFICANCE: Since PDCD4 and HSP27 have previously been associated with cancer and regulation of cell growth and apoptosis, these proteins may have novel implications in CLL cell survival and represent potential therapeutic targets. PDCD4 also represents a

  15. In-depth phosphoproteomic analysis of royal jelly derived from western and eastern honeybee species.

    Science.gov (United States)

    Han, Bin; Fang, Yu; Feng, Mao; Lu, Xiaoshan; Huo, Xinmei; Meng, Lifeng; Wu, Bin; Li, Jianke

    2014-12-01

    The proteins in royal jelly (RJ) play a pivotal role in the nutrition, immune defense, and cast determination of honeybee larvae and have a wide range of pharmacological and health-promoting functions for humans as well. Although the importance of post-translational modifications (PTMs) in protein function is known, investigation of protein phosphorylation of RJ proteins is still very limited. To this end, two complementary phosphopeptide enrichment materials (Ti(4+)-IMAC and TiO2) and high-sensitivity mass spectrometry were applied to establish a detailed phosphoproteome map and to qualitatively and quantitatively compare the phosphoproteomes of RJ produced by Apis mellifera ligustica (Aml) and Apis cerana cerana (Acc). In total, 16 phosphoproteins carrying 67 phosphorylation sites were identified in RJ derived from western bees, and nine proteins phosphorylated on 71 sites were found in RJ produced by eastern honeybees. Of which, eight phosphorylated proteins were common to both RJ samples, and the same motif ([S-x-E]) was extracted, suggesting that the function of major RJ proteins as nutrients and immune agents is evolutionary preserved in both of these honeybee species. All eight overlapping phosphoproteins showed significantly higher abundance in Acc-RJ than in Aml-RJ, and the phosphorylation of Jelleine-II (an antimicrobial peptide, TPFKLSLHL) at S(6) in Acc-RJ had stronger antimicrobial properties than that at T(1) in Aml-RJ even though the overall antimicrobial activity of Jelleine-II was found to decrease after phosphorylation. The differences in phosphosites, peptide abundance, and antimicrobial activity of the phosphorylated RJ proteins indicate that the two major honeybee species employ distinct phosphorylation strategies that align with their different biological characteristics shaped by evolution. The phosphorylation of RJ proteins are potentially driven by the activity of extracellular serine/threonine protein kinase FAM20C-like protein (FAM20C

  16. Analysis of T4SS-induced signaling by H. pylori using quantitative phosphoproteomics

    Directory of Open Access Journals (Sweden)

    Frithjof eGlowinski

    2014-07-01

    Full Text Available Helicobacter pylori is a Gram-negative bacterial pathogen colonizing the human stomach. Infection with H. pylori causes chronic inflammation of the gastric mucosa and may lead to peptic ulceration and/or gastric cancer. A major virulence determinant of H. pylori is the type IV secretion system (T4SS, which is used to inject the virulence factor CagA into the host cell, triggering a wide range of cellular signaling events. Here, we used a phosphoproteomic approach to investigate tyrosine signaling in response to host-pathogen interaction, using stable isotope labeling in cell culture (SILAC of AGS cells to obtain a differential picture between multiple infection conditions. Cells were infected with wild type H. pylori P12, a P12ΔCagA deletion mutant, and a P12ΔT4SS deletion mutant to compare signaling changes over time and in the absence of CagA or the T4SS. Tryptic peptides were enriched for tyrosine (Tyr phosphopeptides and analysed by nano-LC-Orbitrap MS. In total, 58 different phosphosites were found to be regulated following infection. The majority of phosphosites identified were kinases of the MAPK familiy. CagA and the T4SS were found to be key regulators of Tyr phosphosites. Our findings indicate that CagA primarily induces activation of ERK1 and integrin linked factors, whereas the T4SS primarily modulates JNK and p38 activation.

  17. Phosphoproteomic analysis reveals interconnected system-wide responses to perturbations of kinases and phosphatases in yeast.

    Science.gov (United States)

    Bodenmiller, Bernd; Wanka, Stefanie; Kraft, Claudine; Urban, Jörg; Campbell, David; Pedrioli, Patrick G; Gerrits, Bertran; Picotti, Paola; Lam, Henry; Vitek, Olga; Brusniak, Mi-Youn; Roschitzki, Bernd; Zhang, Chao; Shokat, Kevan M; Schlapbach, Ralph; Colman-Lerner, Alejandro; Nolan, Garry P; Nesvizhskii, Alexey I; Peter, Matthias; Loewith, Robbie; von Mering, Christian; Aebersold, Ruedi

    2010-12-21

    The phosphorylation and dephosphorylation of proteins by kinases and phosphatases constitute an essential regulatory network in eukaryotic cells. This network supports the flow of information from sensors through signaling systems to effector molecules and ultimately drives the phenotype and function of cells, tissues, and organisms. Dysregulation of this process has severe consequences and is one of the main factors in the emergence and progression of diseases, including cancer. Thus, major efforts have been invested in developing specific inhibitors that modulate the activity of individual kinases or phosphatases; however, it has been difficult to assess how such pharmacological interventions would affect the cellular signaling network as a whole. Here, we used label-free, quantitative phosphoproteomics in a systematically perturbed model organism (Saccharomyces cerevisiae) to determine the relationships between 97 kinases, 27 phosphatases, and more than 1000 phosphoproteins. We identified 8814 regulated phosphorylation events, describing the first system-wide protein phosphorylation network in vivo. Our results show that, at steady state, inactivation of most kinases and phosphatases affected large parts of the phosphorylation-modulated signal transduction machinery-and not only the immediate downstream targets. The observed cellular growth phenotype was often well maintained despite the perturbations, arguing for considerable robustness in the system. Our results serve to constrain future models of cellular signaling and reinforce the idea that simple linear representations of signaling pathways might be insufficient for drug development and for describing organismal homeostasis.

  18. Quantitative phosphoproteomics to characterize signaling networks

    DEFF Research Database (Denmark)

    Rigbolt, Kristoffer T G; Blagoev, Blagoy

    2012-01-01

    and quantify thousands of phosphorylations, thus providing extensive overviews of the cellular signaling networks. As a result of these developments quantitative phosphoproteomics have been applied to study processes as diverse as immunology, stem cell biology and DNA damage. Here we review the developments......Reversible protein phosphorylation is involved in the regulation of most, if not all, major cellular processes via dynamic signal transduction pathways. During the last decade quantitative phosphoproteomics have evolved from a highly specialized area to a powerful and versatile platform...... for analyzing protein phosphorylation at a system-wide scale and has become the intuitive strategy for comprehensive characterization of signaling networks. Contemporary phosphoproteomics use highly optimized procedures for sample preparation, mass spectrometry and data analysis algorithms to identify...

  19. Phosphoproteome analysis demonstrates the potential role of THRAP3 phosphorylation in androgen-independent prostate cancer cell growth.

    Science.gov (United States)

    Ino, Yoko; Arakawa, Noriaki; Ishiguro, Hitoshi; Uemura, Hiroji; Kubota, Yoshinobu; Hirano, Hisashi; Toda, Tosifusa

    2016-04-01

    Elucidating the androgen-independent growth mechanism is critical for developing effective treatment strategies to combat androgen-independent prostate cancer. We performed a comparative phosphoproteome analysis using a prostate cancer cell line, LNCaP, and an LNCaP-derived androgen-independent cell line, LNCaP-AI, to identify phosphoproteins involved in this mechanism. We performed quantitative comparisons of the phosphopeptide levels in tryptic digests of protein extracts from these cell lines using MS. We found that the levels of 69 phosphopeptides in 66 proteins significantly differed between LNCaP and LNCaP-AI. In particular, we focused on thyroid hormone receptor associated protein 3 (THRAP3), which is a known transcriptional coactivator of the androgen receptor. The phosphorylation level of THRAP3 was significantly lower at S248 and S253 in LNCaP-AI cells. Furthermore, pull-down assays showed that 32 proteins uniquely bound to the nonphosphorylatable mutant form of THRAP3, whereas 31 other proteins uniquely bound to the phosphorylation-mimic form. Many of the differentially interacting proteins were identified as being involved with RNA splicing and processing. These results suggest that the phosphorylation state of THRAP3 at S248 and S253 might be involved in the mechanism of androgen-independent prostate cancer cell growth by changing the interaction partners.

  20. Identifying differentially regulated subnetworks from phosphoproteomic data

    Directory of Open Access Journals (Sweden)

    Tebbe Andreas

    2010-06-01

    Full Text Available Abstract Background Various high throughput methods are available for detecting regulations at the level of transcription, translation or posttranslation (e.g. phosphorylation. Integrating these data with protein networks should make it possible to identify subnetworks that are significantly regulated. Furthermore, such integration can support identification of regulated entities from often noisy high throughput data. In particular, processing mass spectrometry-based phosphoproteomic data in this manner may expose signal transduction pathways and, in the case of experiments with drug-treated cells, reveal the drug's mode of action. Results Here, we introduce SubExtractor, an algorithm that combines phosphoproteomic data with protein network information from STRING to identify differentially regulated subnetworks and individual proteins. The method is based on a Bayesian probabilistic model combined with a genetic algorithm and rigorous significance testing. The Bayesian model accounts for information about both differential regulation and network topology. The method was tested with artificial data and subsequently applied to a comprehensive phosphoproteomics study investigating the mode of action of sorafenib, a small molecule kinase inhibitor. Conclusions SubExtractor reliably identifies differentially regulated subnetworks from phosphoproteomic data by integrating protein networks. The method can also be applied to gene or protein expression data.

  1. Phosphoproteomic Analysis of KSHV-Infected Cells Reveals Roles of ORF45-Activated RSK during Lytic Replication.

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    Denis Avey

    2015-07-01

    Full Text Available Kaposi's Sarcoma-Associated Herpesvirus (KSHV is an oncogenic virus which has adapted unique mechanisms to modulate the cellular microenvironment of its human host. The pathogenesis of KSHV is intimately linked to its manipulation of cellular signaling pathways, including the extracellular signal-regulated kinase (ERK mitogen-activated protein kinase (MAPK pathway. We have previously shown that KSHV ORF45 contributes to the sustained activation of both ERK and p90 ribosomal S6 kinase (RSK, a major functional mediator of ERK/MAPK signaling during KSHV lytic replication. ORF45-activated RSK is required for optimal KSHV lytic gene expression and progeny virion production, though the underlying mechanisms downstream of this activation are still unclear. We hypothesized that the activation of RSK by ORF45 causes differential phosphorylation of cellular and viral substrates, affecting biological processes essential for efficient KSHV lytic replication. Accordingly, we observed widespread and significant differences in protein phosphorylation upon induction of lytic replication. Mass-spectrometry-based phosphoproteomic screening identified putative substrates of ORF45-activated RSK in KSHV-infected cells. Bioinformatic analyses revealed that nuclear proteins, including several transcriptional regulators, were overrepresented among these candidates. We validated the ORF45/RSK-dependent phosphorylation of several putative substrates by employing KSHV BAC mutagenesis, kinase inhibitor treatments, and/or CRISPR-mediated knockout of RSK in KSHV-infected cells. Furthermore, we assessed the consequences of knocking out these substrates on ORF45/RSK-dependent regulation of gene expression and KSHV progeny virion production. Finally, we show data to support that ORF45 regulates the translational efficiency of a subset of viral/cellular genes with complex secondary structure in their 5' UTR. Altogether, these data shed light on the mechanisms by which KSHV ORF45

  2. Targeted quantitative phosphoproteomic analysis of erythrocyte membranes during blood bank storage.

    Science.gov (United States)

    Rinalducci, Sara; Longo, Valentina; Ceci, Luigi R; Zolla, Lello

    2015-02-01

    One of the hallmarks of blood bank stored red blood cells (RBCs) is the irreversible transition from a discoid to a spherocyte-like morphology with membrane perturbation and cytoskeleton disorders. Therefore, identification of the storage-associated modifications in the protein-protein interactions between the cytoskeleton and the lipid bilayer may contribute to enlighten the molecular mechanisms involved in the alterations of mechanical properties of stored RBCs. Here we report the results obtained analyzing RBCs after 0, 21 and 35 days of storage under standard blood banking conditions by label free mass spectrometry (MS)-based experiments. We could quantitatively measure changes in the phosphorylation level of crucial phosphopeptides belonging to β-spectrin, ankyrin-1, α-adducin, dematin, glycophorin A and glycophorin C proteins. Data have been validated by both western blotting and pseudo-Multiple Reaction Monitoring (MRM). Although each phosphopeptide showed a distinctive trend, a sharp increase in the phosphorylation level during the storage duration was observed. Phosphopeptide mapping and structural modeling analysis indicated that the phosphorylated residues localize in protein functional domains fundamental for the maintenance of membrane structural integrity. Along with previous morphological evidence acquired by electron microscopy, our results seem to indicate that 21-day storage may represent a key point for the molecular processes leading to the erythrocyte deformability reduction observed during blood storage. These findings could therefore be helpful in understanding and preventing the morphology-linked mechanisms responsible for the post-transfusion survival of preserved RBCs.

  3. Quantitative phosphoproteomics of CXCL12 (SDF-1 signaling.

    Directory of Open Access Journals (Sweden)

    Jason A Wojcechowskyj

    Full Text Available CXCL12 (SDF-1 is a chemokine that binds to and signals through the seven transmembrane receptor CXCR4. The CXCL12/CXCR4 signaling axis has been implicated in both cancer metastases and human immunodeficiency virus type 1 (HIV-1 infection and a more complete understanding of CXCL12/CXCR4 signaling pathways may support efforts to develop therapeutics for these diseases. Mass spectrometry-based phosphoproteomics has emerged as an important tool in studying signaling networks in an unbiased fashion. We employed stable isotope labeling with amino acids in cell culture (SILAC quantitative phosphoproteomics to examine the CXCL12/CXCR4 signaling axis in the human lymphoblastic CEM cell line. We quantified 4,074 unique SILAC pairs from 1,673 proteins and 89 phosphopeptides were deemed CXCL12-responsive in biological replicates. Several well established CXCL12-responsive phosphosites such as AKT (pS473 and ERK2 (pY204 were confirmed in our study. We also validated two novel CXCL12-responsive phosphosites, stathmin (pS16 and AKT1S1 (pT246 by Western blot. Pathway analysis and comparisons with other phosphoproteomic datasets revealed that genes from CXCL12-responsive phosphosites are enriched for cellular pathways such as T cell activation, epidermal growth factor and mammalian target of rapamycin (mTOR signaling, pathways which have previously been linked to CXCL12/CXCR4 signaling. Several of the novel CXCL12-responsive phosphoproteins from our study have also been implicated with cellular migration and HIV-1 infection, thus providing an attractive list of potential targets for the development of cancer metastasis and HIV-1 therapeutics and for furthering our understanding of chemokine signaling regulation by reversible phosphorylation.

  4. Improving the Phosphoproteome Coverage for Limited Sample Amounts Using TiO2-SIMAC-HILIC (TiSH) Phosphopeptide Enrichment and Fractionation.

    Science.gov (United States)

    Engholm-Keller, Kasper; Larsen, Martin R

    2016-01-01

    Obtaining high phosphoproteome coverage requires specific enrichment of phosphorylated peptides from the often extremely complex peptide mixtures generated by proteolytic digestion of biological samples, as well as extensive chromatographic fractionation prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Due to the sample loss resulting from fractionation, this procedure is mainly performed when large quantities of sample are available. To make large-scale phosphoproteomics applicable to smaller amounts of protein we have recently combined highly specific TiO2-based phosphopeptide enrichment with sequential elution from immobilized metal affinity chromatography (SIMAC) for fractionation of mono- and multi-phosphorylated peptides prior to capillary scale hydrophilic interaction liquid chromatography (HILIC) based fractionation of monophosphorylated peptides. In the following protocol we describe the procedure step by step to allow for comprehensive coverage of the phosphoproteome utilizing only a few hundred micrograms of protein.

  5. Recent findings and technological advances in phosphoproteomics for cells and tissues

    DEFF Research Database (Denmark)

    von Stechow, Louise; Francavilla, Chiara; Olsen, Jesper V

    2015-01-01

    in different diseases, including cancer. Large-scale studies of phosphoproteins - termed phosphoproteomics - strongly rely on the use of high-performance mass spectrometric instrumentation. This powerful technology has been applied to study a great number of phosphorylation-based phenotypes. Nevertheless, many...... technical and biological challenges have to be overcome to identify biologically relevant phosphorylation sites in cells and tissues. This review describes different technological strategies to identify and quantify phosphorylation sites with high accuracy, without significant loss of analysis speed...

  6. In Vivo Phosphoproteomics Analysis Reveals the Cardiac Targets of β-Adrenergic Receptor Signaling

    DEFF Research Database (Denmark)

    Lundby, Alicia; Andersen, Martin N; Steffensen, Annette B;

    2013-01-01

    -X-X-pS/T), and integrative analysis of sequence motifs and interaction networks suggested that the kinases AMPK (adenosine 5'-monophosphate-activated protein kinase), Akt, and mTOR (mammalian target of rapamycin) mediate βAR signaling, in addition to the well-established pathways mediated by PKA (cyclic adenosine...

  7.   Plant Phosphoproteomics: Analysis of Plasma Membrane Transporters by Mass Spectrometry

    DEFF Research Database (Denmark)

    Ye, Juanying; Rudashevskaya, Elena; Young, Clifford

    phosphorylation. Due to the low abundance of phosphoprotein, the specific enrichment prior to MS analysis is necessary. Plant proton pump (H+-ATPase) is an enzyme controls the major transport processes in the plant, such as root nutrient uptake. Moreover, this pump has been proposed to be involved in other......  Phosphorylation is a key regulatory factor in all aspects of eukaryotic biology including the regulation of plant membrane-bound transport proteins. To date, mass spectrometry (MS) has been introduced as powerful technology for study of post translational modifications (PTMs), including protein...

  8. Microarray-Based Phospho-Proteomic Profiling of Complex Biological Systems

    Directory of Open Access Journals (Sweden)

    C. Rory Goodwin

    2016-04-01

    Full Text Available Protein microarray technology has been successfully used for identifying substrates of purified activated kinases. We used protein microarrays to globally interrogate the effects of PTEN and Akt activity on the phospho-kinome of in vitro and in vivo glioma models and validated results in clinical pathological specimens. Whole cell lysates extracted from tumor samples can be applied to human kinome chip microarrays to profile the global kinase phosphorylation patterns in a high-throughput manner and identify novel substrates inherent to the tumor cell and the interactions with tumor microenvironment. Our findings identify a novel microarray-based method for assessing intracellular signaling events applicable to human oncogenesis and other pathophysiologic states.

  9. Phosphoproteome analysis during larval development and metamorphosis in the spionid polychaete Pseudopolydora vexillosa

    KAUST Repository

    Chandramouli, Kondethimmanahalli

    2011-05-25

    Background: The metamorphosis of the spionid polychaete Pseudopolydora vexillosa includes spontaneous settlement onto soft-bottom habitats and morphogenesis that can be completed in a very short time. A previous study on the total changes to the proteome during the various developmental stages of P. vexillosa suggested that little or no de novo protein synthesis occurs during metamorphosis. In this study, we used multicolor fluorescence detection of proteins in 2-D gels for differential analysis of proteins and phosphoproteins to reveal the dynamics of post-translational modification proteins in this species. A combination of affinity chromatography, 2D-PAGE, and mass spectrometry was used to identify the phosphoproteins in pre-competent larvae, competent larvae, and newly metamorphosed juveniles. Results: We reproducibly detected 210, 492, and 172 phosphoproteins in pre-competent larvae, competent larvae, and newly metamorphosed juveniles, respectively. The highest percentage of phosphorylation was observed during the competent larval stage. About 64 stage-specific phosphoprotein spots were detected in the competent stage, and 32 phosphoproteins were found to be significantly differentially expressed in the three stages. We identified 38 phosphoproteins, 10 of which were differentially expressed during metamorphosis. These phosphoproteins belonged to six categories of biological processes: (1) development, (2) cell differentiation and integrity, (3) transcription and translation, (4) metabolism, (5) protein-protein interaction and proteolysis, and (6) receptors and enzymes. Conclusion: This is the first study to report changes in phosphoprotein expression patterns during the metamorphosis of the marine polychaete P. vexillosa. The higher degree of phosphorylation during the process of attaining competence to settle and metamorphose may be due to fast morphological transitions regulated by various mechanisms. Our data are consistent with previous studies showing a

  10. The phosphoproteome of toll-like receptor-activated macrophages

    DEFF Research Database (Denmark)

    Weintz, Gabriele; Olsen, Jesper Velgaard; Frühauf, Katja;

    2010-01-01

    Recognition of microbial danger signals by toll-like receptors (TLR) causes re-programming of macrophages. To investigate kinase cascades triggered by the TLR4 ligand lipopolysaccharide (LPS) on systems level, we performed a global, quantitative and kinetic analysis of the phosphoproteome...

  11. The phosphoproteome in regenerating protoplasts from Physcomitrella patens protonemata shows changes paralleling postembryonic development in higher plants.

    Science.gov (United States)

    Wang, Xiaoqin; Qi, Meiyan; Li, Jingyun; Ji, Zhongzhong; Hu, Yong; Bao, Fang; Mahalingam, Ramamurthy; He, Yikun

    2014-05-01

    The moss Physcomitrella patens is an ideal model plant to study plant developmental processes. To better understand the mechanism of protoplast regeneration, a phosphoproteome analysis was performed. Protoplasts were prepared from protonemata. By 4 d of protoplast regeneration, the first cell divisions had ensued. Through a highly selective titanium dioxide (TiO2)-based phosphopeptide enrichment method and mass spectrometric technology, more than 300 phosphoproteins were identified as protoplast regeneration responsive. Of these, 108 phosphoproteins were present on day 4 but not in fresh protoplasts or those cultured for 2 d. These proteins are catalogued here. They were involved in cell-wall metabolism, transcription, signal transduction, cell growth/division, and cell structure. These protein functions are related to cell morphogenesis, organogenesis, and development adjustment. This study presents a comprehensive analysis of phosphoproteome involved in protoplast regeneration and indicates that the mechanism of plant protoplast regeneration is similar to that of postembryonic development.

  12. Technologies and challenges in large-scale phosphoproteomics

    DEFF Research Database (Denmark)

    Engholm-Keller, Kasper; Larsen, Martin Røssel

    2013-01-01

    , and apoptosis, rely on phosphorylation. This PTM is thus involved in many diseases, rendering localization and assessment of extent of phosphorylation of major scientific interest. MS-based phosphoproteomics, which aims at describing all phosphorylation sites in a specific type of cell, tissue, or organism, has...... with focus on the various challenges and limitations this field currently faces....

  13. Simple and Reproducible Sample Preparation for Single-Shot Phosphoproteomics with High Sensitivity

    DEFF Research Database (Denmark)

    Jersie-Christensen, Rosa R.; Sultan, Abida; Olsen, Jesper V

    2016-01-01

    The traditional sample preparation workflow for mass spectrometry (MS)-based phosphoproteomics is time consuming and usually requires multiple steps, e.g., lysis, protein precipitation, reduction, alkylation, digestion, fractionation, and phosphopeptide enrichment. Each step can introduce chemica...

  14. Strategies for quantitation of phosphoproteomic data

    DEFF Research Database (Denmark)

    Palmisano, Giuseppe; Thingholm, Tine Engberg

    2010-01-01

    Recent developments in phosphoproteomic sample-preparation techniques and sensitive mass spectrometry instrumentation have led to large-scale identifications of phosphoproteins and phosphorylation sites from highly complex samples. This has facilitated the implementation of different quantitation...... will be on different quantitation strategies. Methods for metabolic labeling, chemical modification and label-free quantitation and their applicability or inapplicability in phosphoproteomic studies are discussed.......Recent developments in phosphoproteomic sample-preparation techniques and sensitive mass spectrometry instrumentation have led to large-scale identifications of phosphoproteins and phosphorylation sites from highly complex samples. This has facilitated the implementation of different quantitation...... strategies in order to study the biological role of protein phosphorylation during disease progression, differentiation or during external stimulation of a cellular system. In this article, a brief summary of the most popular strategies for phosphoproteomic studies is given; however, the main focus...

  15. Quantitative phosphoproteomic analysis of early alterations in protein phosphorylation by 2,3,7,8-tetrachlorodibenzo-p-dioxin

    DEFF Research Database (Denmark)

    Schulz, Melanie; Brandner, Stefanie; Eberhagen, Carola;

    2013-01-01

    A comprehensive quantitative analysis of changes in protein phosphorylation preceding or accompanying transcriptional activation by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in 5L rat hepatoma cells was performed using the SILAC approach. Following exposure of the cells to DMSO or 1 nM TCDD for 0.......5 to 2 h, 5648 phosphorylated peptides corresponding to 2156 phosphoproteins were identified. Eight peptides exhibited a statistically significantly altered phosphorylation because of TCDD exposure and 22 showed a regulation factor of ≥ 1.5 in one of the experiments per time point. The vast majority...... of the TCCD-induced phosphorylation changes had not been reported before. The transcription factor ARNT, the obligate partner for gene activation by the TCDD-bound Ah receptor, exhibited an up-regulation of its Ser77 phosphorylation, a modification known to control the differential binding of ARNT homodimers...

  16. Reverse-phase phosphoproteome analysis of signaling pathways induced by Rift valley fever virus in human small airway epithelial cells.

    Directory of Open Access Journals (Sweden)

    Taissia G Popova

    Full Text Available Rift valley fever virus (RVFV infection is an emerging zoonotic disease endemic in many countries of sub-Saharan Africa and in Egypt. In this study we show that human small airway epithelial cells are highly susceptible to RVFV virulent strain ZH-501 and the attenuated strain MP-12. We used the reverse-phase protein arrays technology to identify phosphoprotein signaling pathways modulated during infection of cultured airway epithelium. ZH-501 infection induced activation of MAP kinases (p38, JNK and ERK and downstream transcriptional factors [STAT1 (Y701, ATF2 (T69/71, MSK1 (S360 and CREB (S133]. NF-κB phosphorylation was also increased. Activation of p53 (S15, S46 correlated with the increased levels of cleaved effector caspase-3, -6 and -7, indicating activation of the extrinsic apoptotic pathway. RVFV infection downregulated phosphorylation of a major anti-apoptotic regulator of survival pathways, AKT (S473, along with phosphorylation of FOX 01/03 (T24/31 which controls cell cycle arrest downstream from AKT. Consistent with this, the level of apoptosis inhibitor XIAP was decreased. However, the intrinsic apoptotic pathway marker, caspase-9, demonstrated only a marginal activation accompanied by an increased level of the inhibitor of apoptosome formation, HSP27. Concentration of the autophagy marker, LC3B, which often accompanies the pro-survival signaling, was decreased. Cumulatively, our analysis of RVFV infection in lung epithelium indicated a viral strategy directed toward the control of cell apoptosis through a number of transcriptional factors. Analyses of MP-12 titers in challenged cells in the presence of MAPK inhibitors indicated that activation of p38 represents a protective cell response while ERK activation controls viral replication.

  17. Quantitative phosphoproteomics dissection of seven-transmembrane receptor signaling using full and biased agonists

    DEFF Research Database (Denmark)

    Christensen, Gitte L; Kelstrup, Christian D; Lyngsø, Christina;

    2010-01-01

    , we performed a global quantitative phosphoproteomics analysis of the AT(1)R signaling network. We analyzed ligand-stimulated SILAC (stable isotope labeling by amino acids in cell culture) cells by high resolution (LTQ-Orbitrap) MS and compared the phosphoproteomes of the AT(1)R agonist angiotensin II......(q)-dependent and -independent AT(1)R signaling. This study provides substantial novel insight into angiotensin II signal transduction and is the first study dissecting the differences between a full agonist and a biased agonist from a 7TMR on a systems-wide scale. Importantly, it reveals a previously unappreciated diversity...

  18. Quantitative phosphoproteomics dissection of 7TM receptor signaling using full and biased agonists

    DEFF Research Database (Denmark)

    Christensen, Gitte Lund; Kelstrup, Christian; Lyngsø, Christina;

    2010-01-01

    performed a global quantitative phosphoproteomics analysis of the AT1R signaling network. We analyzed ligand-stimulated SILAC cells by high-resolution mass spectrometry (LTQ Orbitrap MS) and compared the phosphoproteomes of the AT1R agonist Angiotensin II and the biased agonist SII Angiotensin II, which...... site patterns displays a striking distinction between protein kinases activated by Gaq protein-dependent and -independent mechanisms, and we now place protein kinase D as a key protein involved in both Gaq-dependent and independent AT1R signaling.his study provides substantial novel insight...

  19. Recent findings and technological advances in phosphoproteomics for cells and tissues.

    Science.gov (United States)

    von Stechow, Louise; Francavilla, Chiara; Olsen, Jesper V

    2015-01-01

    Site-specific phosphorylation is a fast and reversible covalent post-translational modification that is tightly regulated in cells. The cellular machinery of enzymes that write, erase and read these modifications (kinases, phosphatases and phospho-binding proteins) is frequently deregulated in different diseases, including cancer. Large-scale studies of phosphoproteins - termed phosphoproteomics - strongly rely on the use of high-performance mass spectrometric instrumentation. This powerful technology has been applied to study a great number of phosphorylation-based phenotypes. Nevertheless, many technical and biological challenges have to be overcome to identify biologically relevant phosphorylation sites in cells and tissues. This review describes different technological strategies to identify and quantify phosphorylation sites with high accuracy, without significant loss of analysis speed and reproducibility in tissues and cells. Moreover, computational tools for analysis, integration and biological interpretation of phosphorylation events are discussed.

  20. Automated Immobilized Metal Affinity Chromatography System for Enrichment of Escherichia coli Phosphoproteome

    Energy Technology Data Exchange (ETDEWEB)

    Qu, Yi; Wu, Si; Zhao, Rui; Zink, Erika M.; Orton, Daniel J.; Moore, Ronald J.; Meng, Da; Clauss, Therese RW; Aldrich, Joshua T.; Lipton, Mary S.; Pasa-Tolic, Ljiljana

    2013-06-05

    Enrichment of bacterial phosphopeptides is an essential step prior to bottom-up mass spectrometry-based analysis of the phosphoproteome, which is fundamental to understanding the role of phosphoproteins in cell signaling and regulation of protein activity. We developed an automated IMAC system to enrich strong cation exchange-fractionated phosphopeptides from the soluble proteome of Escherichia coli MG1655 grown on minimal medium. Initial demonstration of the system resulted in identification of 75 phosphopeptides covering 52 phosphoproteins. Consistent with previous studies, many of these phosphoproteins are involved in the carbohydrate portion of central metabolism. The automated system utilizes a large capacity IMAC column that can effectively enrich phosphopeptides from a bacterial sample by increasing peptide loading and reducing the wash time. An additional benefit of the automated IMAC system is reduced labor and associated costs.

  1. Characterization of the Phosphoproteome in SLE Patients.

    Directory of Open Access Journals (Sweden)

    Xinzhou Zhang

    Full Text Available Protein phosphorylation is a complex regulatory event that is involved in the signaling networks that affect virtually every cellular process. The protein phosphorylation may be a novel source for discovering biomarkers and drug targets. However, a systematic analysis of the phosphoproteome in patients with SLE has not been performed. To clarify the pathogenesis of systemic lupus erythematosus (SLE, we compared phosphoprotein expression in PBMCs from SLE patients and normal subjects using proteomics analyses. Phosphopeptides were enriched using TiO₂ from PBMCs isolated from 15 SLE patients and 15 healthy subjects and then analyzed by automated LC-MS/MS analysis. Phosphorylation sites were identified and quantitated by MASCOT and MaxQuant. A total of 1035 phosphorylation sites corresponding to 618 NCBI-annotated genes were identified in SLE patients compared with normal subjects. Differentially expressed proteins, peptides and phosphorylation sites were then subjected to bioinformatics analyses. Gene ontology(GO and pathway analyses showed that nucleic acid metabolism, cellular component organization, transport and multicellular organismal development pathways made up the largest proportions of the differentially expressed genes. Pathway analyses showed that the mitogen-activated protein kinase (MAPK signaling pathway and actin cytoskeleton regulators made up the largest proportions of the metabolic pathways. Network analysis showed that rous sarcoma oncogene (SRC, v-rel reticuloendotheliosis viral oncogene homolog A (RELA, histone deacetylase (HDA1C and protein kinase C, delta (PRKCD play important roles in the stability of the network. These data suggest that aberrant protein phosphorylation may contribute to SLE pathogenesis.

  2. Phosphorylation of proteins during human myometrial contractions: A phosphoproteomic approach.

    Science.gov (United States)

    Hudson, Claire A; López Bernal, Andrés

    2017-01-22

    Phasic myometrial contractility is a key component of human parturition and the contractions are driven by reversible phosphorylation of myosin light chains catalyzed by the calcium (Ca(2+))-dependent enzyme myosin light chain kinase (MYLK). Other yet unknown phosphorylation or de-phosphorylation events may contribute to myometrial contraction and relaxation. In this study we have performed a global phosphoproteomic analysis of human myometrial tissue using tandem mass tagging to detect changes in the phosphorylation status of individual myometrial proteins during spontaneous and oxytocin-driven phasic contractions. We were able to detect 22 individual phosphopeptides whose relative ratio changed (fold > 2 or contraction. The most significant changes in phosphorylation were to MYLK on serine 1760, a site associated with reductions in calmodulin binding and subsequent kinase activity. Phosphorylated MYLK (ser1760) increased significantly during spontaneous (9.83 ± 3.27 fold, P contractions and we were able to validate these data using immunoblotting. Pathway analysis suggested additional proteins involved in calcium signalling, cGMP-PRKG signalling, adrenergic signalling and oxytocin signalling were also phosphorylated during contractions. This study demonstrates that a global phosphoproteomic analysis of myometrial tissue is a sensitive approach to detect changes in the phosphorylation of proteins during myometrial contractions, and provides a platform for further validation of these changes and for identification of their functional significance.

  3. Clinical and Technical Phosphoproteomic Research

    Directory of Open Access Journals (Sweden)

    Ferreira Antonio

    2011-06-01

    Full Text Available Abstract An encouraging approach for the diagnosis and effective therapy of immunological pathologies, which would include cancer, is the identification of proteins and phosphorylated proteins. Disease proteomics, in particular, is a potentially useful method for this purpose. A key role is played by protein phosphorylation in the regulation of normal immunology disorders and targets for several new cancer drugs and drug candidates are cancer cells and protein kinases. Protein phosphorylation is a highly dynamic process. The functioning of new drugs is of major importance as is the selection of those patients who would respond best to a specific treatment regime. In all major aspects of cellular life signalling networks are key elements which play a major role in inter- and intracellular communications. They are involved in diverse processes such as cell-cycle progression, cellular metabolism, cell-cell communication and appropriate response to the cellular environment. A whole range of networks that are involved in the regulation of cell development, differentiation, proliferation, apoptosis, and immunologic responses is contained in the latter. It is so necessary to understand and monitor kinase signalling pathways in order to understand many immunology pathologies. Enrichment of phosphorylated proteins or peptides from tissue or bodily fluid samples is required. The application of technologies such as immunoproteomic techniques, phosphoenrichments and mass spectrometry (MS is crucial for the identification and quantification of protein phosphorylation sites in order to advance in clinical research. Pharmacodynamic readouts of disease states and cellular drug responses in tumour samples will be provided as the field develops. We aim to detail the current and most useful techniques with research examples to isolate and carry out clinical phosphoproteomic studies which may be helpful for immunology and cancer research. Different phosphopeptide

  4. Label-free quantitative phosphoproteomics with novel pairwise abundance normalization reveals synergistic RAS and CIP2A signaling.

    Science.gov (United States)

    Kauko, Otto; Laajala, Teemu Daniel; Jumppanen, Mikael; Hintsanen, Petteri; Suni, Veronika; Haapaniemi, Pekka; Corthals, Garry; Aittokallio, Tero; Westermarck, Jukka; Imanishi, Susumu Y

    2015-08-17

    Hyperactivated RAS drives progression of many human malignancies. However, oncogenic activity of RAS is dependent on simultaneous inactivation of protein phosphatase 2A (PP2A) activity. Although PP2A is known to regulate some of the RAS effector pathways, it has not been systematically assessed how these proteins functionally interact. Here we have analyzed phosphoproteomes regulated by either RAS or PP2A, by phosphopeptide enrichment followed by mass-spectrometry-based label-free quantification. To allow data normalization in situations where depletion of RAS or PP2A inhibitor CIP2A causes a large uni-directional change in the phosphopeptide abundance, we developed a novel normalization strategy, named pairwise normalization. This normalization is based on adjusting phosphopeptide abundances measured before and after the enrichment. The superior performance of the pairwise normalization was verified by various independent methods. Additionally, we demonstrate how the selected normalization method influences the downstream analyses and interpretation of pathway activities. Consequently, bioinformatics analysis of RAS and CIP2A regulated phosphoproteomes revealed a significant overlap in their functional pathways. This is most likely biologically meaningful as we observed a synergistic survival effect between CIP2A and RAS expression as well as KRAS activating mutations in TCGA pan-cancer data set, and synergistic relationship between CIP2A and KRAS depletion in colony growth assays.

  5. TiSH--a robust and sensitive global phosphoproteomics strategy employing a combination of TiO2, SIMAC, and HILIC

    DEFF Research Database (Denmark)

    Engholm-Keller, Kasper; Birck, Pernille; Størling, Joachim

    2012-01-01

    , with an enrichment specificity>94%. When doing network analysis of putative phosphorylation changes it could be noted that the identified protein interaction network centered upon proteins known to be affected by the interferon-¿ pathway, thereby supporting the utility of this global phosphoproteomics strategy......Large scale quantitative phosphoproteomics depends upon multidimensional strategies for peptide fractionation, phosphopeptide enrichment, and mass spectrometric analysis. Previously, most robust comprehensive large-scale phosphoproteomics strategies have relied on milligram amounts of protein. We...... have set up a multi-dimensional phosphoproteomics strategy combining a number of well-established enrichment and fraction methods: An initial TiO(2) phosphopeptide pre-enrichment step is followed by post-fractionation using sequential elution from IMAC (SIMAC) to separate multi- and mono...

  6. Machine learning of global phosphoproteomic profiles enables discrimination of direct versus indirect kinase substrates.

    Science.gov (United States)

    Kanshin, Evgeny; Giguere, Sebastien; Cheng, Jing; Tyers, Michael D; Thibault, Pierre

    2017-03-06

    Mass spectrometry allows quantification of tens of thousands of phosphorylation sites from minute amounts of cellular material. Despite this wealth of information, our understanding of phosphorylation-based signaling is limited, in part because it is not possible to deconvolute substrate phosphorylation that is directly mediated by a particular kinase versus phosphorylation that is mediated by downstream kinases. Here, we describe a framework for assignment of direct in-vivo kinase substrates using a combination of selective chemical inhibition, quantitative phosphoproteomics, and machine learning techniques. Our workflow allows classification of phosphorylation events following inhibition of an analog-sensitive kinase into kinase-independent effects of the inhibitor, direct effects on cognate substrates and indirect effects mediated by downstream kinases or phosphatases. We applied this method to identify many direct targets of Cdc28 and Snf1 kinases in the budding yeast S. cerevisiae. Global phosphoproteome analysis of acute time-series demonstrated that dephosphorylation of direct kinase substrates occurs more rapidly compared to indirect substrates, both after inhibitor treatment and under a physiological nutrient shift in wild-type cells. Mutagenesis experiments revealed a high proportion of functionally relevant phosphorylation sites on Snf1 targets. For example, Snf1 itself was inhibited through autophosphorylation on S391 and new phosphosites were discovered that modulate the activity of the Reg1 regulatory subunit of the Glc7 phosphatase and the Gal83 β-subunit of SNF1 complex. This methodology applies to any kinase for which a functional analog sensitive version can be constructed to facilitate the dissection of the global phosphorylation network.

  7. Search Databases and Statistics: Pitfalls and Best Practices in Phosphoproteomics.

    Science.gov (United States)

    Refsgaard, Jan C; Munk, Stephanie; Jensen, Lars J

    2016-01-01

    Advances in mass spectrometric instrumentation in the past 15 years have resulted in an explosion in the raw data yield from typical phosphoproteomics workflows. This poses the challenge of confidently identifying peptide sequences, localizing phosphosites to proteins and quantifying these from the vast amounts of raw data. This task is tackled by computational tools implementing algorithms that match the experimental data to databases, providing the user with lists for downstream analysis. Several platforms for such automated interpretation of mass spectrometric data have been developed, each having strengths and weaknesses that must be considered for the individual needs. These are reviewed in this chapter. Equally critical for generating highly confident output datasets is the application of sound statistical criteria to limit the inclusion of incorrect peptide identifications from database searches. Additionally, careful filtering and use of appropriate statistical tests on the output datasets affects the quality of all downstream analyses and interpretation of the data. Our considerations and general practices on these aspects of phosphoproteomics data processing are presented here.

  8. SIMAC - A phosphoproteomic strategy for the rapid separation of mono-phosphorylated from multiply phosphorylated peptides

    DEFF Research Database (Denmark)

    Thingholm, Tine E; Jensen, Ole N; Robinson, Phillip J

    2008-01-01

    spectrometric analysis, such as immobilized metal affinity chromatography or titanium dioxide the coverage of the phosphoproteome of a given sample is limited. Here we report a simple and rapid strategy - SIMAC - for sequential separation of mono-phosphorylated peptides and multiply phosphorylated peptides from...... and an optimized titanium dioxide chromatographic method. More than double the total number of identified phosphorylation sites was obtained with SIMAC, primarily from a three-fold increase in recovery of multiply phosphorylated peptides....

  9. Investigating temporal changes in the yeast phosphoproteome upon fatty acid starvation

    DEFF Research Database (Denmark)

    Pultz, Dennis; Bennetzen, Martin; Andersen, Jens S.;

    2011-01-01

    Investigating stemporal changes in the yeast phosphoproteome upon fatty acid starvation Dennis Pultz*, Martin Bennetzen*, Jens S. Andersen and Nils J.Færgeman. Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark, 5230 Reducing food intake to induce...... that the nutrient sensor TOR1 is central in the regulation and orchestration of the downstream cellular response upon fatty acid starvation. By use of mass spectrometry and a SILAC (stable isotope labelling by amino acids in cell culture) based approach we wish to unravel the temporal changes in the phosphoproteome...... coupling fatty acid starvation to autophagy and starvation related cellular responses in general. By use of this approach we have identified approximately 2000 phosphorylation sites of which more than 400 have been identified as being regulated. Here, we present results showing the temporal behaviour...

  10. Sperm phosphoproteome profiling by ultra performance liquid chromatography followed by data independent analysis (LC-MS(E)) reveals altered proteomic signatures in asthenozoospermia.

    Science.gov (United States)

    Parte, Priyanka P; Rao, Parimala; Redij, Shweta; Lobo, Vivian; D'Souza, Serena J; Gajbhiye, Rahul; Kulkarni, Vijay

    2012-10-22

    Sperm motility is an important prerequisite for successful fertilization and is regulated by cyclic AMP activated protein kinase A which phosphorylates flagella proteins like axonemal dynein and initiates motility. Increase in calcium influx reverses this process by dephosphorylation that is mediated by calcineurin. Analyzing the phosphoenriched fractions of spermatozoa lysates from eight normozoospermic-, and asthenozoospermic-samples, respectively, by Nano UPLC-MS(E), the present study investigates the phosphoproteins involved in sperm motility in an attempt to identify the key pathways regulating sperm motility and likely to be altered in spermatozoa of asthenozoospermic individuals. 66 phosphoproteins were differentially regulated in asthenozoospermia. The deregulated proteins comprised predominantly the HSPs, cytoskeletal proteins, proteins associated with the fibrous sheath, and those associated with energy metabolism. EM analysis of these spermatozoa demonstrated significant defects in mitochondria, and fibrous sheath and these defects could be correlated with the altered proteome. Pathway analysis revealed that carbohydrate and energy metabolism, cAMP mediated PKA signaling, PI3K/AKT signaling and pathway regulating actin based motility by Rho were significantly altered indicating that motility in spermatozoa is regulated through the concerted effort of these pathways. The data identified signature molecules that have the potential as biomarkers for diagnosing etiology of asthenozoospermia.

  11. The proteome and phosphoproteome of maize pollen uncovers fertility candidate proteins.

    Science.gov (United States)

    Chao, Qing; Gao, Zhi-Fang; Wang, Yue-Feng; Li, Zhe; Huang, Xia-He; Wang, Ying-Chun; Mei, Ying-Chang; Zhao, Biligen-Gaowa; Li, Liang; Jiang, Yu-Bo; Wang, Bai-Chen

    2016-06-01

    Maize is unique since it is both monoecious and diclinous (separate male and female flowers on the same plant). We investigated the proteome and phosphoproteome of maize pollen containing modified proteins and here we provide a comprehensive pollen proteome and phosphoproteome which contain 100,990 peptides from 6750 proteins and 5292 phosphorylated sites corresponding to 2257 maize phosphoproteins, respectively. Interestingly, among the total 27 overrepresented phosphosite motifs we identified here, 11 were novel motifs, which suggested different modification mechanisms in plants compared to those of animals. Enrichment analysis of pollen phosphoproteins showed that pathways including DNA synthesis/chromatin structure, regulation of RNA transcription, protein modification, cell organization, signal transduction, cell cycle, vesicle transport, transport of ions and metabolisms, which were involved in pollen development, the following germination and pollen tube growth, were regulated by phosphorylation. In this study, we also found 430 kinases and 105 phosphatases in the maize pollen phosphoproteome, among which calcium dependent protein kinases (CDPKs), leucine rich repeat kinase, SNF1 related protein kinases and MAPK family proteins were heavily enriched and further analyzed. From our research, we also uncovered hundreds of male sterility-associated proteins and phosphoproteins that might influence maize productivity and serve as targets for hybrid maize seed production. At last, a putative complex signaling pathway involving CDPKs, MAPKs, ubiquitin ligases and multiple fertility proteins was constructed. Overall, our data provides new insight for further investigation of protein phosphorylation status in mature maize pollen and construction of maize male sterile mutants in the future.

  12. 王浆高产蜜蜂咽下腺磷酸化蛋白质组分析%Phosphoproteome Analysis of Hypopharyngeal Glands of High Royal Jelly Producing Bee (Apis mellifera L.)

    Institute of Scientific and Technical Information of China (English)

    鲁小山; 韩宾; 张兰; 冯毛; 房宇; 李荣丽; 周天娥; 李建科

    2013-01-01

    [Objective]High royal jelly producing bee (Apis mellifera L.) is the unique honeybee resource in China. However, the mechanism of high royal jelly producing has not been clearly addressed. In order to reveal the significance of protein phosphorylation in hypopharyngeal gland for royal jelly synthesis and secretion, the phosphoproteome of hypopharyngeal gland of nurse bee (6-12 day) was analyzed.[Method]IMAC (immobilized metal-affinity chromatography) phosphoprotein enrichment, SCX (strong cation exchange) separation, LC-MS/MS (liquid chromatography-mass/mass) identification and bioinformatics analysis were applied to analyze phosphoproteome of hypopharyngeal gland of nurse bee.[Result]Of the identified 117 proteins in the hypopharyngeal gland, 6 of them were phosphorylated on 6 phosphopeptides and assigned 8 phosphorylated sites. They were related to protein translation and synthesis, such as 60S acidic ribosomal proteins P0, P1, P2 and 60S ribosomal protein L15, and major royal jelly proteins 1 and 7 precursor.[Conclusion]The phosphorylation modifications occurred on ribosome proteins of hypopharyngeal gland mainly contribute to the high efficiency of synthesizing and secreting of royal jelly protein. The phosphorylation of royal jelly proteins 1 and 7 maintain the reasonable ratio of calcium to phosphorus of royal jelly with the increasing yield, thus meeting the nutrition demand of fertile egg-laying queens and developing larvae. Hence, the data obtained in this study will provide new knowledge to deeply understand the mechanism how high royal jelly producing bees could produce higher amount of royal jelly at the level of protein phosphorylation.%[目的]通过对王浆高产蜜蜂(Apis mellifera L.)(浙江浆蜂)哺育蜂(6-12 d)咽下腺的磷酸化蛋白质组分析,以期探明蛋白质磷酸化修饰对王浆分泌的生物学意义。[方法]将哺育蜂咽下腺蛋白质液内酶切后,用固相金属离子亲和层析色谱法(IMAC

  13. Quantitative phosphoproteomics of proteasome inhibition in multiple myeloma cells.

    Directory of Open Access Journals (Sweden)

    Feng Ge

    Full Text Available BACKGROUND: The proteasome inhibitor bortezomib represents an important advance in the treatment of multiple myeloma (MM. Bortezomib inhibits the activity of the 26S proteasome and induces cell death in a variety of tumor cells; however, the mechanism of cytotoxicity is not well understood. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the differential phosphoproteome upon proteasome inhibition by using stable isotope labeling by amino acids in cell culture (SILAC in combination with phosphoprotein enrichment and LC-MS/MS analysis. In total 233 phosphoproteins were identified and 72 phosphoproteins showed a 1.5-fold or greater change upon bortezomib treatment. The phosphoproteins with expression alterations encompass all major protein classes, including a large number of nucleic acid binding proteins. Site-specific phosphopeptide quantitation revealed that Ser38 phosphorylation on stathmin increased upon bortezomib treatment, suggesting new mechanisms associated to bortezomib-induced apoptosis in MM cells. Further studies demonstrated that stathmin phosphorylation profile was modified in response to bortezomib treatment and the regulation of stathmin by phosphorylation at specific Ser/Thr residues participated in the cellular response induced by bortezomib. CONCLUSIONS/SIGNIFICANCE: Our systematic profiling of phosphorylation changes in response to bortezomib treatment not only advanced the global mechanistic understanding of the action of bortezomib on myeloma cells but also identified previously uncharacterized signaling proteins in myeloma cells.

  14. Quantitative phosphoproteomic analysis of postmortem muscle development

    DEFF Research Database (Denmark)

    Huang, Honggang

    Meat quality development is highly dependent on postmortem (PM) metabolism and rigor mortis development in PM muscle. PM glycometabolism and rigor mortis fundamentally determine most of the important qualities of raw meat, such as ultimate pH, tenderness, color and water-holding capacity. Protein...

  15. The Phosphoproteomes of Plasmodium falciparum and Toxoplasma gondii reveal unusual adaptations within and beyond the parasites’ boundaries

    OpenAIRE

    Treeck, Moritz; Sanders, John L.; Elias, Joshua E.; John C Boothroyd

    2011-01-01

    Plasmodium falciparum and Toxoplasma gondii are obligate intracellular apicomplexan parasites that rapidly invade and extensively modify host cells. Protein phosphorylation is one mechanism by which these parasites can control such processes. Here we present a phosphoproteome analysis of peptides enriched from schizont stage P. falciparum and T. gondii tachyzoites that are either “intracellular” or purified away from host material. Using liquid chromatography and tandem mass-spectrometry we i...

  16. TiSH--a robust and sensitive global phosphoproteomics strategy employing a combination of TiO2, SIMAC, and HILIC.

    Science.gov (United States)

    Engholm-Keller, Kasper; Birck, Pernille; Størling, Joachim; Pociot, Flemming; Mandrup-Poulsen, Thomas; Larsen, Martin R

    2012-10-22

    Large scale quantitative phosphoproteomics depends upon multidimensional strategies for peptide fractionation, phosphopeptide enrichment, and mass spectrometric analysis. Previously, most robust comprehensive large-scale phosphoproteomics strategies have relied on milligram amounts of protein. We have set up a multi-dimensional phosphoproteomics strategy combining a number of well-established enrichment and fraction methods: An initial TiO(2) phosphopeptide pre-enrichment step is followed by post-fractionation using sequential elution from IMAC (SIMAC) to separate multi- and mono-phosphorylated peptides, and hydrophilic interaction liquid chromatography (HILIC) of the mono-phosphorylated peptides (collectively abbreviated "TiSH"). The advantages of the strategy include a high specificity and sample preparation workload reduction due to the TiO(2) pre-enrichment step, as well as low adsorptive losses. We demonstrate the capability of this strategy by quantitative investigation of early interferon-γ signaling in low quantities of insulinoma cells. We identified ~6600 unique phosphopeptides from 300 μg of peptides/condition (22 unique phosphopeptides/μg) in a duplex dimethyl labeling experiment, with an enrichment specificity>94%. When doing network analysis of putative phosphorylation changes it could be noted that the identified protein interaction network centered upon proteins known to be affected by the interferon-γ pathway, thereby supporting the utility of this global phosphoproteomics strategy. This strategy thus shows great potential for interrogating signaling networks from low amounts of sample with high sensitivity and specificity.

  17. A novel double-component MOAC honeycomb composite with pollen grains as a template for phosphoproteomics research.

    Science.gov (United States)

    Wang, Jiaxi; Li, Jie; Wang, Yanan; Gao, Mingxia; Zhang, Xiangmin; Deng, Chunhui

    2016-07-01

    The enrichment and separation of phosphopeptides from mixed biological samples is a technologically very significance, but highly challenging work. Current designed materials are mainly based on the broad and effective adsorptive character of metal oxide affinity chromatography (MOAC). Though significant progress has been made in the enrichment of phosphopeptides with MOAC material, there are chances for further development. In this study, a novel pollen-based MOAC honeycomb material was firstly explored in which the suitable hydrophilic channels preferentially enrich much more endogenous phosphopeptides than nonphosphopeptides or proteins while doping binary metal oxides at the atomic level and the ultra-high specific surface area have further allowed it to possess more effective active sites. Based on these unique features, the pollen-based material exhibited high selectivity for β-casein (mass ratio of β-casein/BSA, 1:1500), ultra-low detection limit (0.1fmol), desirable reusability. Moreover, the bionics MOAC composites were investigated in the enrichment of phosphopeptides from nonfat milk, human serum (male and female at the same age) and mice liver, results of which indicate the great potential of the composite for the phosphoproteome analysis of complex biological samples through the cheap and environmentally friendly process.

  18. Phosphoproteomics reveals the effect of ethylene in soybean root under flooding stress.

    Science.gov (United States)

    Yin, Xiaojian; Sakata, Katsumi; Komatsu, Setsuko

    2014-12-05

    Flooding has severe negative effects on soybean growth. To explore the flooding-responsive mechanisms in early-stage soybean, a phosphoproteomic approach was used. Two-day-old soybean plants were treated without or with flooding for 3, 6, 12, and 24 h, and root tip proteins were then extracted and analyzed at each time point. After 3 h of flooding exposure, the fresh weight of soybeans increased, whereas the ATP content of soybean root tips decreased. Using a gel-free proteomic technique, a total of 114 phosphoproteins were identified in the root tip samples, and 34 of the phosphoproteins were significantly changed with respect to phosphorylation status after 3 h of flooding stress. Among these phosphoproteins, eukaryotic translation initiation factors were dephosphorylated, whereas several protein synthesis-related proteins were phosphorylated. The mRNA expression levels of sucrose phosphate synthase 1F and eukaryotic translation initiation factor 4 G were down-regulated, whereas UDP-glucose 6-dehydrogenase mRNA expression was up-regulated during growth but down-regulated under flooding stress. Furthermore, bioinformatic protein interaction analysis of flooding-responsive proteins based on temporal phosphorylation patterns indicated that eukaryotic translation initiation factor 4 G was located in the center of the network during flooding. Soybean eukaryotic translation initiation factor 4 G has homology to programmed cell death 4 protein and is implicated in ethylene signaling. The weight of soybeans was increased with treatment by an ethylene-releasing agent under flooding condition, but it was decreased when plants were exposed to an ethylene receptor antagonist. These results suggest that the ethylene signaling pathway plays an important role, via the protein phosphorylation, in mechanisms of plant tolerance to the initial stages of flooding stress in soybean root tips.

  19. Selection of an Appropriate Protein Extraction Method to Study the Phosphoproteome of Maize Photosynthetic Tissue

    Science.gov (United States)

    Luís, Inês M.; Alexandre, Bruno M.; Oliveira, M. Margarida

    2016-01-01

    Often plant tissues are recalcitrant and, due to that, methods relying on protein precipitation, such as TCA/acetone precipitation and phenol extraction, are usually the methods of choice for protein extraction in plant proteomic studies. However, the addition of precipitation steps to protein extraction methods may negatively impact protein recovery, due to problems associated with protein re-solubilization. Moreover, we show that when working with non-recalcitrant plant tissues, such as young maize leaves, protein extraction methods with precipitation steps compromise the maintenance of some labile post-translational modifications (PTMs), such as phosphorylation. Therefore, a critical issue when studying PTMs in plant proteins is to ensure that the protein extraction method is the most appropriate, both at qualitative and quantitative levels. In this work, we compared five methods for protein extraction of the C4-photosynthesis related proteins, in the tip of fully expanded third-leaves. These included: TCA/Acetone Precipitation; Phenol Extraction; TCA/Acetone Precipitation followed by Phenol Extraction; direct extraction in Lysis Buffer (a urea-based buffer); and direct extraction in Lysis Buffer followed by Cleanup with a commercial kit. Protein extraction in Lysis Buffer performed better in comparison to the other methods. It gave one of the highest protein yields, good coverage of the extracted proteome and phosphoproteome, high reproducibility, and little protein degradation. This was also the easiest and fastest method, warranting minimal sample handling. We also show that this method is adequate for the successful extraction of key enzymes of the C4-photosynthetic metabolism, such as PEPC, PPDK, PEPCK, and NADP-ME. This was confirmed by MALDI-TOF/TOF MS analysis of excised spots of 2DE analyses of the extracted protein pools. Staining for phosphorylated proteins in 2DE revealed the presence of several phosphorylated isoforms of PEPC, PPDK, and PEPCK. PMID

  20. Phosphoproteomic profiling of selenate-treated Alzheimer's disease model cells.

    Directory of Open Access Journals (Sweden)

    Ping Chen

    Full Text Available The reversible phosphorylation of proteins regulates most biological processes, while abnormal phosphorylation is a cause or consequence of many diseases including Alzheimer's disease (AD. One of the hallmarks of AD is the formation of neurofibrillary tangles (NFTs, which is composed of hyperphosphorylated tau proteins. Sodium selenate has been recently found to reduce tau hyperphosphorylation and NFTs formation, and to improve spatial learning and motor performance in AD mice. In the current study, the phosphoproteomics of N2aSW cells treated with selenate were investigated. To avoid missing low-abundance phosphoproteins, both the total proteins of cells and the phosphor-enriched proteins were extracted and subjected to the two-dimensional gel electrophoresis with Pro-Q diamond staining and then LC-MS/MS analysis. A total of 65 proteins were altered in phosphorylation level, of which 39 were up-regulated and 26 were down-regulated. All identified phosphoproteins were bioinformatically annotated according to their physiochemical features, subcellular location, and biological function. Most of these significantly changed phosphoproteins are involved in crucial neural processes such as protesome activity, oxidative stress, cysteine and methionine metabolism, and energy metabolism. Furthermore, decreases were found in homocysteine, phosphor-tau and amyloid β upon selenate treatment. Our results suggest that selenate may intervene in the pathological process of AD by altering the phosphorylation of some key proteins involved in oxidative stress, energy metabolism and protein degradation, thus play important roles in maintaining redox homeostasis, generating ATP, and clearing misfolded proteins and aggregates. The present paper provides some new clues to the mechanism of selenate in AD prevention.

  1. Phosphoproteomic profiling of human myocardial tissues distinguishes ischemic from non-ischemic end stage heart failure.

    Science.gov (United States)

    Schechter, Matthew A; Hsieh, Michael K H; Njoroge, Linda W; Thompson, J Will; Soderblom, Erik J; Feger, Bryan J; Troupes, Constantine D; Hershberger, Kathleen A; Ilkayeva, Olga R; Nagel, Whitney L; Landinez, Gina P; Shah, Kishan M; Burns, Virginia A; Santacruz, Lucia; Hirschey, Matthew D; Foster, Matthew W; Milano, Carmelo A; Moseley, M Arthur; Piacentino, Valentino; Bowles, Dawn E

    2014-01-01

    The molecular differences between ischemic (IF) and non-ischemic (NIF) heart failure are poorly defined. A better understanding of the molecular differences between these two heart failure etiologies may lead to the development of more effective heart failure therapeutics. In this study extensive proteomic and phosphoproteomic profiles of myocardial tissue from patients diagnosed with IF or NIF were assembled and compared. Proteins extracted from left ventricular sections were proteolyzed and phosphopeptides were enriched using titanium dioxide resin. Gel- and label-free nanoscale capillary liquid chromatography coupled to high resolution accuracy mass tandem mass spectrometry allowed for the quantification of 4,436 peptides (corresponding to 450 proteins) and 823 phosphopeptides (corresponding to 400 proteins) from the unenriched and phospho-enriched fractions, respectively. Protein abundance did not distinguish NIF from IF. In contrast, 37 peptides (corresponding to 26 proteins) exhibited a ≥ 2-fold alteration in phosphorylation state (pfailure etiology examined. Proteins exhibiting phosphorylation alterations were grouped into functional categories: transcriptional activation/RNA processing; cytoskeleton structure/function; molecular chaperones; cell adhesion/signaling; apoptosis; and energetic/metabolism. Phosphoproteomic analysis demonstrated profound post-translational differences in proteins that are involved in multiple cellular processes between different heart failure phenotypes. Understanding the roles these phosphorylation alterations play in the development of NIF and IF has the potential to generate etiology-specific heart failure therapeutics, which could be more effective than current therapeutics in addressing the growing concern of heart failure.

  2. Characterization of early autophagy signaling by quantitative phosphoproteomics

    DEFF Research Database (Denmark)

    Rigbolt, Kristoffer Tg; Zarei, Mostafa; Sprenger, Adrian;

    2014-01-01

    Under conditions of nutrient shortage autophagy is the primary cellular mechanism ensuring availability of substrates for continuous biosynthesis. Subjecting cells to starvation or rapamycin efficiently induces autophagy by inhibiting the MTOR signaling pathway triggering increased autophagic flux....... To elucidate the regulation of early signaling events upon autophagy induction, we applied quantitative phosphoproteomics characterizing the temporal phosphorylation dynamics after starvation and rapamycin treatment. We obtained a comprehensive atlas of phosphorylation kinetics within the first 30 min upon...

  3. Towards single-cell LC-MS phosphoproteomics

    OpenAIRE

    Polat, Ayşe Nur; Özlü, Nurhan

    2014-01-01

    Protein phosphorylation is a ubiquitous posttranslational modification, which is heavily involved in signal transduction. Misregulation of protein phosphorylation is often associated with a decrease in cell viability and complex diseases such as cancer. The dynamic and low abundant nature of phosphorylated proteins makes studying phosphoproteome a challenging task. In this review, we summarize state of the art proteomic techniques to study and quantify peptide phosphorylation in biological sy...

  4. Dissection of the insulin signaling pathway via quantitative phosphoproteomics

    DEFF Research Database (Denmark)

    Krüger, Marcus; Kratchmarova, Irina; Blagoev, Blagoy;

    2008-01-01

    The insulin signaling pathway is of pivotal importance in metabolic diseases, such as diabetes, and in cellular processes, such as aging. Insulin activates a tyrosine phosphorylation cascade that branches to create a complex network affecting multiple biological processes. To understand the full ...... the calcium transporting ATPase SERCA2, supporting a connection to calcium signaling. The combination of quantitative phosphoproteomics with cell culture models provides a powerful strategy to dissect the insulin signaling pathways in intact cells....

  5. Characterization of the human plasma phosphoproteome using linear ion trap mass spectrometry and multiple search engines.

    Science.gov (United States)

    Carrascal, Montserrat; Gay, Marina; Ovelleiro, David; Casas, Vanessa; Gelpí, Emilio; Abian, Joaquin

    2010-02-05

    Major plasma protein families play different roles in blood physiology and hemostasis and in immunodefense. Other proteins in plasma can be involved in signaling as chemical messengers or constitute biological markers of the status of distant tissues. In this respect, the plasma phosphoproteome holds potentially relevant information on the mechanisms modulating these processes through the regulation of protein activity. In this work we describe for the first time a collection of phosphopeptides identified in human plasma using immunoaffinity separation of the seven major serum protein families from other plasma proteins, SCX fractionation, and TiO(2) purification prior to LC-MS/MS analysis. One-hundred and twenty-seven phosphosites in 138 phosphopeptides mapping 70 phosphoproteins were identified with FDR < 1%. A high-confidence collection of phosphosites was obtained using a combined search with the OMSSA, SEQUEST, and Phenyx search engines.

  6. Phosphoproteome Profiling Reveals Circadian Clock Regulation of Posttranslational Modifications in the Murine Hippocampus

    Science.gov (United States)

    Chiang, Cheng-Kang; Xu, Bo; Mehta, Neel; Mayne, Janice; Sun, Warren Y. L.; Cheng, Kai; Ning, Zhibin; Dong, Jing; Zou, Hanfa; Cheng, Hai-Ying Mary; Figeys, Daniel

    2017-01-01

    The circadian clock is an endogenous oscillator that drives daily rhythms in physiology, behavior, and gene expression. The underlying mechanisms of circadian timekeeping are cell-autonomous and involve oscillatory expression of core clock genes that is driven by interconnecting transcription–translation feedback loops (TTFLs). Circadian clock TTFLs are further regulated by posttranslational modifications, in particular, phosphorylation. The hippocampus plays an important role in spatial memory and the conversion of short- to long-term memory. Several studies have reported the presence of a peripheral oscillator in the hippocampus and have highlighted the importance of circadian regulation in memory formation. Given the general importance of phosphorylation in circadian clock regulation, we performed global quantitative proteome and phosphoproteome analyses of the murine hippocampus across the circadian cycle, applying spiked-in labeled reference and high accuracy mass spectrometry (MS). Of the 3,052 proteins and 2,868 phosphosites on 1,368 proteins that were accurately quantified, 1.7% of proteins and 5.2% of phosphorylation events exhibited time-of-day-dependent expression profiles. The majority of circadian phosphopeptides displayed abrupt fluctuations at mid-to-late day without underlying rhythms of protein abundance. Bioinformatic analysis of cyclic phosphorylation events revealed their diverse distribution in different biological pathways, most notably, cytoskeletal organization and neuronal morphogenesis. This study provides the first large-scale, quantitative MS analysis of the circadian phosphoproteome and proteome of the murine hippocampus and highlights the significance of rhythmic regulation at the posttranslational level in this peripheral oscillator. In addition to providing molecular insights into the hippocampal circadian clock, our results will assist in the understanding of genetic factors that underlie rhythms-associated pathological states of

  7. A quantitative map of the liver mitochondrial phosphoproteome reveals posttranslational control of ketogenesis.

    Science.gov (United States)

    Grimsrud, Paul A; Carson, Joshua J; Hebert, Alex S; Hubler, Shane L; Niemi, Natalie M; Bailey, Derek J; Jochem, Adam; Stapleton, Donald S; Keller, Mark P; Westphall, Michael S; Yandell, Brian S; Attie, Alan D; Coon, Joshua J; Pagliarini, David J

    2012-11-07

    Mitochondria are dynamic organelles that play a central role in a diverse array of metabolic processes. Elucidating mitochondrial adaptations to changing metabolic demands and the pathogenic alterations that underlie metabolic disorders represent principal challenges in cell biology. Here, we performed multiplexed quantitative mass spectrometry-based proteomics to chart the remodeling of the mouse liver mitochondrial proteome and phosphoproteome during both acute and chronic physiological transformations in more than 50 mice. Our analyses reveal that reversible phosphorylation is widespread in mitochondria, and is a key mechanism for regulating ketogenesis during the onset of obesity and type 2 diabetes. Specifically, we have demonstrated that phosphorylation of a conserved serine on Hmgcs2 (S456) significantly enhances its catalytic activity in response to increased ketogenic demand. Collectively, our work describes the plasticity of this organelle at high resolution and provides a framework for investigating the roles of proteome restructuring and reversible phosphorylation in mitochondrial adaptation.

  8. Phosphoproteomic profiling of human myocardial tissues distinguishes ischemic from non-ischemic end stage heart failure.

    Directory of Open Access Journals (Sweden)

    Matthew A Schechter

    Full Text Available The molecular differences between ischemic (IF and non-ischemic (NIF heart failure are poorly defined. A better understanding of the molecular differences between these two heart failure etiologies may lead to the development of more effective heart failure therapeutics. In this study extensive proteomic and phosphoproteomic profiles of myocardial tissue from patients diagnosed with IF or NIF were assembled and compared. Proteins extracted from left ventricular sections were proteolyzed and phosphopeptides were enriched using titanium dioxide resin. Gel- and label-free nanoscale capillary liquid chromatography coupled to high resolution accuracy mass tandem mass spectrometry allowed for the quantification of 4,436 peptides (corresponding to 450 proteins and 823 phosphopeptides (corresponding to 400 proteins from the unenriched and phospho-enriched fractions, respectively. Protein abundance did not distinguish NIF from IF. In contrast, 37 peptides (corresponding to 26 proteins exhibited a ≥ 2-fold alteration in phosphorylation state (p<0.05 when comparing IF and NIF. The degree of protein phosphorylation at these 37 sites was specifically dependent upon the heart failure etiology examined. Proteins exhibiting phosphorylation alterations were grouped into functional categories: transcriptional activation/RNA processing; cytoskeleton structure/function; molecular chaperones; cell adhesion/signaling; apoptosis; and energetic/metabolism. Phosphoproteomic analysis demonstrated profound post-translational differences in proteins that are involved in multiple cellular processes between different heart failure phenotypes. Understanding the roles these phosphorylation alterations play in the development of NIF and IF has the potential to generate etiology-specific heart failure therapeutics, which could be more effective than current therapeutics in addressing the growing concern of heart failure.

  9. Celiac Anti-Type 2 Transglutaminase Antibodies Induce Phosphoproteome Modification in Intestinal Epithelial Caco-2 Cells

    Science.gov (United States)

    Marabotti, Anna; Lepretti, Marilena; Salzano, Anna Maria; Scaloni, Andrea; Vitale, Monica; Zambrano, Nicola; Sblattero, Daniele; Esposito, Carla

    2013-01-01

    Background Celiac disease is an inflammatory condition of the small intestine that affects genetically predisposed individuals after dietary wheat gliadin ingestion. Type 2-transglutaminase (TG2) activity seems to be responsible for a strong autoimmune response in celiac disease, TG2 being the main autoantigen. Several studies support the concept that celiac anti-TG2 antibodies may contribute to disease pathogenesis. Our recent findings on the ability of anti-TG2 antibodies to induce a rapid intracellular mobilization of calcium ions, as well as extracellular signal-regulated kinase phosphorylation, suggest that they potentially act as signaling molecules. In line with this concept, we have investigated whether anti-TG2 antibodies can induce phosphoproteome modification in an intestinal epithelial cell line. Methods and Principal Findings We studied phosphoproteome modification in Caco-2 cells treated with recombinant celiac anti-TG2 antibodies. We performed a two-dimensional electrophoresis followed by specific staining of phosphoproteins and mass spectrometry analysis of differentially phosphorylated proteins. Of 14 identified proteins (excluding two uncharacterized proteins), three were hypophosphorylated and nine were hyperphosphorylated. Bioinformatics analyses confirmed the presence of phosphorylation sites in all the identified proteins and highlighted their involvement in several fundamental biological processes, such as cell cycle progression, cell stress response, cytoskeletal organization and apoptosis. Conclusions Identification of differentially phosphorylated proteins downstream of TG2-antibody stimulation suggests that in Caco-2 cells these antibodies perturb cell homeostasis by behaving as signaling molecules. We hypothesize that anti-TG2 autoantibodies may destabilize the integrity of the intestinal mucosa in celiac individuals, thus contributing to celiac disease establishment and progression. Since several proteins here identified in this study

  10. Celiac anti-type 2 transglutaminase antibodies induce phosphoproteome modification in intestinal epithelial Caco-2 cells.

    Directory of Open Access Journals (Sweden)

    Gaetana Paolella

    Full Text Available BACKGROUND: Celiac disease is an inflammatory condition of the small intestine that affects genetically predisposed individuals after dietary wheat gliadin ingestion. Type 2-transglutaminase (TG2 activity seems to be responsible for a strong autoimmune response in celiac disease, TG2 being the main autoantigen. Several studies support the concept that celiac anti-TG2 antibodies may contribute to disease pathogenesis. Our recent findings on the ability of anti-TG2 antibodies to induce a rapid intracellular mobilization of calcium ions, as well as extracellular signal-regulated kinase phosphorylation, suggest that they potentially act as signaling molecules. In line with this concept, we have investigated whether anti-TG2 antibodies can induce phosphoproteome modification in an intestinal epithelial cell line. METHODS AND PRINCIPAL FINDINGS: We studied phosphoproteome modification in Caco-2 cells treated with recombinant celiac anti-TG2 antibodies. We performed a two-dimensional electrophoresis followed by specific staining of phosphoproteins and mass spectrometry analysis of differentially phosphorylated proteins. Of 14 identified proteins (excluding two uncharacterized proteins, three were hypophosphorylated and nine were hyperphosphorylated. Bioinformatics analyses confirmed the presence of phosphorylation sites in all the identified proteins and highlighted their involvement in several fundamental biological processes, such as cell cycle progression, cell stress response, cytoskeletal organization and apoptosis. CONCLUSIONS: Identification of differentially phosphorylated proteins downstream of TG2-antibody stimulation suggests that in Caco-2 cells these antibodies perturb cell homeostasis by behaving as signaling molecules. We hypothesize that anti-TG2 autoantibodies may destabilize the integrity of the intestinal mucosa in celiac individuals, thus contributing to celiac disease establishment and progression. Since several proteins here

  11. The current state of the art of quantitative phosphoproteomics and its applications to diabetes research

    Energy Technology Data Exchange (ETDEWEB)

    Chan, Chi Yuet X’avia; Gritsenko, Marina A.; Smith, Richard D.; Qian, Wei-Jun

    2016-03-17

    Protein phosphorylation is a fundamental regulatory mechanism in many cellular processes and aberrant perturbation of phosphorylation has been revealed in various human diseases. Kinases and their cognate inhibitors have been hotspot for drug development. Therefore, the emerging tools, which enable a system-wide quantitative profiling of phosphoproteome, would offer a powerful impetus in unveiling novel signaling pathways, drug targets and/or biomarkers for the disease of interest. In this review, we will highlight recent advances in phosphoproteomics, the current state-of-the-art of the technologies, and the challenges and future perspectives of this research area. Finally, we will underscore some exemplary applications of phosphoproteomics in diabetes research.

  12. The current state of the art of quantitative phosphoproteomics and its applications to diabetes research.

    Science.gov (United States)

    Chan, Chi Yuet X'avia; Gritsenko, Marina A; Smith, Richard D; Qian, Wei-Jun

    2016-01-01

    Protein phosphorylation is a fundamental regulatory mechanism in many cellular processes and aberrant perturbation of phosphorylation has been implicated in various human diseases. Kinases and their cognate inhibitors have been considered as hotspots for drug development. Therefore, the emerging tools, which enable a system-wide quantitative profiling of phosphoproteome, would offer a powerful impetus in unveiling novel signaling pathways, drug targets and/or biomarkers for diseases of interest. This review highlights recent advances in phosphoproteomics, the current state of the art of the technologies and the challenges and future perspectives of this research area. Finally, some exemplary applications of phosphoproteomics in diabetes research are underscored.

  13. Comparative phosphoproteomics of zebrafish Fyn/Yes morpholino knockdown embryos.

    Science.gov (United States)

    Lemeer, Simone; Jopling, Chris; Gouw, Joost; Mohammed, Shabaz; Heck, Albert J R; Slijper, Monique; den Hertog, Jeroen

    2008-11-01

    The coordinated movement of cells is indispensable for normal vertebrate gastrulation. Several important players and signaling pathways have been identified in convergence and extension (CE) cell movements during gastrulation, including non-canonical Wnt signaling. Fyn and Yes, members of the Src family of kinases, are key regulators of CE movements as well. Here we investigated signaling pathways in early development by comparison of the phosphoproteome of wild type zebrafish embryos with Fyn/Yes knockdown embryos that display specific CE cell movement defects. For quantitation we used differential stable isotope labeling by reductive amination of peptides. Equal amounts of labeled peptides from wild type and Fyn/Yes knockdown embryos were mixed and analyzed by on-line reversed phase TiO(2)-reversed phase LC-MS/MS. Phosphorylated and non-phosphorylated peptides were quantified, and significant changes in protein expression and/or phosphorylation were detected. We identified 348 phosphoproteins of which 69 showed a decrease in phosphorylation in Fyn/Yes knockdown embryos and 72 showed an increase in phosphorylation. Among these phosphoproteins were known regulators of cell movements, including Adducin and PDLIM5. Our results indicate that quantitative phosphoproteomics combined with morpholino-mediated knockdowns can be used to identify novel signaling pathways that act in zebrafish development in vivo.

  14. Phosphoproteomics profiling of human skin fibroblast cells reveals pathways and proteins affected by low doses of ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Feng; Waters, Katrina M.; Miller, John H.; Gritsenko, Marina A.; Zhao, Rui; Du, Xiuxia; Livesay, Eric A.; Purvine, Samuel O.; Monroe, Matthew E.; Wang, Yingchun; Camp, David G.; Smith, Richard D.; Stenoien, David L.

    2010-11-30

    Background: High doses of ionizing radiation result in biological damage, however the precise relationships between long term health effects, including cancer, and low dose exposures remain poorly understood and are currently extrapolated using high dose exposure data. Identifying the signaling pathways and individual proteins affected at the post-translational level by radiation should shed valuable insight into the molecular mechanisms that regulate dose dependent responses to radiation. Principle Findings: We have identified 6845 unique phosphopeptides (2566 phosphoproteins) from control and irradiated (2 and 50 cGy) primary human skin fibroblasts one hour post-exposure. Dual statistical analyses based on spectral counts and peak intensities identified 287 phosphopeptides (from 231 proteins) and 244 phosphopeptides (from 182 proteins) that varied significantly following exposure to 2 and 50 cGy respectively. This screen identified phosphorylation sites on proteins with known roles in radiation responses including TP53BP1 as well as previously unidentified radiation responsive proteins such as the candidate tumor suppressor SASH1. Bioinformatics analyses suggest that low and high doses of radiation affect both overlapping and unique biological processes and suggest a role of MAP kinase and protein kinase A (PKA) signaling in the radiation response as well as differential regulation of p53 networks at low and high doses of radiation. Conlcusions: Our results represent the most comprehensive analysis of the phosphoproteomes of human primary fibroblasts exposed to multiple doses of ionizing radiation published to date and provides a basis for the systems level identification of biological processes, molecular pathways and individual proteins regulated in a dose dependent manner by ionizing radiation. Further study of these modified proteins and affected networks should help to define the molecular mechanisms that regulate biological responses to radiation at

  15. Quantitative Label-Free Phosphoproteomics Reveals Differentially Regulated Protein Phosphorylation Involved in West Nile Virus-Induced Host Inflammatory Response.

    Science.gov (United States)

    Zhang, Hao; Sun, Jun; Ye, Jing; Ashraf, Usama; Chen, Zheng; Zhu, Bibo; He, Wen; Xu, Qiuping; Wei, Yanming; Chen, Huanchun; Fu, Zhen F; Liu, Rong; Cao, Shengbo

    2015-12-01

    West Nile virus (WNV) can cause neuro-invasive and febrile illness that may be fatal to humans. The production of inflammatory cytokines is key to mediating WNV-induced immunopathology in the central nervous system. Elucidating the host factors utilized by WNV for productive infection would provide valuable insights into the evasion strategies used by this virus. Although attempts have been made to determine these host factors, proteomic data depicting WNV-host protein interactions are limited. We applied liquid chromatography-tandem mass spectrometry for label-free, quantitative phosphoproteomics to systematically investigate the global phosphorylation events induced by WNV infection. Quantifiable changes to 1,657 phosphoproteins were found; of these, 626 were significantly upregulated and 227 were downregulated at 12 h postinfection. The phosphoproteomic data were subjected to gene ontology enrichment analysis, which returned the inflammation-related spliceosome, ErbB, mitogen-activated protein kinase, nuclear factor kappa B, and mechanistic target of rapamycin signaling pathways. We used short interfering RNAs to decrease the levels of glycogen synthase kinase-3 beta, bifunctional polynucleotide phosphatase/kinase, and retinoblastoma 1 and found that the activity of nuclear factor kappa B (p65) is significantly decreased in WNV-infected U251 cells, which in turn led to markedly reduced inflammatory cytokine production. Our results provide a better understanding of the host response to WNV infection and highlight multiple targets for the development of antiviral and anti-inflammatory therapies.

  16. Up-to-Date Workflow for Plant (Phospho)proteomics Identifies Differential Drought-Responsive Phosphorylation Events in Maize Leaves.

    Science.gov (United States)

    Vu, Lam Dai; Stes, Elisabeth; Van Bel, Michiel; Nelissen, Hilde; Maddelein, Davy; Inzé, Dirk; Coppens, Frederik; Martens, Lennart; Gevaert, Kris; De Smet, Ive

    2016-12-02

    Protein phosphorylation is one of the most common post-translational modifications (PTMs), which can regulate protein activity and localization as well as protein-protein interactions in numerous cellular processes. Phosphopeptide enrichment techniques enable plant researchers to acquire insight into phosphorylation-controlled signaling networks in various plant species. Most phosphoproteome analyses of plant samples still involve stable isotope labeling, peptide fractionation, and demand a lot of mass spectrometry (MS) time. Here, we present a simple workflow to probe, map, and catalogue plant phosphoproteomes, requiring relatively low amounts of starting material, no labeling, no fractionation, and no excessive analysis time. Following optimization of the different experimental steps on Arabidopsis thaliana samples, we transferred our workflow to maize, a major monocot crop, to study signaling upon drought stress. In addition, we included normalization to protein abundance to identify true phosphorylation changes. Overall, we identified a set of new phosphosites in both Arabidopsis thaliana and maize, some of which are differentially phosphorylated upon drought. All data are available via ProteomeXchange with identifier PXD003634, but to provide easy access to our model plant and crop data sets, we created an online database, Plant PTM Viewer ( bioinformatics.psb.ugent.be/webtools/ptm_viewer/ ), where all phosphosites identified in our study can be consulted.

  17. Online Nanoflow Multidimensional Fractionation for High Efficiency Phosphopeptide Analysis*

    OpenAIRE

    Ficarro, Scott B.; Zhang, Yi; Carrasco-Alfonso, Marlene J.; Garg, Brijesh; Adelmant, Guillaume; Webber, James T.; Luckey, C. John; Marto, Jarrod A.

    2011-01-01

    Despite intense, continued interest in global analyses of signaling cascades through mass spectrometry-based studies, the large-scale, systematic production of phosphoproteomics data has been hampered in-part by inefficient fractionation strategies subsequent to phosphopeptide enrichment. Here we explore two novel multidimensional fractionation strategies for analysis of phosphopeptides. In the first technique we utilize aliphatic ion pairing agents to improve retention of phosphopeptides at ...

  18. Evidence for a Minimal Eukaryotic Phosphoproteome?

    NARCIS (Netherlands)

    Diks, Sander H.; Parikh, Kaushal; van der Sijde, Marijke; Joore, Jos; Ritsema, Tita; Peppelenbosch, Maikel P.

    2007-01-01

    Background. Reversible phosphorylation catalysed by kinases is probably the most important regulatory mechanism in eukaryotes. Methodology/Principal Findings. We studied the in vitro phosphorylation of peptide arrays exhibiting the majority of PhosphoBase-deposited protein sequences, by factors in c

  19. Quantitative proteomics and phosphoproteomics on serial tumor biopsies from a sorafenib-treated HCC patient

    Science.gov (United States)

    Dazert, Eva; Colombi, Marco; Boldanova, Tujana; Moes, Suzette; Adametz, David; Quagliata, Luca; Roth, Volker; Terracciano, Luigi; Heim, Markus H.; Jenoe, Paul; Hall, Michael N.

    2016-01-01

    Compensatory signaling pathways in tumors confer resistance to targeted therapy, but the pathways and their mechanisms of activation remain largely unknown. We describe a procedure for quantitative proteomics and phosphoproteomics on snap-frozen biopsies of hepatocellular carcinoma (HCC) and matched nontumor liver tissue. We applied this procedure to monitor signaling pathways in serial biopsies taken from an HCC patient before and during treatment with the multikinase inhibitor sorafenib. At diagnosis, the patient had an advanced HCC. At the time of the second biopsy, abdominal imaging revealed progressive disease despite sorafenib treatment. Sorafenib was confirmed to inhibit MAPK signaling in the tumor, as measured by reduced ribosomal protein S6 kinase phosphorylation. Hierarchical clustering and enrichment analysis revealed pathways broadly implicated in tumor progression and resistance, such as epithelial-to-mesenchymal transition and cell adhesion pathways. Thus, we describe a protocol for quantitative analysis of oncogenic pathways in HCC biopsies and obtained first insights into the effect of sorafenib in vivo. This protocol will allow elucidation of mechanisms of resistance and enable precision medicine. PMID:26787912

  20. Newly fabricated magnetic lanthanide oxides core-shell nanoparticles in phosphoproteomics.

    Science.gov (United States)

    Jabeen, Fahmida; Najam-Ul-Haq, Muhammad; Rainer, Matthias; Güzel, Yüksel; Huck, Christian W; Bonn, Guenther K

    2015-01-01

    Metal oxides show high selectivity and sensitivity toward mass spectrometry based enrichment strategies. Phosphopeptides/phosphoproteins enrichment from biological samples is cumbersome because of their low abundance. Phosphopeptides are of interest in enzymes and phosphorylation pathways which lead to the clinical links of a disease. Magnetic core-shell lanthanide oxide nanoparticles (Fe3O4@SiO2-La2O3 and Fe3O4@SiO2-Sm2O3) are fabricated, characterized by SEM, FTIR, and EDX and employed in the enrichment of phosphopeptides. The nanoparticles enrich phosphopeptides from casein variants, nonfat milk, egg yolk, human serum and HeLa cell extract. The materials and enrichment protocols are designed in a way that there are almost no nonspecific bindings. The selectivity is achieved up to 1:8500 using β-casein/BSA mixture and sensitivity down to 1 atto-mole. Batch-to-batch reproducibility is high with the reuse of core-shell nanoparticles up to four cycles. The enrichment followed by MALDI-MS analyses is carried out for the identification of phosphopeptides from serum digest and HeLa cell extract. Characteristic phosphopeptides of phosphoproteins are identified from human serum after the enrichment, which have the diagnostic potential toward prostate cancer. Thus, the lanthanide based magnetic core-shell materials offer a highly selective and sensitive workflow in phosphoproteomics.

  1. Quantitative Proteomic and Phosphoproteomic Approaches for Deciphering the Signaling Pathway for Tension Wood Formation in Poplar.

    Science.gov (United States)

    Mauriat, Mélanie; Leplé, Jean-Charles; Claverol, Stéphane; Bartholomé, Jérôme; Negroni, Luc; Richet, Nicolas; Lalanne, Céline; Bonneu, Marc; Coutand, Catherine; Plomion, Christophe

    2015-08-07

    Trees adjust their growth following forced changes in orientation to re-establish a vertical position. In angiosperms, this adjustment involves the differential regulation of vascular cambial activity between the lower (opposite wood) and upper (tension wood) sides of the leaning stem. We investigated the molecular mechanisms leading to the formation of differential wood types through a quantitative proteomic and phosphoproteomic analysis on poplar subjected to a gravitropic stimulus. We identified and quantified 675 phosphopeptides, corresponding to 468 phosphoproteins, and 3 763 nonphosphorylated peptides, corresponding to 1 155 proteins, in the differentiating xylem of straight-growing trees (control) and trees subjected to a gravitational stimulus during 8 weeks. About 1% of the peptides were specific to a wood type (straight, opposite, or tension wood). Proteins quantified in more than one type of wood were more numerous: a mixed linear model showed 389 phosphopeptides and 556 proteins to differ in abundance between tension wood and opposite wood. Twenty-one percent of the phosphoproteins identified here were described in their phosphorylated form for the first time. Our analyses revealed remarkable developmental molecular plasticity, with wood type-specific phosphorylation events, and highlighted the involvement of different proteins in the biosynthesis of cell wall components during the formation of the three types of wood.

  2. Phosphoproteomics and bioinformatics analyses of spinal cord proteins in rats with morphine tolerance.

    Directory of Open Access Journals (Sweden)

    Wen-Jinn Liaw

    Full Text Available INTRODUCTION: Morphine is the most effective pain-relieving drug, but it can cause unwanted side effects. Direct neuraxial administration of morphine to spinal cord not only can provide effective, reliable pain relief but also can prevent the development of supraspinal side effects. However, repeated neuraxial administration of morphine may still lead to morphine tolerance. METHODS: To better understand the mechanism that causes morphine tolerance, we induced tolerance in rats at the spinal cord level by giving them twice-daily injections of morphine (20 µg/10 µL for 4 days. We confirmed tolerance by measuring paw withdrawal latencies and maximal possible analgesic effect of morphine on day 5. We then carried out phosphoproteomic analysis to investigate the global phosphorylation of spinal proteins associated with morphine tolerance. Finally, pull-down assays were used to identify phosphorylated types and sites of 14-3-3 proteins, and bioinformatics was applied to predict biological networks impacted by the morphine-regulated proteins. RESULTS: Our proteomics data showed that repeated morphine treatment altered phosphorylation of 10 proteins in the spinal cord. Pull-down assays identified 2 serine/threonine phosphorylated sites in 14-3-3 proteins. Bioinformatics further revealed that morphine impacted on cytoskeletal reorganization, neuroplasticity, protein folding and modulation, signal transduction and biomolecular metabolism. CONCLUSIONS: Repeated morphine administration may affect multiple biological networks by altering protein phosphorylation. These data may provide insight into the mechanism that underlies the development of morphine tolerance.

  3. Comparative Proteome and Phosphoproteome Analyses during Cyprid Development of the Barnacle Balanus ( =Amphibalanus ) amphitrite

    KAUST Repository

    Zhang, Yu

    2010-06-04

    The barnacle Balanus amphitrite (=Amphibalanus amphitrite) is a major marine biofouling invertebrate worldwide. It has a complex life cycle during which the larva (called a nauplius) molts six times before transforming into the cyprid stage. The cyprid stage in B. amphitrite is the critical stage for the larval decision to attach and metamorphose. In this study, proteome and phosphoproteome alterations during cyprid development/aging and upon treatment with the antifouling agent butenolide were examined with a two-dimensional electrophoresis (2-DE) multiplexed fluorescent staining approach. Optimized protein separation strategies, including solution-phase isoelectric fractionation and narrow-pH-range 2-DE, were used in a proteomic analysis. Our results show that the differential regulation of the target proteins is highly dynamic on the levels of both protein expression and posttranslational modification. Two groups of proteins, stress-associated and energy metabolism-related proteins, are differentially expressed during cyprid development. Comparison of the control and treatment groups suggests that butenolide exerts its effects by sustaining the expression levels of these proteins. Altogether, our data suggest that proteins involved in stress regulation and energy metabolism play crucial roles in regulating larval attachment and metamorphosis of B. amphitrite. © 2010 American Chemical Society.

  4. The differential hippocampal phosphoproteome of Apodemus sylvaticus paralleling spatial memory retrieval in the Barnes maze.

    Science.gov (United States)

    Li, Lin; Csaszar, Edina; Szodorai, Edit; Patil, Sudarshan; Pollak, Arnold; Lubec, Gert

    2014-05-01

    Protein phosphorylation is a well-known and well-documented mechanism in memory processes. Although a large series of protein kinases involved in memory processes have been reported, information on phosphoproteins is limited. It was therefore the aim of the study to determine a partial and differential phosphoproteome along with the corresponding network in hippocampus of a wild caught mouse strain with excellent performance in several paradigms of spatial memory. Apodemus sylvaticus mice were trained in the Barnes maze, a non-invasive test system for spatial memory and untrained mice served as controls. Animals were sacrificed 6h following memory retrieval, hippocampi were taken, proteins extracted and in-solution digestion was carried out with subsequent iTRAQ double labelling. Phosphopeptides were enriched by a TiO2-based method and semi-quantified using two fragmentation principles on the LTQ-orbitrap Velos. In hippocampi of trained animals phosphopeptide levels representing signalling, neuronal, synaptosomal, cytoskeletal and metabolism proteins were at least twofold reduced or increased. Furthermore, a network revealing a link to pathways of ubiquitination, the androgen receptor, small GTPase Rab5 and MAPK signaling as well as synucleins was constructed. This work is relevant for interpretation of previous work and the design of future studies on protein phosphorylation in spatial memory.

  5. Phosphoproteomics Profiling of Tobacco Mature Pollen and Pollen Activated in vitro.

    Science.gov (United States)

    Fíla, Jan; Radau, Sonja; Matros, Andrea; Hartmann, Anja; Scholz, Uwe; Feciková, Jana; Mock, Hans-Peter; Čapková, Věra; Zahedi, René Peiman; Honys, David

    2016-04-01

    Tobacco mature pollen has extremely desiccated cytoplasm, and is metabolically quiescent. Upon re-hydration it becomes metabolically active and that results in later emergence of rapidly growing pollen tube. These changes in cytoplasm hydration and metabolic activity are accompanied by protein phosphorylation. In this study, we subjected mature pollen, 5-min-activated pollen, and 30-min-activated pollen to TCA/acetone protein extraction, trypsin digestion and phosphopeptide enrichment by titanium dioxide. The enriched fraction was subjected to nLC-MS/MS. We identified 471 phosphopeptides that carried 432 phosphorylation sites, position of which was exactly matched by mass spectrometry. These 471 phosphopeptides were assigned to 301 phosphoproteins, because some proteins carried more phosphorylation sites. Of the 13 functional groups, the majority of proteins were put into these categories: transcription, protein synthesis, protein destination and storage, and signal transduction. Many proteins were of unknown function, reflecting the fact that male gametophyte contains many specific proteins that have not been fully functionally annotated. The quantitative data highlighted the dynamics of protein phosphorylation during pollen activation; the identified phosphopeptides were divided into seven groups based on the regulatory trends. The major group comprised mature pollen-specific phosphopeptides that were dephosphorylated during pollen activation. Several phosphopeptides representing the same phosphoprotein had different regulation, which pinpointed the complexity of protein phosphorylation and its clear functional context. Collectively, we showed the first phosphoproteomics data on activated pollen where the position of phosphorylation sites was clearly demonstrated and regulatory kinetics was resolved.

  6. The proteome and phosphoproteome of Neurospora crassa in response to cellulose, sucrose and carbon starvation.

    Science.gov (United States)

    Xiong, Yi; Coradetti, Samuel T; Li, Xin; Gritsenko, Marina A; Clauss, Therese; Petyuk, Vlad; Camp, David; Smith, Richard; Cate, Jamie H D; Yang, Feng; Glass, N Louise

    2014-11-01

    Improving cellulolytic enzyme production by plant biomass degrading fungi holds great potential in reducing costs associated with production of next-generation biofuels generated from lignocellulose. How fungi sense cellulosic materials and respond by secreting enzymes has mainly been examined by assessing function of transcriptional regulators and via transcriptional profiling. Here, we obtained global proteomic and phosphoproteomic profiles of the plant biomass degrading filamentous fungus Neurospora crassa grown on different carbon sources, i.e. sucrose, no carbon, and cellulose, by performing isobaric tags for relative and absolute quantification (iTRAQ)-based LC-MS/MS analyses. A comparison between proteomes and transcriptomes under identical carbon conditions suggests that extensive post-transcriptional regulation occurs in N. crassa in response to exposure to cellulosic material. Several hundred amino acid residues with differential phosphorylation levels on crystalline cellulose (Avicel) or carbon-free medium vs sucrose medium were identified, including phosphorylation sites in a major transcriptional activator for cellulase genes, CLR1, as well as a cellobionic acid transporter, CBT1. Mutation of phosphorylation sites on CLR1 did not have a major effect on transactivation of cellulase production, while mutation of phosphorylation sites in CBT1 increased its transporting capacity. Our data provides rich information at both the protein and phosphorylation levels of the early cellular responses to carbon starvation and cellulosic induction and aids in a greater understanding of the underlying post-transcriptional regulatory mechanisms in filamentous fungi.

  7. Exploring the human leukocyte phosphoproteome using a microfluidic reversed-phase-TiO2-reversed-phase high-performance liquid chromatography phosphochip coupled to a quadrupole time-of-flight mass spectrometer.

    Science.gov (United States)

    Raijmakers, Reinout; Kraiczek, Karsten; de Jong, Ad P; Mohammed, Shabaz; Heck, Albert J R

    2010-02-01

    The study of protein phosphorylation events is one of the most important challenges in proteome analysis. Despite the importance of phosphorylation for many regulatory processes in cells and many years of phosphoprotein and phosphopeptide research, the identification and characterization of phosphorylation by mass spectrometry is still a challenging task. Recently, we introduced an approach that facilitates the analysis of phosphopeptides by performing automated, online, TiO(2) enrichment of phosphopeptides prior to mass spectrometry (MS) analysis. The implementation of that method on a "plug-and-play" microfluidic high-performance liquid chromatography (HPLC) chip design will potentially open up efficient phosphopeptide enrichment methods enabling phosphoproteomics analyses by a broader research community. Following our initial proof of principle, whereby the device was coupled to an ion trap, we now show that this so-called phosphochip is capable of the enrichment of large numbers of phosphopeptides from complex cellular lysates, which can be more readily identified when coupled to a higher resolution quadrupole time-of-flight (Q-TOF) mass spectrometer. We use the phosphochip-Q-TOF setup to explore the phosphoproteome of nonstimulated primary human leukocytes where we identify 1012 unique phosphopeptides corresponding to 960 different phosphorylation sites providing for the first time an overview of the phosphoproteome of these important circulating white blood cells.

  8. Phosphoproteomic profiling of in vivo signaling in liver by the mammalian target of rapamycin complex 1 (mTORC1.

    Directory of Open Access Journals (Sweden)

    Gokhan Demirkan

    Full Text Available Our understanding of signal transduction networks in the physiological context of an organism remains limited, partly due to the technical challenge of identifying serine/threonine phosphorylated peptides from complex tissue samples. In the present study, we focused on signaling through the mammalian target of rapamycin (mTOR complex 1 (mTORC1, which is at the center of a nutrient- and growth factor-responsive cell signaling network. Though studied extensively, the mechanisms involved in many mTORC1 biological functions remain poorly understood.We developed a phosphoproteomic strategy to purify, enrich and identify phosphopeptides from rat liver homogenates. Using the anticancer drug rapamycin, the only known target of which is mTORC1, we characterized signaling in liver from rats in which the complex was maximally activated by refeeding following 48 hr of starvation. Using protein and peptide fractionation methods, TiO(2 affinity purification of phosphopeptides and mass spectrometry, we reproducibly identified and quantified over four thousand phosphopeptides. Along with 5 known rapamycin-sensitive phosphorylation events, we identified 62 new rapamycin-responsive candidate phosphorylation sites. Among these were PRAS40, gephyrin, and AMP kinase 2. We observed similar proportions of increased and reduced phosphorylation in response to rapamycin. Gene ontology analysis revealed over-representation of mTOR pathway components among rapamycin-sensitive phosphopeptide candidates.In addition to identifying potential new mTORC1-mediated phosphorylation events, and providing information relevant to the biology of this signaling network, our experimental and analytical approaches indicate the feasibility of large-scale phosphoproteomic profiling of tissue samples to study physiological signaling events in vivo.

  9. Comprehensive analysis of signal transduction in three-dimensional ECM-based tumor cell cultures

    Directory of Open Access Journals (Sweden)

    Iris Eke

    2015-11-01

    Full Text Available Analysis of signal transduction and protein phosphorylation is fundamental to understand physiological and pathological cell behavior as well as identification of novel therapeutic targets. Despite the fact that more physiological three-dimensional cell culture assays are increasingly used, particularly proteomics and phosphoproteomics remain challenging due to easy, robust and reproducible sample preparation. Here, we present an easy-to-perform, reliable and time-efficient method for the production of 3D cell lysates without compromising cell adhesion before cell lysis. The samples can be used for Western blotting as well as phosphoproteome array technology. This technique would be of interest for researchers working in all fields of biology and drug development.

  10. Analysis of protein phosphorylation using mass spectrometry: deciphering the phosphoproteome

    DEFF Research Database (Denmark)

    Mann, Matthias; Ong, Shao En; Grønborg, Mads

    2002-01-01

    In signal transduction in eukaryotes, protein phosphorylation is a key event. To understand signaling processes, we must first acquire an inventory of phosphoproteins and their phosphorylation sites under different conditions. Because phosphorylation is a dynamic process, elucidation of signaling...

  11. Enhanced Phosphoproteomic Profiling Workflow For Growth Factor Signaling Analysis

    DEFF Research Database (Denmark)

    Sylvester, Marc; Burbridge, Mike; Leclerc, Gregory;

    2010-01-01

    Background Our understanding of complex signaling networks is still fragmentary. Isolated processes have been studied extensively but cross-talk is omnipresent and precludes intuitive predictions of signaling outcomes. The need for quantitative data on dynamic systems is apparent especially for our...... A549 lung carcinoma cells were used as a model and stimulated with hepatocyte growth factor, epidermal growth factor or fibroblast growth factor. We employed a quick protein digestion workflow with spin filters without using urea. Phosphopeptides in general were enriched by sequential elution from...... transfer dissociation adds confidence in modification site assignment. The workflow is relatively simple but the integration of complementary techniques leads to a deeper insight into cellular signaling networks and the potential pharmacological intervention thereof....

  12. Quantitative Proteomic and Phosphoproteomic Analysis of Trypanosoma cruzi Amastigogenesis

    DEFF Research Database (Denmark)

    Queiroz, Rayner M L; Charneau, Sebastien; Mandacaru, Samuel C;

    2014-01-01

    Chagas disease is a tropical neglected disease endemic in Latin America and it is caused by the protozoan Trypanosoma cruzi. The parasite has four major life stages: epimastigote, metacyclic trypomastigote, bloodstream trypomastigote and amastigote. The differentiation from infective trypomastigo......Chagas disease is a tropical neglected disease endemic in Latin America and it is caused by the protozoan Trypanosoma cruzi. The parasite has four major life stages: epimastigote, metacyclic trypomastigote, bloodstream trypomastigote and amastigote. The differentiation from infective...

  13. Identification of BCAP-{sub L} as a negative regulator of the TLR signaling-induced production of IL-6 and IL-10 in macrophages by tyrosine phosphoproteomics

    Energy Technology Data Exchange (ETDEWEB)

    Matsumura, Takayuki [Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, Shinjuku-ku, Tokyo 162-0041 (Japan); Department of Life Science and Medical Bio-Science, Waseda University, Shinjuku-ku, Tokyo 162-8480 (Japan); Oyama, Masaaki; Kozuka-Hata, Hiroko [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639 (Japan); Ishikawa, Kosuke; Inoue, Takafumi [Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, Shinjuku-ku, Tokyo 162-0041 (Japan); Department of Life Science and Medical Bio-Science, Waseda University, Shinjuku-ku, Tokyo 162-8480 (Japan); Muta, Tatsushi [Laboratory of Cell Recognition and Response, Graduate School of Life Sciences, Tohoku University, Sendai 980-8578 (Japan); Semba, Kentaro, E-mail: ksemba@waseda.jp [Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, Shinjuku-ku, Tokyo 162-0041 (Japan); Department of Life Science and Medical Bio-Science, Waseda University, Shinjuku-ku, Tokyo 162-8480 (Japan); Inoue, Jun-ichiro, E-mail: jun-i@ims.u-tokyo.ac.jp [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639 (Japan); Division of Cellular and Molecular Biology, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639 (Japan)

    2010-09-17

    Research highlights: {yields} Twenty five tyrosine-phosphorylated proteins in LPS-stimulated macrophages were determined. {yields} BCAP is a novel tyrosine-phosphorylated protein in LPS-stimulated macrophages. {yields} BCAP-{sub L} inhibits IL-6 and IL-10 production in LPS-stimulated macrophages. -- Abstract: Toll-like receptor (TLR) signaling in macrophages is essential for anti-pathogen responses such as cytokine production and antigen presentation. Although numerous reports suggest that protein tyrosine kinases (PTKs) are involved in cytokine induction in response to lipopolysaccharides (LPS; TLR4 ligand) in macrophages, the PTK-mediated signal transduction pathway has yet to be analyzed in detail. Here, we carried out a comprehensive and quantitative dynamic tyrosine phosphoproteomic analysis on the TLR4-mediated host defense system in RAW264.7 macrophages using stable isotope labeling by amino acids in cell culture (SILAC). We determined the temporal profiles of 25 proteins based on SILAC-encoded peptide(s). Of these, we focused on the tyrosine phosphorylation of B-cell adaptor for phosphatidylinositol 3-kinase (BCAP) because the function of BCAP remains unknown in TLR signaling in macrophages. Furthermore, Bcap has two distinct transcripts, a full-length (Bcap-{sub L}) and an alternatively initiated or spliced (Bcap-{sub S}) mRNA, and little is known about the differential functions of the BCAP-{sub L} and BCAP-{sub S} proteins. Our study showed, for the first time, that RNAi-mediated selective depletion of BCAP-{sub L} enhanced IL-6 and IL-10 production but not TNF-{alpha} production in TLR ligand-stimulated macrophages. We propose that BCAP-{sub L} (but not BCAP-{sub S}) is a negative regulator of the TLR-mediated host defense system in macrophages.

  14. Comparative phosphoproteomics reveals components of host cell invasion and post-transcriptional regulation during Francisella infection

    Energy Technology Data Exchange (ETDEWEB)

    Nakayasu, Ernesto S.; Tempel, Rebecca; Cambronne, Xiaolu A.; Petyuk, Vladislav A.; Jones, Marcus B.; Gritsenko, Marina A.; Monroe, Matthew E.; Yang, Feng; Smith, Richard D.; Adkins, Joshua N.; Heffron, Fred

    2013-09-22

    Francisella tularensis is a facultative intracellular bacterium that causes the deadly disease tularemia. Most evidence suggests that Francisella is not well recognized by the innate immune system that normally leads to cytokine expression and cell death. In previous work, we identified new bacterial factors that were hyper-cytotoxic to macrophages. Four of the identified hyper-cytotoxic strains (lpcC, manB, manC and kdtA) had an impaired lipopolysaccharide (LPS) synthesis and produced an exposed lipid A lacking the O-antigen. These mutants were not only hyper-cytotoxic but also were phagocytosed at much higher rates compared to the wild type parent strain. To elucidate the cellular signaling underlying this enhanced phagocytosis and cell death, we performed a large-scale comparative phosphoproteomic analysis of cells infected with wild-type and delta-lpcC F. novicida. Our data suggest that not only actin but also intermediate filaments and microtubules are important for F. novicida entry into the host cells. In addition, we observed differential phosphorylation of tristetraprolin (TTP), a key component of the mRNA-degrading machinery that controls the expression of a variety of genes including many cytokines. Infection with the delta-lpcC mutant induced the hyper-phosphorylation and inhibition of TTP, leading to the production of cytokines such as IL-1beta and TNF-alpha which may kill the host cells by triggering apoptosis. Together, our data provide new insights for Francisella invasion and a post-transcriptional mechanism that prevents the expression of host immune response factors that controls infection by this pathogen.

  15. Comparative Phosphoproteomics Reveals Components of Host Cell Invasion and Post-transcriptional Regulation During Francisella Infection*

    Science.gov (United States)

    Nakayasu, Ernesto S.; Tempel, Rebecca; Cambronne, Xiaolu A.; Petyuk, Vladislav A.; Jones, Marcus B.; Gritsenko, Marina A.; Monroe, Matthew E.; Yang, Feng; Smith, Richard D.; Adkins, Joshua N.; Heffron, Fred

    2013-01-01

    Francisella tularensis is a facultative intracellular bacterium that causes the deadly disease tularemia. Most evidence suggests that Francisella is not well recognized by the innate immune system that normally leads to cytokine expression and cell death. In previous work, we identified new bacterial factors that were hyper-cytotoxic to macrophages. Four of the identified hyper-cytotoxic strains (lpcC, manB, manC, and kdtA) had an impaired lipopolysaccharide (LPS) synthesis and produced an exposed lipid A lacking the O-antigen. These mutants were not only hyper-cytotoxic but also were phagocytosed at much higher rates compared with the wild type parent strain. To elucidate the cellular signaling underlying this enhanced phagocytosis and cell death, we performed a large-scale comparative phosphoproteomic analysis of cells infected with wild-type and delta-lpcC F. novicida. Our data suggest that not only actin but also intermediate filaments and microtubules are important for F. novicida entry into the host cells. In addition, we observed differential phosphorylation of tristetraprolin, a key component of the mRNA-degrading machinery that controls the expression of a variety of genes including many cytokines. Infection with the delta-lpcC mutant induced the hyper-phosphorylation and inhibition of tristetraprolin, leading to the production of cytokines such as IL-1beta and TNF-alpha that may kill the host cells by triggering apoptosis. Together, our data provide new insights for Francisella invasion and a post-transcriptional mechanism that prevents the expression of host immune response factors that control infection by this pathogen. PMID:23970565

  16. Comparative phosphoproteomics reveals components of host cell invasion and post-transcriptional regulation during Francisella infection.

    Science.gov (United States)

    Nakayasu, Ernesto S; Tempel, Rebecca; Cambronne, Xiaolu A; Petyuk, Vladislav A; Jones, Marcus B; Gritsenko, Marina A; Monroe, Matthew E; Yang, Feng; Smith, Richard D; Adkins, Joshua N; Heffron, Fred

    2013-11-01

    Francisella tularensis is a facultative intracellular bacterium that causes the deadly disease tularemia. Most evidence suggests that Francisella is not well recognized by the innate immune system that normally leads to cytokine expression and cell death. In previous work, we identified new bacterial factors that were hyper-cytotoxic to macrophages. Four of the identified hyper-cytotoxic strains (lpcC, manB, manC, and kdtA) had an impaired lipopolysaccharide (LPS) synthesis and produced an exposed lipid A lacking the O-antigen. These mutants were not only hyper-cytotoxic but also were phagocytosed at much higher rates compared with the wild type parent strain. To elucidate the cellular signaling underlying this enhanced phagocytosis and cell death, we performed a large-scale comparative phosphoproteomic analysis of cells infected with wild-type and delta-lpcC F. novicida. Our data suggest that not only actin but also intermediate filaments and microtubules are important for F. novicida entry into the host cells. In addition, we observed differential phosphorylation of tristetraprolin, a key component of the mRNA-degrading machinery that controls the expression of a variety of genes including many cytokines. Infection with the delta-lpcC mutant induced the hyper-phosphorylation and inhibition of tristetraprolin, leading to the production of cytokines such as IL-1beta and TNF-alpha that may kill the host cells by triggering apoptosis. Together, our data provide new insights for Francisella invasion and a post-transcriptional mechanism that prevents the expression of host immune response factors that control infection by this pathogen.

  17. Salt-induced changes in cardiac phosphoproteome in a rat model of chronic renal failure.

    Directory of Open Access Journals (Sweden)

    Zhengxiu Su

    Full Text Available Heart damage is widely present in patients with chronic kidney disease. Salt diet is the most important environmental factor affecting development of chronic renal failure and cardiovascular diseases. The proteins involved in chronic kidney disease -induced heart damage, especially their posttranslational modifications, remain largely unknown to date. Sprague-Dawley rats underwent 5/6 nephrectomy (chronic renal failure model or sham operation were treated for 2 weeks with a normal-(0.4% NaCl, or high-salt (4% NaCl diet. We employed TiO2 enrichment, iTRAQ labeling and liquid-chromatography tandem mass spectrometry strategy for phosphoproteomic profiling of left ventricular free walls in these animals. A total of 1724 unique phosphopeptides representing 2551 non-redundant phosphorylation sites corresponding to 763 phosphoproteins were identified. During normal salt feeding, 89 (54% phosphopeptides upregulated and 76 (46% phosphopeptides downregulated in chronic renal failure rats relative to sham rats. In chronic renal failure rats, high salt intake induced upregulation of 84 (49% phosphopeptides and downregulation of 88 (51% phosphopeptides. Database searches revealed that most of the identified phospholproteins were important signaling molecules such as protein kinases, receptors and phosphatases. These phospholproteins were involved in energy metabolism, cell communication, cell differentiation, cell death and other biological processes. The Search Tool for the Retrieval of Interacting Genes analysis revealed functional links among 15 significantly regulated phosphoproteins in chronic renal failure rats compared to sham group, and 23 altered phosphoproteins induced by high salt intake. The altered phosphorylation levels of two proteins involved in heart damage, lamin A and phospholamban were validated. Expression of the downstream genes of these two proteins, desmin and SERCA2a, were also analyzed.

  18. Salt-induced changes in cardiac phosphoproteome in a rat model of chronic renal failure.

    Science.gov (United States)

    Su, Zhengxiu; Zhu, Hongguo; Zhang, Menghuan; Wang, Liangliang; He, Hanchang; Jiang, Shaoling; Hou, Fan Fan; Li, Aiqing

    2014-01-01

    Heart damage is widely present in patients with chronic kidney disease. Salt diet is the most important environmental factor affecting development of chronic renal failure and cardiovascular diseases. The proteins involved in chronic kidney disease -induced heart damage, especially their posttranslational modifications, remain largely unknown to date. Sprague-Dawley rats underwent 5/6 nephrectomy (chronic renal failure model) or sham operation were treated for 2 weeks with a normal-(0.4% NaCl), or high-salt (4% NaCl) diet. We employed TiO2 enrichment, iTRAQ labeling and liquid-chromatography tandem mass spectrometry strategy for phosphoproteomic profiling of left ventricular free walls in these animals. A total of 1724 unique phosphopeptides representing 2551 non-redundant phosphorylation sites corresponding to 763 phosphoproteins were identified. During normal salt feeding, 89 (54%) phosphopeptides upregulated and 76 (46%) phosphopeptides downregulated in chronic renal failure rats relative to sham rats. In chronic renal failure rats, high salt intake induced upregulation of 84 (49%) phosphopeptides and downregulation of 88 (51%) phosphopeptides. Database searches revealed that most of the identified phospholproteins were important signaling molecules such as protein kinases, receptors and phosphatases. These phospholproteins were involved in energy metabolism, cell communication, cell differentiation, cell death and other biological processes. The Search Tool for the Retrieval of Interacting Genes analysis revealed functional links among 15 significantly regulated phosphoproteins in chronic renal failure rats compared to sham group, and 23 altered phosphoproteins induced by high salt intake. The altered phosphorylation levels of two proteins involved in heart damage, lamin A and phospholamban were validated. Expression of the downstream genes of these two proteins, desmin and SERCA2a, were also analyzed.

  19. A comparison of the chicken and turkey proteomes and phosphoproteomes in the development of poultry-specific immuno-metabolism kinome peptide arrays

    Directory of Open Access Journals (Sweden)

    Ryan J Arsenault

    2014-11-01

    Full Text Available The use of species-specific peptide arrays for the study of animal kinomes has a proven track record of success. This technique has been used in a variety of species for the study of host-pathogen interactions and metabolism. Species-specific peptide arrays have been designed previously for use with chicken, but a turkey array has never been attempted. In addition, arrays designed around individual cellular functions have been designed and utilized, but cross-function immuno-metabolic arrays have not been considered previously. Antecedent to designing separate chicken and turkey immuno-metabolic kinome peptide arrays, we show that while the chicken and turkey genomes are quite similar, the two species are much more distinct at the proteome and phosphoproteome levels. Despite a genome identity of approximately 90%, we observe that only 83% of chicken and turkey orthologous proteins display sequence matches between the two species. Further, less than 70% of kinase recognition target sequences are exact matches between chicken and turkey. Thus, our analysis shows that, at the proteome and kinome level, these two species must be considered separately in the design of novel peptide arrays. Our ultimate array design covers numerous immune and metabolic processes including innate and adaptive immunity, inflammatory responses, carbohydrate, protein and fat metabolism, and response to hormones. We have shown the proteomic and phosphoproteomic diversity of chicken and turkey and have designed a valuable research tool for the study of immuno-metabolism within these two species.

  20. Hippocampal phosphoproteomics of F344 rats exposed to 1-bromopropane

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Zhenlie [Guangdong Provincial Key Laboratory of Occupational Disease Prevention and Treatment, Guangdong Province Hospital for Occupational Disease Prevention and Treatment, Guangzhou 510-300 (China); Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550 (Japan); Ichihara, Sahoko [Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507 (Japan); Oikawa, Shinji [Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Mie 514-8507 (Japan); Chang, Jie [Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550 (Japan); Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507 (Japan); Zhang, Lingyi [Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550 (Japan); Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510 (Japan); Hu, Shijie [Guangdong Provincial Key Laboratory of Occupational Disease Prevention and Treatment, Guangdong Province Hospital for Occupational Disease Prevention and Treatment, Guangzhou 510-300 (China); Huang, Hanlin, E-mail: huanghl@gdoh.org [Guangdong Provincial Key Laboratory of Occupational Disease Prevention and Treatment, Guangdong Province Hospital for Occupational Disease Prevention and Treatment, Guangzhou 510-300 (China); Ichihara, Gaku, E-mail: gak@rs.tus.ac.jp [Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550 (Japan); Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510 (Japan)

    2015-01-15

    1-Bromopropane (1-BP) is neurotoxic in both experimental animals and human. To identify phosphorylated modification on the unrecognized post-translational modifications of proteins and investigate their role in 1-BP-induced neurotoxicity, changes in hippocampal phosphoprotein expression levels were analyzed quantitatively in male F344 rats exposed to 1-BP inhalation at 0, 400, or 1000 ppm for 8 h/day for 1 or 4 weeks. Hippocampal protein extracts were analyzed qualitatively and quantitatively by Pro-Q Diamond gel staining and SYPRO Ruby staining coupled with two-dimensional difference in gel electrophoresis (2D-DIGE), respectively, as well as by matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to identify phosphoproteins. Changes in selected proteins were further confirmed by Manganese II (Mn{sup 2+})-Phos-tag SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Bax and cytochrome c protein levels were determined by western blotting. Pro-Q Diamond gel staining combined with 2D-DIGE identified 26 phosphoprotein spots (p < 0.05), and MALDI-TOF/MS identified 18 up-regulated proteins and 8 down-regulated proteins. These proteins are involved in the biological process of response to stimuli, metabolic processes, and apoptosis signaling. Changes in the expression of phosphorylated 14-3-3 θ were further confirmed by Mn{sup 2+}-Phos-tag SDS-PAGE. Western blotting showed overexpression of Bax protein in the mitochondria with down-regulation in the cytoplasm, whereas cytochrome c expression was high in the cytoplasm but low in the mitochondria after 1-BP exposure. Our results suggest that the pathogenesis of 1-BP-induced hippocampal damage involves inhibition of antiapoptosis process. Phosphoproteins identified in this study can potentially serve as biomarkers for 1-BP-induced neurotoxicity. - Highlights: • 1-BP modified hippocampal phosphoproteome in rat and 23 altered proteins were identified. • 1-BP changed phosphorylation

  1. Quantitative phosphoproteomics in nuclei of vasopressin-sensitive renal collecting duct cells

    OpenAIRE

    Bolger, Steven J.; Hurtado, Patricia A. Gonzales; Hoffert, Jason D.; Saeed, Fahad; Pisitkun, Trairak; Knepper, Mark A.

    2012-01-01

    Vasopressin regulates transport across the collecting duct epithelium in part via effects on gene transcription. Transcriptional regulation occurs partially via changes in phosphorylation of transcription factors, transcriptional coactivators, and protein kinases in the nucleus. To test whether vasopressin alters the nuclear phosphoproteome of vasopressin-sensitive cultured mouse mpkCCD cells, we used stable isotope labeling and mass spectrometry to quantify thousands of phosphorylation sites...

  2. Phosphoproteome of the cyanobacterium Synechocystis sp. PCC 6803 and its dynamics during nitrogen starvation.

    Directory of Open Access Journals (Sweden)

    Philipp eSpät

    2015-03-01

    Full Text Available Cyanobacteria have shaped the earth’s biosphere as the first oxygenic photoautotrophs and still play an important role in many ecosystems. The ability to adapt to changing environmental conditions is an essential characteristic in order to ensure survival. To this end, numerous studies have shown that bacteria use protein post-translational modifications such as Ser/Thr/Tyr phosphorylation in cell signalling, adaptation and regulation. Nevertheless, our knowledge of cyanobacterial phosphoproteomes and their dynamic response to environmental stimuli is relatively limited. In this study, we applied gel-free methods and high accuracy mass spectrometry towards the unbiased detection of Ser/Thr/Tyr phosphorylation events in the model cyanobacterium Synechocystis sp. PCC 6803. We could identify over 300 phosphorylation events in cultures grown on nitrate as exclusive nitrogen source. Chemical dimethylation labelling was applied to investigate proteome and phosphoproteome dynamics during nitrogen starvation. Our dataset describes the most comprehensive (phosphoproteome of Synechocystis to date, identifying 2,382 proteins and 183 phosphorylation events and quantifying 2,111 proteins and 148 phosphorylation events during nitrogen starvation. Global protein phosphorylation levels were increased in response to nitrogen depletion after 24 hours. Among the proteins with increased phosphorylation, the PII signalling protein showed the highest fold-change, serving as positive control. Other proteins with increased phosphorylation levels comprised functions in photosynthesis and in carbon and nitrogen metabolism. This study reveals dynamics of Synechocystis phosphoproteome in response to environmental stimuli and suggests an important role of protein Ser/Thr/Tyr phosphorylation in fundamental mechanisms of homeostatic control in cyanobacteria.

  3. Comparative Phosphoproteomics Reveals an Important Role of MKK2 in Banana (Musa spp.) Cold Signal Network

    Science.gov (United States)

    Gao, Jie; Zhang, Sheng; He, Wei-Di; Shao, Xiu-Hong; Li, Chun-Yu; Wei, Yue-Rong; Deng, Gui-Ming; Kuang, Rui-Bin; Hu, Chun-Hua; Yi, Gan-Jun; Yang, Qiao-Song

    2017-01-01

    Low temperature is one of the key environmental stresses, which greatly affects global banana production. However, little is known about the global phosphoproteomes in Musa spp. and their regulatory roles in response to cold stress. In this study, we conducted a comparative phosphoproteomic profiling of cold-sensitive Cavendish Banana and relatively cold tolerant Dajiao under cold stress. Phosphopeptide abundances of five phosphoproteins involved in MKK2 interaction network, including MKK2, HY5, CaSR, STN7 and kinesin-like protein, show a remarkable difference between Cavendish Banana and Dajiao in response to cold stress. Western blotting of MKK2 protein and its T31 phosphorylated peptide verified the phosphoproteomic results of increased T31 phosphopeptide abundance with decreased MKK2 abundance in Daojiao for a time course of cold stress. Meanwhile increased expression of MKK2 with no detectable T31 phosphorylation was found in Cavendish Banana. These results suggest that the MKK2 pathway in Dajiao, along with other cold-specific phosphoproteins, appears to be associated with the molecular mechanisms of high tolerance to cold stress in Dajiao. The results also provide new evidence that the signaling pathway of cellular MKK2 phosphorylation plays an important role in abiotic stress tolerance that likely serves as a universal plant cold tolerance mechanism. PMID:28106078

  4. Reactive Oxygen Species (ROS)-Activated ATM-Dependent Phosphorylation of Cytoplasmic Substrates Identified by Large-Scale Phosphoproteomics Screen.

    Science.gov (United States)

    Kozlov, Sergei V; Waardenberg, Ashley J; Engholm-Keller, Kasper; Arthur, Jonathan W; Graham, Mark E; Lavin, Martin

    2016-03-01

    Ataxia-telangiectasia, mutated (ATM) protein plays a central role in phosphorylating a network of proteins in response to DNA damage. These proteins function in signaling pathways designed to maintain the stability of the genome and minimize the risk of disease by controlling cell cycle checkpoints, initiating DNA repair, and regulating gene expression. ATM kinase can be activated by a variety of stimuli, including oxidative stress. Here, we confirmed activation of cytoplasmic ATM by autophosphorylation at multiple sites. Then we employed a global quantitative phosphoproteomics approach to identify cytoplasmic proteins altered in their phosphorylation state in control and ataxia-telangiectasia (A-T) cells in response to oxidative damage. We demonstrated that ATM was activated by oxidative damage in the cytoplasm as well as in the nucleus and identified a total of 9,833 phosphorylation sites, including 6,686 high-confidence sites mapping to 2,536 unique proteins. A total of 62 differentially phosphorylated peptides were identified; of these, 43 were phosphorylated in control but not in A-T cells, and 19 varied in their level of phosphorylation. Motif enrichment analysis of phosphopeptides revealed that consensus ATM serine glutamine sites were overrepresented. When considering phosphorylation events, only observed in control cells (not observed in A-T cells), with predicted ATM sites phosphoSerine/phosphoThreonine glutamine, we narrowed this list to 11 candidate ATM-dependent cytoplasmic proteins. Two of these 11 were previously described as ATM substrates (HMGA1 and UIMCI/RAP80), another five were identified in a whole cell extract phosphoproteomic screens, and the remaining four proteins had not been identified previously in DNA damage response screens. We validated the phosphorylation of three of these proteins (oxidative stress responsive 1 (OSR1), HDGF, and ccdc82) as ATM dependent after H2O2 exposure, and another protein (S100A11) demonstrated ATM

  5. Nuclear Phosphoproteomic Screen Uncovers ACLY as Mediator of IL-2-induced Proliferation of CD4+ T lymphocytes.

    Science.gov (United States)

    Osinalde, Nerea; Mitxelena, Jone; Sánchez-Quiles, Virginia; Akimov, Vyacheslav; Aloria, Kerman; Arizmendi, Jesus M; Zubiaga, Ana M; Blagoev, Blagoy; Kratchmarova, Irina

    2016-06-01

    Anti-cancer immunotherapies commonly rely on the use of interleukin-2 (IL-2) to promote the expansion of T lymphocytes. IL-2- dependent proliferation is the culmination of a complex network of phosphorylation-driven signaling events that impact on gene transcription through mechanisms that are not clearly understood. To study the role of IL-2 in the regulation of nuclear protein function we have performed an unbiased mass spectrometry-based study of the nuclear phosphoproteome of resting and IL-2-treated CD4(+) T lymphocytes. We detected 8521distinct phosphosites including many that are not yet reported in curated phosphorylation databases. Although most phosphorylation sites remained unaffected upon IL-2 treatment, 391 sites corresponding to 288 gene products showed robust IL-2-dependent regulation. Importantly, we show that ATP-citrate lyase (ACLY) is a key phosphoprotein effector of IL-2-mediated T-cell responses. ACLY becomes phosphorylated on serine 455 in T lymphocytes upon IL-2-driven activation of AKT, and depletion or inactivation of ACLY compromises IL-2-promoted T-cell growth. Mechanistically, we demonstrate that ACLY is required for enhancing histone acetylation levels and inducing the expression of cell cycle regulating genes in response to IL-2. Thus, the metabolic enzyme ACLY emerges as a bridge between cytokine signaling and proliferation of T lymphocytes, and may be an attractive candidate target for the development of more efficient anti-cancer immunotherapies.

  6. Quantitative phosphoproteomics of murine Fmr1-KO cell lines provides new insights into FMRP-dependent signal transduction mechanisms.

    Science.gov (United States)

    Matic, Katarina; Eninger, Timo; Bardoni, Barbara; Davidovic, Laetitia; Macek, Boris

    2014-10-03

    Fragile X mental retardation protein (FMRP) is an RNA-binding protein that has a major effect on neuronal protein synthesis. Transcriptional silencing of the FMR1 gene leads to loss of FMRP and development of Fragile X syndrome (FXS), the most common known hereditary cause of intellectual impairment and autism. Here we utilize SILAC-based quantitative phosphoproteomics to analyze murine FMR1(-) and FMR1(+) fibroblastic cell lines derived from FMR1-KO embryos to identify proteins and phosphorylation sites dysregulated as a consequence of FMRP loss. We quantify FMRP-related changes in the levels of 5,023 proteins and 6,133 phosphorylation events and map them onto major signal transduction pathways. Our study confirms global downregulation of the MAPK/ERK pathway and decrease in phosphorylation level of ERK1/2 in the absence of FMRP, which is connected to attenuation of long-term potentiation. We detect differential expression of several key proteins from the p53 pathway, pointing to the involvement of p53 signaling in dysregulated cell cycle control in FXS. Finally, we detect differential expression and phosphorylation of proteins involved in pre-mRNA processing and nuclear transport, as well as Wnt and calcium signaling, such as PLC, PKC, NFAT, and cPLA2. We postulate that calcium homeostasis is likely affected in molecular pathogenesis of FXS.

  7. System-wide temporal characterization of the proteome and phosphoproteome of human embryonic stem cell differentiation

    DEFF Research Database (Denmark)

    Rigbolt, Kristoffer T.G.; Prokhorova, Tatyana; Akimov, Vyacheslav;

    2011-01-01

    To elucidate cellular events underlying the pluripotency of human embryonic stem cells (hESCs), we performed parallel quantitative proteomic and phosphoproteomic analyses of hESCs during differentiation initiated by a diacylglycerol analog or transfer to media that had not been conditioned...... by feeder cells. We profiled 6521 proteins and 23,522 phosphorylation sites, of which almost 50% displayed dynamic changes in phosphorylation status during 24 hours of differentiation. These data are a resource for studies of the events associated with the maintenance of hESC pluripotency and those...

  8. Picking the right tool for the job--Phosphoproteomics of egg activation.

    Science.gov (United States)

    Wessel, Gary M

    2015-12-01

    Eggs are the rarest cell in the human body, yet their study is essential for the fields of fertility, reproduction, and fetal health. Guo et al. (Proteomics 2015, 15, 4080-4095) use a "surrogate" animal to discover the phosphoproteomic pathways involved in egg activation. With datasets of several thousand phosphosites on 2500 different proteins, these investigators have defined new pathways, connections to pathways, and priorities in searches for how eggs are activated at fertilization. These results in a sea urchin are now transposable to mammals for testing on a per candidate strategy.

  9. Identification of novel protein functions and signaling mechanisms by genetics and quantitative phosphoproteomics in Caenorhabditis elegans

    DEFF Research Database (Denmark)

    Fredens, Julius; Engholm-Keller, Kasper; Møller-Jensen, Jakob;

    2014-01-01

    knockdown by feeding the nematode on pre-labeled lysine auxotroph Escherichia coli. In this chapter, we describe in details the generation of the E. coli strain, incorporation of heavy isotope-labeled lysine in C. elegans, and the procedure for a comprehensive global phosphoproteomic experiment.......Stable isotope labeling by amino acids combined with mass spectrometry is a widely used methodology for measuring relative changes in protein and phosphorylation levels at a global level. We have applied this method to the model organism Caenorhabditis elegans in combination with RNAi-mediated gene...

  10. Projectile Base Flow Analysis

    Science.gov (United States)

    2007-11-02

    S) AND ADDRESS(ES) DCW Industries, Inc. 5354 Palm Drive La Canada, CA 91011 8. PERFORMING ORGANIZATION...REPORT NUMBER DCW -38-R-05 9. SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) U. S. Army Research Office...Turbulence Modeling for CFD, Second Edition, DCW Industries, Inc., La Cañada, CA. Wilcox, D. C. (2001), “Projectile Base Flow Analysis,” DCW

  11. HOPE-fixation of lung tissue allows retrospective proteome and phosphoproteome studies.

    Science.gov (United States)

    Shevchuk, Olga; Abidi, Nada; Klawonn, Frank; Wissing, Josef; Nimtz, Manfred; Kugler, Christian; Steinert, Michael; Goldmann, Torsten; Jänsch, Lothar

    2014-11-07

    Hepes-glutamic acid buffer-mediated organic solvent protection effect (HOPE)-fixation has been introduced as an alternative to formalin fixation of clinical samples. Beyond preservation of morphological structures for histology, HOPE-fixation was demonstrated to be compatible with recent methods for RNA and DNA sequencing. However, the suitability of HOPE-fixed materials for the inspection of proteomes by mass spectrometry so far remained undefined. This is of particular interest, since proteins constitute a prime resource for drug research and can give valuable insights into the activity status of signaling pathways. In this study, we extracted proteins from human lung tissue and tested HOPE-treated and snap-frozen tissues comparatively by proteome and phosphoproteome analyses. High confident data from accurate mass spectrometry allowed the identification of 2603 proteins and 3036 phosphorylation sites. HOPE-fixation did not hinder the representative extraction of proteins, and investigating their biochemical properties, covered subcellular localizations, and cellular processes revealed no bias caused by the type of fixation. In conclusion, proteome as well as phosphoproteome data of HOPE lung samples were qualitatively equivalent to results obtained from snap-frozen tissues. Thus, HOPE-treated tissues match clinical demands in both histology and retrospective proteome analyses of patient samples by proteomics.

  12. Proteomic and phosphoproteomic analyses of chromatin-associated proteins from Arabidopsis thaliana

    KAUST Repository

    Bigeard, Jean

    2014-07-10

    The nucleus is the organelle where basically all DNA-related processes take place in eukaryotes, such as replication, transcription, and splicing as well as epigenetic regulation. The identification and description of the nuclear proteins is one of the requisites toward a comprehensive understanding of the biological functions accomplished in the nucleus. Many of the regulatory mechanisms of protein functions rely on their PTMs among which phosphorylation is probably one of the most important properties affecting enzymatic activity, interaction with other molecules, localization, or stability. So far, the nuclear and subnuclear proteome and phosphoproteome of the model plant Arabidopsis thaliana have been the subject of very few studies. In this work, we developed a purification protocol of Arabidopsis chromatin-associated proteins and performed proteomic and phosphoproteomic analyses identifying a total of 879 proteins of which 198 were phosphoproteins that were mainly involved in chromatin remodeling, transcriptional regulation, and RNA processing. From 230 precisely localized phosphorylation sites (phosphosites), 52 correspond to hitherto unidentified sites. This protocol and data thereby obtained should be a valuable resource for many domains of plant research.

  13. NeuCode Labeling in Nematodes: Proteomic and Phosphoproteomic Impact of Ascaroside Treatment in Caenorhabditis elegans*

    Science.gov (United States)

    Rhoads, Timothy W.; Prasad, Aman; Kwiecien, Nicholas W.; Merrill, Anna E.; Zawack, Kelson; Westphall, Michael S.; Schroeder, Frank C.; Kimble, Judith; Coon, Joshua J.

    2015-01-01

    The nematode Caenorhabditis elegans is an important model organism for biomedical research. We previously described NeuCode stable isotope labeling by amino acids in cell culture (SILAC), a method for accurate proteome quantification with potential for multiplexing beyond the limits of traditional stable isotope labeling by amino acids in cell culture. Here we apply NeuCode SILAC to profile the proteomic and phosphoproteomic response of C. elegans to two potent members of the ascaroside family of nematode pheromones. By consuming labeled E. coli as part of their diet, C. elegans nematodes quickly and easily incorporate the NeuCode heavy lysine isotopologues by the young adult stage. Using this approach, we report, at high confidence, one of the largest proteomic and phosphoproteomic data sets to date in C. elegans: 6596 proteins at a false discovery rate ≤ 1% and 6620 phosphorylation isoforms with localization probability ≥75%. Our data reveal a post-translational signature of pheromone sensing that includes many conserved proteins implicated in longevity and response to stress. PMID:26392051

  14. NeuCode Labeling in Nematodes: Proteomic and Phosphoproteomic Impact of Ascaroside Treatment in Caenorhabditis elegans.

    Science.gov (United States)

    Rhoads, Timothy W; Prasad, Aman; Kwiecien, Nicholas W; Merrill, Anna E; Zawack, Kelson; Westphall, Michael S; Schroeder, Frank C; Kimble, Judith; Coon, Joshua J

    2015-11-01

    The nematode Caenorhabditis elegans is an important model organism for biomedical research. We previously described NeuCode stable isotope labeling by amino acids in cell culture (SILAC), a method for accurate proteome quantification with potential for multiplexing beyond the limits of traditional stable isotope labeling by amino acids in cell culture. Here we apply NeuCode SILAC to profile the proteomic and phosphoproteomic response of C. elegans to two potent members of the ascaroside family of nematode pheromones. By consuming labeled E. coli as part of their diet, C. elegans nematodes quickly and easily incorporate the NeuCode heavy lysine isotopologues by the young adult stage. Using this approach, we report, at high confidence, one of the largest proteomic and phosphoproteomic data sets to date in C. elegans: 6596 proteins at a false discovery rate ≤ 1% and 6620 phosphorylation isoforms with localization probability ≥75%. Our data reveal a post-translational signature of pheromone sensing that includes many conserved proteins implicated in longevity and response to stress.

  15. Improving the Phosphoproteome Coverage for Limited Sample Amounts Using TiO2-SIMAC-HILIC (TiSH) Phosphopeptide Enrichment and Fractionation

    DEFF Research Database (Denmark)

    Engholm-Keller, Kasper; Larsen, Martin R

    2016-01-01

    Obtaining high phosphoproteome coverage requires specific enrichment of phosphorylated peptides from the often extremely complex peptide mixtures generated by proteolytic digestion of biological samples, as well as extensive chromatographic fractionation prior to liquid chromatography-tandem mass...

  16. Quantitative Phosphoproteomics Unravels Biased Phosphorylation of Serotonin 2A Receptor at Ser280 by Hallucinogenic versus Nonhallucinogenic Agonists*

    Science.gov (United States)

    Karaki, Samah; Becamel, Carine; Murat, Samy; Mannoury la Cour, Clotilde; Millan, Mark J.; Prézeau, Laurent; Bockaert, Joël; Marin, Philippe; Vandermoere, Franck

    2014-01-01

    The serotonin 5-HT2A receptor is a primary target of psychedelic hallucinogens such as lysergic acid diethylamine, mescaline, and psilocybin, which reproduce some of the core symptoms of schizophrenia. An incompletely resolved paradox is that only some 5-HT2A receptor agonists exhibit hallucinogenic activity, whereas structurally related agonists with comparable affinity and activity lack such a psychoactive activity. Using a strategy combining stable isotope labeling by amino acids in cell culture with enrichment in phosphorylated peptides by means of hydrophilic interaction liquid chromatography followed by immobilized metal affinity chromatography, we compared the phosphoproteome in HEK-293 cells transiently expressing the 5-HT2A receptor and exposed to either vehicle or the synthetic hallucinogen 1-[2,5-dimethoxy-4-iodophenyl]-2-aminopropane (DOI) or the nonhallucinogenic 5-HT2A agonist lisuride. Among the 5995 identified phosphorylated peptides, 16 sites were differentially phosphorylated upon exposure of cells to DOI versus lisuride. These include a serine (Ser280) located in the third intracellular loop of the 5-HT2A receptor, a region important for its desensitization. The specific phosphorylation of Ser280 by hallucinogens was further validated by quantitative mass spectrometry analysis of immunopurified receptor digests and by Western blotting using a phosphosite specific antibody. The administration of DOI, but not of lisuride, to mice, enhanced the phosphorylation of 5-HT2A receptors at Ser280 in the prefrontal cortex. Moreover, hallucinogens induced a less pronounced desensitization of receptor-operated signaling in HEK-293 cells and neurons than did nonhallucinogenic agonists. The mutation of Ser280 to aspartic acid (to mimic phosphorylation) reduced receptor desensitization by nonhallucinogenic agonists, whereas its mutation to alanine increased the ability of hallucinogens to desensitize the receptor. This study reveals a biased phosphorylation of

  17. Phosphoproteomics Profiling of Human Skin Fibroblast Cells Reveals Pathways and Proteins Affected by Low Doses of Ionizing Radiation

    Science.gov (United States)

    Yang, Feng; Waters, Katrina M.; Miller, John H.; Gritsenko, Marina A.; Zhao, Rui; Du, Xiuxia; Livesay, Eric A.; Purvine, Samuel O.; Monroe, Matthew E.; Wang, Yingchun; Camp, David G.; Smith, Richard D.; Stenoien, David L.

    2010-01-01

    Background High doses of ionizing radiation result in biological damage; however, the precise relationships between long-term health effects, including cancer, and low-dose exposures remain poorly understood and are currently extrapolated using high-dose exposure data. Identifying the signaling pathways and individual proteins affected at the post-translational level by radiation should shed valuable insight into the molecular mechanisms that regulate dose-dependent responses to radiation. Principal Findings We have identified 7117 unique phosphopeptides (2566 phosphoproteins) from control and irradiated (2 and 50 cGy) primary human skin fibroblasts 1 h post-exposure. Semi-quantitative label-free analyses were performed to identify phosphopeptides that are apparently altered by radiation exposure. This screen identified phosphorylation sites on proteins with known roles in radiation responses including TP53BP1 as well as previously unidentified radiation-responsive proteins such as the candidate tumor suppressor SASH1. Bioinformatic analyses suggest that low and high doses of radiation affect both overlapping and unique biological processes and suggest a role for MAP kinase and protein kinase A (PKA) signaling in the radiation response as well as differential regulation of p53 networks at low and high doses of radiation. Conclusions Our results represent the most comprehensive analysis of the phosphoproteomes of human primary fibroblasts exposed to multiple doses of ionizing radiation published to date and provide a basis for the systems-level identification of biological processes, molecular pathways and individual proteins regulated in a dose dependent manner by ionizing radiation. Further study of these modified proteins and affected networks should help to define the molecular mechanisms that regulate biological responses to radiation at different radiation doses and elucidate the impact of low-dose radiation exposure on human health. PMID:21152398

  18. Phosphoproteomics profiling of human skin fibroblast cells reveals pathways and proteins affected by low doses of ionizing radiation.

    Directory of Open Access Journals (Sweden)

    Feng Yang

    Full Text Available BACKGROUND: High doses of ionizing radiation result in biological damage; however, the precise relationships between long-term health effects, including cancer, and low-dose exposures remain poorly understood and are currently extrapolated using high-dose exposure data. Identifying the signaling pathways and individual proteins affected at the post-translational level by radiation should shed valuable insight into the molecular mechanisms that regulate dose-dependent responses to radiation. PRINCIPAL FINDINGS: We have identified 7117 unique phosphopeptides (2566 phosphoproteins from control and irradiated (2 and 50 cGy primary human skin fibroblasts 1 h post-exposure. Semi-quantitative label-free analyses were performed to identify phosphopeptides that are apparently altered by radiation exposure. This screen identified phosphorylation sites on proteins with known roles in radiation responses including TP53BP1 as well as previously unidentified radiation-responsive proteins such as the candidate tumor suppressor SASH1. Bioinformatic analyses suggest that low and high doses of radiation affect both overlapping and unique biological processes and suggest a role for MAP kinase and protein kinase A (PKA signaling in the radiation response as well as differential regulation of p53 networks at low and high doses of radiation. CONCLUSIONS: Our results represent the most comprehensive analysis of the phosphoproteomes of human primary fibroblasts exposed to multiple doses of ionizing radiation published to date and provide a basis for the systems-level identification of biological processes, molecular pathways and individual proteins regulated in a dose dependent manner by ionizing radiation. Further study of these modified proteins and affected networks should help to define the molecular mechanisms that regulate biological responses to radiation at different radiation doses and elucidate the impact of low-dose radiation exposure on human health.

  19. Quantitative phosphoproteomics applied to the yeast pheromone signaling pathway

    DEFF Research Database (Denmark)

    Gruhler, Albrecht; Olsen, Jesper Velgaard; Mohammed, Shabaz

    2005-01-01

    . Phosphopeptide fractions were analyzed by LC-MS using a linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer. MS/MS and neutral loss-directed MS/MS/MS analysis allowed detection and sequencing of phosphopeptides with exceptional accuracy and specificity. Of more than 700 identified...

  20. A phosphoproteomics view at human pluripotent stem cells

    NARCIS (Netherlands)

    Zoumaro-Djayoon, A.D.

    2013-01-01

    Reversible protein phosphorylation has been one of the most investigated post-translational modifications (PTM) in the decade, made possible by advances in enrichment technologies and MS-based proteomics. PTMs are generally involved in the regulation of many biological processes, including processes

  1. Sequential enrichment with titania-coated magnetic mesoporous hollow silica microspheres and zirconium arsenate-modified magnetic nanoparticles for the study of phosphoproteome of HL60 cells.

    Science.gov (United States)

    Yu, Qiong-Wei; Li, Xiao-Shui; Xiao, Yongsheng; Guo, Lei; Zhang, Fan; Cai, Qian; Feng, Yu-Qi; Yuan, Bi-Feng; Wang, Yinsheng

    2014-10-24

    As one of the most important types of post-translational modifications, reversible phosphorylation of proteins plays crucial roles in a large number of biological processes. However, owing to the relatively low abundance and dynamic nature of phosphorylation and the presence of the unphosphorylated peptides in large excess, phosphopeptide enrichment is indispensable in large-scale phosphoproteomic analysis. Metal oxides including titanium dioxide have become prominent affinity materials to enrich phosphopeptides prior to their analysis using liquid chromatography-mass spectrometry (LC-MS). In the current study, we established a novel strategy, which encompassed strong cation exchange chromatography, sequential enrichment of phosphopeptides using titania-coated magnetic mesoporous hollow silica microspheres (TiO2/MHMSS) and zirconium arsenate-modified magnetic nanoparticles (ZrAs-Fe3O4@SiO2), and LC-MS/MS analysis, for the proteome-wide identification of phosphosites of proteins in HL60 cells. In total, we were able to identify 11,579 unique phosphorylation sites in 3432 unique proteins. Additionally, our results suggested that TiO2/MHMSS and ZrAs-Fe3O4@SiO2 are complementary in phosphopeptide enrichment, where the two types of materials displayed preferential binding of peptides carrying multiple and single phosphorylation sites, respectively.

  2. 哺乳动物精子磷酸化蛋白质组学研究进展%Resent Advances on Phosphoproteomics Researches of Mammalian Sperm Capacitation

    Institute of Scientific and Technical Information of China (English)

    张媛媛; 胡启蒙; 王亮亮; 李新红

    2012-01-01

    Studies on protein expression, modilicalion and interaction have become important in proteomics in the post-genomics era. Phosphorylation/dephosphorylation of proteins is the main mechanism of signal transduction and regulation of enzymes in sperm cells, what' s more, it plays a key role during the sperm-egg recognition and fertilization process. Researches about function of phosphorylated proteins contribute to the understanding of the molecular mechanism of sperm capacitation, hyperactive motility and acrosome reaction process. The progress in the researches of mammalian sperm phosphoproteomics was reviewed that include the phosphoproteomics technology, phosphorylated proteins identification and quantitation, function analysis benefits in finding new important fertilization related biological markers and revealing the sperm development, reproductive potential changes and molecular mechanism of fertilization.%蛋白质的表达、修饰及相互作用的研究已成为后基因组学时代蛋白质组学中的重要内容.蛋白质磷酸化和去磷酸化作为最普遍的翻译后修饰之一,是精子细胞信号转导和酶调控、表达的主要分子机制,亦是精子、卵细胞信号识别及完成受精作用的关键环节.对精子磷酸化蛋白功能的研究有助于深入理解精子的获能、超激活运动的维持、发生顶体反应及精卵结合等受精过程的分子调控机理.对哺乳动物精子磷酸化蛋白质组学的研究进展,包括动物精子磷酸化蛋白质组学研究的技术方法、磷酸化蛋白质种类的鉴定、定量及其功能分析进行了综述,为进一步发掘与受精相关的重要生物标志物,揭示精子发育、繁殖潜能变化及受精分子机理奠定基础.

  3. Trajectory Based Traffic Analysis

    DEFF Research Database (Denmark)

    Krogh, Benjamin Bjerre; Andersen, Ove; Lewis-Kelham, Edwin

    2013-01-01

    -and-click analysis, due to a novel and efficient indexing structure. With the web-site daisy.aau.dk/its/spqdemo/we will demonstrate several analyses, using a very large real-world data set consisting of 1.9 billion GPS records (1.5 million trajectories) recorded from more than 13000 vehicles, and touching most...

  4. Phosphoproteomics Reveals HMGA1, a CK2 Substrate, as a Drug-Resistant Target in Non-Small Cell Lung Cancer

    Science.gov (United States)

    Wang, Yi-Ting; Pan, Szu-Hua; Tsai, Chia-Feng; Kuo, Ting-Chun; Hsu, Yuan-Ling; Yen, Hsin-Yung; Choong, Wai-Kok; Wu, Hsin-Yi; Liao, Yen-Chen; Hong, Tse-Ming; Sung, Ting-Yi; Yang, Pan-Chyr; Chen, Yu-Ju

    2017-01-01

    Although EGFR tyrosine kinase inhibitors (TKIs) have demonstrated good efficacy in non-small-cell lung cancer (NSCLC) patients harboring EGFR mutations, most patients develop intrinsic and acquired resistance. We quantitatively profiled the phosphoproteome and proteome of drug-sensitive and drug-resistant NSCLC cells under gefitinib treatment. The construction of a dose-dependent responsive kinase-substrate network of 1548 phosphoproteins and 3834 proteins revealed CK2-centric modules as the dominant core network for the potential gefitinib resistance-associated proteins. CK2 knockdown decreased cell survival in gefitinib-resistant NSCLCs. Using motif analysis to identify the CK2 core sub-network, we verified that elevated phosphorylation level of a CK2 substrate, HMGA1 was a critical node contributing to EGFR-TKI resistance in NSCLC cell. Both HMGA1 knockdown or mutation of the CK2 phosphorylation site, S102, of HMGA1 reinforced the efficacy of gefitinib in resistant NSCLC cells through reactivation of the downstream signaling of EGFR. Our results delineate the TKI resistance-associated kinase-substrate network, suggesting a potential therapeutic strategy for overcoming TKI-induced resistance in NSCLC. PMID:28290473

  5. The beginnings of crop phosphoproteomics: exploring early warning systems of stress.

    Directory of Open Access Journals (Sweden)

    Christof eRampitsch

    2012-07-01

    Full Text Available This review examines why a knowledge of plant protein phosphorylation events is important in devising strategies to protect crops from both biotic and abiotic stresses, and why proteomics should be included when studying stress pathways. Most of the achievements in elucidating phospho-signalling pathways in biotic and abiotic stress are reported from model systems: while these are discussed, this review attempts mainly to focus on work done with crops, with examples of achievements reported from rice, maize, wheat, grape, Brassica, tomato and soy bean after cold acclimation, hormonal and oxidative H2O2 treatment, salt stress, mechanical wounding or pathogen challenge. The challenges that remain to transfer this information into a format that can be used to protect crops against biotic and abiotic stresses are enormous. The tremendous increase in the speed and ease of DNA sequencing is poised to reveal the whole genomes of many crop species in the near future, which will facilitate phosphoproteomics and phosphogenomics research.

  6. Site-Specific Ser/Thr/Tyr Phosphoproteome of Sinorhizobium meliloti at Stationary Phase.

    Science.gov (United States)

    Liu, Tao; Tian, Chang Fu; Chen, Wen Xin

    2015-01-01

    Sinorhizobium meliloti, a facultative microsymbiont of alfalfa, should fine-tune its cellular processes to live saprophytically in soils characterized with limited nutrients and diverse stresses. In this study, TiO2 enrichment and LC-MS/MS were used to uncover the site-specific Ser/Thr/Tyr phosphoproteome of S. meliloti in minimum medium at stationary phase. There are a total of 96 unique phosphorylated sites, with a Ser/Thr/Tyr distribution of 63:28:5, in 77 proteins. Phosphoproteins identified in S. meliloti showed a wide distribution pattern regarding to functional categories, such as replication, transcription, translation, posttranslational modification, transport and metabolism of amino acids, carbohydrate, inorganic ion, succinoglycan etc. Ser/Thr/Tyr phosphosites identified within the conserved motif in proteins of key cellular function indicate a crucial role of phosphorylation in modulating cellular physiology. Moreover, phosphorylation in proteins involved in processes related to rhizobial adaptation was also discussed, such as those identified in SMa0114 and PhaP2 (polyhydroxybutyrate synthesis), ActR (pH stress and microaerobic adaption), SupA (potassium stress), chaperonin GroEL2 (viability and potentially symbiosis), and ExoP (succinoglycan synthesis and secretion). These Ser/Thr/Tyr phosphosites identified herein would be helpful for our further investigation and understanding of the role of phosphorylation in rhizobial physiology.

  7. Site-Specific Ser/Thr/Tyr Phosphoproteome of Sinorhizobium meliloti at Stationary Phase.

    Directory of Open Access Journals (Sweden)

    Tao Liu

    Full Text Available Sinorhizobium meliloti, a facultative microsymbiont of alfalfa, should fine-tune its cellular processes to live saprophytically in soils characterized with limited nutrients and diverse stresses. In this study, TiO2 enrichment and LC-MS/MS were used to uncover the site-specific Ser/Thr/Tyr phosphoproteome of S. meliloti in minimum medium at stationary phase. There are a total of 96 unique phosphorylated sites, with a Ser/Thr/Tyr distribution of 63:28:5, in 77 proteins. Phosphoproteins identified in S. meliloti showed a wide distribution pattern regarding to functional categories, such as replication, transcription, translation, posttranslational modification, transport and metabolism of amino acids, carbohydrate, inorganic ion, succinoglycan etc. Ser/Thr/Tyr phosphosites identified within the conserved motif in proteins of key cellular function indicate a crucial role of phosphorylation in modulating cellular physiology. Moreover, phosphorylation in proteins involved in processes related to rhizobial adaptation was also discussed, such as those identified in SMa0114 and PhaP2 (polyhydroxybutyrate synthesis, ActR (pH stress and microaerobic adaption, SupA (potassium stress, chaperonin GroEL2 (viability and potentially symbiosis, and ExoP (succinoglycan synthesis and secretion. These Ser/Thr/Tyr phosphosites identified herein would be helpful for our further investigation and understanding of the role of phosphorylation in rhizobial physiology.

  8. Quantitative phosphoproteomic analyses of the inferior parietal lobule from three different pathological stages of Alzheimer's disease.

    Science.gov (United States)

    Triplett, Judy C; Swomley, Aaron M; Cai, Jian; Klein, Jon B; Butterfield, D Allan

    2015-01-01

    Alzheimer's disease (AD), the most common age-related neurodegenerative disorder, is clinically characterized by progressive neuronal loss resulting in loss of memory and dementia. AD is histopathologically characterized by the extensive distribution of senile plaques and neurofibrillary tangles, and synapse loss. Amnestic mild cognitive impairment (MCI) is generally accepted to be an early stage of AD. MCI subjects have pathology and symptoms that fall on the scale intermediately between 'normal' cognition with little or no pathology and AD. A rare number of individuals, who exhibit normal cognition on psychometric tests but whose brains show widespread postmortem AD pathology, are classified as 'asymptomatic' or 'preclinical' AD (PCAD). In this study, we evaluated changes in protein phosphorylation states in the inferior parietal lobule of subjects with AD, MCI, PCAD, and control brain using a 2-D PAGE proteomics approach in conjunction with Pro-Q Diamond phosphoprotein staining. Statistically significant changes in phosphorylation levels were found in 19 proteins involved in energy metabolism, neuronal plasticity, signal transduction, and oxidative stress response. Changes in the disease state phosphoproteome may provide insights into underlying mechanisms for the preservation of memory with expansive AD pathology in PCAD and the progressive memory loss in amnestic MCI that escalates to the dementia and the characteristic pathology of AD brain.

  9. Quantitative phosphoproteomics identifies filaggrin and other targets of ionizing radiation in a human skin model.

    Science.gov (United States)

    Yang, Feng; Waters, Katrina M; Webb-Robertson, Bobbie-Jo; Sowa, Marianne B; von Neubeck, Claere; Aldrich, Josh T; Markillie, Lye Meng; Wirgau, Rachel M; Gritsenko, Marina A; Zhao, Rui; Camp, David G; Smith, Richard D; Stenoien, David L

    2012-05-01

    Our objective here was to perform a quantitative phosphoproteomic study on a reconstituted human skin tissue to identify low- and high-dose ionizing radiation-dependent signalling in a complex three-dimensional setting. Application of an isobaric labelling strategy using sham and three radiation doses (3, 10, 200 cGy) resulted in the identification of 1052 unique phosphopeptides. Statistical analyses identified 176 phosphopeptides showing significant changes in response to radiation and radiation dose. Proteins responsible for maintaining skin structural integrity including keratins and desmosomal proteins (desmoglein, desmoplakin, plakophilin 1, 2 and 3) had altered phosphorylation levels following exposure to both low and high doses of radiation. Altered phosphorylation of multiple sites in profilaggrin linker domains coincided with altered profilaggrin processing suggesting a role for linker phosphorylation in human profilaggrin regulation. These studies demonstrate that the reconstituted human skin system undergoes a coordinated response to both low and high doses of ionizing radiation involving multiple layers of the stratified epithelium that serve to maintain tissue integrity and mitigate effects of radiation exposure.

  10. Effects of poly-ether B on proteome and phosphoproteome expression in biofouling Balanus amphitrite cyprids

    KAUST Repository

    Dash, Swagatika

    2012-04-01

    Biofouling is ubiquitous in marine environments, and the barnacle Balanus amphitrite is one of the most recalcitrant and aggressive biofoulers in tropical waters. Several natural antifoulants that were claimed to be non-toxic have been isolated in recent years, although the mechanism by which they inhibit fouling is yet to be investigated. Poly-ether B has shown promise in the non-toxic inhibition of larval barnacle attachment. Hence, in this study, multiplex two-dimensional electrophoresis (2-DE) was applied in conjunction with mass spectrometry to investigate the effects of poly-ether B on barnacle larvae at the molecular level. The cyprid proteome response to poly-ether B treatment was analyzed at the total proteome and phosphoproteome levels, with 65 protein and 19 phosphoprotein spots found to be up- or down-regulated. The proteins were found to be related to energy-metabolism, oxidative stress, and molecular chaperones, thus indicating that poly-ether B may interfere with the redox-regulatory mechanisms governing the settlement of barnacle larvae. The results of this study demonstrate the usefulness of the proteomic technique in revealing the working mechanisms of antifouling compounds. © 2012 Copyright Taylor and Francis Group, LLC.

  11. Phosphoproteomics Identifies CK2 as a Negative Regulator of Beige Adipocyte Thermogenesis and Energy Expenditure.

    Science.gov (United States)

    Shinoda, Kosaku; Ohyama, Kana; Hasegawa, Yutaka; Chang, Hsin-Yi; Ogura, Mayu; Sato, Ayaka; Hong, Haemin; Hosono, Takashi; Sharp, Louis Z; Scheel, David W; Graham, Mark; Ishihama, Yasushi; Kajimura, Shingo

    2015-12-01

    Catecholamines promote lipolysis both in brown and white adipocytes, whereas the same stimuli preferentially activate thermogenesis in brown adipocytes. Molecular mechanisms for the adipose-selective activation of thermogenesis remain poorly understood. Here, we employed quantitative phosphoproteomics to map global and temporal phosphorylation profiles in brown, beige, and white adipocytes under β3-adrenenoceptor activation and identified kinases responsible for the adipose-selective phosphorylation profiles. We found that casein kinase2 (CK2) activity is preferentially higher in white adipocytes than brown/beige adipocytes. Genetic or pharmacological blockade of CK2 in white adipocytes activates the thermogenic program in response to cAMP stimuli. Such activation is largely through reduced CK2-mediated phosphorylation of class I HDACs. Notably, inhibition of CK2 promotes beige adipocyte biogenesis and leads to an increase in whole-body energy expenditure and ameliorates diet-induced obesity and insulin resistance. These results indicate that CK2 is a plausible target to rewire the β3-adrenenoceptor signaling cascade that promotes thermogenesis in adipocytes.

  12. Deoxygenation affects tyrosine phosphoproteome of red cell membrane from patients with sickle cell disease.

    Science.gov (United States)

    Siciliano, Angela; Turrini, Franco; Bertoldi, Mariarita; Matte, Alessandro; Pantaleo, Antonella; Olivieri, Oliviero; De Franceschi, Lucia

    2010-04-15

    Sickle cell disease (SCD) is a worldwide distributed hereditary red cell disorder related to the production of a defective form of hemoglobin, hemoglobin S (HbS). One of the hallmarks of SCD is the presence of dense, dehydrate highly adhesive sickle red blood cells (RBCs) that result from persistent membrane damage associated with HbS polymerization, abnormal activation of membrane cation transports and generation of distorted and rigid red cells with membrane perturbation and cytoskeleton dysfunction. Although modulation of phosphorylation state of the proteins from membrane and cytoskeleton networks has been proposed to participate in red cell homeostasis, much still remains to be investigated in normal and diseased red cells. Here, we report that tyrosine (Tyr-) phosphoproteome of sickle red cells was different from normal controls and was affected by deoxygenation. We found proteins, p55 and band 4.1, from the junctional complex, differently Tyr-phosphorylated in SCD RBCs compared to normal RBCs under normoxia and modulated by deoxygenation, while band 4.2 was similarly Tyr-phosphorylated in both conditions. In SCD RBCs we identified the phosphopeptides for protein 4.1R located in the protein FERM domain (Tyr-13) and for alpha-spectrin located near or in a linker region (Tyr-422 and Tyr-1498) involving protein areas crucial for their functions in the context of red cell membrane properties, suggesting that Tyr-phosphorylation may be part of the events involved in maintaining membrane mechanical stability in SCD red cells.

  13. SILAC-based quantitative proteomic analysis of human lung cell response to copper oxide nanoparticles.

    Science.gov (United States)

    Edelmann, Mariola J; Shack, Leslie A; Naske, Caitlin D; Walters, Keisha B; Nanduri, Bindu

    2014-01-01

    Copper (II) oxide (CuO) nanoparticles (NP) are widely used in industry and medicine. In our study we evaluated the response of BEAS-2B human lung cells to CuO NP, using Stable isotope labeling by amino acids in cell culture (SILAC)-based proteomics and phosphoproteomics. Pathway modeling of the protein differential expression showed that CuO NP affect proteins relevant in cellular function and maintenance, protein synthesis, cell death and survival, cell cycle and cell morphology. Some of the signaling pathways represented by BEAS-2B proteins responsive to the NP included mTOR signaling, protein ubiquitination pathway, actin cytoskeleton signaling and epithelial adherens junction signaling. Follow-up experiments showed that CuO NP altered actin cytoskeleton, protein phosphorylation and protein ubiquitination level.

  14. SILAC-based quantitative proteomic analysis of human lung cell response to copper oxide nanoparticles.

    Directory of Open Access Journals (Sweden)

    Mariola J Edelmann

    Full Text Available Copper (II oxide (CuO nanoparticles (NP are widely used in industry and medicine. In our study we evaluated the response of BEAS-2B human lung cells to CuO NP, using Stable isotope labeling by amino acids in cell culture (SILAC-based proteomics and phosphoproteomics. Pathway modeling of the protein differential expression showed that CuO NP affect proteins relevant in cellular function and maintenance, protein synthesis, cell death and survival, cell cycle and cell morphology. Some of the signaling pathways represented by BEAS-2B proteins responsive to the NP included mTOR signaling, protein ubiquitination pathway, actin cytoskeleton signaling and epithelial adherens junction signaling. Follow-up experiments showed that CuO NP altered actin cytoskeleton, protein phosphorylation and protein ubiquitination level.

  15. Hand-Based Biometric Analysis

    Science.gov (United States)

    Bebis, George (Inventor); Amayeh, Gholamreza (Inventor)

    2015-01-01

    Hand-based biometric analysis systems and techniques are described which provide robust hand-based identification and verification. An image of a hand is obtained, which is then segmented into a palm region and separate finger regions. Acquisition of the image is performed without requiring particular orientation or placement restrictions. Segmentation is performed without the use of reference points on the images. Each segment is analyzed by calculating a set of Zernike moment descriptors for the segment. The feature parameters thus obtained are then fused and compared to stored sets of descriptors in enrollment templates to arrive at an identity decision. By using Zernike moments, and through additional manipulation, the biometric analysis is invariant to rotation, scale, or translation or an in put image. Additionally, the analysis utilizes re-use of commonly-seen terms in Zernike calculations to achieve additional efficiencies over traditional Zernike moment calculation.

  16. Phosphoproteomics of collagen receptor networks reveals SHP-2 phosphorylation downstream of wild-type DDR2 and its lung cancer mutants.

    Science.gov (United States)

    Iwai, Leo K; Payne, Leo S; Luczynski, Maciej T; Chang, Francis; Xu, Huifang; Clinton, Ryan W; Paul, Angela; Esposito, Edward A; Gridley, Scott; Leitinger, Birgit; Naegle, Kristen M; Huang, Paul H

    2013-09-15

    Collagen is an important extracellular matrix component that directs many fundamental cellular processes including differentiation, proliferation and motility. The signalling networks driving these processes are propagated by collagen receptors such as the β1 integrins and the DDRs (discoidin domain receptors). To gain an insight into the molecular mechanisms of collagen receptor signalling, we have performed a quantitative analysis of the phosphorylation networks downstream of collagen activation of integrins and DDR2. Temporal analysis over seven time points identified 424 phosphorylated proteins. Distinct DDR2 tyrosine phosphorylation sites displayed unique temporal activation profiles in agreement with in vitro kinase data. Multiple clustering analysis of the phosphoproteomic data revealed several DDR2 candidate downstream signalling nodes, including SHP-2 (Src homology 2 domain-containing protein tyrosine phosphatase 2), NCK1 (non-catalytic region of tyrosine kinase adaptor protein 1), LYN, SHIP-2 [SH2 (Src homology 2)-domain-containing inositol phosphatase 2], PIK3C2A (phosphatidylinositol-4-phosphate 3-kinase, catalytic subunit type 2α) and PLCL2 (phospholipase C-like 2). Biochemical validation showed that SHP-2 tyrosine phosphorylation is dependent on DDR2 kinase activity. Targeted proteomic profiling of a panel of lung SCC (squamous cell carcinoma) DDR2 mutants demonstrated that SHP-2 is tyrosine-phosphorylated by the L63V and G505S mutants. In contrast, the I638F kinase domain mutant exhibited diminished DDR2 and SHP-2 tyrosine phosphorylation levels which have an inverse relationship with clonogenic potential. Taken together, the results of the present study indicate that SHP-2 is a key signalling node downstream of the DDR2 receptor which may have therapeutic implications in a subset of DDR2 mutations recently uncovered in genome-wide lung SCC sequencing screens.

  17. Chronic low-dose-rate ionising radiation affects the hippocampal phosphoproteome in the ApoE−/− Alzheimer mouse model

    DEFF Research Database (Denmark)

    Kempf, Stefan; Janik, Dirk; Barjaktarovic, Zarko

    2017-01-01

    Accruing data indicate that radiation-induced consequences resemble pathologies of neurodegenerative diseases such as Alzheimer´s. The aim of this study was to elucidate the effect on hippocampus of chronic low-dose-rate radiation exposure (1 mGy/day or 20 mGy/day) given over 300 days with cumula......Accruing data indicate that radiation-induced consequences resemble pathologies of neurodegenerative diseases such as Alzheimer´s. The aim of this study was to elucidate the effect on hippocampus of chronic low-dose-rate radiation exposure (1 mGy/day or 20 mGy/day) given over 300 days...... with cumulative doses of 0.3 Gy and 6.0 Gy, respectively. ApoE deficient mutant C57Bl/6 mouse was used as an Alzheimer´s model. Using mass spectrometry, a marked alteration in the phosphoproteome was found at both dose rates. The radiation-induced changes in the phosphoproteome were associated with the control...... that several molecular targets induced by chronic low-dose-rate radiation overlap with those of Alzheimer´s pathology. It may suggest that ionising radiation functions as a contributing risk factor to this neurodegenerative disease....

  18. Searching for Novel Cdk5 Substrates in Brain by Comparative Phosphoproteomics of Wild Type and Cdk5−/− Mice

    Science.gov (United States)

    Contreras-Vallejos, Erick; Utreras, Elías; Bórquez, Daniel A.; Prochazkova, Michaela; Terse, Anita; Jaffe, Howard; Toledo, Andrea; Arruti, Cristina; Pant, Harish C.; Kulkarni, Ashok B.; González-Billault, Christian

    2014-01-01

    Protein phosphorylation is the most common post-translational modification that regulates several pivotal functions in cells. Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase which is mostly active in the nervous system. It regulates several biological processes such as neuronal migration, cytoskeletal dynamics, axonal guidance and synaptic plasticity among others. In search for novel substrates of Cdk5 in the brain we performed quantitative phosphoproteomics analysis, isolating phosphoproteins from whole brain derived from E18.5 Cdk5+/+ and Cdk5−/− embryos, using an Immobilized Metal-Ion Affinity Chromatography (IMAC), which specifically binds to phosphorylated proteins. The isolated phosphoproteins were eluted and isotopically labeled for relative and absolute quantitation (iTRAQ) and mass spectrometry identification. We found 40 proteins that showed decreased phosphorylation at Cdk5−/− brains. In addition, out of these 40 hypophosphorylated proteins we characterized two proteins, :MARCKS (Myristoylated Alanine-Rich protein Kinase C substrate) and Grin1 (G protein regulated inducer of neurite outgrowth 1). MARCKS is known to be phosphorylated by Cdk5 in chick neural cells while Grin1 has not been reported to be phosphorylated by Cdk5. When these proteins were overexpressed in N2A neuroblastoma cell line along with p35, serine phosphorylation in their Cdk5 motifs was found to be increased. In contrast, treatments with roscovitine, the Cdk5 inhibitor, resulted in an opposite effect on serine phosphorylation in N2A cells and primary hippocampal neurons transfected with MARCKS. In summary, the results presented here identify Grin 1 as novel Cdk5 substrate and confirm previously identified MARCKS as a a bona fide Cdk5 substrate. PMID:24658276

  19. Proteomic and Phosphoproteomic Insights into a Signaling Hub Role for Cdc14 in Asexual Development and Multiple Stress Responses in Beauveria bassiana.

    Science.gov (United States)

    Wang, Zhi-Kang; Wang, Jie; Liu, Jing; Ying, Sheng-Hua; Peng, Xiao-Jun; Feng, Ming-Guang

    2016-01-01

    Cdc14 is a dual-specificity phosphatase that regulates nuclear behavior by dephosphorylating phosphotyrosine and phosphoserine/phosphothreonine in fungi. Previously, Cdc14 was shown to act as a positive regulator of cytokinesis, asexual development and multiple stress responses in Beauveria bassiana, a fungal insect pathogen. This study seeks to gain deep insight into a pivotal role of Cdc14 in the signaling network of B. bassiana by analyzing the Cdc14-specific proteome and phosphoproteome generated by the 8-plex iTRAQ labeling and MS/MS analysis of peptides and phosphopeptides. Under normal conditions, 154 proteins and 86 phosphorylation sites in 67 phosphoproteins were upregulated in Δcdc14 versus wild-type, whereas 117 proteins and 85 phosphorylation sites in 58 phosphoproteins were significantly downregulated. Co-cultivation of Δcdc14 with NaCl (1 M), H2O2 (3 mM) and Congo red (0.15 mg/ml) resulted in the upregulation / downregulation of 23/63, 41/39 and 79/79 proteins and of 127/112, 52/47 and 105/226 phosphorylation sites in 85/92, 45/36 and 79/146 phosphoproteins, respectively. Bioinformatic analyses revealed that Cdc14 could participate in many biological and cellular processes, such as carbohydrate metabolism, glycerophospholipid metabolism, the MAP Kinase signaling pathway, and DNA conformation, by regulating protein expression and key kinase phosphorylation in response to different environmental cues. These indicate that in B. bassiana, Cdc14 is a vital regulator of not only protein expression but also many phosphorylation events involved in developmental and stress-responsive pathways. Fourteen conserved and novel motifs were identified in the fungal phosphorylation events.

  20. Exploring signal transduction networks using mass spectrometry-based proteomics

    NARCIS (Netherlands)

    Meijer, L.A.T.

    2012-01-01

    Mass spectrometry (MS)-based proteomics can be used to answer a diversity of biological questions. In this thesis, we describe the application of several MS-based proteomics approaches to get insight into several aspects of signal transduction. In Chapter 2, quantitative global phosphoproteomics are

  1. Video-Based Motion Analysis

    Science.gov (United States)

    French, Paul; Peterson, Joel; Arrighi, Julie

    2005-04-01

    Video-based motion analysis has recently become very popular in introductory physics classes. This paper outlines general recommendations regarding equipment and software; videography issues such as scaling, shutter speed, lighting, background, and camera distance; as well as other methodological aspects. Also described are the measurement and modeling of the gravitational, drag, and Magnus forces on 1) a spherical projectile undergoing one-dimensional motion and 2) a spinning spherical projectile undergoing motion within a plane. Measurement and correction methods are devised for four common, major sources of error: parallax, lens distortion, discretization, and improper scaling.

  2. Certification-Based Process Analysis

    Science.gov (United States)

    Knight, Russell L.

    2013-01-01

    Space mission architects are often challenged with knowing which investment in technology infusion will have the highest return. Certification-based analysis (CBA) gives architects and technologists a means to communicate the risks and advantages of infusing technologies at various points in a process. Various alternatives can be compared, and requirements based on supporting streamlining or automation can be derived and levied on candidate technologies. CBA is a technique for analyzing a process and identifying potential areas of improvement. The process and analysis products are used to communicate between technologists and architects. Process means any of the standard representations of a production flow; in this case, any individual steps leading to products, which feed into other steps, until the final product is produced at the end. This sort of process is common for space mission operations, where a set of goals is reduced eventually to a fully vetted command sequence to be sent to the spacecraft. Fully vetting a product is synonymous with certification. For some types of products, this is referred to as verification and validation, and for others it is referred to as checking. Fundamentally, certification is the step in the process where one insures that a product works as intended, and contains no flaws.

  3. Chemical phosphoproteomics and development of bisubstrate based inhibitors of protein kinase C isozymes

    NARCIS (Netherlands)

    Poot, A.J.

    2010-01-01

    Protein phosphorylation is one of the most abundant post-translational modifications. Phosphorylation is important for the function of the protein, for example it regulates enzyme activity, signal transduction and cell division. Protein phosphorylation plays a central role in virtually all crucial c

  4. Discovery of Novel Virulence Factors of Biothreat Agents: Validation of the Phosphoproteome-Based Approach

    Science.gov (United States)

    2008-06-30

    the Qiagen Mini- Xpress in vitro protein transpcription and translation system. We found this amount to be insufficient and so we obtained an...experiment generated fascinating results, which will contribute to the field of Francisella LPS studies. Finally, by performing a more global comparison...1948 FTT1275 mglA sspA FTT1275 mglA sspA global transcriptional regulation, possibly through binding RNA polymerase allow survival within

  5. Phosphoproteomic analysis of cells treated with longevity-related autophagy inducers

    DEFF Research Database (Denmark)

    Bennetzen, Martin V; Mariño, Guillermo; Pultz, Dennis

    2012-01-01

    Macroautophagy is a self-cannibalistic process that enables cells to adapt to various stresses and maintain energy homeostasis. Additionally, autophagy is an important route for turnover of misfolded proteins and damaged organelles, with important implications in cancer, neurodegenerative disease...

  6. Phosphoproteomics analysis of postmortem porcine muscle with pH decline rate and time difference

    DEFF Research Database (Denmark)

    Huang, Honggang; Larsen, Martin R; Karlsson, Anders H;

    2012-01-01

    the reverse case. The phosphorylation level of 12 bands in sarcoplasmic fraction and 3 bands in myofibrillar fraction were significantly affected by the synergy effects of pH and time (ppyruvate kinase, triosephosphate isomerase-1...

  7. Kinomic and phospho-proteomic analysis of breast cancer stem-like cells

    DEFF Research Database (Denmark)

    Leth-Larsen, Rikke; Christensen, Anne Geske Lindhard; Ehmsen, Sidse

    /CD24-/low compartment of human breast cancer is enriched in tumor-initiating cells; however the functional heterogeneity within this subpopulation remains poorly defined. From a triple-negative breast cancer cell line we isolated and cloned CD44hi single-cells that exhibited functional heterogeneity...

  8. Phosphoproteomic analysis of differentiating Leishmania parasites reveals a unique stage-specific phosphorylation motif

    OpenAIRE

    Tsigankov, Polina; Gherardini, Pier Federico; Helmer-Citterich, Manuela; Späth, Gerald F; Zilberstein, Dan

    2013-01-01

    International audience; Protists of the genus Leishmania are obligatory intracellular parasites that cause a wide range of cutaneous, mucocutaneous, and visceral diseases in humans. They cycle between phagolysosomes of mammalian macrophages and the sand fly midgut, proliferating as intracellular amastigotes and extracellular promastigotes, respectively. Exposure to a lysosomal environment, i.e. acidic pH and body temperature, signals promastigotes to differentiate into amastigotes. Time cours...

  9. An Augmented Multiple-Protease-Based Human Phosphopeptide Atlas

    NARCIS (Netherlands)

    Giansanti, Piero; Aye, Thin Thin; van den Toorn, Henk; Peng, Mao; van Breukelen, Bas; Heck, Albert J R

    2015-01-01

    Although mass-spectrometry-based screens enable thousands of protein phosphorylation sites to be monitored simultaneously, they often do not cover important regulatory sites. Here, we hypothesized that this is due to the fact that nearly all large-scale phosphoproteome studies are initiated by tryps

  10. Probabilistic Model-Based Safety Analysis

    CERN Document Server

    Güdemann, Matthias; 10.4204/EPTCS.28.8

    2010-01-01

    Model-based safety analysis approaches aim at finding critical failure combinations by analysis of models of the whole system (i.e. software, hardware, failure modes and environment). The advantage of these methods compared to traditional approaches is that the analysis of the whole system gives more precise results. Only few model-based approaches have been applied to answer quantitative questions in safety analysis, often limited to analysis of specific failure propagation models, limited types of failure modes or without system dynamics and behavior, as direct quantitative analysis is uses large amounts of computing resources. New achievements in the domain of (probabilistic) model-checking now allow for overcoming this problem. This paper shows how functional models based on synchronous parallel semantics, which can be used for system design, implementation and qualitative safety analysis, can be directly re-used for (model-based) quantitative safety analysis. Accurate modeling of different types of proba...

  11. Extensions in model-based system analysis

    OpenAIRE

    Graham, Matthew R.

    2007-01-01

    Model-based system analysis techniques provide a means for determining desired system performance prior to actual implementation. In addition to specifying desired performance, model-based analysis techniques require mathematical descriptions that characterize relevant behavior of the system. The developments of this dissertation give ex. tended formulations for control- relevant model estimation as well as model-based analysis conditions for performance requirements specified as frequency do...

  12. Posttranslational regulation of self-renewal capacity: insights from proteome and phosphoproteome analyses of stem cell leukemia

    Science.gov (United States)

    Trost, Matthias; Sauvageau, Martin; Hérault, Olivier; Deleris, Paul; Pomiès, Christelle; Chagraoui, Jalila; Mayotte, Nadine; Meloche, Sylvain; Sauvageau, Guy; Thibault, Pierre

    2017-01-01

    We recently generated 2 phenotypically similar Hoxa9+Meis1 overexpressing acute myeloid leukemias that differ by their in vivo biologic behavior. The first leukemia, named FLA2, shows a high frequency of leukemia stem cells (LSCs; 1 in 1.4 cells), whereas the second, FLB1, is more typical with a frequency of LSCs in the range of 1 per several hundred cells. To gain insights into possible mechanisms that determine LSC self-renewal, we profiled and compared the abundance of nuclear and cytoplasmic proteins and phosphoproteins from these leukemias using quantitative proteomics. These analyses revealed differences in proteins associated with stem cell fate, including a hyperactive p38 MAP kinase in FLB1 and a differentially localized Polycomb group protein Ezh2, which is mostly nuclear in FLA2 and predominantly cytoplasmic in FLB1. Together, these newly documented proteomes and phosphoproteomes represent a unique resource with more than 440 differentially expressed proteins and 11 543 unique phosphopeptides, of which 80% are novel and 7% preferentially phosphorylated in the stem cell–enriched leukemia. PMID:22802335

  13. Changes in the proteome and phosphoproteome expression in the bryozoan Bugula neritina larvae in response to the antifouling agent butenolide

    KAUST Repository

    Qian, Pei Yuan

    2010-09-08

    Larval attachment and metamorphosis, commonly referred to as larval settlement, of marine sessile invertebrates can be triggered or blocked by chemical cues and affected by changes in overall protein expression pattern and phosphorylation dynamics. This study focuses on the effects of butenolide, an effective larval settlement inhibitor, on larval settlement at the proteome level in the bryozoan Bugula neritina. Liquid-phase IEF sample prefractionation combined with 2-DE and MALDI-TOF MS was used to identify the differentially expressed proteins. Substantial changes occurred both in protein abundance and in phosphorylation status during larval settlement and when settling larvae were challenged with butenolide. The proteins that responded to treatment were identified as structural proteins, molecular chaperones, mitochondrial peptidases and calcium-binding proteins. Compared with our earlier results, both genistein and butenolide inhibited larval settlement of B. neritina primarily by changes in protein abundance and the phosphorylation status of proteins but have different protein targets in the same species. Clearly, to design potent antifouling compounds and to understand the mode of action of compounds, more studies on the effects of different compounds on proteome and phosphoproteome of different larval species are required. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.

  14. Desalting of phosphopeptides by tandem polypyrrole-c18 reverse phase micropipette tip (TMTip{sub PPY-C18}) based on hybrid electrostatic, {Pi}-{Pi} stacking and hydrophobic interactions for mass spectrometric analysis

    Energy Technology Data Exchange (ETDEWEB)

    Zheng Shi; Wang Xiaoli; Fu Jieying; Hu Xuejiao; Xiao Xiao; Huang Lulu; Zhou Youe [Key Laboratory of Pesticides and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079 (China); Zhong Hongying, E-mail: hyzhong@mail.ccnu.edu.cn [Key Laboratory of Pesticides and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079 (China)

    2012-04-29

    Highlights: Black-Right-Pointing-Pointer A new micropipette tip TMTip{sub PPY-C18} was developed for desalting of phosphopeptides. Black-Right-Pointing-Pointer TMTip{sub PPY-C18} is based on polypyrrole in tandem with C18 chromatographic material. Black-Right-Pointing-Pointer TMTip{sub PPY-C18} combines electrostatic, {Pi}-{Pi} stacking and hydrophobic interactions. Black-Right-Pointing-Pointer TMTip{sub PPY-C18} can be used in both acidic and basic experimental conditions. - Abstract: Desalting and concentration of peptides using reverse phase (RP) C18 chromatographic material based on hydrophobic interaction is a routine approach used in mass spectrometry (MS)-based proteomics. However, MS detection of small hydrophilic peptides, in particular, phosphopeptides that bear multiple negative charges, is challenging due to the insufficient binding to C18 stationary phase. We described here the development of a new desalting method that takes the unique properties of polypyrrole (PPY). The presence of positively charged nitrogen atoms under acidic conditions and polyunsaturated bonds in polypyrrole provide a prospect for enhanced adsorption of phosphopeptides or hydrophilic peptides through extra electrostatic and {Pi}-{Pi} stacking interactions in addition to hydrophobic interactions. In tandem with reversed phase C18 chromatographic material, the new type of desalting method termed as TMTip{sub PPY-C18} can significantly improve the MS detection of phosphopeptides with multiple phosphate groups and other small hydrophilic peptides. It has been applied to not only tryptic digest of model proteins but also the analysis of complex lysates of zebrafish eggs. The number of detected phosphate groups on a peptide ranged from 1 to 6. Particularly, polypyrrole based method can also be used in basic condition. Thus it provides a useful means to handle peptides that may not be detectable in acidic condition. It can be envisioned that the TMTip{sub PPY-C18} should be able to

  15. Ovarian Cancer Proteomic, Phosphoproteomic, and Glycoproteomic Data Released - Office of Cancer Clinical Proteomics Research

    Science.gov (United States)

    National Cancer Institute (NCI) Clinical Proteomic Tumor Analysis Consortium (CPTAC) scientists have just released a comprehensive dataset of the proteomic analysis of high grade serous ovarian tumor samples,

  16. Towards a Judgement-Based Statistical Analysis

    Science.gov (United States)

    Gorard, Stephen

    2006-01-01

    There is a misconception among social scientists that statistical analysis is somehow a technical, essentially objective, process of decision-making, whereas other forms of data analysis are judgement-based, subjective and far from technical. This paper focuses on the former part of the misconception, showing, rather, that statistical analysis…

  17. Reachability Analysis of Sampling Based Planners

    NARCIS (Netherlands)

    Geraerts, R.J.; Overmars, M.H.

    2005-01-01

    The last decade, sampling based planners like the Probabilistic Roadmap Method have proved to be successful in solving complex motion planning problems. We give a reachability based analysis for these planners which leads to a better understanding of the success of the approach and enhancements of t

  18. Analysis of Enhanced Associativity Based Routing Protocol

    Directory of Open Access Journals (Sweden)

    Said A. Shaar

    2006-01-01

    Full Text Available This study introduces an analysis to the performance of the Enhanced Associativity Based Routing protocol (EABR based on two factors; Operation complexity (OC and Communication Complexity (CC. OC can be defined as the number of steps required in performing a protocol operation, while CC can be defined as the number of messages exchanged in performing a protocol operation[1]. The values represent the worst-case analysis. The EABR has been analyzed based on CC and OC and the results have been compared with another routing technique called ABR. The results have shown that EABR can perform better than ABR in many circumstances during the route reconstruction.

  19. Triple quad ICPMS (ICPQQQ) as a new tool for absolute quantitative proteomics and phosphoproteomics.

    Science.gov (United States)

    Diez Fernández, Silvia; Sugishama, Naoki; Ruiz Encinar, Jorge; Sanz-Medel, Alfredo

    2012-07-17

    It is clear that sensitive and interference-free quantification of ICP-detectable elements naturally present in proteins will boost the role of ICPMS in proteomics. In this study, a completely new way of polyatomic interference removal in ICPMS for detection of sulfur (present in the majority of proteins as methionine or cysteine) and phosphorus (present in phosphorylated proteins) is presented. It is based on the concept of tandem mass spectrometry (QQQ) typically used in molecular MS. Briefly, the first quadrupole can be operated as 1 amu window band-pass mass filter to select target analyte ions ((31)P, (32)S, and their on-mass polyatomic interferences). In this way, only selected ions enter the cell and react with O(2), reducing the interferences produced by matrix ions as well as background noise. After optimization of the cell conditions, product ions formed for the targets, (47)PO(+) and (48)SO(+), could be detected with enhanced sensitivity and selectivity. The coupling to capillary HPLC allowed analysis of S- and P-containing species with the lowest detection limits ever published (11 and 6.6 fmol, respectively). The potential of the approach for proteomics studies was demonstrated for the highly sensitive simultaneous absolute quantification of different S-containing peptides and phosphopeptides.

  20. Quantitative Site-Specific Phosphoproteomics of Trichoderma reesei Signaling Pathways upon Induction of Hydrolytic Enzyme Production.

    Science.gov (United States)

    Nguyen, Elizabeth V; Imanishi, Susumu Y; Haapaniemi, Pekka; Yadav, Avinash; Saloheimo, Markku; Corthals, Garry L; Pakula, Tiina M

    2016-02-01

    The filamentous fungus Trichoderma reesei is used for industrial production of secreted enzymes including carbohydrate active enzymes, such as cellulases and hemicellulases. The production of many of these enzymes by T. reesei is influenced by the carbon source it grows on, where the regulation system controlling hydrolase genes involves various signaling pathways. T. reesei was cultivated in the presence of sorbitol, a carbon source that does not induce the production of cellulases and hemicellulases, and then exposed to either sophorose or spent-grain extract, which are efficient inducers of the enzyme production. Specific changes at phosphorylation sites were investigated in relation to the production of cellulases and hemicellulases using an MS-based framework. Proteome-wide phosphorylation following carbon source exchange was investigated in the early stages of induction: 0, 2, 5, and 10 min. The workflow involved sequential trypsin digestion, TiO2 enrichment, and MS analysis using a Q Exactive mass spectrometer. We report on the identification and quantitation of 1721 phosphorylation sites. Investigation of the data revealed a complex signaling network activated upon induction involving components related to light-mediated cellulase induction, osmoregulation, and carbon sensing. Changes in protein phosphorylation were detected in the glycolytic pathway, suggesting an inhibition of glucose catabolism at 10 min after the addition of sophorose and as early as 2 min after the addition of spent-grain extract. Differential phosphorylation of factors related to carbon storage, intracellular trafficking, cytoskeleton, and cellulase gene regulation were also observed.

  1. Combined Phosphoproteomics and Bioinformatics Strategy in Deciphering Drug Resistant Related Pathways in Triple Negative Breast Cancer

    Directory of Open Access Journals (Sweden)

    Xinyu Deng

    2014-01-01

    Full Text Available Because of the absence of a clear therapeutic target for triple negative breast cancer (TNBC, conventional chemotherapy is the only available systemic treatment option for these patients. Despite chemotherapy treatment, TNBC patients still have worse prognosis when compared with other breast cancer patients. The study is to investigate unique phosphorylated proteins expressed in chemoresistant TNBC cell lines. In the current study, twelve TNBC cell lines were subjected to drug sensitivity assays against chemotherapy drugs docetaxel, doxorubicin, gemcitabine, and cisplatin. Based on their half maximal inhibitory concentrations, four resistant and two sensitive cell lines were selected for further analysis. The phosphopeptides from these cells were enriched with TiO2 beads and fractionated using strong cation exchange. 1,645 phosphoprotein groups and 9,585 unique phosphopeptides were identified by a high throughput LC-MS/MS system LTQ-Orbitrap. The phosphopeptides were further filtered with Ascore system and 1,340 phosphoprotein groups, 2,760 unique phosphopeptides, and 4,549 unique phosphosites were identified. Our study suggested that differentially phosphorylated Cdk5, PML, AP-1, and HSF-1 might work together to promote vimentin induced epithelial to mesenchymal transition (EMT in the drug resistant cells. EGFR and HGF were also shown to be involved in this process.

  2. Quantitative label-free phosphoproteomics of six different life stages of the late blight pathogen Phytophthora infestans reveals abundant phosphorylation of members of the CRN effector family.

    Science.gov (United States)

    Resjö, Svante; Ali, Ashfaq; Meijer, Harold J G; Seidl, Michael F; Snel, Berend; Sandin, Marianne; Levander, Fredrik; Govers, Francine; Andreasson, Erik

    2014-04-04

    The oomycete Phytophthora infestans is the causal agent of late blight in potato and tomato. Since the underlying processes that govern pathogenicity and development in P. infestans are largely unknown, we have performed a large-scale phosphoproteomics study of six different P. infestans life stages. We have obtained quantitative data for 2922 phosphopeptides and compared their abundance. Life-stage-specific phosphopeptides include ATP-binding cassette transporters and a kinase that only occurs in appressoria. In an extended data set, we identified 2179 phosphorylation sites and deduced 22 phosphomotifs. Several of the phosphomotifs matched consensus sequences of kinases that occur in P. infestans but not Arabidopsis. In addition, we detected tyrosine phosphopeptides that are potential targets of kinases resembling mammalian tyrosine kinases. Among the phosphorylated proteins are members of the RXLR and Crinkler effector families. The latter are phosphorylated in several life stages and at multiple positions, in sites that are conserved between different members of the Crinkler family. This indicates that proteins in the Crinkler family have functions beyond their putative role as (necrosis-inducing) effectors. This phosphoproteomics data will be instrumental for studies on oomycetes and host-oomycete interactions. The data sets have been deposited to ProteomeXchange (identifier PXD000433).

  3. Integrated phosphoproteomic and metabolomic profiling reveals NPM-ALK-mediated phosphorylation of PKM2 and metabolic reprogramming in anaplastic large cell lymphoma.

    Science.gov (United States)

    McDonnell, Scott R P; Hwang, Steven R; Rolland, Delphine; Murga-Zamalloa, Carlos; Basrur, Venkatesha; Conlon, Kevin P; Fermin, Damian; Wolfe, Thomas; Raskind, Alexander; Ruan, Chunhai; Jiang, Jian-Kang; Thomas, Craig J; Hogaboam, Cory M; Burant, Charles F; Elenitoba-Johnson, Kojo S J; Lim, Megan S

    2013-08-01

    The mechanisms underlying the pathogenesis of the constitutively active tyrosine kinase nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) expressing anaplastic large cell lymphoma are not completely understood. Here we show using an integrated phosphoproteomic and metabolomic strategy that NPM-ALK induces a metabolic shift toward aerobic glycolysis, increased lactate production, and biomass production. The metabolic shift is mediated through the anaplastic lymphoma kinase (ALK) phosphorylation of the tumor-specific isoform of pyruvate kinase (PKM2) at Y105, resulting in decreased enzymatic activity. Small molecule activation of PKM2 or expression of Y105F PKM2 mutant leads to reversal of the metabolic switch with increased oxidative phosphorylation and reduced lactate production coincident with increased cell death, decreased colony formation, and reduced tumor growth in an in vivo xenograft model. This study provides comprehensive profiling of the phosphoproteomic and metabolomic consequences of NPM-ALK expression and reveals a novel role of ALK in the regulation of multiple components of cellular metabolism. Our studies show that PKM2 is a novel substrate of ALK and plays a critical role in mediating the metabolic shift toward biomass production and tumorigenesis.

  4. Epoch-based analysis of speech signals

    Indian Academy of Sciences (India)

    B Yegnanarayana; Suryakanth V Gangashetty

    2011-10-01

    Speech analysis is traditionally performed using short-time analysis to extract features in time and frequency domains. The window size for the analysis is fixed somewhat arbitrarily, mainly to account for the time varying vocal tract system during production. However, speech in its primary mode of excitation is produced due to impulse-like excitation in each glottal cycle. Anchoring the speech analysis around the glottal closure instants (epochs) yields significant benefits for speech analysis. Epoch-based analysis of speech helps not only to segment the speech signals based on speech production characteristics, but also helps in accurate analysis of speech. It enables extraction of important acoustic-phonetic features such as glottal vibrations, formants, instantaneous fundamental frequency, etc. Epoch sequence is useful to manipulate prosody in speech synthesis applications. Accurate estimation of epochs helps in characterizing voice quality features. Epoch extraction also helps in speech enhancement and multispeaker separation. In this tutorial article, the importance of epochs for speech analysis is discussed, and methods to extract the epoch information are reviewed. Applications of epoch extraction for some speech applications are demonstrated.

  5. Identifying Proper Names Based on Association Analysis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The issue of proper names recognition in Chinese text was discussed. An automatic approach based on association analysis to extract rules from corpus was presented. The method tries to discover rules relevant to external evidence by association analysis, without additional manual effort. These rules can be used to recognize the proper nouns in Chinese texts. The experimental result shows that our method is practical in some applications.Moreover, the method is language independent.

  6. Cloud Based Development Issues: A Methodical Analysis

    Directory of Open Access Journals (Sweden)

    Sukhpal Singh

    2012-11-01

    Full Text Available Cloud based development is a challenging task for various software engineering projects, especifically for those which demand extraordinary quality, reusability and security along with general architecture. In this paper we present a report on a methodical analysis of cloud based development problems published in major computer science and software engineering journals and conferences organized by various researchers. Research papers were collected from different scholarly databases using search engines within a particular period of time. A total of 89 research papers were analyzed in this methodical study and we categorized into four classes according to the problems addressed by them. The majority of the research papers focused on quality (24 papers associated with cloud based development and 16 papers focused on analysis and design. By considering the areas focused by existing authors and their gaps, untouched areas of cloud based development can be discovered for future research works.

  7. Security Analysis of Discrete Logarithm Based Cryptosystems

    Institute of Scientific and Technical Information of China (English)

    WANG Yuzhu; LIAO Xiaofeng

    2006-01-01

    Discrete logarithm based cryptosystems have subtle problems that make the schemes vulnerable. This paper gives a comprehensive listing of security issues in the systems and analyzes three classes of attacks which are based on mathematical structure of the group which is used in the schemes, the disclosed information of the subgroup and implementation details respectively. The analysis will, in turn, allow us to motivate protocol design and implementation decisions.

  8. Social Network Analysis Based on Network Motifs

    OpenAIRE

    2014-01-01

    Based on the community structure characteristics, theory, and methods of frequent subgraph mining, network motifs findings are firstly introduced into social network analysis; the tendentiousness evaluation function and the importance evaluation function are proposed for effectiveness assessment. Compared with the traditional way based on nodes centrality degree, the new approach can be used to analyze the properties of social network more fully and judge the roles of the nodes effectively. I...

  9. Abstraction based Analysis and Arbiter Synthesis

    DEFF Research Database (Denmark)

    Ernits, Juhan-Peep; Yi, Wang

    2004-01-01

    The work focuses on the analysis of an example of synchronous systems containing FIFO buffers, registers and memory interconnected by several private and shared busses. The example used in this work is based on a Terma radar system memory interface case study from the IST AMETIST project....

  10. Network-based analysis of proteomic profiles

    KAUST Repository

    Wong, Limsoon

    2016-01-26

    Mass spectrometry (MS)-based proteomics is a widely used and powerful tool for profiling systems-wide protein expression changes. It can be applied for various purposes, e.g. biomarker discovery in diseases and study of drug responses. Although RNA-based high-throughput methods have been useful in providing glimpses into the underlying molecular processes, the evidences they provide are indirect. Furthermore, RNA and corresponding protein levels have been known to have poor correlation. On the other hand, MS-based proteomics tend to have consistency issues (poor reproducibility and inter-sample agreement) and coverage issues (inability to detect the entire proteome) that need to be urgently addressed. In this talk, I will discuss how these issues can be addressed by proteomic profile analysis techniques that use biological networks (especially protein complexes) as the biological context. In particular, I will describe several techniques that we have been developing for network-based analysis of proteomics profile. And I will present evidence that these techniques are useful in identifying proteomics-profile analysis results that are more consistent, more reproducible, and more biologically coherent, and that these techniques allow expansion of the detected proteome to uncover and/or discover novel proteins.

  11. Workflow-based approaches to neuroimaging analysis.

    Science.gov (United States)

    Fissell, Kate

    2007-01-01

    Analysis of functional and structural magnetic resonance imaging (MRI) brain images requires a complex sequence of data processing steps to proceed from raw image data to the final statistical tests. Neuroimaging researchers have begun to apply workflow-based computing techniques to automate data analysis tasks. This chapter discusses eight major components of workflow management systems (WFMSs): the workflow description language, editor, task modules, data access, verification, client, engine, and provenance, and their implementation in the Fiswidgets neuroimaging workflow system. Neuroinformatics challenges involved in applying workflow techniques in the domain of neuroimaging are discussed.

  12. Texture-based analysis of COPD

    DEFF Research Database (Denmark)

    Sørensen, Lauge; Nielsen, Mads; Lo, Pechin Chien Pau

    2012-01-01

    This study presents a fully automatic, data-driven approach for texture-based quantitative analysis of chronic obstructive pulmonary disease (COPD) in pulmonary computed tomography (CT) images. The approach uses supervised learning where the class labels are, in contrast to previous work, based...... on measured lung function instead of on manually annotated regions of interest (ROIs). A quantitative measure of COPD is obtained by fusing COPD probabilities computed in ROIs within the lung fields where the individual ROI probabilities are computed using a k nearest neighbor (kNN ) classifier. The distance...... and subsequently applied to classify 200 independent images from the same screening trial. The texture-based measure was significantly better at discriminating between subjects with and without COPD than were the two most common quantitative measures of COPD in the literature, which are based on density...

  13. Proteomic and phosphoproteomic comparison of human ES and iPS cells.

    Science.gov (United States)

    Phanstiel, Douglas H; Brumbaugh, Justin; Wenger, Craig D; Tian, Shulan; Probasco, Mitchell D; Bailey, Derek J; Swaney, Danielle L; Tervo, Mark A; Bolin, Jennifer M; Ruotti, Victor; Stewart, Ron; Thomson, James A; Coon, Joshua J

    2011-01-01

    Combining high-mass-accuracy mass spectrometry, isobaric tagging and software for multiplexed, large-scale protein quantification, we report deep proteomic coverage of four human embryonic stem cell and four induced pluripotent stem cell lines in biological triplicate. This 24-sample comparison resulted in a very large set of identified proteins and phosphorylation sites in pluripotent cells. The statistical analysis afforded by our approach revealed subtle but reproducible differences in protein expression and protein phosphorylation between embryonic stem cells and induced pluripotent cells. Merging these results with RNA-seq analysis data, we found functionally related differences across each tier of regulation. We also introduce the Stem Cell-Omics Repository (SCOR), a resource to collate and display quantitative information across multiple planes of measurement, including mRNA, protein and post-translational modifications.

  14. Web-based pre-Analysis Tools

    CERN Document Server

    Moskalets, Tetiana

    2014-01-01

    The project consists in the initial development of a web based and cloud computing services to allow students and researches to perform fast and very useful cut-based pre-analysis on a browser, using real data and official Monte-Carlo simulations (MC). Several tools are considered: ROOT files filter, JavaScript Multivariable Cross-Filter, JavaScript ROOT browser and JavaScript Scatter-Matrix Libraries. Preliminary but satisfactory results have been deployed online for test and future upgrades.

  15. Measuring Class Cohesion Based on Dependence Analysis

    Institute of Scientific and Technical Information of China (English)

    Zhen-Qiang Chen; Bao-Wen Xu; Yu-Ming Zhou

    2004-01-01

    Classes are the basic modules in object-oriented (OO) software, which consist of attributes and methods. Thus, in OO environment, the cohesion is mainly about the tightness of the attributes and methods of classes. This paper discusses the relationships between attributes and attributes, attributes and methods, methods and methods of a class based on dependence analysis. Then the paper presents methods to compute these dependencies. Based on these, the paper proposes a method to measure the class cohesion, which satisfies the properties that a good measurement should have. The approach overcomes the limitations of previous class cohesion measures, which consider only one or two of the three relationships in a class.

  16. TEST COVERAGE ANALYSIS BASED ON PROGRAM SLICING

    Institute of Scientific and Technical Information of China (English)

    Chen Zhenqiang; Xu Baowen; Guanjie

    2003-01-01

    Coverage analysis is a structural testing technique that helps to eliminate gaps in atest suite and determines when to stop testing. To compute test coverage, this letter proposes anew concept coverage about variables, based on program slicing. By adding powers accordingto their importance, the users can focus on the important variables to obtain higher test coverage.The letter presents methods to compute basic coverage based on program structure graphs. Inmost cases, the coverage obtained in the letter is bigger than that obtained by a traditionalmeasure, because the coverage about a variable takes only the related codes into account.

  17. Musical Structural Analysis Database Based on GTTM

    OpenAIRE

    Hamanaka, Masatoshi; Hirata, Keiji; Tojo, Satoshi

    2014-01-01

    This paper, we present the publication of our analysis data and analyzing tool based on the generative theory of tonal music (GTTM). Musical databases such as score databases, instrument sound databases, and musical pieces with standard MIDI files and annotated data are key to advancements in the field of music information technology. We started implementing the GTTM on a computer in 2004 and ever since have collected and publicized test data by musicologists in a step-by-step manner. In our ...

  18. Battle through signaling between wheat and the fungal pathogen Septoria tritici revealed by proteomics and phosphoproteomics

    DEFF Research Database (Denmark)

    Yang, Fen; Melo-Braga, Marcella N; Larsen, Martin Røssel;

    2013-01-01

    compatible and incompatible interactions. However, differential regulation of the phosphorylation status of signaling proteins, transcription and translation regulators, and membrane-associated proteins was observed between two interactions. The proteomic data were correlated with a more rapid or stronger......-binding proteins, 14-3-3 proteins, and calcium-binding proteins. Quantitative PCR analysis showed the expression of fungal signaling genes and genes encoding a superoxide dismutase and cell-wall degrading enzymes. These results indicate roles of signaling, antioxidative stress mechanisms, and nutrient acquisition...

  19. Chapter 11. Community analysis-based methods

    Energy Technology Data Exchange (ETDEWEB)

    Cao, Y.; Wu, C.H.; Andersen, G.L.; Holden, P.A.

    2010-05-01

    Microbial communities are each a composite of populations whose presence and relative abundance in water or other environmental samples are a direct manifestation of environmental conditions, including the introduction of microbe-rich fecal material and factors promoting persistence of the microbes therein. As shown by culture-independent methods, different animal-host fecal microbial communities appear distinctive, suggesting that their community profiles can be used to differentiate fecal samples and to potentially reveal the presence of host fecal material in environmental waters. Cross-comparisons of microbial communities from different hosts also reveal relative abundances of genetic groups that can be used to distinguish sources. In increasing order of their information richness, several community analysis methods hold promise for MST applications: phospholipid fatty acid (PLFA) analysis, denaturing gradient gel electrophoresis (DGGE), terminal restriction fragment length polymorphism (TRFLP), cloning/sequencing, and PhyloChip. Specific case studies involving TRFLP and PhyloChip approaches demonstrate the ability of community-based analyses of contaminated waters to confirm a diagnosis of water quality based on host-specific marker(s). The success of community-based MST for comprehensively confirming fecal sources relies extensively upon using appropriate multivariate statistical approaches. While community-based MST is still under evaluation and development as a primary diagnostic tool, results presented herein demonstrate its promise. Coupled with its inherently comprehensive ability to capture an unprecedented amount of microbiological data that is relevant to water quality, the tools for microbial community analysis are increasingly accessible, and community-based approaches have unparalleled potential for translation into rapid, perhaps real-time, monitoring platforms.

  20. Global phosphoproteomic profiling reveals perturbed signaling in a mouse model of dilated cardiomyopathy

    Science.gov (United States)

    Kuzmanov, Uros; Guo, Hongbo; Buchsbaum, Diana; Cosme, Jake; Abbasi, Cynthia; Isserlin, Ruth; Sharma, Parveen; Gramolini, Anthony O.; Emili, Andrew

    2016-01-01

    Phospholamban (PLN) plays a central role in Ca2+ homeostasis in cardiac myocytes through regulation of the sarco(endo)plasmic reticulum Ca2+-ATPase 2A (SERCA2A) Ca2+ pump. An inherited mutation converting arginine residue 9 in PLN to cysteine (R9C) results in dilated cardiomyopathy (DCM) in humans and transgenic mice, but the downstream signaling defects leading to decompensation and heart failure are poorly understood. Here we used precision mass spectrometry to study the global phosphorylation dynamics of 1,887 cardiac phosphoproteins in early affected heart tissue in a transgenic R9C mouse model of DCM compared with wild-type littermates. Dysregulated phosphorylation sites were quantified after affinity capture and identification of 3,908 phosphopeptides from fractionated whole-heart homogenates. Global statistical enrichment analysis of the differential phosphoprotein patterns revealed selective perturbation of signaling pathways regulating cardiovascular activity in early stages of DCM. Strikingly, dysregulated signaling through the Notch-1 receptor, recently linked to cardiomyogenesis and embryonic cardiac stem cell development and differentiation but never directly implicated in DCM before, was a prominently perturbed pathway. We verified alterations in Notch-1 downstream components in early symptomatic R9C transgenic mouse cardiomyocytes compared with wild type by immunoblot analysis and confocal immunofluorescence microscopy. These data reveal unexpected connections between stress-regulated cell signaling networks, specific protein kinases, and downstream effectors essential for proper cardiac function. PMID:27742792

  1. Novel aspects of grapevine response to phytoplasma infection investigated by a proteomic and phospho-proteomic approach with data integration into functional networks

    Directory of Open Access Journals (Sweden)

    Margaria Paolo

    2013-01-01

    Full Text Available Abstract Background Translational and post-translational protein modifications play a key role in the response of plants to pathogen infection. Among the latter, phosphorylation is critical in modulating protein structure, localization and interaction with other partners. In this work, we used a multiplex staining approach with 2D gels to study quantitative changes in the proteome and phosphoproteome of Flavescence dorée-affected and recovered ‘Barbera’ grapevines, compared to healthy plants. Results We identified 48 proteins that differentially changed in abundance, phosphorylation, or both in response to Flavescence dorée phytoplasma infection. Most of them did not show any significant difference in recovered plants, which, by contrast, were characterized by changes in abundance, phosphorylation, or both for 17 proteins not detected in infected plants. Some enzymes involved in the antioxidant response that were up-regulated in infected plants, such as isocitrate dehydrogenase and glutathione S-transferase, returned to healthy-state levels in recovered plants. Others belonging to the same functional category were even down-regulated in recovered plants (oxidoreductase GLYR1 and ascorbate peroxidase. Our proteomic approach thus agreed with previously published biochemical and RT-qPCR data which reported down-regulation of scavenging enzymes and accumulation of H2O2 in recovered plants, possibly suggesting a role for this molecule in remission from infection. Fifteen differentially phosphorylated proteins (| ratio | > 2, p  Conclusions Proteomic data were integrated into biological networks and their interactions were represented through a hypothetical model, showing the effects of protein modulation on primary metabolic ways and related secondary pathways. By following a multiplex-staining approach, we obtained new data on grapevine proteome pathways that specifically change at the phosphorylation level during phytoplasma infection

  2. Differences in Beef Quality between Angus (Bos taurus taurus) and Nellore (Bos taurus indicus) Cattle through a Proteomic and Phosphoproteomic Approach

    Science.gov (United States)

    Chizzotti, Mario Luiz; Vital, Camilo Elber; Baracat-Pereira, Maria Cristina; Barros, Edvaldo; Busato, Karina Costa; Gomes, Rafael Aparecido; Ladeira, Márcio Machado; Martins, Taiane da Silva

    2017-01-01

    Proteins are the major constituents of muscle and are key molecules regulating the metabolic changes during conversion of muscle to meat. Brazil is one of the largest exporters of beef and most Brazilian cattle are composed by zebu (Nellore) genotype. Bos indicus beef is generally leaner and tougher than Bos taurus such as Angus. The aim of this study was to compare the muscle proteomic and phosphoproteomic profile of Angus and Nellore. Seven animals of each breed previously subjected the same growth management were confined for 84 days. Proteins were extracted from Longissimus lumborum samples collected immediately after slaughter and separated by two-dimensional electrophoresis. Pro-Q Diamond stain was used in phosphoproteomics. Proteins identification was performed using matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Tropomyosin alpha-1 chain, troponin-T, myosin light chain-1 fragment, cytoplasmic malate dehydrogenase, alpha-enolase and 78 kDa glucose-regulated protein were more abundant in Nellore, while myosin light chain 3, prohibitin, mitochondrial stress-70 protein and heat shock 70 kDa protein 6 were more abundant in Angus (P<0.05). Nellore had higher phosphorylation of myosin regulatory light chain-2, alpha actin-1, triosephosphate isomerase and 14-3-3 protein epsilon. However, Angus had greater phosphorylation of phosphoglucomutase-1 and troponin-T (P<0.05). Therefore, proteins involved in contraction and muscle organization, myofilaments expressed in fast or slow-twitch fibers and heat shock proteins localized in mitochondria or sarcoplasmic reticulum and involved in cell flux of calcium and apoptosis might be associated with differences in beef quality between Angus and Nellore. Furthermore, prohibitin appears to be a potential biomarker of intramuscular fat in cattle. Additionally, differences in phosphorylation of myofilaments and glycolytic enzymes could be involved with differences in muscle contraction force

  3. Signal transduction in cerebral arteries after subarachnoid hemorrhage-a phosphoproteomic approach

    DEFF Research Database (Denmark)

    Parker, Benjamin; Larsen, Martin Røssel; Povlsen, Gro Klitgaard

    2013-01-01

    After subarachnoid hemorrhage (SAH), pathologic changes in cerebral arteries contribute to delayed cerebral ischemia and poor outcome. We hypothesize such changes are triggered by early intracellular signals, targeting of which may prevent SAH-induced vasculopathy. We performed an unbiased quanti......-induced signaling components downstream and upstream of ERK1/2.Journal of Cerebral Blood Flow & Metabolism advance online publication, 29 May 2013; doi:10.1038/jcbfm.2013.78....... quantitative analysis of early SAH-induced phosphorylations in cerebral arteries and evaluated identified signaling components as targets for prevention of delayed vasculopathy and ischemia. Labeled phosphopeptides from rat cerebral arteries were quantified by high-resolution tandem mass spectrometry. Selected...

  4. Phosphoproteomics of the Arabidopsis plasma membrane and a new phosphorylation site database

    DEFF Research Database (Denmark)

    Nühse, Thomas S; Stensballe, Allan; Jensen, Ole N;

    2004-01-01

    Functional genomic technologies are generating vast amounts of data describing the presence of transcripts or proteins in plant cells. Together with classical genetics, these approaches broaden our understanding of the gene products required for specific responses. Looking to the future, the focus...... of research must shift to the dynamic aspects of biology: molecular mechanisms of function and regulation. Phosphorylation is a key regulatory factor in all aspects of plant biology; but it is difficult, if not impossible, for most researchers to identify in vivo phosphorylation sites within their proteins...... will be a valuable resource for many fields of plant biology and overcome a major impediment to the elucidation of signal transduction pathways. We present an analysis of the characteristics of phosphorylation sites, their conservation among orthologs and paralogs, and the existence of putative motifs surrounding...

  5. Global investigation of interleukin-1β signaling in primary β-cells using quantitative phosphoproteomics

    DEFF Research Database (Denmark)

    Engholm-Keller, Kasper; Størling, Joachim; Pociot, Flemming

    to culture medium with or without IL-1 β for 10 min. After cellular lysis in 8M urea, the proteome was digested using Lys-C and trypsin, and stable isotope-labeled by reductive dimethylation for subsequent mass spectrometry-based quantification. Phosphopeptides were enriched by TiO2 chromatography...... in the HCD-cell. Preliminary Data We performed three biological replicates of control versus 10 min. IL-1β stimulation of 600, 500, and 800 randomly picked rat islets of Langerhans (approximately 110, 90, and 150 µg of protein per condition, respectively) using quantification by stable isotope labeling via...... and separated into monophosphorylated and multiphosphorylated peptide pools using Sequential elution from IMAC (SIMAC). The monophosphorylated sample was fractionated by microscale HILIC HPLC and all fractions analyzed by nanoLC-ESI-MS/MS on an LTQ-Orbitrap Velos using beam-type collision-induced dissociation...

  6. Gait Correlation Analysis Based Human Identification

    Directory of Open Access Journals (Sweden)

    Jinyan Chen

    2014-01-01

    Full Text Available Human gait identification aims to identify people by a sequence of walking images. Comparing with fingerprint or iris based identification, the most important advantage of gait identification is that it can be done at a distance. In this paper, silhouette correlation analysis based human identification approach is proposed. By background subtracting algorithm, the moving silhouette figure can be extracted from the walking images sequence. Every pixel in the silhouette has three dimensions: horizontal axis (x, vertical axis (y, and temporal axis (t. By moving every pixel in the silhouette image along these three dimensions, we can get a new silhouette. The correlation result between the original silhouette and the new one can be used as the raw feature of human gait. Discrete Fourier transform is used to extract features from this correlation result. Then, these features are normalized to minimize the affection of noise. Primary component analysis method is used to reduce the features’ dimensions. Experiment based on CASIA database shows that this method has an encouraging recognition performance.

  7. Electric Equipment Diagnosis based on Wavelet Analysis

    Directory of Open Access Journals (Sweden)

    Stavitsky Sergey A.

    2016-01-01

    Full Text Available Due to electric equipment development and complication it is necessary to have a precise and intense diagnosis. Nowadays there are two basic ways of diagnosis: analog signal processing and digital signal processing. The latter is more preferable. The basic ways of digital signal processing (Fourier transform and Fast Fourier transform include one of the modern methods based on wavelet transform. This research is dedicated to analyzing characteristic features and advantages of wavelet transform. This article shows the ways of using wavelet analysis and the process of test signal converting. In order to carry out this analysis, computer software Mathcad was used and 2D wavelet spectrum for a complex function was created.

  8. Similarity-based pattern analysis and recognition

    CERN Document Server

    Pelillo, Marcello

    2013-01-01

    This accessible text/reference presents a coherent overview of the emerging field of non-Euclidean similarity learning. The book presents a broad range of perspectives on similarity-based pattern analysis and recognition methods, from purely theoretical challenges to practical, real-world applications. The coverage includes both supervised and unsupervised learning paradigms, as well as generative and discriminative models. Topics and features: explores the origination and causes of non-Euclidean (dis)similarity measures, and how they influence the performance of traditional classification alg

  9. Constructing storyboards based on hierarchical clustering analysis

    Science.gov (United States)

    Hasebe, Satoshi; Sami, Mustafa M.; Muramatsu, Shogo; Kikuchi, Hisakazu

    2005-07-01

    There are growing needs for quick preview of video contents for the purpose of improving accessibility of video archives as well as reducing network traffics. In this paper, a storyboard that contains a user-specified number of keyframes is produced from a given video sequence. It is based on hierarchical cluster analysis of feature vectors that are derived from wavelet coefficients of video frames. Consistent use of extracted feature vectors is the key to avoid a repetition of computationally-intensive parsing of the same video sequence. Experimental results suggest that a significant reduction in computational time is gained by this strategy.

  10. Arabic Interface Analysis Based on Cultural Markers

    CERN Document Server

    Khanum, Mohammadi Akheela; Chaurasia, Mousmi A

    2012-01-01

    This study examines the Arabic interface design elements that are largely influenced by the cultural values. Cultural markers are examined in websites from educational, business, and media. Cultural values analysis is based on Geert Hofstede's cultural dimensions. The findings show that there are cultural markers which are largely influenced by the culture and that the Hofstede's score for Arab countries is partially supported by the website design components examined in this study. Moderate support was also found for the long term orientation, for which Hoftsede has no score.

  11. Motion Analysis Based on Invertible Rapid Transform

    Directory of Open Access Journals (Sweden)

    J. Turan

    1999-06-01

    Full Text Available This paper presents the results of a study on the use of invertible rapid transform (IRT for the motion estimation in a sequence of images. Motion estimation algorithms based on the analysis of the matrix of states (produced in the IRT calculation are described. The new method was used experimentally to estimate crowd and traffic motion from the image data sequences captured at railway stations and at high ways in large cities. The motion vectors may be used to devise a polar plot (showing velocity magnitude and direction for moving objects where the dominant motion tendency can be seen. The experimental results of comparison of the new motion estimation methods with other well known block matching methods (full search, 2D-log, method based on conventional (cross correlation (CC function or phase correlation (PC function for application of crowd motion estimation are also presented.

  12. Visibility Graph Based Time Series Analysis

    Science.gov (United States)

    Stephen, Mutua; Gu, Changgui; Yang, Huijie

    2015-01-01

    Network based time series analysis has made considerable achievements in the recent years. By mapping mono/multivariate time series into networks, one can investigate both it’s microscopic and macroscopic behaviors. However, most proposed approaches lead to the construction of static networks consequently providing limited information on evolutionary behaviors. In the present paper we propose a method called visibility graph based time series analysis, in which series segments are mapped to visibility graphs as being descriptions of the corresponding states and the successively occurring states are linked. This procedure converts a time series to a temporal network and at the same time a network of networks. Findings from empirical records for stock markets in USA (S&P500 and Nasdaq) and artificial series generated by means of fractional Gaussian motions show that the method can provide us rich information benefiting short-term and long-term predictions. Theoretically, we propose a method to investigate time series from the viewpoint of network of networks. PMID:26571115

  13. Identification of novel PAMP-triggered phosphorylation and dephosphorylation events in arabidopsis thaliana by quantitative phosphoproteomic analysis

    KAUST Repository

    Rayapuram, Naganand

    2014-04-04

    Signaling cascades rely strongly on protein kinase-mediated substrate phosphorylation. Currently a major challenge in signal transduction research is to obtain high confidence substrate phosphorylation sites and assign them to specific kinases. In response to bacterial flagellin, a pathogen-associated molecular pattern (PAMP), we searched for rapidly phosphorylated proteins in Arabidopsis thaliana by combining multistage activation (MSA) and electron transfer dissociation (ETD) fragmentation modes, which generate complementary spectra and identify phosphopeptide sites with increased reliability. Of a total of 825 phosphopeptides, we identified 58 to be differentially phosphorylated. These peptides harbor kinase motifs of mitogen-activated protein kinases (MAPKs) and calcium-dependent protein kinases (CDPKs), as well as yet unknown protein kinases. Importantly, 12 of the phosphopeptides show reduced phosphorylation upon flagellin treatment. Since protein abundance levels did not change, these results indicate that flagellin induces not only various protein kinases but also protein phosphatases, even though a scenario of inhibited kinase activity may also be possible. © 2014 American Chemical Society.

  14. Watermark Resistance Analysis Based On Linear Transformation

    Directory of Open Access Journals (Sweden)

    N.Karthika Devi

    2012-06-01

    Full Text Available Generally, digital watermark can be embedded in any copyright image whose size is not larger than it. The watermarking schemes can be classified into two categories: spatial domain approach or transform domain approach. Previous works have shown that the transform domain scheme is typically more robust to noise, common image processing, and compression when compared with the spatial transform scheme. Improvements in performance of watermarking schemes can be obtained by exploiting the characteristics of the human visual system (HVS in the watermarking process. We propose a linear transformation based watermarking algorithm. The watermarking bits are embedded into cover image to produce watermarked image. The efficiency of watermark is checked using pre-defined attacks. Attack resistance analysis is done using BER (Bit Error Rate calculation. Finally, the Quality of the watermarked image can be obtained.

  15. Web Template Extraction Based on Hyperlink Analysis

    Directory of Open Access Journals (Sweden)

    Julián Alarte

    2015-01-01

    Full Text Available Web templates are one of the main development resources for website engineers. Templates allow them to increase productivity by plugin content into already formatted and prepared pagelets. For the final user templates are also useful, because they provide uniformity and a common look and feel for all webpages. However, from the point of view of crawlers and indexers, templates are an important problem, because templates usually contain irrelevant information such as advertisements, menus, and banners. Processing and storing this information is likely to lead to a waste of resources (storage space, bandwidth, etc.. It has been measured that templates represent between 40% and 50% of data on the Web. Therefore, identifying templates is essential for indexing tasks. In this work we propose a novel method for automatic template extraction that is based on similarity analysis between the DOM trees of a collection of webpages that are detected using menus information. Our implementation and experiments demonstrate the usefulness of the technique.

  16. Visual Similarity Based Document Layout Analysis

    Institute of Scientific and Technical Information of China (English)

    Di Wen; Xiao-Qing Ding

    2006-01-01

    In this paper, a visual similarity based document layout analysis (DLA) scheme is proposed, which by using clustering strategy can adaptively deal with documents in different languages, with different layout structures and skew angles. Aiming at a robust and adaptive DLA approach, the authors first manage to find a set of representative filters and statistics to characterize typical texture patterns in document images, which is through a visual similarity testing process.Texture features are then extracted from these filters and passed into a dynamic clustering procedure, which is called visual similarity clustering. Finally, text contents are located from the clustered results. Benefit from this scheme, the algorithm demonstrates strong robustness and adaptability in a wide variety of documents, which previous traditional DLA approaches do not possess.

  17. Voxel-Based LIDAR Analysis and Applications

    Science.gov (United States)

    Hagstrom, Shea T.

    One of the greatest recent changes in the field of remote sensing is the addition of high-quality Light Detection and Ranging (LIDAR) instruments. In particular, the past few decades have been greatly beneficial to these systems because of increases in data collection speed and accuracy, as well as a reduction in the costs of components. These improvements allow modern airborne instruments to resolve sub-meter details, making them ideal for a wide variety of applications. Because LIDAR uses active illumination to capture 3D information, its output is fundamentally different from other modalities. Despite this difference, LIDAR datasets are often processed using methods appropriate for 2D images and that do not take advantage of its primary virtue of 3-dimensional data. It is this problem we explore by using volumetric voxel modeling. Voxel-based analysis has been used in many applications, especially medical imaging, but rarely in traditional remote sensing. In part this is because the memory requirements are substantial when handling large areas, but with modern computing and storage this is no longer a significant impediment. Our reason for using voxels to model scenes from LIDAR data is that there are several advantages over standard triangle-based models, including better handling of overlapping surfaces and complex shapes. We show how incorporating system position information from early in the LIDAR point cloud generation process allows radiometrically-correct transmission and other novel voxel properties to be recovered. This voxelization technique is validated on simulated data using the Digital Imaging and Remote Sensing Image Generation (DIRSIG) software, a first-principles based ray-tracer developed at the Rochester Institute of Technology. Voxel-based modeling of LIDAR can be useful on its own, but we believe its primary advantage is when applied to problems where simpler surface-based 3D models conflict with the requirement of realistic geometry. To

  18. Interactive analysis of geodata based intelligence

    Science.gov (United States)

    Wagner, Boris; Eck, Ralf; Unmüessig, Gabriel; Peinsipp-Byma, Elisabeth

    2016-05-01

    When a spatiotemporal events happens, multi-source intelligence data is gathered to understand the problem, and strategies for solving the problem are investigated. The difficulties arising from handling spatial and temporal intelligence data represent the main problem. The map might be the bridge to visualize the data and to get the most understand model for all stakeholders. For the analysis of geodata based intelligence data, a software was developed as a working environment that combines geodata with optimized ergonomics. The interaction with the common operational picture (COP) is so essentially facilitated. The composition of the COP is based on geodata services, which are normalized by international standards of the Open Geospatial Consortium (OGC). The basic geodata are combined with intelligence data from images (IMINT) and humans (HUMINT), stored in a NATO Coalition Shared Data Server (CSD). These intelligence data can be combined with further information sources, i.e., live sensors. As a result a COP is generated and an interaction suitable for the specific workspace is added. This allows the users to work interactively with the COP, i.e., searching with an on board CSD client for suitable intelligence data and integrate them into the COP. Furthermore, users can enrich the scenario with findings out of the data of interactive live sensors and add data from other sources. This allows intelligence services to contribute effectively to the process by what military and disaster management are organized.

  19. Operating cost analysis of anaesthesia: Activity based costing (ABC analysis

    Directory of Open Access Journals (Sweden)

    Majstorović Branislava M.

    2011-01-01

    Full Text Available Introduction. Cost of anaesthesiology represent defined measures to determine a precise profile of expenditure estimation of surgical treatment, which is important regarding planning of healthcare activities, prices and budget. Objective. In order to determine the actual value of anaestesiological services, we started with the analysis of activity based costing (ABC analysis. Methods. Retrospectively, in 2005 and 2006, we estimated the direct costs of anestesiological services (salaries, drugs, supplying materials and other: analyses and equipment. of the Institute of Anaesthesia and Resuscitation of the Clinical Centre of Serbia. The group included all anesthetized patients of both sexes and all ages. We compared direct costs with direct expenditure, “each cost object (service or unit” of the Republican Health-care Insurance. The Summary data of the Departments of Anaesthesia documented in the database of the Clinical Centre of Serbia. Numerical data were utilized and the numerical data were estimated and analyzed by computer programs Microsoft Office Excel 2003 and SPSS for Windows. We compared using the linear model of direct costs and unit costs of anaesthesiological services from the Costs List of the Republican Health-care Insurance. Results. Direct costs showed 40% of costs were spent on salaries, (32% on drugs and supplies, and 28% on other costs, such as analyses and equipment. The correlation of the direct costs of anaestesiological services showed a linear correlation with the unit costs of the Republican Healthcare Insurance. Conclusion. During surgery, costs of anaesthesia would increase by 10% the surgical treatment cost of patients. Regarding the actual costs of drugs and supplies, we do not see any possibility of costs reduction. Fixed elements of direct costs provide the possibility of rationalization of resources in anaesthesia.

  20. Automatic malware analysis an emulator based approach

    CERN Document Server

    Yin, Heng

    2012-01-01

    Malicious software (i.e., malware) has become a severe threat to interconnected computer systems for decades and has caused billions of dollars damages each year. A large volume of new malware samples are discovered daily. Even worse, malware is rapidly evolving becoming more sophisticated and evasive to strike against current malware analysis and defense systems. Automatic Malware Analysis presents a virtualized malware analysis framework that addresses common challenges in malware analysis. In regards to this new analysis framework, a series of analysis techniques for automatic malware analy

  1. Node-based analysis of species distributions

    DEFF Research Database (Denmark)

    Borregaard, Michael Krabbe; Rahbek, Carsten; Fjeldså, Jon;

    2014-01-01

    The integration of species distributions and evolutionary relationships is one of the most rapidly moving research fields today and has led to considerable advances in our understanding of the processes underlying biogeographical patterns. Here, we develop a set of metrics, the specific overrepre......The integration of species distributions and evolutionary relationships is one of the most rapidly moving research fields today and has led to considerable advances in our understanding of the processes underlying biogeographical patterns. Here, we develop a set of metrics, the specific...... with case studies on two groups with well-described biogeographical histories: a local-scale community data set of hummingbirds in the North Andes, and a large-scale data set of the distribution of all species of New World flycatchers. The node-based analysis of these two groups generates a set...... of intuitively interpretable patterns that are consistent with current biogeographical knowledge.Importantly, the results are statistically tractable, opening many possibilities for their use in analyses of evolutionary, historical and spatial patterns of species diversity. The method is implemented...

  2. A Translation Case Analysis Based on Skopos Theory

    Institute of Scientific and Technical Information of China (English)

    盖孟姣

    2015-01-01

    This paper is a translation case analysis based on Skopos Theory.This paper choose President Xi’s New Year congratulations of 2015 as analysis text and gives the case analysis.This paper focuses on translating the text based on Skopos Theory.

  3. Using Willie's Acid-Base Box for Blood Gas Analysis

    Science.gov (United States)

    Dietz, John R.

    2011-01-01

    In this article, the author describes a method developed by Dr. William T. Lipscomb for teaching blood gas analysis of acid-base status and provides three examples using Willie's acid-base box. Willie's acid-base box is constructed using three of the parameters of standard arterial blood gas analysis: (1) pH; (2) bicarbonate; and (3) CO[subscript…

  4. A high-resolution tissue-specific proteome and phosphoproteome atlas of maize primary roots reveals functional gradients along the root axes.

    Science.gov (United States)

    Marcon, Caroline; Malik, Waqas Ahmed; Walley, Justin W; Shen, Zhouxin; Paschold, Anja; Smith, Laurie G; Piepho, Hans-Peter; Briggs, Steven P; Hochholdinger, Frank

    2015-05-01

    A high-resolution proteome and phosphoproteome atlas of four maize (Zea mays) primary root tissues, the cortex, stele, meristematic zone, and elongation zone, was generated. High-performance liquid chromatography coupled with tandem mass spectrometry identified 11,552 distinct nonmodified and 2,852 phosphorylated proteins across the four root tissues. Two gradients reflecting the abundance of functional protein classes along the longitudinal root axis were observed. While the classes RNA, DNA, and protein peaked in the meristematic zone, cell wall, lipid metabolism, stress, transport, and secondary metabolism culminated in the differentiation zone. Functional specialization of tissues is underscored by six of 10 cortex-specific proteins involved in flavonoid biosynthesis. Comparison of this data set with high-resolution seed and leaf proteome studies revealed 13% (1,504/11,552) root-specific proteins. While only 23% of the 1,504 root-specific proteins accumulated in all four root tissues, 61% of all 11,552 identified proteins accumulated in all four root tissues. This suggests a much higher degree of tissue-specific functionalization of root-specific proteins. In summary, these data illustrate the remarkable plasticity of the proteomic landscape of maize primary roots and thus provide a starting point for gaining a better understanding of their tissue-specific functions.

  5. ANALYSIS OF CIRCUIT TOLERANCE BASED ON RANDOM SET THEORY

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Monte Carlo Analysis has been an accepted method for circuit tolerance analysis,but the heavy computational complexity has always prevented its applications.Based on random set theory,this paper presents a simple and flexible tolerance analysis method to estimate circuit yield.It is the alternative to Monte Carlo analysis,but reduces the number of calculations dramatically.

  6. Image based performance analysis of thermal imagers

    Science.gov (United States)

    Wegner, D.; Repasi, E.

    2016-05-01

    Due to advances in technology, modern thermal imagers resemble sophisticated image processing systems in functionality. Advanced signal and image processing tools enclosed into the camera body extend the basic image capturing capability of thermal cameras. This happens in order to enhance the display presentation of the captured scene or specific scene details. Usually, the implemented methods are proprietary company expertise, distributed without extensive documentation. This makes the comparison of thermal imagers especially from different companies a difficult task (or at least a very time consuming/expensive task - e.g. requiring the execution of a field trial and/or an observer trial). For example, a thermal camera equipped with turbulence mitigation capability stands for such a closed system. The Fraunhofer IOSB has started to build up a system for testing thermal imagers by image based methods in the lab environment. This will extend our capability of measuring the classical IR-system parameters (e.g. MTF, MTDP, etc.) in the lab. The system is set up around the IR- scene projector, which is necessary for the thermal display (projection) of an image sequence for the IR-camera under test. The same set of thermal test sequences might be presented to every unit under test. For turbulence mitigation tests, this could be e.g. the same turbulence sequence. During system tests, gradual variation of input parameters (e. g. thermal contrast) can be applied. First ideas of test scenes selection and how to assembly an imaging suite (a set of image sequences) for the analysis of imaging thermal systems containing such black boxes in the image forming path is discussed.

  7. A Requirements Analysis Model Based on QFD

    Institute of Scientific and Technical Information of China (English)

    TANG Zhi-wei; Nelson K.H.Tang

    2004-01-01

    The enterprise resource planning (ERP) system has emerged to offer an integrated IT solution and more and more enterprises are increasing by adopting this system and regarding it as an important innovation. However, there is already evidence of high failure risks in ERP project implementation, one major reason is poor analysis of the requirements for system implementation. In this paper, the importance of requirements analysis for ERP project implementation is highlighted, and a requirements analysis model by applying quality function deployment (QFD) is presented, which will support to conduct requirements analysis for ERP project.

  8. Scope-Based Method Cache Analysis

    DEFF Research Database (Denmark)

    Huber, Benedikt; Hepp, Stefan; Schoeberl, Martin

    2014-01-01

    , as it requests memory transfers at well-defined instructions only. In this article, we present a new cache analysis framework that generalizes and improves work on cache persistence analysis. The analysis demonstrates that a global view on the cache behavior permits the precise analyses of caches which are hard......The quest for time-predictable systems has led to the exploration of new hardware architectures that simplify analysis and reasoning in the temporal domain, while still providing competitive performance. For the instruction memory, the method cache is a conceptually attractive solution...

  9. Web Based Distributed Coastal Image Analysis System Project

    Data.gov (United States)

    National Aeronautics and Space Administration — This project develops Web based distributed image analysis system processing the Moderate Resolution Imaging Spectroradiometer (MODIS) data to provide decision...

  10. Canonical analysis based on mutual information

    DEFF Research Database (Denmark)

    Nielsen, Allan Aasbjerg; Vestergaard, Jacob Schack

    2015-01-01

    combinations with the information theoretical measure mutual information (MI). We term this type of analysis canonical information analysis (CIA). MI allows for the actual joint distribution of the variables involved and not just second order statistics. While CCA is ideal for Gaussian data, CIA facilitates...

  11. Phospho-Network Analysis Identifies and Quantifies Hepatitis C Virus (HCV)-induced Hepatocellular Carcinoma (HCC) Proteins Regulating Viral-mediated Tumor Growth

    OpenAIRE

    Lu, Nu T.; Liu, Natalie M.; VU, JAMES Q; PATEL, DARSHIL; COHN, WHITAKER; CAPRI, JOE; ZIEGLER, MARY; Patel, Nikita; Tramontano, Angela; Williams, Roger; Whitelegge, Julian; French, Samuel W.

    2016-01-01

    Background: Patients with chronic hepatitis C virus (HCV) infection are at risk of serious complications of cirrhosis and hepatocellular carcinoma (HCC). Mass spectrometry (MS) is a versatile methodology that produces a global proteomic landscape for analysis of cancer mechanisms. Materials and Methods: Using multiplex peptide stable isotopic labeling and immobilized metal affinity chromatography (IMAC), we enriched and quantified the phosphoproteome of HCC, with and without HCV. While raw da...

  12. Decision making based on data analysis methods

    OpenAIRE

    Sirola, Miki; Sulkava, Mika

    2016-01-01

    This technical report is based on four our recent articles:"Data fusion of pre-election gallups and polls for improved support estimates", "Analyzing parliamentary elections based on voting advice application data", "The Finnish car rejection reasons shown in an interactive SOM visualization tool", and "Network visualization of car inspection data using graph layout". Neural methods are applied in political and technical decision making. We introduce decision support schemes based on Self-Org...

  13. Pathway-Based Functional Analysis of Metagenomes

    Science.gov (United States)

    Bercovici, Sivan; Sharon, Itai; Pinter, Ron Y.; Shlomi, Tomer

    Metagenomic data enables the study of microbes and viruses through their DNA as retrieved directly from the environment in which they live. Functional analysis of metagenomes explores the abundance of gene families, pathways, and systems, rather than their taxonomy. Through such analysis researchers are able to identify those functional capabilities most important to organisms in the examined environment. Recently, a statistical framework for the functional analysis of metagenomes was described that focuses on gene families. Here we describe two pathway level computational models for functional analysis that take into account important, yet unaddressed issues such as pathway size, gene length and overlap in gene content among pathways. We test our models over carefully designed simulated data and propose novel approaches for performance evaluation. Our models significantly improve over current approach with respect to pathway ranking and the computations of relative abundance of pathways in environments.

  14. Analysis of Task-based Syllabus

    Institute of Scientific and Technical Information of China (English)

    马进胜

    2011-01-01

    Task-based language teaching is very popular in the modem English teaching.It is based on the Task-based Syllabus.Taskbased Syllabus focuses on the learners' communicative competence,which stresses learning by doing.From the theoretical assumption and definitions of the task,the paper analysizes the components of the task,then points out the merits and demerits of the syllabus.By this means the paper may give some tips to teachers and students when they use the tsk-based language teaching.

  15. FUZZY PRINCIPAL COMPONENT ANALYSIS AND ITS KERNEL BASED MODEL

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Principal Component Analysis (PCA) is one of the most important feature extraction methods, and Kernel Principal Component Analysis (KPCA) is a nonlinear extension of PCA based on kernel methods. In real world, each input data may not be fully assigned to one class and it may partially belong to other classes. Based on the theory of fuzzy sets, this paper presents Fuzzy Principal Component Analysis (FPCA) and its nonlinear extension model, i.e., Kernel-based Fuzzy Principal Component Analysis (KFPCA). The experimental results indicate that the proposed algorithms have good performances.

  16. Transect based analysis versus area based analysis to quantify shoreline displacement: spatial resolution issues.

    Science.gov (United States)

    Anfuso, Giorgio; Bowman, Dan; Danese, Chiara; Pranzini, Enzo

    2016-10-01

    Field surveys, aerial photographs, and satellite images are the most commonly employed sources of data to analyze shoreline position, which are further compared by area based analysis (ABA) or transect based analysis (TBA) methods. The former is performed by computing the mean shoreline displacement for the identified coastal segments, i.e., dividing the beach area variation by the segment length; the latter is based on the measurement of the distance between each shoreline at set points along transects. The present study compares, by means of GIS tools, the ABA and TBA methods by computing shoreline displacements recorded on two stretches of the Tuscany coast (Italy): the beaches of Punta Ala, a linear coast without shore protection structures, and the one at Follonica, which is irregular due to the presence of groins and detached breakwaters. Surveys were carried out using a differential global positioning system (DGPS) in RTK mode. For each site, a 4800-m-long coastal segment was analyzed and divided into ninety-six 50-m-long sectors for which changes were computed using both the ABA and TBA methods. Sectors were progressively joined to have a length of 100, 200, 400, and 800 m to examine how this influenced results. ABA and TBA results are highly correlated for transect distance and sector length up to 100 m at both investigated locations. If longer transects are considered, the two methods still produce good correlated data on the smooth shoreline (i.e. at Punta Ala), but correlation became significantly lower on the irregular shoreline (i.e., at Follonica).

  17. Description-based and experience-based decisions: individual analysis

    Directory of Open Access Journals (Sweden)

    Andrey Kudryavtsev

    2012-05-01

    Full Text Available We analyze behavior in two basic classes of decision tasks: description-based and experience-based. In particular, we compare the prediction power of a number of decision learning models in both kinds of tasks. Unlike most previous studies, we focus on individual, rather than aggregate, behavioral characteristics. We carry out an experiment involving a battery of both description- and experience-based choices between two mixed binary prospects made by each of the participants, and employ a number of formal models for explaining and predicting participants' choices: Prospect theory (PT (Kahneman and Tversky, 1979; Expectancy-Valence model (EVL (Busemeyer and Stout, 2002; and three combinations of these well-established models. We document that the PT and the EVL models are best for predicting people's decisions in description- and experience-based tasks, respectively, which is not surprising as these two models are designed specially for these kinds of tasks. Furthermore, we find that models involving linear weighting of gains and losses perform better in both kinds of tasks, from the point of view of generalizability and individual parameter consistency. We therefore, conclude that, overall, when both prospects are mixed, the assumption of diminishing sensitivity does not improve models' prediction power for individual decision-makers. Finally, for some of the models' parameters, we document consistency at the individual level between description- and experience-based tasks.

  18. Performance Analysis Based on Timing Simulation

    DEFF Research Database (Denmark)

    Nielsen, Christian Dalsgaard; Kishinevsky, Michael

    1994-01-01

    Determining the cycle time and a critical cycle is a fundamental problem in the analysis of concurrent systems. We solve this problemusing timing simulation of an underlying Signal Graph (an extension of Marked Graphs). For a Signal Graph with n vertices and m arcs our algorithm has the polynomia...... time complexity O(b2m), where b is the number of vertices with initially marked in-arcs (typically b≪n). The algorithm has a clear semantic and a low descriptive complexity. We illustrate the use of the algorithm by applying it to performance analysis of asynchronous circuits.......Determining the cycle time and a critical cycle is a fundamental problem in the analysis of concurrent systems. We solve this problemusing timing simulation of an underlying Signal Graph (an extension of Marked Graphs). For a Signal Graph with n vertices and m arcs our algorithm has the polynomial...

  19. Pathway-based analysis tools for complex diseases: a review.

    Science.gov (United States)

    Jin, Lv; Zuo, Xiao-Yu; Su, Wei-Yang; Zhao, Xiao-Lei; Yuan, Man-Qiong; Han, Li-Zhen; Zhao, Xiang; Chen, Ye-Da; Rao, Shao-Qi

    2014-10-01

    Genetic studies are traditionally based on single-gene analysis. The use of these analyses can pose tremendous challenges for elucidating complicated genetic interplays involved in complex human diseases. Modern pathway-based analysis provides a technique, which allows a comprehensive understanding of the molecular mechanisms underlying complex diseases. Extensive studies utilizing the methods and applications for pathway-based analysis have significantly advanced our capacity to explore large-scale omics data, which has rapidly accumulated in biomedical fields. This article is a comprehensive review of the pathway-based analysis methods-the powerful methods with the potential to uncover the biological depths of the complex diseases. The general concepts and procedures for the pathway-based analysis methods are introduced and then, a comprehensive review of the major approaches for this analysis is presented. In addition, a list of available pathway-based analysis software and databases is provided. Finally, future directions and challenges for the methodological development and applications of pathway-based analysis techniques are discussed. This review will provide a useful guide to dissect complex diseases.

  20. Pathway-based Analysis Tools for Complex Diseases: A Review

    Directory of Open Access Journals (Sweden)

    Lv Jin

    2014-10-01

    Full Text Available Genetic studies are traditionally based on single-gene analysis. The use of these analyses can pose tremendous challenges for elucidating complicated genetic interplays involved in complex human diseases. Modern pathway-based analysis provides a technique, which allows a comprehensive understanding of the molecular mechanisms underlying complex diseases. Extensive studies utilizing the methods and applications for pathway-based analysis have significantly advanced our capacity to explore large-scale omics data, which has rapidly accumulated in biomedical fields. This article is a comprehensive review of the pathway-based analysis methods—the powerful methods with the potential to uncover the biological depths of the complex diseases. The general concepts and procedures for the pathway-based analysis methods are introduced and then, a comprehensive review of the major approaches for this analysis is presented. In addition, a list of available pathway-based analysis software and databases is provided. Finally, future directions and challenges for the methodological development and applications of pathway-based analysis techniques are discussed. This review will provide a useful guide to dissect complex diseases.

  1. Improving Cluster Analysis with Automatic Variable Selection Based on Trees

    Science.gov (United States)

    2014-12-01

    ANALYSIS WITH AUTOMATIC VARIABLE SELECTION BASED ON TREES by Anton D. Orr December 2014 Thesis Advisor: Samuel E. Buttrey Second Reader...DATES COVERED Master’s Thesis 4. TITLE AND SUBTITLE IMPROVING CLUSTER ANALYSIS WITH AUTOMATIC VARIABLE SELECTION BASED ON TREES 5. FUNDING NUMBERS 6...2006 based on classification and regression trees to address problems with determining dissimilarity. Current algorithms do not simultaneously address

  2. Quantitative phosphoproteomics reveals the role of the AMPK plant ortholog SnRK1 as a metabolic master regulator under energy deprivation

    Science.gov (United States)

    Nukarinen, Ella; Nägele, Thomas; Pedrotti, Lorenzo; Wurzinger, Bernhard; Mair, Andrea; Landgraf, Ramona; Börnke, Frederik; Hanson, Johannes; Teige, Markus; Baena-Gonzalez, Elena; Dröge-Laser, Wolfgang; Weckwerth, Wolfram

    2016-01-01

    Since years, research on SnRK1, the major cellular energy sensor in plants, has tried to define its role in energy signalling. However, these attempts were notoriously hampered by the lethality of a complete knockout of SnRK1. Therefore, we generated an inducible amiRNA::SnRK1α2 in a snrk1α1 knock out background (snrk1α1/α2) to abolish SnRK1 activity to understand major systemic functions of SnRK1 signalling under energy deprivation triggered by extended night treatment. We analysed the in vivo phosphoproteome, proteome and metabolome and found that activation of SnRK1 is essential for repression of high energy demanding cell processes such as protein synthesis. The most abundant effect was the constitutively high phosphorylation of ribosomal protein S6 (RPS6) in the snrk1α1/α2 mutant. RPS6 is a major target of TOR signalling and its phosphorylation correlates with translation. Further evidence for an antagonistic SnRK1 and TOR crosstalk comparable to the animal system was demonstrated by the in vivo interaction of SnRK1α1 and RAPTOR1B in the cytosol and by phosphorylation of RAPTOR1B by SnRK1α1 in kinase assays. Moreover, changed levels of phosphorylation states of several chloroplastic proteins in the snrk1α1/α2 mutant indicated an unexpected link to regulation of photosynthesis, the main energy source in plants. PMID:27545962

  3. Encounter-based worms: Analysis and Defense

    CERN Document Server

    Tanachaiwiwat, Sapon

    2007-01-01

    Encounter-based network is a frequently-disconnected wireless ad-hoc network requiring immediate neighbors to store and forward aggregated data for information disseminations. Using traditional approaches such as gateways or firewalls for deterring worm propagation in encounter-based networks is inappropriate. We propose the worm interaction approach that relies upon automated beneficial worm generation aiming to alleviate problems of worm propagations in such networks. To understand the dynamic of worm interactions and its performance, we mathematically model worm interactions based on major worm interaction factors including worm interaction types, network characteristics, and node characteristics using ordinary differential equations and analyze their effects on our proposed metrics. We validate our proposed model using extensive synthetic and trace-driven simulations. We find that, all worm interaction factors significantly affect the pattern of worm propagations. For example, immunization linearly decrea...

  4. Model Based Analysis of Insider Threats

    DEFF Research Database (Denmark)

    Chen, Taolue; Han, Tingting; Kammueller, Florian

    2016-01-01

    In order to detect malicious insider attacks it is important to model and analyse infrastructures and policies of organisations and the insiders acting within them. We extend formal approaches that allow modelling such scenarios by quantitative aspects to enable a precise analysis of security...... designs. Our framework enables evaluating the risks of an insider attack to happen quantitatively. The framework first identifies an insider's intention to perform an inside attack, using Bayesian networks, and in a second phase computes the probability of success for an inside attack by this actor, using...... probabilistic model checking. We provide prototype tool support using Matlab for Bayesian networks and PRISM for the analysis of Markov decision processes, and validate the framework with case studies....

  5. Structural Analysis of Plate Based Tensegrity Structures

    DEFF Research Database (Denmark)

    Hald, Frederik; Kirkegaard, Poul Henning; Damkilde, Lars

    2013-01-01

    Plate tensegrity structures combine tension cables with a cross laminated timber plate and can then form e.g. a roof structure. The topology of plate tensegrity structures is investigated through a parametric investigation. Plate tensegrity structures are investigated, and a method...... for determination of the structures pre-stresses is used. A parametric investigation is performed to determine a more optimized form of the plate based tensegrity structure. Conclusions of the use of plate based tensegrity in civil engineering and further research areas are discussed....

  6. Symbolic Analysis of OTRAs-Based Circuits

    Directory of Open Access Journals (Sweden)

    C. Sánchez-López

    2011-04-01

    Full Text Available A new nullor-based model to describe the behavior of Operational Transresistance Amplifiers (OTRAs is introduced.The new model is composed of four nullors and three grounded resistors. As a consequence, standard nodal analysiscan be applied to compute fully-symbolic small-signal characteristics of OTRA-based analog circuits, and the nullorbasedOTRAs model can be used in CAD tools. In this manner, the fully-symbolic transfer functions of severalapplication circuits, such as filters and oscillators can easily be approximated.

  7. Crime prevention: more evidence-based analysis.

    Science.gov (United States)

    Garrido Genovés, Vicente; Farrington, David P; Welsh, Brandon C

    2008-02-01

    This paper introduces a new section of Psicothema dedicated to the evidence-based approach to crime prevention. Along with an original sexual-offender-treatment programme implemented in Spain, this section presents four systematic reviews of important subjects in the criminological arena, such as sexual offender treatment, the well-known programme, the effectiveness of custodial versus non-custodial sanctions in reoffending and the fight against terrorism. We also highlight some of the focal points that scientists, practitioners and governments should take into account in order to support this evidence-based viewpoint of crime prevention.

  8. Thanatophoric dysplasia: case-based bioethical analysis

    Directory of Open Access Journals (Sweden)

    Edgar Abarca López

    2014-04-01

    Full Text Available This paper presents a case report of thanatophoric displasia diagnosed in the prenatal period using ultrasound standards. The course of the case pregnancy, birth process, and postnatal period is described. This report invites bioethical analysis using its principles, appealing to human dignity, diversity and otherness, particularly in the mother-child dyad and their family. An early diagnosis allows parental support as they face the course of this condition and its potentially fatal outcome.

  9. Movement Pattern Analysis Based on Sequence Signatures

    Directory of Open Access Journals (Sweden)

    Seyed Hossein Chavoshi

    2015-09-01

    Full Text Available Increased affordability and deployment of advanced tracking technologies have led researchers from various domains to analyze the resulting spatio-temporal movement data sets for the purpose of knowledge discovery. Two different approaches can be considered in the analysis of moving objects: quantitative analysis and qualitative analysis. This research focuses on the latter and uses the qualitative trajectory calculus (QTC, a type of calculus that represents qualitative data on moving point objects (MPOs, and establishes a framework to analyze the relative movement of multiple MPOs. A visualization technique called sequence signature (SESI is used, which enables to map QTC patterns in a 2D indexed rasterized space in order to evaluate the similarity of relative movement patterns of multiple MPOs. The applicability of the proposed methodology is illustrated by means of two practical examples of interacting MPOs: cars on a highway and body parts of a samba dancer. The results show that the proposed method can be effectively used to analyze interactions of multiple MPOs in different domains.

  10. Chip based electroanalytical systems for cell analysis

    DEFF Research Database (Denmark)

    Spegel, C.; Heiskanen, A.; Skjolding, L.H.D.

    2008-01-01

    This review with 239 references has as its aim to give the reader an introduction to the kinds of methods used for developing microchip based electrode systems as well as to cover the existing literature on electroanalytical systems where microchips play a crucial role for 'nondestructive...

  11. Analysis of Cloud-Based Database Systems

    Science.gov (United States)

    2015-06-01

    University San Luis Obispo, 2009 Submitted in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE IN COMPUTER SCIENCE...for a query to complete on average for the production system was 136,746 xvi microseconds. On our cloud-based system, the average was 198,875

  12. Web-Based Statistical Sampling and Analysis

    Science.gov (United States)

    Quinn, Anne; Larson, Karen

    2016-01-01

    Consistent with the Common Core State Standards for Mathematics (CCSSI 2010), the authors write that they have asked students to do statistics projects with real data. To obtain real data, their students use the free Web-based app, Census at School, created by the American Statistical Association (ASA) to help promote civic awareness among school…

  13. In vivo quantitative phosphoproteomic profiling identifies novel regulators of castration-resistant prostate cancer growth

    DEFF Research Database (Denmark)

    Jiang, Nan; Hjorth-Jensen, Kim; Hekmat, Omid

    2015-01-01

    pathways in castration-resistant tumors, a notion that was confirmed by tumor transcriptome analysis. Further analysis demonstrated that the activation of mTORC1, PAK2 and the increased levels of YAP1 in castration-resistant tumors can be explained by the loss of androgen inhibitory actions. The analysis...

  14. An SQL-based approach to physics analysis

    Science.gov (United States)

    Limper, Maaike, Dr

    2014-06-01

    As part of the CERN openlab collaboration a study was made into the possibility of performing analysis of the data collected by the experiments at the Large Hadron Collider (LHC) through SQL-queries on data stored in a relational database. Currently LHC physics analysis is done using data stored in centrally produced "ROOT-ntuple" files that are distributed through the LHC computing grid. The SQL-based approach to LHC physics analysis presented in this paper allows calculations in the analysis to be done at the database and can make use of the database's in-built parallelism features. Using this approach it was possible to reproduce results for several physics analysis benchmarks. The study shows the capability of the database to handle complex analysis tasks but also illustrates the limits of using row-based storage for storing physics analysis data, as performance was limited by the I/O read speed of the system.

  15. Desiccant-Based Preconditioning Market Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Fischer, J.

    2001-01-11

    A number of important conclusions can be drawn as a result of this broad, first-phase market evaluation. The more important conclusions include the following: (1) A very significant market opportunity will exist for specialized outdoor air-handling units (SOAHUs) as more construction and renovation projects are designed to incorporate the recommendations made by the ASHRAE 62-1989 standard. Based on this investigation, the total potential market is currently $725,000,000 annually (see Table 6, Sect. 3). Based on the market evaluations completed, it is estimated that approximately $398,000,000 (55%) of this total market could be served by DBC systems if they were made cost-effective through mass production. Approximately $306,000,000 (42%) of the total can be served by a non-regenerated, desiccant-based total recovery approach, based on the information provided by this investigation. Approximately $92,000,000 (13%) can be served by a regenerated desiccant-based cooling approach (see Table 7, Sect. 3). (2) A projection of the market selling price of various desiccant-based SOAHU systems was prepared using prices provided by Trane for central-station, air-handling modules currently manufactured. The wheel-component pricing was added to these components by SEMCO. This resulted in projected pricing for these systems that is significantly less than that currently offered by custom suppliers (see Table 4, Sect. 2). Estimated payback periods for all SOAHU approaches were quite short when compared with conventional over-cooling and reheat systems. Actual paybacks may vary significantly depending on site-specific considerations. (3) In comparing cost vs benefit of each SOAHU approach, it is critical that the total system design be evaluated. For example, the cost premium of a DBC system is very significant when compared to a conventional air handling system, yet the reduced chiller, boiler, cooling tower, and other expense often equals or exceeds this premium, resulting in a

  16. Google glass based immunochromatographic diagnostic test analysis

    Science.gov (United States)

    Feng, Steve; Caire, Romain; Cortazar, Bingen; Turan, Mehmet; Wong, Andrew; Ozcan, Aydogan

    2015-03-01

    Integration of optical imagers and sensors into recently emerging wearable computational devices allows for simpler and more intuitive methods of integrating biomedical imaging and medical diagnostics tasks into existing infrastructures. Here we demonstrate the ability of one such device, the Google Glass, to perform qualitative and quantitative analysis of immunochromatographic rapid diagnostic tests (RDTs) using a voice-commandable hands-free software-only interface, as an alternative to larger and more bulky desktop or handheld units. Using the built-in camera of Glass to image one or more RDTs (labeled with Quick Response (QR) codes), our Glass software application uploads the captured image and related information (e.g., user name, GPS, etc.) to our servers for remote analysis and storage. After digital analysis of the RDT images, the results are transmitted back to the originating Glass device, and made available through a website in geospatial and tabular representations. We tested this system on qualitative human immunodeficiency virus (HIV) and quantitative prostate-specific antigen (PSA) RDTs. For qualitative HIV tests, we demonstrate successful detection and labeling (i.e., yes/no decisions) for up to 6-fold dilution of HIV samples. For quantitative measurements, we activated and imaged PSA concentrations ranging from 0 to 200 ng/mL and generated calibration curves relating the RDT line intensity values to PSA concentration. By providing automated digitization of both qualitative and quantitative test results, this wearable colorimetric diagnostic test reader platform on Google Glass can reduce operator errors caused by poor training, provide real-time spatiotemporal mapping of test results, and assist with remote monitoring of various biomedical conditions.

  17. Model Based Analysis and Test Generation for Flight Software

    Science.gov (United States)

    Pasareanu, Corina S.; Schumann, Johann M.; Mehlitz, Peter C.; Lowry, Mike R.; Karsai, Gabor; Nine, Harmon; Neema, Sandeep

    2009-01-01

    We describe a framework for model-based analysis and test case generation in the context of a heterogeneous model-based development paradigm that uses and combines Math- Works and UML 2.0 models and the associated code generation tools. This paradigm poses novel challenges to analysis and test case generation that, to the best of our knowledge, have not been addressed before. The framework is based on a common intermediate representation for different modeling formalisms and leverages and extends model checking and symbolic execution tools for model analysis and test case generation, respectively. We discuss the application of our framework to software models for a NASA flight mission.

  18. Discontinuous deformation analysis based on complementary theory

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The contact between blocks is treated by the open-close iteration in the conventional discontinuous deformation analysis (DDA),which needs to introduce spurious springs between two blocks in contact and to assume the normal stiffness and the tangential stiffness (the penalty factors). Unreasonable values of stiffness would result in numerical problems. To avoid the penalty factors and the open-close iteration,we reformulate the DDA as a mixed complementary problem (MiCP) and then choose the path Newton method (PNM) to solve the problem. Some examples including those originally designed by Shi are reanalyzed,which proves feasibility of the proposed procedure.

  19. Knowledge-based analysis of phenotypes

    KAUST Repository

    Hoendorf, Robert

    2016-01-27

    Phenotypes are the observable characteristics of an organism, and they are widely recorded in biology and medicine. To facilitate data integration, ontologies that formally describe phenotypes are being developed in several domains. I will describe a formal framework to describe phenotypes. A formalized theory of phenotypes is not only useful for domain analysis, but can also be applied to assist in the diagnosis of rare genetic diseases, and I will show how our results on the ontology of phenotypes is now applied in biomedical research.

  20. Dynamic Analysis of Multilayers Based MEMS Resonators

    Directory of Open Access Journals (Sweden)

    Hassen M. Ouakad

    2017-01-01

    Full Text Available The dynamic behavior of a microelectromechanical system (MEMS parallel and electrically coupled double-layers (microbeams based resonator is investigated. Two numerical methods were used to solve the dynamical problem: the reduced-order modeling (ROM and the perturbation method. The ROM was derived using the so-called Galerkin expansion with considering the linear undamped mode shapes of straight beam as the basis functions. The perturbation method was generated using the method of multiple scales by direct attack of the equations of motion. Dynamic analyses, assuming the above two numerical methods were performed, and a comparison of the results showed good agreement. Finally, a parametric study was performed using the perturbation on different parameters and the results revealed different interesting features, which hopefully can be useful for some MEMS based applications.

  1. Nondestructive Damage Detection Based on Modal Analysis

    Directory of Open Access Journals (Sweden)

    T. Plachý

    2004-01-01

    Full Text Available Three studies of damage identification and localization based on methods using experimentally estimated modal characteristics are presented. The results of an experimental investigation of simple structural elements (three RC-beams and three RC-slabs obtained in the laboratory are compared with the results obtained on a real structure (a composite bridge – a concrete deck supported by steel girders in situ. 

  2. Risk-Based Explosive Safety Analysis

    Science.gov (United States)

    2016-11-30

    Risk-based analyses can also be used for risk management purposes and comparative studies when evaluating test programs that utilize energetic...liquids or propellants. 15. SUBJECT TERMS N/A 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES 19a. NAME OF...analyses can also be used for risk management purposes and comparative studies when evaluating test programs that utilize energetic liquids or

  3. Model-based methods for linkage analysis.

    Science.gov (United States)

    Rice, John P; Saccone, Nancy L; Corbett, Jonathan

    2008-01-01

    The logarithm of an odds ratio (LOD) score method originated in a seminal article by Newton Morton in 1955. The method is broadly concerned with issues of power and the posterior probability of linkage, ensuring that a reported linkage has a high probability of being a true linkage. In addition, the method is sequential so that pedigrees or LOD curves may be combined from published reports to pool data for analysis. This approach has been remarkably successful for 50 years in identifying disease genes for Mendelian disorders. After discussing these issues, we consider the situation for complex disorders where the maximum LOD score statistic shares some of the advantages of the traditional LOD score approach, but is limited by unknown power and the lack of sharing of the primary data needed to optimally combine analytic results. We may still learn from the LOD score method as we explore new methods in molecular biology and genetic analysis to utilize the complete human DNA sequence and the cataloging of all human genes.

  4. Face Recognition Based on Principal Component Analysis

    Directory of Open Access Journals (Sweden)

    Ali Javed

    2013-02-01

    Full Text Available The purpose of the proposed research work is to develop a computer system that can recognize a person by comparing the characteristics of face to those of known individuals. The main focus is on frontal two dimensional images that are taken in a controlled environment i.e. the illumination and the background will be constant. All the other methods of person’s identification and verification like iris scan or finger print scan require high quality and costly equipment’s but in face recognition we only require a normal camera giving us a 2-D frontal image of the person that will be used for the process of the person’s recognition. Principal Component Analysis technique has been used in the proposed system of face recognition. The purpose is to compare the results of the technique under the different conditions and to find the most efficient approach for developing a facial recognition system

  5. Mental EEG Analysis Based on Infomax Algorithm

    Institute of Scientific and Technical Information of China (English)

    WUXiao-pei; GuoXiao-jing; ZANGDao-xin; SHENQian

    2004-01-01

    The patterns of EEG will change with mental tasks performed by the subject. In the field of EEG signal analysis and application, the study to get the patterns of mental EEG and then to use them to classify mental tasks has the significant scientific meaning and great application value. But for the reasons of different artifacts existing in EEG, the pattern detection of EEG under normal mental states is a very difficult problem. In this paper, Independent Component Analysisis applied to EEG signals collected from performing different mental tasks. The experiment results show that when one subject performs a single mental task in different trials, the independent components of EEG are very similar. It means that the independent components can be used as the mental EEG patterns to classify the different mental tasks.

  6. Network Anomaly Detection Based on Wavelet Analysis

    Directory of Open Access Journals (Sweden)

    Ali A. Ghorbani

    2008-11-01

    Full Text Available Signal processing techniques have been applied recently for analyzing and detecting network anomalies due to their potential to find novel or unknown intrusions. In this paper, we propose a new network signal modelling technique for detecting network anomalies, combining the wavelet approximation and system identification theory. In order to characterize network traffic behaviors, we present fifteen features and use them as the input signals in our system. We then evaluate our approach with the 1999 DARPA intrusion detection dataset and conduct a comprehensive analysis of the intrusions in the dataset. Evaluation results show that the approach achieves high-detection rates in terms of both attack instances and attack types. Furthermore, we conduct a full day's evaluation in a real large-scale WiFi ISP network where five attack types are successfully detected from over 30 millions flows.

  7. Wavelet Based Fractal Analysis of Airborne Pollen

    CERN Document Server

    Degaudenzi, M E

    1999-01-01

    The most abundant biological particles in the atmosphere are pollen grains and spores. Self protection of pollen allergy is possible through the information of future pollen contents in the air. In spite of the importance of airborne pol len concentration forecasting, it has not been possible to predict the pollen concentrations with great accuracy, and about 25% of the daily pollen forecasts have resulted in failures. Previous analysis of the dynamic characteristics of atmospheric pollen time series indicate that the system can be described by a low dimensional chaotic map. We apply the wavelet transform to study the multifractal characteristics of an a irborne pollen time series. We find the persistence behaviour associated to low pollen concentration values and to the most rare events of highest pollen co ncentration values. The information and the correlation dimensions correspond to a chaotic system showing loss of information with time evolution.

  8. Sandia National Laboratories analysis code data base

    Energy Technology Data Exchange (ETDEWEB)

    Peterson, C.W.

    1994-11-01

    Sandia National Laboratories, mission is to solve important problems in the areas of national defense, energy security, environmental integrity, and industrial technology. The Laboratories` strategy for accomplishing this mission is to conduct research to provide an understanding of the important physical phenomena underlying any problem, and then to construct validated computational models of the phenomena which can be used as tools to solve the problem. In the course of implementing this strategy, Sandia`s technical staff has produced a wide variety of numerical problem-solving tools which they use regularly in the design, analysis, performance prediction, and optimization of Sandia components, systems and manufacturing processes. This report provides the relevant technical and accessibility data on the numerical codes used at Sandia, including information on the technical competency or capability area that each code addresses, code ``ownership`` and release status, and references describing the physical models and numerical implementation.

  9. LEARNING DIFFICULTIES: AN ANALYSIS BASED ON VIGOTSKY

    Directory of Open Access Journals (Sweden)

    Adriane Cenci

    2010-06-01

    Full Text Available We aimed, along the text, to bring a reflection upon learning difficulties based on Socio-Historical Theory, relating what is observed in schools to what has been discussed about learning difficulties and the theory proposed by Vygotsky in the early XX century. We understand that children enter school carrying experiences and knowledge from their cultural group and that school ignores such knowledge very often. Then, it is in such disengagement that emerges what we started to call learning difficulties. One cannot forget to see a child as a whole – a student is a social being constituted by culture, language and specific values to which one must be attentive.

  10. Building Extraction from LIDAR Based Semantic Analysis

    Institute of Scientific and Technical Information of China (English)

    YU Jie; YANG Haiquan; TAN Ming; ZHANG Guoning

    2006-01-01

    Extraction of buildings from LIDAR data has been an active research field in recent years. A scheme for building detection and reconstruction from LIDAR data is presented with an object-oriented method which is based on the buildings' semantic rules. Two key steps are discussed: how to group the discrete LIDAR points into single objects and how to establish the buildings' semantic rules. In the end, the buildings are reconstructed in 3D form and three common parametric building models (flat, gabled, hipped) are implemented.

  11. Advanced overlay analysis through design based metrology

    Science.gov (United States)

    Ji, Sunkeun; Yoo, Gyun; Jo, Gyoyeon; Kang, Hyunwoo; Park, Minwoo; Kim, Jungchan; Park, Chanha; Yang, Hyunjo; Yim, Donggyu; Maruyama, Kotaro; Park, Byungjun; Yamamoto, Masahiro

    2015-03-01

    As design rule shrink, overlay has been critical factor for semiconductor manufacturing. However, the overlay error which is determined by a conventional measurement with an overlay mark based on IBO and DBO often does not represent the physical placement error in the cell area. The mismatch may arise from the size or pitch difference between the overlay mark and the cell pattern. Pattern distortion caused by etching or CMP also can be a source of the mismatch. In 2014, we have demonstrated that method of overlay measurement in the cell area by using DBM (Design Based Metrology) tool has more accurate overlay value than conventional method by using an overlay mark. We have verified the reproducibility by measuring repeatable patterns in the cell area, and also demonstrated the reliability by comparing with CD-SEM data. We have focused overlay mismatching between overlay mark and cell area until now, further more we have concerned with the cell area having different pattern density and etch loading. There appears a phenomenon which has different overlay values on the cells with diverse patterning environment. In this paper, the overlay error was investigated from cell edge to center. For this experiment, we have verified several critical layers in DRAM by using improved(Better resolution and speed) DBM tool, NGR3520.

  12. The Route Analysis Based On Flight Plan

    Science.gov (United States)

    Feriyanto, Nur; Saleh, Chairul; Fauzi, Achmad; Rachman Dzakiyullah, Nur; Riza Iwaputra, Kahfi

    2016-02-01

    Economic development effects use of air transportation since the business process in every aspect was increased. Many people these days was prefer using airplane because it can save time and money. This situation also effects flight routes, many airlines offer new routes to deal with competition. Managing flight routes is one of the problems that must be faced in order to find the efficient and effective routes. This paper investigates the best routes based on flight performance by determining the amount of block fuel for the Jakarta-Denpasar flight route. Moreover, in this work compares a two kinds of aircraft and tracks by calculating flight distance, flight time and block fuel. The result shows Jakarta-Denpasar in the Track II has effective and efficient block fuel that can be performed by Airbus 320-200 aircraft. This study can contribute to practice in making an effective decision, especially helping executive management of company due to selecting appropriate aircraft and the track in the flight plan based on the block fuel consumption for business operation.

  13. IMAGE ANALYSIS BASED ON EDGE DETECTION TECHNIQUES

    Institute of Scientific and Technical Information of China (English)

    纳瑟; 刘重庆

    2002-01-01

    A method that incorporates edge detection technique, Markov Random field (MRF), watershed segmentation and merging techniques was presented for performing image segmentation and edge detection tasks. It first applies edge detection technique to obtain a Difference In Strength (DIS) map. An initial segmented result is obtained based on K-means clustering technique and the minimum distance. Then the region process is modeled by MRF to obtain an image that contains different intensity regions. The gradient values are calculated and then the watershed technique is used. DIS calculation is used for each pixel to define all the edges (weak or strong) in the image. The DIS map is obtained. This help as priority knowledge to know the possibility of the region segmentation by the next step (MRF), which gives an image that has all the edges and regions information. In MRF model,gray level l, at pixel location i, in an image X, depends on the gray levels of neighboring pixels. The segmentation results are improved by using watershed algorithm. After all pixels of the segmented regions are processed, a map of primitive region with edges is generated. The edge map is obtained using a merge process based on averaged intensity mean values. A common edge detectors that work on (MRF) segmented image are used and the results are compared. The segmentation and edge detection result is one closed boundary per actual region in the image.

  14. Choice-Based Conjoint Analysis: Classification vs. Discrete Choice Models

    Science.gov (United States)

    Giesen, Joachim; Mueller, Klaus; Taneva, Bilyana; Zolliker, Peter

    Conjoint analysis is a family of techniques that originated in psychology and later became popular in market research. The main objective of conjoint analysis is to measure an individual's or a population's preferences on a class of options that can be described by parameters and their levels. We consider preference data obtained in choice-based conjoint analysis studies, where one observes test persons' choices on small subsets of the options. There are many ways to analyze choice-based conjoint analysis data. Here we discuss the intuition behind a classification based approach, and compare this approach to one based on statistical assumptions (discrete choice models) and to a regression approach. Our comparison on real and synthetic data indicates that the classification approach outperforms the discrete choice models.

  15. Adding trend data to Depletion-Based Stock Reduction Analysis

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — A Bayesian model of Depletion-Based Stock Reduction Analysis (DB-SRA), informed by a time series of abundance indexes, was developed, using the Sampling Importance...

  16. Method for detecting software anomalies based on recurrence plot analysis

    OpenAIRE

    Michał Mosdorf

    2012-01-01

    Presented paper evaluates method for detecting software anomalies based on recurrence plot analysis of trace log generated by software execution. Described method for detecting software anomalies is based on windowed recurrence quantification analysis for selected measures (e.g. Recurrence rate - RR or Determinism - DET). Initial results show that proposed method is useful in detecting silent software anomalies that do not result in typical crashes (e.g. exceptions).

  17. Method for detecting software anomalies based on recurrence plot analysis

    Directory of Open Access Journals (Sweden)

    Michał Mosdorf

    2012-03-01

    Full Text Available Presented paper evaluates method for detecting software anomalies based on recurrence plot analysis of trace log generated by software execution. Described method for detecting software anomalies is based on windowed recurrence quantification analysis for selected measures (e.g. Recurrence rate - RR or Determinism - DET. Initial results show that proposed method is useful in detecting silent software anomalies that do not result in typical crashes (e.g. exceptions.

  18. Discrete Discriminant analysis based on tree-structured graphical models

    DEFF Research Database (Denmark)

    Perez de la Cruz, Gonzalo; Eslava, Guillermina

    The purpose of this paper is to illustrate the potential use of discriminant analysis based on tree{structured graphical models for discrete variables. This is done by comparing its empirical performance using estimated error rates for real and simulated data. The results show that discriminant...... analysis based on tree{structured graphical models is a simple nonlinear method competitive with, and sometimes superior to, other well{known linear methods like those assuming mutual independence between variables and linear logistic regression....

  19. Sentiment Analysis of Document Based on Annotation

    CERN Document Server

    Shukla, Archana

    2011-01-01

    I present a tool which tells the quality of document or its usefulness based on annotations. Annotation may include comments, notes, observation, highlights, underline, explanation, question or help etc. comments are used for evaluative purpose while others are used for summarization or for expansion also. Further these comments may be on another annotation. Such annotations are referred as meta-annotation. All annotation may not get equal weightage. My tool considered highlights, underline as well as comments to infer the collective sentiment of annotators. Collective sentiments of annotators are classified as positive, negative, objectivity. My tool computes collective sentiment of annotations in two manners. It counts all the annotation present on the documents as well as it also computes sentiment scores of all annotation which includes comments to obtain the collective sentiments about the document or to judge the quality of document. I demonstrate the use of tool on research paper.

  20. Analysis of Vehicle-Based Security Operations

    Energy Technology Data Exchange (ETDEWEB)

    Carter, Jason M [ORNL; Paul, Nate R [ORNL

    2015-01-01

    Vehicle-to-vehicle (V2V) communications promises to increase roadway safety by providing each vehicle with 360 degree situational awareness of other vehicles in proximity, and by complementing onboard sensors such as radar or camera in detecting imminent crash scenarios. In the United States, approximately three hundred million automobiles could participate in a fully deployed V2V system if Dedicated Short-Range Communication (DSRC) device use becomes mandatory. The system s reliance on continuous communication, however, provides a potential means for unscrupulous persons to transmit false data in an attempt to cause crashes, create traffic congestion, or simply render the system useless. V2V communications must be highly scalable while retaining robust security and privacy preserving features to meet the intra-vehicle and vehicle-to-infrastructure communication requirements for a growing vehicle population. Oakridge National Research Laboratory is investigating a Vehicle-Based Security System (VBSS) to provide security and privacy for a fully deployed V2V and V2I system. In the VBSS an On-board Unit (OBU) generates short-term certificates and signs Basic Safety Messages (BSM) to preserve privacy and enhance security. This work outlines a potential VBSS structure and its operational concepts; it examines how a vehicle-based system might feasibly provide security and privacy, highlights remaining challenges, and explores potential mitigations to address those challenges. Certificate management alternatives that attempt to meet V2V security and privacy requirements have been examined previously by the research community including privacy-preserving group certificates, shared certificates, and functional encryption. Due to real-world operational constraints, adopting one of these approaches for VBSS V2V communication is difficult. Timely misbehavior detection and revocation are still open problems for any V2V system. We explore the alternative approaches that may be

  1. Simulation based analysis of laser beam brazing

    Science.gov (United States)

    Dobler, Michael; Wiethop, Philipp; Schmid, Daniel; Schmidt, Michael

    2016-03-01

    Laser beam brazing is a well-established joining technology in car body manufacturing with main applications in the joining of divided tailgates and the joining of roof and side panels. A key advantage of laser brazed joints is the seam's visual quality which satisfies highest requirements. However, the laser beam brazing process is very complex and process dynamics are only partially understood. In order to gain deeper knowledge of the laser beam brazing process, to determine optimal process parameters and to test process variants, a transient three-dimensional simulation model of laser beam brazing is developed. This model takes into account energy input, heat transfer as well as fluid and wetting dynamics that lead to the formation of the brazing seam. A validation of the simulation model is performed by metallographic analysis and thermocouple measurements for different parameter sets of the brazing process. These results show that the multi-physical simulation model not only can be used to gain insight into the laser brazing process but also offers the possibility of process optimization in industrial applications. The model's capabilities in determining optimal process parameters are exemplarily shown for the laser power. Small deviations in the energy input can affect the brazing results significantly. Therefore, the simulation model is used to analyze the effect of the lateral laser beam position on the energy input and the resulting brazing seam.

  2. Product Lifecycle Management Architecture: A Model Based Systems Engineering Analysis.

    Energy Technology Data Exchange (ETDEWEB)

    Noonan, Nicholas James [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2015-07-01

    This report is an analysis of the Product Lifecycle Management (PLM) program. The analysis is centered on a need statement generated by a Nuclear Weapons (NW) customer. The need statement captured in this report creates an opportunity for the PLM to provide a robust service as a solution. Lifecycles for both the NW and PLM are analyzed using Model Based System Engineering (MBSE).

  3. PYTHON-based Physics Analysis Environment for LHCb

    CERN Document Server

    Belyaev, I; Mato, P; Barrand, G; Tsaregorodtsev, A; de Oliveira, E

    2004-01-01

    BENDER is the PYTHON based physics analysis application for LHCb. It combines the best features of the underlying GAUDI software architecture with the flexibility of the PYTHON scripting language and provides end-users with a friendly physics analysis oriented environment.

  4. Geographic Object-Based Image Analysis: Towards a new paradigm

    NARCIS (Netherlands)

    Blaschke, T.; Hay, G.J.; Kelly, M.; Lang, S.; Hofmann, P.; Addink, E.A.; Queiroz Feitosa, R.; van der Meer, F.D.; van der Werff, H.M.A.; van Coillie, F.; Tiede, A.

    2014-01-01

    The amount of scientific literature on (Geographic) Object-based Image Analysis – GEOBIA has been and still is sharply increasing. These approaches to analysing imagery have antecedents in earlier research on image segmentation and use GIS-like spatial analysis within classification and feature extr

  5. Web-Based Trainer for Electrical Circuit Analysis

    Science.gov (United States)

    Weyten, L.; Rombouts, P.; De Maeyer, J.

    2009-01-01

    A Web-based system for training electric circuit analysis is presented in this paper. It is centered on symbolic analysis techniques and it not only verifies the student's final answer, but it also tracks and coaches him/her through all steps of his/her reasoning path. The system mimics homework assignments, enhanced by immediate personalized…

  6. Lists2Networks: Integrated analysis of gene/protein lists

    Directory of Open Access Journals (Sweden)

    Ma'ayan Avi

    2010-02-01

    Full Text Available Abstract Background Systems biologists are faced with the difficultly of analyzing results from large-scale studies that profile the activity of many genes, RNAs and proteins, applied in different experiments, under different conditions, and reported in different publications. To address this challenge it is desirable to compare the results from different related studies such as mRNA expression microarrays, genome-wide ChIP-X, RNAi screens, proteomics and phosphoproteomics experiments in a coherent global framework. In addition, linking high-content multilayered experimental results with prior biological knowledge can be useful for identifying functional themes and form novel hypotheses. Results We present Lists2Networks, a web-based system that allows users to upload lists of mammalian genes/proteins onto a server-based program for integrated analysis. The system includes web-based tools to manipulate lists with different set operations, to expand lists using existing mammalian networks of protein-protein interactions, co-expression correlation, or background knowledge co-annotation correlation, as well as to apply gene-list enrichment analyses against many gene-list libraries of prior biological knowledge such as pathways, gene ontology terms, kinase-substrate, microRNA-mRAN, and protein-protein interactions, metabolites, and protein domains. Such analyses can be applied to several lists at once against many prior knowledge libraries of gene-lists associated with specific annotations. The system also contains features that allow users to export networks and share lists with other users of the system. Conclusions Lists2Networks is a user friendly web-based software system expected to significantly ease the computational analysis process for experimental systems biologists employing high-throughput experiments at multiple layers of regulation. The system is freely available at http://www.lists2networks.org.

  7. Surveillance data bases, analysis, and standardization program

    Energy Technology Data Exchange (ETDEWEB)

    Kam, F.B.K.

    1990-09-26

    The traveler presented a paper at the Seventh ASTM-EURATOM Symposium on Reactor Dosimetry and co-chaired an oral session on Computer Codes and Methods. Papers of considerable interest to the NRC Surveillance Dosimetry Program involved statistically based adjustment procedures and uncertainties. The information exchange meetings with Czechoslovakia and Hungary were very enlightening. Lack of large computers have hindered their surveillance program. They depended very highly on information from their measurement programs which were somewhat limited because of the lack of sophisticated electronics. The Nuclear Research Institute at Rez had to rely on expensive mockups of power reactor configurations to test their fluence exposures. Computers, computer codes, and updated nuclear data would advance their technology rapidly, and they were not hesitant to admit this fact. Both eastern-bloc countries said that IBM is providing an IBM 3090 for educational purposes but research and development studies would have very limited access. They were very apologetic that their currencies were not convertible, and any exchange means that they could provide services or pay for US scientists in their respective countries, but funding for their scientists in the United States, or expenses that involved payment in dollars, must come from us.

  8. Preprocessing and Analysis of LC-MS-Based Proteomic Data.

    Science.gov (United States)

    Tsai, Tsung-Heng; Wang, Minkun; Ressom, Habtom W

    2016-01-01

    Liquid chromatography coupled with mass spectrometry (LC-MS) has been widely used for profiling protein expression levels. This chapter is focused on LC-MS data preprocessing, which is a crucial step in the analysis of LC-MS based proteomics. We provide a high-level overview, highlight associated challenges, and present a step-by-step example for analysis of data from LC-MS based untargeted proteomic study. Furthermore, key procedures and relevant issues with the subsequent analysis by multiple reaction monitoring (MRM) are discussed.

  9. An Evidence-Based Videotaped Running Biomechanics Analysis.

    Science.gov (United States)

    Souza, Richard B

    2016-02-01

    Running biomechanics play an important role in the development of injuries. Performing a running biomechanics analysis on injured runners can help to develop treatment strategies. This article provides a framework for a systematic video-based running biomechanics analysis plan based on the current evidence on running injuries, using 2-dimensional (2D) video and readily available tools. Fourteen measurements are proposed in this analysis plan from lateral and posterior video. Identifying simple 2D surrogates for 3D biomechanic variables of interest allows for widespread translation of best practices, and have the best opportunity to impact the highly prevalent problem of the injured runner.

  10. A Contrastive Analysis on Web-based Intercultural Peer Feedback

    Directory of Open Access Journals (Sweden)

    Qin Wang

    2013-11-01

    Full Text Available This paper made a contrastive analysis on peer feedback generated by Chinese EFL learners and Native Speakers of English, who participated in a web-based Cross-Pacific Writing Exchange program. The analysis mainly focused on differences in terms of commenting size, nature and function; the pragmatic differences between two groups of learners were investigated as well. The present study afforded us lessons for peer review training program and provided pedagogical implications for L2 writing. Keywords: peer feedback; web-based; intercultural; contrastive analysis

  11. Reliability-Based Robustness Analysis for a Croatian Sports Hall

    DEFF Research Database (Denmark)

    Čizmar, Dean; Kirkegaard, Poul Henning; Sørensen, John Dalsgaard;

    2011-01-01

    This paper presents a probabilistic approach for structural robustness assessment for a timber structure built a few years ago. The robustness analysis is based on a structural reliability based framework for robustness and a simplified mechanical system modelling of a timber truss system. A comp...

  12. Differential Regulatory Analysis Based on Coexpression Network in Cancer Research

    Directory of Open Access Journals (Sweden)

    Junyi Li

    2016-01-01

    Full Text Available With rapid development of high-throughput techniques and accumulation of big transcriptomic data, plenty of computational methods and algorithms such as differential analysis and network analysis have been proposed to explore genome-wide gene expression characteristics. These efforts are aiming to transform underlying genomic information into valuable knowledges in biological and medical research fields. Recently, tremendous integrative research methods are dedicated to interpret the development and progress of neoplastic diseases, whereas differential regulatory analysis (DRA based on gene coexpression network (GCN increasingly plays a robust complement to regular differential expression analysis in revealing regulatory functions of cancer related genes such as evading growth suppressors and resisting cell death. Differential regulatory analysis based on GCN is prospective and shows its essential role in discovering the system properties of carcinogenesis features. Here we briefly review the paradigm of differential regulatory analysis based on GCN. We also focus on the applications of differential regulatory analysis based on GCN in cancer research and point out that DRA is necessary and extraordinary to reveal underlying molecular mechanism in large-scale carcinogenesis studies.

  13. Vehicle passes detector based on multi-sensor analysis

    Science.gov (United States)

    Bocharov, D.; Sidorchuk, D.; Konovalenko, I.; Koptelov, I.

    2015-02-01

    The study concerned deals with a new approach to the problem of detecting vehicle passes in vision-based automatic vehicle classification system. Essential non-affinity image variations and signals from induction loop are the events that can be considered as detectors of an object presence. We propose several vehicle detection techniques based on image processing and induction loop signal analysis. Also we suggest a combined method based on multi-sensor analysis to improve vehicle detection performance. Experimental results in complex outdoor environments show that the proposed multi-sensor algorithm is effective for vehicles detection.

  14. Online Fault Diagnosis Method Based on Nonlinear Spectral Analysis

    Institute of Scientific and Technical Information of China (English)

    WEI Rui-xuan; WU Li-xun; WANG Yong-chang; HAN Chong-zhao

    2005-01-01

    The fault diagnosis based on nonlinear spectral analysis is a new technique for the nonlinear fault diagnosis, but its online application could be limited because of the enormous compution requirements for the estimation of general frequency response functions. Based on the fully decoupled Volterra identification algorithm, a new online fault diagnosis method based on nonlinear spectral analysis is presented, which can availably reduce the online compution requirements of general frequency response functions. The composition and working principle of the method are described, the test experiments have been done for damping spring of a vehicle suspension system by utilizing the new method, and the results indicate that the method is efficient.

  15. Kernel-Based Nonlinear Discriminant Analysis for Face Recognition

    Institute of Scientific and Technical Information of China (English)

    LIU QingShan (刘青山); HUANG Rui (黄锐); LU HanQing (卢汉清); MA SongDe (马颂德)

    2003-01-01

    Linear subspace analysis methods have been successfully applied to extract features for face recognition. But they are inadequate to represent the complex and nonlinear variations of real face images, such as illumination, facial expression and pose variations, because of their linear properties. In this paper, a nonlinear subspace analysis method, Kernel-based Nonlinear Discriminant Analysis (KNDA), is presented for face recognition, which combines the nonlinear kernel trick with the linear subspace analysis method - Fisher Linear Discriminant Analysis (FLDA).First, the kernel trick is used to project the input data into an implicit feature space, then FLDA is performed in this feature space. Thus nonlinear discriminant features of the input data are yielded. In addition, in order to reduce the computational complexity, a geometry-based feature vectors selection scheme is adopted. Another similar nonlinear subspace analysis is Kernel-based Principal Component Analysis (KPCA), which combines the kernel trick with linear Principal Component Analysis (PCA). Experiments are performed with the polynomial kernel, and KNDA is compared with KPCA and FLDA. Extensive experimental results show that KNDA can give a higher recognition rate than KPCA and FLDA.

  16. Spectral Efficiency Analysis for Multicarrier Based 4G Systems

    DEFF Research Database (Denmark)

    Silva, Nuno; Rahman, Muhammad Imadur; Frederiksen, Flemming Bjerge;

    2006-01-01

    In this paper, a spectral efficiency definition is proposed. Spectral efficiency for multicarrier based multiaccess techniques, such as OFDMA, MC-CDMA and OFDMA-CDM, is analyzed. Simulations for different indoor and outdoor scenarios are carried out. Based on the simulations, we have discussed ho...... different wireless channel’s condition affects the performance of a system in terms of spectral efficiency. Based on our analysis, we have also recommended different access techniques for different scenarios....

  17. Earthquake Analysis of Structure by Base Isolation Technique in SAP

    OpenAIRE

    T. Subramani; J. Jothi

    2014-01-01

    This paper presents an overview of the present state of base isolation techniques with special emphasis and a brief on other techniques developed world over for mitigating earthquake forces on the structures. The dynamic analysis procedure for isolated structures is briefly explained. The provisions of FEMA 450 for base isolated structures are highlighted. The effects of base isolation on structures located on soft soils and near active faults are given in brief. Simple case s...

  18. Abnormal traffic flow data detection based on wavelet analysis

    Directory of Open Access Journals (Sweden)

    Xiao Qian

    2016-01-01

    Full Text Available In view of the traffic flow data of non-stationary, the abnormal data detection is difficult.proposed basing on the wavelet analysis and least squares method of abnormal traffic flow data detection in this paper.First using wavelet analysis to make the traffic flow data of high frequency and low frequency component and separation, and then, combined with least square method to find abnormal points in the reconstructed signal data.Wavelet analysis and least square method, the simulation results show that using wavelet analysis of abnormal traffic flow data detection, effectively reduce the detection results of misjudgment rate and false negative rate.

  19. Adaptive Fourier Decomposition Based Time-Frequency Analysis

    Institute of Scientific and Technical Information of China (English)

    Li-Ming Zhang

    2014-01-01

    The attempt to represent a signal simultaneously in time and frequency domains is full of challenges. The recently proposed adaptive Fourier decomposition (AFD) offers a practical approach to solve this problem. This paper presents the principles of the AFD based time-frequency analysis in three aspects: instantaneous frequency analysis, frequency spectrum analysis, and the spectrogram analysis. An experiment is conducted and compared with the Fourier transform in convergence rate and short-time Fourier transform in time-frequency distribution. The proposed approach performs better than both the Fourier transform and short-time Fourier transform.

  20. Noise analysis for sensitivity-based structural damage detection

    Institute of Scientific and Technical Information of China (English)

    YIN Tao; ZHU Hong-ping; YU Ling

    2007-01-01

    As vibration-based structural damage detection methods are easily affected by environmental noise, a new statistic-based noise analysis method is proposed together with the Monte Carlo technique to investigate the influence of experimental noise of modal data on sensitivity-based damage detection methods. Different from the commonly used random perturbation technique, the proposed technique is deduced directly by Moore-Penrose generalized inverse of the sensitivity matrix, which does not only make the analysis process more efficient but also can analyze the influence of noise on both frequencies and mode shapes for three commonly used sensitivity-based damage detection methods in a similar way. A one-story portal frame is adopted to evaluate the efficiency of the proposed noise analysis technique.

  1. Analysis of security protocols based on challenge-response

    Institute of Scientific and Technical Information of China (English)

    LUO JunZhou; YANG Ming

    2007-01-01

    Security protocol is specified as the procedure of challenge-response, which uses applied cryptography to confirm the existence of other principals and fulfill some data negotiation such as session keys. Most of the existing analysis methods,which either adopt theorem proving techniques such as state exploration or logic reasoning techniques such as authentication logic, face the conflicts between analysis power and operability. To solve the problem, a new efficient method is proposed that provides SSM semantics-based definition of secrecy and authentication goals and applies authentication logic as fundamental analysis techniques,in which secrecy analysis is split into two parts: Explicit-Information-Leakage and Implicit-Information-Leakage, and correspondence analysis is concluded as the analysis of the existence relationship of Strands and the agreement of Strand parameters. This new method owns both the power of the Strand Space Model and concision of authentication logic.

  2. Static Analysis for Event-Based XML Processing

    DEFF Research Database (Denmark)

    Møller, Anders

    2008-01-01

    Event-based processing of XML data - as exemplified by the popular SAX framework - is a powerful alternative to using W3C's DOM or similar tree-based APIs. The event-based approach is a streaming fashion with minimal memory consumption. This paper discusses challenges for creating program analyses...... for SAX applications. In particular, we consider the problem of statically guaranteeing the a given SAX program always produces only well-formed and valid XML output. We propose an analysis technique based on ecisting anglyses of Servlets, string operations, and XML graphs....

  3. A Framework for Web-Based Mechanical Design and Analysis

    Institute of Scientific and Technical Information of China (English)

    Chiaming; Yen; Wujeng; Li

    2002-01-01

    In this paper, a Web-based Mechanical Design and A na lysis Framework (WMDAF) is proposed. This WMADF allows designers to develop web -based computer aided programs in a systematic way during the collaborative mec hanical system design and analysis process. This system is based on an emerg ing web-based Content Management System (CMS) called eXtended Object Oriented P ortal System (XOOPS). Due to the Open Source Status of the XOOPS CMS, programs d eveloped with this framework can be further customized to ...

  4. Tikhonov regularization-based operational transfer path analysis

    Science.gov (United States)

    Cheng, Wei; Lu, Yingying; Zhang, Zhousuo

    2016-06-01

    To overcome ill-posed problems in operational transfer path analysis (OTPA), and improve the stability of solutions, this paper proposes a novel OTPA based on Tikhonov regularization, which considers both fitting degrees and stability of solutions. Firstly, fundamental theory of Tikhonov regularization-based OTPA is presented, and comparative studies are provided to validate the effectiveness on ill-posed problems. Secondly, transfer path analysis and source contribution evaluations for numerical cases studies on spherical radiating acoustical sources are comparatively studied. Finally, transfer path analysis and source contribution evaluations for experimental case studies on a test bed with thin shell structures are provided. This study provides more accurate transfer path analysis for mechanical systems, which can benefit for vibration reduction by structural path optimization. Furthermore, with accurate evaluation of source contributions, vibration monitoring and control by active controlling vibration sources can be effectively carried out.

  5. Matrix-based introduction to multivariate data analysis

    CERN Document Server

    Adachi, Kohei

    2016-01-01

    This book enables readers who may not be familiar with matrices to understand a variety of multivariate analysis procedures in matrix forms. Another feature of the book is that it emphasizes what model underlies a procedure and what objective function is optimized for fitting the model to data. The author believes that the matrix-based learning of such models and objective functions is the fastest way to comprehend multivariate data analysis. The text is arranged so that readers can intuitively capture the purposes for which multivariate analysis procedures are utilized: plain explanations of the purposes with numerical examples precede mathematical descriptions in almost every chapter. This volume is appropriate for undergraduate students who already have studied introductory statistics. Graduate students and researchers who are not familiar with matrix-intensive formulations of multivariate data analysis will also find the book useful, as it is based on modern matrix formulations with a special emphasis on ...

  6. Data Warehouse Requirements Analysis Framework: Business-Object Based Approach

    Directory of Open Access Journals (Sweden)

    Anirban Sarkar

    2012-01-01

    Full Text Available Detailed requirements analysis plays a key role towards the design of successful Data Warehouse (DW system. The requirements analysis specifications are used as the prime input for the construction of conceptual level multidimensional data model. This paper has proposed a Business Object based requirements analysis framework for DW system which is supported with abstraction mechanism and reuse capability. It also facilitate the stepwise mapping of requirements descriptions into high level design components of graph semantic based conceptual level object oriented multidimensional data model. The proposed framework starts with the identification of the analytical requirements using business process driven approach and finally refine the requirements in further detail to map into the conceptual level DW design model using either Demand-driven of Mixed-driven approach for DW requirements analysi

  7. NONLINEAR DATA RECONCILIATION METHOD BASED ON KERNEL PRINCIPAL COMPONENT ANALYSIS

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    In the industrial process situation, principal component analysis (PCA) is a general method in data reconciliation.However, PCA sometime is unfeasible to nonlinear feature analysis and limited in application to nonlinear industrial process.Kernel PCA (KPCA) is extension of PCA and can be used for nonlinear feature analysis.A nonlinear data reconciliation method based on KPCA is proposed.The basic idea of this method is that firstly original data are mapped to high dimensional feature space by nonlinear function, and PCA is implemented in the feature space.Then nonlinear feature analysis is implemented and data are reconstructed by using the kernel.The data reconciliation method based on KPCA is applied to ternary distillation column.Simulation results show that this method can filter the noise in measurements of nonlinear process and reconciliated data can represent the true information of nonlinear process.

  8. Basic gait analysis based on continuous wave radar.

    Science.gov (United States)

    Zhang, Jun

    2012-09-01

    A gait analysis method based on continuous wave (CW) radar is proposed in this paper. Time-frequency analysis is used to analyze the radar micro-Doppler echo from walking humans, and the relationships between the time-frequency spectrogram and human biological gait are discussed. The methods for extracting the gait parameters from the spectrogram are studied in depth and experiments on more than twenty subjects have been performed to acquire the radar gait data. The gait parameters are calculated and compared. The gait difference between men and women are presented based on the experimental data and extracted features. Gait analysis based on CW radar will provide a new method for clinical diagnosis and therapy.

  9. Intelligent Hybrid Cluster Based Classification Algorithm for Social Network Analysis

    Directory of Open Access Journals (Sweden)

    S. Muthurajkumar

    2014-05-01

    Full Text Available In this paper, we propose an hybrid clustering based classification algorithm based on mean approach to effectively classify to mine the ordered sequences (paths from weblog data in order to perform social network analysis. In the system proposed in this work for social pattern analysis, the sequences of human activities are typically analyzed by switching behaviors, which are likely to produce overlapping clusters. In this proposed system, a robust Modified Boosting algorithm is proposed to hybrid clustering based classification for clustering the data. This work is useful to provide connection between the aggregated features from the network data and traditional indices used in social network analysis. Experimental results show that the proposed algorithm improves the decision results from data clustering when combined with the proposed classification algorithm and hence it is proved that of provides better classification accuracy when tested with Weblog dataset. In addition, this algorithm improves the predictive performance especially for multiclass datasets which can increases the accuracy.

  10. Study of engine noise based on independent component analysis

    Institute of Scientific and Technical Information of China (English)

    HAO Zhi-yong; JIN Yan; YANG Chen

    2007-01-01

    Independent component analysis was applied to analyze the acoustic signals from diesel engine. First the basic principle of independent component analysis (ICA) was reviewed. Diesel engine acoustic signal was decomposed into several independent components (Ics); Fourier transform and continuous wavelet transform (CWT) were applied to analyze the independent components. Different noise sources of the diesel engine were separated, based on the characteristics of different component in time-frequency domain.

  11. 13C-based metabolic flux analysis: fundamentals and practice.

    Science.gov (United States)

    Yang, Tae Hoon

    2013-01-01

    Isotope-based metabolic flux analysis is one of the emerging technologies applied to system level metabolic phenotype characterization in metabolic engineering. Among the developed approaches, (13)C-based metabolic flux analysis has been established as a standard tool and has been widely applied to quantitative pathway characterization of diverse biological systems. To implement (13)C-based metabolic flux analysis in practice, comprehending the underlying mathematical and computational modeling fundamentals is of importance along with carefully conducted experiments and analytical measurements. Such knowledge is also crucial when designing (13)C-labeling experiments and properly acquiring key data sets essential for in vivo flux analysis implementation. In this regard, the modeling fundamentals of (13)C-labeling systems and analytical data processing are the main topics we will deal with in this chapter. Along with this, the relevant numerical optimization techniques are addressed to help implementation of the entire computational procedures aiming at (13)C-based metabolic flux analysis in vivo.

  12. Rare variant detection using family-based sequencing analysis.

    Science.gov (United States)

    Peng, Gang; Fan, Yu; Palculict, Timothy B; Shen, Peidong; Ruteshouser, E Cristy; Chi, Aung-Kyaw; Davis, Ronald W; Huff, Vicki; Scharfe, Curt; Wang, Wenyi

    2013-03-05

    Next-generation sequencing is revolutionizing genomic analysis, but this analysis can be compromised by high rates of missing true variants. To develop a robust statistical method capable of identifying variants that would otherwise not be called, we conducted sequence data simulations and both whole-genome and targeted sequencing data analysis of 28 families. Our method (Family-Based Sequencing Program, FamSeq) integrates Mendelian transmission information and raw sequencing reads. Sequence analysis using FamSeq reduced the number of false negative variants by 14-33% as assessed by HapMap sample genotype confirmation. In a large family affected with Wilms tumor, 84% of variants uniquely identified by FamSeq were confirmed by Sanger sequencing. In children with early-onset neurodevelopmental disorders from 26 families, de novo variant calls in disease candidate genes were corrected by FamSeq as mendelian variants, and the number of uniquely identified variants in affected individuals increased proportionally as additional family members were included in the analysis. To gain insight into maximizing variant detection, we studied factors impacting actual improvements of family-based calling, including pedigree structure, allele frequency (common vs. rare variants), prior settings of minor allele frequency, sequence signal-to-noise ratio, and coverage depth (∼20× to >200×). These data will help guide the design, analysis, and interpretation of family-based sequencing studies to improve the ability to identify new disease-associated genes.

  13. Using the DOE Knowledge Base for Special Event Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Armstrong, H.M.; Harris, J.M.; Young, C.J.

    1998-10-20

    The DOE Knowledge Base is a library of detailed information whose purpose is to support the United States National Data Center (USNDC) in its mission to monitor compliance with the Comprehensive Test Ban Treaty (CTBT). One of the important tasks which the USNDC must accomplish is to periodically perform detailed analysis of events of high interest, so-called "Special Events", to provide the national authority with information needed to make policy decisions. In this paper we investigate some possible uses of the Knowledge Base for Special Event Analysis (SEA), and make recommendations for improving Knowledge Base support for SEA. To analyze an event in detail, there are two basic types of data which must be used sensor-derived data (wave- forms, arrivals, events, etc.) and regiohalized contextual data (known sources, geological characteristics, etc.). Cur- rently there is no single package which can provide full access to both types of data, so for our study we use a separate package for each MatSeis, the Sandia Labs-developed MATLAB-based seismic analysis package, for wave- form data analysis, and ArcView, an ESRI product, for contextual data analysis. Both packages are well-suited to pro- totyping because they provide a rich set of currently available functionality and yet are also flexible and easily extensible, . Using these tools and Phase I Knowledge Base data sets, we show how the Knowledge Base can improve both the speed and the quality of SEA. Empirically-derived interpolated correction information can be accessed to improve both location estimates and associated error estimates. This information can in turn be used to identi~ any known nearby sources (e.g. mines, volcanos), which may then trigger specialized processing of the sensor data. Based on the location estimate, preferred magnitude formulas and discriminants can be retrieved, and any known blockages can be identified to prevent miscalculations. Relevant historic events can be identilled either by

  14. Segmentation of Stick Text Based on Sub Connected Area Analysis

    Institute of Scientific and Technical Information of China (English)

    高静波; 李新友; 等

    1998-01-01

    A new stick text segmentation method based on the sub connected area analysis is introduced in this paper.The foundation of this method is the sub connected area representation of text image that can represent all connected areas in an image efficiently.This method consists mainly of four steps:sub connected area classification,finding initial boundary following point,finding optimal segmentation point by boundary tracing,and text segmentaton.This method is similar to boundary analysis method but is more efficient than boundary analysis.

  15. AN HMM BASED ANALYSIS FRAMEWORK FOR SEMANTIC VIDEO EVENTS

    Institute of Scientific and Technical Information of China (English)

    You Junyong; Liu Guizhong; Zhang Yaxin

    2007-01-01

    Semantic video analysis plays an important role in the field of machine intelligence and pattern recognition. In this paper, based on the Hidden Markov Model (HMM), a semantic recognition framework on compressed videos is proposed to analyze the video events according to six low-level features. After the detailed analysis of video events, the pattern of global motion and five features in foreground--the principal parts of videos, are employed as the observations of the Hidden Markov Model to classify events in videos. The applications of the proposed framework in some video event detections demonstrate the promising success of the proposed framework on semantic video analysis.

  16. RULE-BASED SENTIMENT ANALYSIS OF UKRAINIAN REVIEWS

    Directory of Open Access Journals (Sweden)

    Mariana Romanyshyn

    2013-07-01

    Full Text Available Last decade witnessed a lot of research in the field of sentiment analysis. Understanding the attitude and the emotions that people express in written text proved to be really important and helpful in sociology, political science, psychology, market research, and, of course, artificial intelligence. This paper demonstrates a rule-based approach to clause-level sentiment analysis of reviews in Ukrainian. The general architecture of the implemented sentiment analysis system is presented, the current stage of research is described and further work is explained. The main emphasis is made on the design of rules for computing sentiments.

  17. Stability analysis of underground engineering based on multidisciplinary design optimization

    Institute of Scientific and Technical Information of China (English)

    MA Rong; ZHOU Ke-ping; GAO Feng

    2008-01-01

    Aiming at characteristics of underground engineering,analyzed the feasibility of Multidisciplinary Design Optimization (MDO) used in underground engineering,and put forward a modularization-based MDO method and the idea of MDO to resolve problems in stability analysis,proving the validity and feasibility of using MDO in underground engineering.Characteristics of uncertainty,complexity and nonlinear become bottle-neck to carry on underground engineering stability analysis by MDO.Therefore,the application of MDO in underground engineering stability analysis is still at a stage of exploration,which need some deep research.

  18. Stability analysis of underground engineering based on multidisciplinary design optimization

    Institute of Scientific and Technical Information of China (English)

    MA Rong; ZHOU Ke-ping; GAO Feng

    2008-01-01

    Aiming at characteristics of underground engineering, analyzed the feasibility of Multidisciplinary Design Optimization (MDO) used in underground engineering, and put forward a modularization-based MDO method and the idea of MDO to resolve problems in stability analysis, proving the validity and feasibility of using MDO in underground engi-neering. Characteristics of uncertainty, complexity and nonlinear become bottle-neck to carry on underground engineering stability analysis by MDO. Therefore, the application of MDO in underground engineering stability analysis is still at a stage of exploration, which need some deep research.

  19. Open access for ALICE analysis based on virtualization technology

    CERN Document Server

    Buncic, P; Schutz, Y

    2015-01-01

    Open access is one of the important leverages for long-term data preservation for a HEP experiment. To guarantee the usability of data analysis tools beyond the experiment lifetime it is crucial that third party users from the scientific community have access to the data and associated software. The ALICE Collaboration has developed a layer of lightweight components built on top of virtualization technology to hide the complexity and details of the experiment-specific software. Users can perform basic analysis tasks within CernVM, a lightweight generic virtual machine, paired with an ALICE specific contextualization. Once the virtual machine is launched, a graphical user interface is automatically started without any additional configuration. This interface allows downloading the base ALICE analysis software and running a set of ALICE analysis modules. Currently the available tools include fully documented tutorials for ALICE analysis, such as the measurement of strange particle production or the nuclear modi...

  20. Research of second harmonic generation images based on texture analysis

    Science.gov (United States)

    Liu, Yao; Li, Yan; Gong, Haiming; Zhu, Xiaoqin; Huang, Zufang; Chen, Guannan

    2014-09-01

    Texture analysis plays a crucial role in identifying objects or regions of interest in an image. It has been applied to a variety of medical image processing, ranging from the detection of disease and the segmentation of specific anatomical structures, to differentiation between healthy and pathological tissues. Second harmonic generation (SHG) microscopy as a potential noninvasive tool for imaging biological tissues has been widely used in medicine, with reduced phototoxicity and photobleaching. In this paper, we clarified the principles of texture analysis including statistical, transform, structural and model-based methods and gave examples of its applications, reviewing studies of the technique. Moreover, we tried to apply texture analysis to the SHG images for the differentiation of human skin scar tissues. Texture analysis method based on local binary pattern (LBP) and wavelet transform was used to extract texture features of SHG images from collagen in normal and abnormal scars, and then the scar SHG images were classified into normal or abnormal ones. Compared with other texture analysis methods with respect to the receiver operating characteristic analysis, LBP combined with wavelet transform was demonstrated to achieve higher accuracy. It can provide a new way for clinical diagnosis of scar types. At last, future development of texture analysis in SHG images were discussed.

  1. Comparative proteomic and phosphoproteomic profiling of pancreatic adenocarcinoma cells treated with CB1 or CB2 agonists.

    Science.gov (United States)

    Brandi, Jessica; Dando, Ilaria; Palmieri, Marta; Donadelli, Massimo; Cecconi, Daniela

    2013-05-01

    The pancreatic adenocarcinoma cell line Panc1 was treated with cannabinoid receptor ligands (arachidonylcyclopropylamide or GW405833) in order to elucidate the molecular mechanism of their anticancer effect. A proteomic approach was used to analyze the protein and phosphoprotein profiles. Western blot and functional data mining were also employed in order to validate results, classify proteins, and explore their potential relationships. We demonstrated that the two cannabinoids act through a widely common mechanism involving up- and down-regulation of proteins related to energetic metabolism and cell growth regulation. Overall, the results reported might contribute to the development of a therapy based on cannabinoids for pancreatic adenocarcinoma.

  2. Modeling and Grid impedance Variation Analysis of Parallel Connected Grid Connected Inverter based on Impedance Based Harmonic Analysis

    DEFF Research Database (Denmark)

    Kwon, JunBum; Wang, Xiongfei; Bak, Claus Leth;

    2014-01-01

    This paper addresses the harmonic compensation error problem existing with parallel connected inverter in the same grid interface conditions by means of impedance-based analysis and modeling. Unlike the single grid connected inverter, it is found that multiple parallel connected inverters and grid...... impedance can make influence to each other if they each have a harmonic compensation function. The analysis method proposed in this paper is based on the relationship between the overall output impedance and input impedance of parallel connected inverter, where controller gain design method, which can...

  3. A Genre-based Analysis of English Refusal Letters

    Institute of Scientific and Technical Information of China (English)

    费伟

    2014-01-01

    The present study analyzes ten English refusal letters of two subcategories based on Swales ’genre analysis model and finds that difference exists in the generic features of the two subcategorized types. Teachers should reveal the underlying rationale behind linguistic features of a specific genre so that students can not only identify genre but also apply it appropriately.

  4. Linear feature selection in texture analysis - A PLS based method

    DEFF Research Database (Denmark)

    Marques, Joselene; Igel, Christian; Lillholm, Martin

    2013-01-01

    We present a texture analysis methodology that combined uncommitted machine-learning techniques and partial least square (PLS) in a fully automatic framework. Our approach introduces a robust PLS-based dimensionality reduction (DR) step to specifically address outliers and high-dimensional featur...

  5. Digital Simulation-Based Training: A Meta-Analysis

    Science.gov (United States)

    Gegenfurtner, Andreas; Quesada-Pallarès, Carla; Knogler, Maximilian

    2014-01-01

    This study examines how design characteristics in digital simulation-based learning environments moderate self-efficacy and transfer of learning. Drawing on social cognitive theory and the cognitive theory of multimedia learning, the meta-analysis psychometrically cumulated k?=?15 studies of 25 years of research with a total sample size of…

  6. Graph- versus Vector-Based Analysis of a Consensus Protocol

    NARCIS (Netherlands)

    Delzanno, Giorgio; Rensink, Arend; Traverso, Riccardo; Bošnački, Dragan; Edelkamp, Stefan; Lluch Lafuente, Alberto; Wijs, Anton

    2014-01-01

    The Paxos distributed consensus algorithm is a challenging case-study for standard, vector-based model checking techniques. Due to asynchronous communication, exhaustive analysis may generate very large state spaces already for small model instances. In this paper, we show the advantages of graph tr

  7. A Corpus-based Analysis of English Noun Suffixes

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    This paper provides a brief analysis of English suffixes. First, make a classification of the English noun suffixes etymologically; then, obtain the frequencies of each English noun suffixes in sub-corpus FR88 and WSJ88, and last draw a conclusion based on the statistics. That is from the word origins we can see its influences on English vocabulary.

  8. Spinoza II: Conceptual Case-Based Natural Language Analysis.

    Science.gov (United States)

    Schank, Roger C.; And Others

    This paper presents the theoretical changes that have developed in Conceptual Dependency Theory and their ramifications in computer analysis of natural language. The major items of concern are: the elimination of reliance on "grammar rules" for parsing with the emphasis given to conceptual rule based parsing; the development of a conceptual case…

  9. Computer-Based Interaction Analysis with DEGREE Revisited

    Science.gov (United States)

    Barros, B.; Verdejo, M. F.

    2016-01-01

    We review our research with "DEGREE" and analyse how our work has impacted the collaborative learning community since 2000. Our research is framed within the context of computer-based interaction analysis and the development of computer-supported collaborative learning (CSCL) tools. We identify some aspects of our work which have been…

  10. LES based POD analysis of Jet in Cross Flow

    DEFF Research Database (Denmark)

    Cavar, Dalibor; Meyer, Knud Erik; Jakirlic, S.

    2010-01-01

    The paper presents results of a POD investigation of the LES based numerical simulation of the jet-in-crossflow (JICF) flowfield. LES results are firstly compared to the pointwise LDA measurements. 2D POD analysis is then used as a comparison basis for PIV measurements and LES, and finally 3D POD...

  11. Project-Based Language Learning: An Activity Theory Analysis

    Science.gov (United States)

    Gibbes, Marina; Carson, Lorna

    2014-01-01

    This paper reports on an investigation of project-based language learning (PBLL) in a university language programme. Learner reflections of project work were analysed through Activity Theory, where tool-mediated activity is understood as the central unit of analysis for human interaction. Data were categorised according to the components of human…

  12. Advancing School-Based Interventions through Economic Analysis

    Science.gov (United States)

    Olsson, Tina M.; Ferrer-Wreder, Laura; Eninger, Lilianne

    2014-01-01

    Commentators interested in school-based prevention programs point to the importance of economic issues for the future of prevention efforts. Many of the processes and aims of prevention science are dependent upon prevention resources. Although economic analysis is an essential tool for assessing resource use, the attention given economic analysis…

  13. Situational Analysis: A Framework for Evidence-Based Practice

    Science.gov (United States)

    Annan, Jean

    2005-01-01

    Situational analysis is a framework for professional practice and research in educational psychology. The process is guided by a set of practice principles requiring that psychologists' work is evidence-based, ecological, collaborative and constructive. The framework is designed to provide direction for psychologists who wish to tailor their…

  14. Utilizing Problem-Based Learning in Qualitative Analysis Lab Experiments

    Science.gov (United States)

    Hicks, Randall W.; Bevsek, Holly M.

    2012-01-01

    A series of qualitative analysis (QA) laboratory experiments utilizing a problem-based learning (PBL) module has been designed and implemented. The module guided students through the experiments under the guise of cleaning up a potentially contaminated water site as employees of an environmental chemistry laboratory. The main goal was the…

  15. System of gait analysis based on ground reaction force assessment

    Directory of Open Access Journals (Sweden)

    František Vaverka

    2015-12-01

    Full Text Available Background: Biomechanical analysis of gait employs various methods used in kinematic and kinetic analysis, EMG, and others. One of the most frequently used methods is kinetic analysis based on the assessment of the ground reaction forces (GRF recorded on two force plates. Objective: The aim of the study was to present a method of gait analysis based on the assessment of the GRF recorded during the stance phase of two steps. Methods: The GRF recorded with a force plate on one leg during stance phase has three components acting in directions: Fx - mediolateral, Fy - anteroposterior, and Fz - vertical. A custom-written MATLAB script was used for gait analysis in this study. This software displays instantaneous force data for both legs as Fx(t, Fy(t and Fz(t curves, automatically determines the extremes of functions and sets the visual markers defining the individual points of interest. Positions of these markers can be easily adjusted by the rater, which may be necessary if the GRF has an atypical pattern. The analysis is fully automated and analyzing one trial takes only 1-2 minutes. Results: The method allows quantification of temporal variables of the extremes of the Fx(t, Fy(t, Fz(t functions, durations of the braking and propulsive phase, duration of the double support phase, the magnitudes of reaction forces in extremes of measured functions, impulses of force, and indices of symmetry. The analysis results in a standardized set of 78 variables (temporal, force, indices of symmetry which can serve as a basis for further research and diagnostics. Conclusions: The resulting set of variable offers a wide choice for selecting a specific group of variables with consideration to a particular research topic. The advantage of this method is the standardization of the GRF analysis, low time requirements allowing rapid analysis of a large number of trials in a short time, and comparability of the variables obtained during different research measurements.

  16. Geographical classification of Epimedium based on HPLC fingerprint analysis combined with multi-ingredients quantitative analysis.

    Science.gov (United States)

    Xu, Ning; Zhou, Guofu; Li, Xiaojuan; Lu, Heng; Meng, Fanyun; Zhai, Huaqiang

    2017-05-01

    A reliable and comprehensive method for identifying the origin and assessing the quality of Epimedium has been developed. The method is based on analysis of HPLC fingerprints, combined with similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA) and multi-ingredient quantitative analysis. Nineteen batches of Epimedium, collected from different areas in the western regions of China, were used to establish the fingerprints and 18 peaks were selected for the analysis. Similarity analysis, HCA and PCA all classified the 19 areas into three groups. Simultaneous quantification of the five major bioactive ingredients in the Epimedium samples was also carried out to confirm the consistency of the quality tests. These methods were successfully used to identify the geographical origin of the Epimedium samples and to evaluate their quality.

  17. Teaching-Learning Activity Modeling Based on Data Analysis

    Directory of Open Access Journals (Sweden)

    Kyungrog Kim

    2015-03-01

    Full Text Available Numerous studies are currently being carried out on personalized services based on data analysis to find and provide valuable information about information overload. Furthermore, the number of studies on data analysis of teaching-learning activities for personalized services in the field of teaching-learning is increasing, too. This paper proposes a learning style recency-frequency-durability (LS-RFD model for quantified analysis on the level of activities of learners, to provide the elements of teaching-learning activities according to the learning style of the learner among various parameters for personalized service. This is to measure preferences as to teaching-learning activity according to recency, frequency and durability of such activities. Based on the results, user characteristics can be classified into groups for teaching-learning activity by categorizing the level of preference and activity of the learner.

  18. Control volume based hydrocephalus research; analysis of human data

    Science.gov (United States)

    Cohen, Benjamin; Wei, Timothy; Voorhees, Abram; Madsen, Joseph; Anor, Tomer

    2010-11-01

    Hydrocephalus is a neuropathophysiological disorder primarily diagnosed by increased cerebrospinal fluid volume and pressure within the brain. To date, utilization of clinical measurements have been limited to understanding of the relative amplitude and timing of flow, volume and pressure waveforms; qualitative approaches without a clear framework for meaningful quantitative comparison. Pressure volume models and electric circuit analogs enforce volume conservation principles in terms of pressure. Control volume analysis, through the integral mass and momentum conservation equations, ensures that pressure and volume are accounted for using first principles fluid physics. This approach is able to directly incorporate the diverse measurements obtained by clinicians into a simple, direct and robust mechanics based framework. Clinical data obtained for analysis are discussed along with data processing techniques used to extract terms in the conservation equation. Control volume analysis provides a non-invasive, physics-based approach to extracting pressure information from magnetic resonance velocity data that cannot be measured directly by pressure instrumentation.

  19. Flow cytometry-based DNA hybridization and polymorphism analysis

    Energy Technology Data Exchange (ETDEWEB)

    Cai, H.; Kommander, K.; White, P.S.; Nolan, J.P.

    1998-07-01

    Functional analysis of the humane genome, including the quantification of differential gene expression and the identification of polymorphic sites and disease genes, is an important element of the Human Genome Project. Current methods of analysis are mainly gel-based assays that are not well-suited to rapid genome-scale analyses. To analyze DNA sequence on a large scale, robust and high throughput assays are needed. The authors are developing a suite of microsphere-based approaches employing fluorescence detection to screen and analyze genomic sequence. The approaches include competitive DNA hybridization to measure DNA or RNA targets in unknown samples, and oligo ligation or extension assays to analyze single-nucleotide polymorphisms. Apart from the advances of sensitivity, simplicity, and low sample consumption, these flow cytometric approaches have the potential for high throughput multiplexed analysis using multicolored microspheres and automated sample handling.

  20. [Electroencephalogram Feature Selection Based on Correlation Coefficient Analysis].

    Science.gov (United States)

    Zhou, Jinzhi; Tang, Xiaofang

    2015-08-01

    In order to improve the accuracy of classification with small amount of motor imagery training data on the development of brain-computer interface (BCD systems, we proposed an analyzing method to automatically select the characteristic parameters based on correlation coefficient analysis. Throughout the five sample data of dataset IV a from 2005 BCI Competition, we utilized short-time Fourier transform (STFT) and correlation coefficient calculation to reduce the number of primitive electroencephalogram dimension, then introduced feature extraction based on common spatial pattern (CSP) and classified by linear discriminant analysis (LDA). Simulation results showed that the average rate of classification accuracy could be improved by using correlation coefficient feature selection method than those without using this algorithm. Comparing with support vector machine (SVM) optimization features algorithm, the correlation coefficient analysis can lead better selection parameters to improve the accuracy of classification.

  1. Unified HMM-based layout analysis framework and algorithm

    Institute of Scientific and Technical Information of China (English)

    陈明; 丁晓青; 吴佑寿

    2003-01-01

    To manipulate the layout analysis problem for complex or irregular document image, a Unified HMM-based Layout Analysis Framework is presented in this paper. Based on the multi-resolution wavelet analysis results of the document image, we use HMM method in both inner-scale image model and trans-scale context model to classify the pixel region properties, such as text, picture or background. In each scale, a HMM direct segmentation method is used to get better inner-scale classification result. Then another HMM method is used to fuse the inner-scale result in each scale and then get better final seg- mentation result. The optimized algorithm uses a stop rule in the coarse to fine multi-scale segmentation process, so the speed is improved remarkably. Experiments prove the efficiency of proposed algorithm.

  2. Environmental Assessment: General Plan-Based Environmental Impact Analysis Process, Laughlin Air Force Base

    Science.gov (United States)

    2007-05-01

    BASED ENVIROMENTAL IMPACT ANALYSIS PROCESS LAUGHLIN AIR FORCE BASE, TEXAS AGENCY: 47th Flying Training Wing (FTW), Laughlin Air Force Base (AFB), Texas...activities that take place in a particular area and generally refers to human modification of land, often for residential or economic purposes. It also...is asphaltic and is generally not economical to drill. There are some small natural gas deposits being tapped in the northwest part of the county

  3. ODVBA: optimally-discriminative voxel-based analysis.

    Science.gov (United States)

    Zhang, Tianhao; Davatzikos, Christos

    2011-08-01

    Gaussian smoothing of images prior to applying voxel-based statistics is an important step in voxel-based analysis and statistical parametric mapping (VBA-SPM) and is used to account for registration errors, to Gaussianize the data and to integrate imaging signals from a region around each voxel. However, it has also become a limitation of VBA-SPM based methods, since it is often chosen empirically and lacks spatial adaptivity to the shape and spatial extent of the region of interest, such as a region of atrophy or functional activity. In this paper, we propose a new framework, named optimally-discriminative voxel-based analysis (ODVBA), for determining the optimal spatially adaptive smoothing of images, followed by applying voxel-based group analysis. In ODVBA, nonnegative discriminative projection is applied regionally to get the direction that best discriminates between two groups, e.g., patients and controls; this direction is equivalent to local filtering by an optimal kernel whose coefficients define the optimally discriminative direction. By considering all the neighborhoods that contain a given voxel, we then compose this information to produce the statistic for each voxel. Finally, permutation tests are used to obtain a statistical parametric map of group differences. ODVBA has been evaluated using simulated data in which the ground truth is known and with data from an Alzheimer's disease (AD) study. The experimental results have shown that the proposed ODVBA can precisely describe the shape and location of structural abnormality.

  4. Environmental analysis of the eastern shale hydroretorting data base

    Energy Technology Data Exchange (ETDEWEB)

    Rex, R.C. Jr.; Lynch, P.A.

    1984-05-01

    The purpose of this study is to perform a preliminary environmental analysis of certain chemical aspects of Eastern shale hydroretorting utilizing the data from the twenty-one (21) bench scale unit runs conducted during the HYTORT Feasibility Study. The report contained herein primarily addresses the potential types and quantities of pollutants emanating directly from the hydroretorting of oil shale (i.e., the retort paper). The following areas are discussed in detail: nitrogen distribution; sulfur distribution; gas trace constituents; sour water constituents; and shale leachates. The results of the analysis have not identified any potential pollutants or quantities which cannot be brought to conformance with currently promulgated environmental standards using existing technology. Additional analysis of the process chemistry portion of the HYTORT data base, coupled with the process and mechanical design information, can provide a methodology for dealing with the identified environmental concerns as they pertain to a commercial facility. Section 5.0 of the report delineates the areas which should be addressed in a continuing analysis of environmental concerns. The suggested program divides naturally into three phases, of which Phase 1 has been completed: Phase 1 - Environmental Analysis of the Eastern Shale Hydroretorting Data Base; Phase 2 - Generic (non-site-specific) Environmental Analysis; and Phase 3 - Site-Specific Environmental Analysis. Phase 2 details the anticipated emissions from all areas of a commercial HYTORT facility operating on a typical Eastern shale using the results of this Phase 1 effort and the HYTORT data base. Phase 3 utilizes this information to assess the effects of plant emissions on chosen sites in the context of applicable laws and regulations. 7 references, 18 figures, 2 tables.

  5. Content-based analysis and indexing of sports video

    Science.gov (United States)

    Luo, Ming; Bai, Xuesheng; Xu, Guang-you

    2001-12-01

    An explosion of on-line image and video data in digital form is already well underway. With the exponential rise in interactive information exploration and dissemination through the World-Wide Web, the major inhibitors of rapid access to on-line video data are the management of capture and storage, and content-based intelligent search and indexing techniques. This paper proposes an approach for content-based analysis and event-based indexing of sports video. It includes a novel method to organize shots - classifying shots as close shots and far shots, an original idea of blur extent-based event detection, and an innovative local mutation-based algorithm for caption detection and retrieval. Results on extensive real TV programs demonstrate the applicability of our approach.

  6. Research on supplier evaluation and selection based on fuzzy hierarchy analysis and grey relational analysis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Considering the disadvantages of selecting evaluation index of supplier based on old purchase relation and in view of transformation of relation between manufacture and supplier under the dynamic, cooperative, competitive and quickly response environment, research on supplier selection evaluation was presented based on enterprise capability, cooperation degree and service level from the perspective of cooperative partnership and coordination, and the evaluation index system was established. A more objective and veracious supplier selection and evaluation method based on fuzzy analysis hierarchy process and grey relational analysis was developed, and then empirical research on electric equipment manufacturer was explored to analyze the supplier selection and evaluation.

  7. Finding Suitable Variability Abstractions for Family-Based Analysis

    DEFF Research Database (Denmark)

    Dimovski, Aleksandar; Brabrand, Claus; Wasowski, Andrzej

    2016-01-01

    For program families (Software Product Lines), specially designed variability-aware static (dataflow) analyses allow analyzing all variants (products) of the family, simultaneously, in a single run without generating any of the variants explicitly. They are also known as lifted or family......-based analyses. The variability-aware analyses may be too costly or even infeasible for families with a large number of variants. In order to make them computationally cheaper, we can apply variability abstractions which aim to tame the combinatorial explosion of the number of variants (configurations...... suitable variability abstractions from a large family of abstractions for a variability-aware static analysis. The idea is to use a pre-analysis to estimate the impact of variability-specific parts of the program family on the analysis's precision. Then we use the pre-analysis results to find out when...

  8. Incorporating Semantics into Data Driven Workflows for Content Based Analysis

    Science.gov (United States)

    Argüello, M.; Fernandez-Prieto, M. J.

    Finding meaningful associations between text elements and knowledge structures within clinical narratives in a highly verbal domain, such as psychiatry, is a challenging goal. The research presented here uses a small corpus of case histories and brings into play pre-existing knowledge, and therefore, complements other approaches that use large corpus (millions of words) and no pre-existing knowledge. The paper describes a variety of experiments for content-based analysis: Linguistic Analysis using NLP-oriented approaches, Sentiment Analysis, and Semantically Meaningful Analysis. Although it is not standard practice, the paper advocates providing automatic support to annotate the functionality as well as the data for each experiment by performing semantic annotation that uses OWL and OWL-S. Lessons learnt can be transmitted to legacy clinical databases facing the conversion of clinical narratives according to prominent Electronic Health Records standards.

  9. Design and Implementation of Hardware Based Entropy Analysis

    Directory of Open Access Journals (Sweden)

    S. Saravanan

    2012-07-01

    Full Text Available The aim of this study is hardware implementation of the Entropy analysis. Designing and verifying entropy analysis is the major finding of this study aper. Entropy tells how much amount of data can be compressed. Entropy analysis plays a major role in scan based SoC testing. Size and complexity have been the major issues for current scenario of System-on-a-Chip (SoC testing. Test data compression is a must for such cases. Entropy analysis is taken for both specified and unspecified bits (don’t care bits. Unspecified bits are specified using Zero and One fill algorithms. The X-filling technique is applied for fixed to fixed codes. The proposed method is successfully tested on ISCAS89 benchmark circuits.

  10. Physics-Based Simulator for NEO Exploration Analysis & Simulation

    Science.gov (United States)

    Balaram, J.; Cameron, J.; Jain, A.; Kline, H.; Lim, C.; Mazhar, H.; Myint, S.; Nayar, H.; Patton, R.; Pomerantz, M.; Quadrelli, M.; Shakkotai, P.; Tso, K.

    2011-01-01

    As part of the Space Exploration Analysis and Simulation (SEAS) task, the National Aeronautics and Space Administration (NASA) is using physics-based simulations at NASA's Jet Propulsion Laboratory (JPL) to explore potential surface and near-surface mission operations at Near Earth Objects (NEOs). The simulator is under development at JPL and can be used to provide detailed analysis of various surface and near-surface NEO robotic and human exploration concepts. In this paper we describe the SEAS simulator and provide examples of recent mission systems and operations concepts investigated using the simulation. We also present related analysis work and tools developed for both the SEAS task as well as general modeling, analysis and simulation capabilites for asteroid/small-body objects.

  11. Thermodynamics-based Metabolite Sensitivity Analysis in metabolic networks.

    Science.gov (United States)

    Kiparissides, A; Hatzimanikatis, V

    2017-01-01

    The increasing availability of large metabolomics datasets enhances the need for computational methodologies that can organize the data in a way that can lead to the inference of meaningful relationships. Knowledge of the metabolic state of a cell and how it responds to various stimuli and extracellular conditions can offer significant insight in the regulatory functions and how to manipulate them. Constraint based methods, such as Flux Balance Analysis (FBA) and Thermodynamics-based flux analysis (TFA), are commonly used to estimate the flow of metabolites through genome-wide metabolic networks, making it possible to identify the ranges of flux values that are consistent with the studied physiological and thermodynamic conditions. However, unless key intracellular fluxes and metabolite concentrations are known, constraint-based models lead to underdetermined problem formulations. This lack of information propagates as uncertainty in the estimation of fluxes and basic reaction properties such as the determination of reaction directionalities. Therefore, knowledge of which metabolites, if measured, would contribute the most to reducing this uncertainty can significantly improve our ability to define the internal state of the cell. In the present work we combine constraint based modeling, Design of Experiments (DoE) and Global Sensitivity Analysis (GSA) into the Thermodynamics-based Metabolite Sensitivity Analysis (TMSA) method. TMSA ranks metabolites comprising a metabolic network based on their ability to constrain the gamut of possible solutions to a limited, thermodynamically consistent set of internal states. TMSA is modular and can be applied to a single reaction, a metabolic pathway or an entire metabolic network. This is, to our knowledge, the first attempt to use metabolic modeling in order to provide a significance ranking of metabolites to guide experimental measurements.

  12. Biopipe: a flexible framework for protocol-based bioinformatics analysis.

    Science.gov (United States)

    Hoon, Shawn; Ratnapu, Kiran Kumar; Chia, Jer-Ming; Kumarasamy, Balamurugan; Juguang, Xiao; Clamp, Michele; Stabenau, Arne; Potter, Simon; Clarke, Laura; Stupka, Elia

    2003-08-01

    We identify several challenges facing bioinformatics analysis today. Firstly, to fulfill the promise of comparative studies, bioinformatics analysis will need to accommodate different sources of data residing in a federation of databases that, in turn, come in different formats and modes of accessibility. Secondly, the tsunami of data to be handled will require robust systems that enable bioinformatics analysis to be carried out in a parallel fashion. Thirdly, the ever-evolving state of bioinformatics presents new algorithms and paradigms in conducting analysis. This means that any bioinformatics framework must be flexible and generic enough to accommodate such changes. In addition, we identify the need for introducing an explicit protocol-based approach to bioinformatics analysis that will lend rigorousness to the analysis. This makes it easier for experimentation and replication of results by external parties. Biopipe is designed in an effort to meet these goals. It aims to allow researchers to focus on protocol design. At the same time, it is designed to work over a compute farm and thus provides high-throughput performance. A common exchange format that encapsulates the entire protocol in terms of the analysis modules, parameters, and data versions has been developed to provide a powerful way in which to distribute and reproduce results. This will enable researchers to discuss and interpret the data better as the once implicit assumptions are now explicitly defined within the Biopipe framework.

  13. A Knowledge-based Environment for Software Process Performance Analysis

    Directory of Open Access Journals (Sweden)

    Natália Chaves Lessa Schots

    2015-08-01

    Full Text Available Background: Process performance analysis is a key step for implementing continuous improvement in software organizations. However, the knowledge to execute such analysis is not trivial and the person responsible to executing it must be provided with appropriate support. Aim: This paper presents a knowledge-based environment, named SPEAKER, proposed for supporting software organizations during the execution of process performance analysis. SPEAKER comprises a body of knowledge and a set of activities and tasks for software process performance analysis along with supporting tools to executing these activities and tasks. Method: We conducted an informal literature reviews and a systematic mapping study, which provided basic requirements for the proposed environment. We implemented the SPEAKER environment integrating supporting tools for the execution of activities and tasks of performance analysis and the knowledge necessary to execute them, in order to meet the variability presented by the characteristics of these activities. Results: In this paper, we describe each SPEAKER module and the individual evaluations of these modules, and also present an example of use comprising how the environment can guide the user through a specific performance analysis activity. Conclusion: Although we only conducted individual evaluations of SPEAKER’s modules, the example of use indicates the feasibility of the proposed environment. Therefore, the environment as a whole will be further evaluated to verify if it attains its goal of assisting in the execution of process performance analysis by non-specialist people.

  14. Cryptographic protocol security analysis based on bounded constructing algorithm

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    An efficient approach to analyzing cryptographic protocols is to develop automatic analysis tools based on formal methods. However, the approach has encountered the high computational complexity problem due to reasons that participants of protocols are arbitrary, their message structures are complex and their executions are concurrent. We propose an efficient automatic verifying algorithm for analyzing cryptographic protocols based on the Cryptographic Protocol Algebra (CPA) model proposed recently, in which algebraic techniques are used to simplify the description of cryptographic protocols and their executions. Redundant states generated in the analysis processes are much reduced by introducing a new algebraic technique called Universal Polynomial Equation and the algorithm can be used to verify the correctness of protocols in the infinite states space. We have implemented an efficient automatic analysis tool for cryptographic protocols, called ACT-SPA, based on this algorithm, and used the tool to check more than 20 cryptographic protocols. The analysis results show that this tool is more efficient, and an attack instance not offered previously is checked by using this tool.

  15. Analysis of system trustworthiness based on information flow noninterference theory

    Institute of Scientific and Technical Information of China (English)

    Xiangying Kong; Yanhui Chen; Yi Zhuang

    2015-01-01

    The trustworthiness analysis and evaluation are the bases of the trust chain transfer. In this paper the formal method of trustworthiness analysis of a system based on the noninterfer-ence (NI) theory of the information flow is studied. Firstly, existing methods cannot analyze the impact of the system states on the trustworthiness of software during the process of trust chain trans-fer. To solve this problem, the impact of the system state on trust-worthiness of software is investigated, the run-time mutual interfer-ence behavior of software entities is described and an interference model of the access control automaton of a system is established. Secondly, based on the intransitive noninterference (INI) theory, a formal analytic method of trustworthiness for trust chain transfer is proposed, providing a theoretical basis for the analysis of dynamic trustworthiness of software during the trust chain transfer process. Thirdly, a prototype system with dynamic trustworthiness on a plat-form with dual core architecture is constructed and a verification algorithm of the system trustworthiness is provided. Final y, the monitor hypothesis is extended to the dynamic monitor hypothe-sis, a theorem of static judgment rule of system trustworthiness is provided, which is useful to prove dynamic trustworthiness of a system at the beginning of system construction. Compared with previous work in this field, this research proposes not only a formal analytic method for the determination of system trustworthiness, but also a modeling method and an analysis algorithm that are feasible for practical implementation.

  16. Consistency analysis of accelerated degradation mechanism based on gray theory

    Institute of Scientific and Technical Information of China (English)

    Yunxia Chen; Hongxia Chen; Zhou Yang; Rui Kang; Yi Yang

    2014-01-01

    A fundamental premise of an accelerated testing is that the failure mechanism under elevated and normal stress levels should remain the same. Thus, verification of the consistency of failure mechanisms is essential during an accelerated testing. A new consistency analysis method based on the gray theory is pro-posed for complex products. First of al , existing consistency ana-lysis methods are reviewed with a focus on the comparison of the differences among them. Then, the proposed consistency ana-lysis method is introduced. Two effective gray prediction models, gray dynamic model and new information and equal dimensional (NIED) model, are adapted in the proposed method. The process to determine the dimension of NIED model is also discussed, and a decision rule is expanded. Based on that, the procedure of ap-plying the new consistent analysis method is developed. Final y, a case study of the consistency analysis of a reliability enhancement testing is conducted to demonstrate and validate the proposed method.

  17. Risk-based planning analysis for a single levee

    Science.gov (United States)

    Hui, Rui; Jachens, Elizabeth; Lund, Jay

    2016-04-01

    Traditional risk-based analysis for levee planning focuses primarily on overtopping failure. Although many levees fail before overtopping, few planning studies explicitly include intermediate geotechnical failures in flood risk analysis. This study develops a risk-based model for two simplified levee failure modes: overtopping failure and overall intermediate geotechnical failure from through-seepage, determined by the levee cross section represented by levee height and crown width. Overtopping failure is based only on water level and levee height, while through-seepage failure depends on many geotechnical factors as well, mathematically represented here as a function of levee crown width using levee fragility curves developed from professional judgment or analysis. These levee planning decisions are optimized to minimize the annual expected total cost, which sums expected (residual) annual flood damage and annualized construction costs. Applicability of this optimization approach to planning new levees or upgrading existing levees is demonstrated preliminarily for a levee on a small river protecting agricultural land, and a major levee on a large river protecting a more valuable urban area. Optimized results show higher likelihood of intermediate geotechnical failure than overtopping failure. The effects of uncertainty in levee fragility curves, economic damage potential, construction costs, and hydrology (changing climate) are explored. Optimal levee crown width is more sensitive to these uncertainties than height, while the derived general principles and guidelines for risk-based optimal levee planning remain the same.

  18. Dynamic Chest Image Analysis: Model-Based Perfusion Analysis in Dynamic Pulmonary Imaging

    Directory of Open Access Journals (Sweden)

    Kiuru Aaro

    2003-01-01

    Full Text Available The "Dynamic Chest Image Analysis" project aims to develop model-based computer analysis and visualization methods for showing focal and general abnormalities of lung ventilation and perfusion based on a sequence of digital chest fluoroscopy frames collected with the dynamic pulmonary imaging technique. We have proposed and evaluated a multiresolutional method with an explicit ventilation model for ventilation analysis. This paper presents a new model-based method for pulmonary perfusion analysis. According to perfusion properties, we first devise a novel mathematical function to form a perfusion model. A simple yet accurate approach is further introduced to extract cardiac systolic and diastolic phases from the heart, so that this cardiac information may be utilized to accelerate the perfusion analysis and improve its sensitivity in detecting pulmonary perfusion abnormalities. This makes perfusion analysis not only fast but also robust in computation; consequently, perfusion analysis becomes computationally feasible without using contrast media. Our clinical case studies with 52 patients show that this technique is effective for pulmonary embolism even without using contrast media, demonstrating consistent correlations with computed tomography (CT and nuclear medicine (NM studies. This fluoroscopical examination takes only about 2 seconds for perfusion study with only low radiation dose to patient, involving no preparation, no radioactive isotopes, and no contrast media.

  19. Real Time Engineering Analysis Based on a Generative Component Implementation

    DEFF Research Database (Denmark)

    Kirkegaard, Poul Henning; Klitgaard, Jens

    2007-01-01

    The present paper outlines the idea of a conceptual design tool with real time engineering analysis which can be used in the early conceptual design phase. The tool is based on a parametric approach using Generative Components with embedded structural analysis. Each of these components uses...... without jumping from aesthetics to structural digital design tools and back, but to work with both simultaneously and real time. The engineering level of knowledge is incorporated at a conceptual thinking level, i.e. qualitative information is used in stead of using quantitative information. An example...

  20. APL-based flexibility analysis of manufacturing grid

    Institute of Scientific and Technical Information of China (English)

    LIU; Li-lan; SUN; Xue-hua; CAI; Hong-xia; CHAI; Jian-fei

    2009-01-01

    With the characteristics of diversity, randomness, concurrency and decomposability, tasks in manufacturing field are very complicated, and so manufacturing grid(MG)should have considerable flexibility to deal with this problem. With the definition of node and arc, MG structure is converted into a small-world network. Given construction cost constraint, the problem of shortest task waiting time is transformed into the constrained optimization problem, and a corresponding flexibility analysis model based on average path length(APL)is proposed, and the premise of arc-length and node-distance are defined. The results of application example show that the analysis model is effective.

  1. Applying Content Analysis to Web-based Content

    OpenAIRE

    Kim, Inhwa; Kuljis, Jasna

    2010-01-01

    Using Content Analysis onWeb-based content, in particular the content available onWeb 2.0 sites, is investigated. The relative strengths and limitations of the method are described. To illustrate how content analysis may be used, we provide a brief overview of a case study that investigates cultural impacts on the use of design features with regard to self-disclosure on the blogs of South Korean and United Kingdom’s users. In this study we took a standard approach to conducting the content an...

  2. System modeling based measurement error analysis of digital sun sensors

    Institute of Scientific and Technical Information of China (English)

    WEI; M; insong; XING; Fei; WANG; Geng; YOU; Zheng

    2015-01-01

    Stringent attitude determination accuracy is required for the development of the advanced space technologies and thus the accuracy improvement of digital sun sensors is necessary.In this paper,we presented a proposal for measurement error analysis of a digital sun sensor.A system modeling including three different error sources was built and employed for system error analysis.Numerical simulations were also conducted to study the measurement error introduced by different sources of error.Based on our model and study,the system errors from different error sources are coupled and the system calibration should be elaborately designed to realize a digital sun sensor with extra-high accuracy.

  3. Towards Performance Measurement And Metrics Based Analysis of PLA Applications

    CERN Document Server

    Ahmed, Zeeshan

    2010-01-01

    This article is about a measurement analysis based approach to help software practitioners in managing the additional level complexities and variabilities in software product line applications. The architecture of the proposed approach i.e. ZAC is designed and implemented to perform preprocessesed source code analysis, calculate traditional and product line metrics and visualize results in two and three dimensional diagrams. Experiments using real time data sets are performed which concluded with the results that the ZAC can be very helpful for the software practitioners in understanding the overall structure and complexity of product line applications. Moreover the obtained results prove strong positive correlation between calculated traditional and product line measures.

  4. Logistics Enterprise Evaluation Model Based On Fuzzy Clustering Analysis

    Science.gov (United States)

    Fu, Pei-hua; Yin, Hong-bo

    In this thesis, we introduced an evaluation model based on fuzzy cluster algorithm of logistics enterprises. First of all,we present the evaluation index system which contains basic information, management level, technical strength, transport capacity,informatization level, market competition and customer service. We decided the index weight according to the grades, and evaluated integrate ability of the logistics enterprises using fuzzy cluster analysis method. In this thesis, we introduced the system evaluation module and cluster analysis module in detail and described how we achieved these two modules. At last, we gave the result of the system.

  5. FARO base case post-test analysis by COMETA code

    Energy Technology Data Exchange (ETDEWEB)

    Annunziato, A.; Addabbo, C. [Joint Research Centre, Ispra (Italy)

    1995-09-01

    The paper analyzes the COMETA (Core Melt Thermal-Hydraulic Analysis) post test calculations of FARO Test L-11, the so-called Base Case Test. The FARO Facility, located at JRC Ispra, is used to simulate the consequences of Severe Accidents in Nuclear Power Plants under a variety of conditions. The COMETA Code has a 6 equations two phase flow field and a 3 phases corium field: the jet, the droplets and the fused-debris bed. The analysis shown that the code is able to pick-up all the major phenomena occurring during the fuel-coolant interaction pre-mixing phase.

  6. A CONTENT ANALYSIS ON PROBLEM-BASED LEARNING APPROACH

    OpenAIRE

    BİBER, Mahir; Esen ERSOY; KÖSE BİBER, Sezer

    2014-01-01

    Problem Based Learning is one of the learning models that contain the general principles of active learning and students can use scientific process skills. Within this research it was aimed to investigate in detail the postgraduate thesis held in Turkey about PBL approach. The content analysis method was used in the research. The study sample was consisted of a total of 64 masters and PhD thesis made between the years 2012-2013 and reached over the web. A “Content Analysis Template” prepared ...

  7. A CONTENT ANALYSIS ON PROBLEM-BASED LEARNING APPROACH

    OpenAIRE

    BİBER, Mahir; Esen ERSOY; KÖSE BİBER, Sezer

    2015-01-01

    Problem Based Learning is one of the learning models that contain the general principles of active learning and students can use scientific process skills. Within this research it was aimed to investigate in detail the postgraduate thesis held in Turkey about PBL approach. The content analysis method was used in the research. The study sample was consisted of a total of 64 masters and PhD thesis made between the years 2012-2013 and reached over the web. A “Content Analysis Template” prepared ...

  8. Transit Traffic Analysis Zone Delineating Method Based on Thiessen Polygon

    Directory of Open Access Journals (Sweden)

    Shuwei Wang

    2014-04-01

    Full Text Available A green transportation system composed of transit, busses and bicycles could be a significant in alleviating traffic congestion. However, the inaccuracy of current transit ridership forecasting methods is imposing a negative impact on the development of urban transit systems. Traffic Analysis Zone (TAZ delineating is a fundamental and essential step in ridership forecasting, existing delineating method in four-step models have some problems in reflecting the travel characteristics of urban transit. This paper aims to come up with a Transit Traffic Analysis Zone delineation method as supplement of traditional TAZs in transit service analysis. The deficiencies of current TAZ delineating methods were analyzed, and the requirements of Transit Traffic Analysis Zone (TTAZ were summarized. Considering these requirements, Thiessen Polygon was introduced into TTAZ delineating. In order to validate its feasibility, Beijing was then taken as an example to delineate TTAZs, followed by a spatial analysis of office buildings within a TTAZ and transit station departure passengers. Analysis result shows that the TTAZs based on Thiessen polygon could reflect the transit travel characteristic and is of in-depth research value.

  9. Sentiment analysis framework organization based on twitter corpus data

    Directory of Open Access Journals (Sweden)

    Adela Beres

    2012-06-01

    Full Text Available Since its inception in 2006, Twitter has gathered millions of users. They post daily tweets about news, events or conversations. These tweets express their opinion about the topic they are discussing. Twitter is a large database of content that can be semantically exploited to extract opinions and based on these opinions to classify the users. This paper presents the organization of a sentiment analysis framework based on Twitter corpus data, including crawling tweets and opinion mining of the tweets, making it easy for its users to create portfolios of trustful Twitter accounts.

  10. Coverage analysis for sensor networks based on Clifford algebra

    Institute of Scientific and Technical Information of China (English)

    XIE WeiXin; CAO WenMing; MENG Shan

    2008-01-01

    The coverage performance is the foundation of information acquisition in distrib-uted sensor networks. The previously proposed coverage work was mostly based on unit disk coverage model or ball coverage model in 2D or 3D space, respectively. However, most methods cannot give a homogeneous coverage model for targets with hybrid types. This paper presents a coverage analysis approach for sensor networks based on Clifford algebra and establishes a homogeneous coverage model for sensor networks with hybrid types of targets. The effectiveness of the approach is demonstrated with examples.

  11. Image registration based on matrix perturbation analysis using spectral graph

    Institute of Scientific and Technical Information of China (English)

    Chengcai Leng; Zheng Tian; Jing Li; Mingtao Ding

    2009-01-01

    @@ We present a novel perspective on characterizing the spectral correspondence between nodes of the weighted graph with application to image registration.It is based on matrix perturbation analysis on the spectral graph.The contribution may be divided into three parts.Firstly, the perturbation matrix is obtained by perturbing the matrix of graph model.Secondly, an orthogonal matrix is obtained based on an optimal parameter, which can better capture correspondence features.Thirdly, the optimal matching matrix is proposed by adjusting signs of orthogonal matrix for image registration.Experiments on both synthetic images and real-world images demonstrate the effectiveness and accuracy of the proposed method.

  12. Image Analysis of Fabric Pilling Based on Light Projection

    Institute of Scientific and Technical Information of China (English)

    陈霞; 黄秀宝

    2003-01-01

    The objective assessment of fabric pilling based on light projection and image analysis has been exploited recently.The device for capturing the cross-sectional images of the pilled fabrics with light projection is elaborated.The detection of the profile line and integration of the sequential cross-sectional pilled image are discussed.The threshold based on Gaussian model is recommended for pill segmentation.The results show that the installed system is capable of eliminating the interference with pill information from the fabric color and pattern.

  13. Plug-in Based Analysis Framework for LHC Post-Mortem Analysis

    CERN Document Server

    Gorbonosov, R; Zerlauth, M; Baggiolini, V

    2014-01-01

    Plug-in based software architectures [1] are extensible, enforce modularity and allow several teams to work in parallel. But they have certain technical and organizational challenges, which we discuss in this paper. We gained our experience when developing the Post-Mortem Analysis (PMA) system, which is a mission critical system for the Large Hadron Collider (LHC). We used a plugin-based architecture with a general-purpose analysis engine, for which physicists and equipment experts code plugins containing the analysis algorithms. We have over 45 analysis plugins developed by a dozen of domain experts. This paper focuses on the design challenges we faced in order to mitigate the risks of executing third-party code: assurance that even a badly written plugin doesn't perturb the work of the overall application; plugin execution control which allows to detect plugin misbehaviour and react; robust communication mechanism between plugins, diagnostics facilitation in case of plugin failure; testing of the plugins be...

  14. Classification analysis of microarray data based on ontological engineering

    Institute of Scientific and Technical Information of China (English)

    LI Guo-qi; SHENG Huan-ye

    2007-01-01

    Background knowledge is important for data mining, especially in complicated situation. Ontological engineering is the successor of knowledge engineering. The sharable knowledge bases built on ontology can be used to provide background knowledge to direct the process of data mining. This paper gives a common introduction to the method and presents a practical analysis example using SVM (support vector machine) as the classifier. Gene Ontology and the accompanying annotations compose a big knowledge base, on which many researches have been carried out. Microarray dataset is the output of DNA chip.With the help of Gene Ontology we present a more elaborate analysis on microarray data than former researchers. The method can also be used in other fields with similar scenario.

  15. GNSS Spoofing Detection Based on Signal Power Measurements: Statistical Analysis

    Directory of Open Access Journals (Sweden)

    V. Dehghanian

    2012-01-01

    Full Text Available A threat to GNSS receivers is posed by a spoofing transmitter that emulates authentic signals but with randomized code phase and Doppler values over a small range. Such spoofing signals can result in large navigational solution errors that are passed onto the unsuspecting user with potentially dire consequences. An effective spoofing detection technique is developed in this paper, based on signal power measurements and that can be readily applied to present consumer grade GNSS receivers with minimal firmware changes. An extensive statistical analysis is carried out based on formulating a multihypothesis detection problem. Expressions are developed to devise a set of thresholds required for signal detection and identification. The detection processing methods developed are further manipulated to exploit incidental antenna motion arising from user interaction with a GNSS handheld receiver to further enhance the detection performance of the proposed algorithm. The statistical analysis supports the effectiveness of the proposed spoofing detection technique under various multipath conditions.

  16. INNOVATION ANALYSIS BASED ON SCORES AT THE FIRM LEVEL

    Directory of Open Access Journals (Sweden)

    Cătălin George ALEXE

    2014-04-01

    Full Text Available Innovation analysis based on scores (Innovation Scorecard is a simple way to get a quick diagnosis on the potential of innovation of a firm in its intention to achieve the innovation capability. It aims to identify and remedy the deficient aspects related to innovation management being used as a measuring tool for the innovation initiatives over time within the innovation audit. The paper aims to present the advantages and disadvantages of using the method, and the three approaches developed over time. Therefore, the model proposed by the consulting firm Arthur D. Little in collaboration with the European Business School, Eckelmann's model and AGGB's local model are summarized and compared. At the end of the paper, several possible solutions are proposed to improve the way of analysis based on scores.

  17. Protein analysis based on molecular beacon probes and biofunctionalized nanoparticles

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    With the completion of the human genome-sequencing project, there has been a resulting change in the focus of studies from genomics to proteomics. By utilizing the inherent advantages of molecular beacon probes and biofunctionalized nanoparticles, a series of novel principles, methods and techniques have been exploited for bioanalytical and biomedical studies. This review mainly discusses the applications of molecular beacon probes and biofunctionalized nanoparticles-based technologies for realtime, in-situ, highly sensitive and highly selective protein analysis, including the nonspecific or specific protein detection and separation, protein/DNA interaction studies, cell surface protein recognition, and antigen-antibody binding process-based bacteria assays. The introduction of molecular beacon probes and biofunctionalized nanoparticles into the protein analysis area would necessarily advance the proteomics research.

  18. Model-Based Dependability Analysis of Physical Systems with Modelica

    Directory of Open Access Journals (Sweden)

    Andrea Tundis

    2017-01-01

    Full Text Available Modelica is an innovative, equation-based, and acausal language that allows modeling complex physical systems, which are made of mechanical, electrical, and electrotechnical components, and evaluates their design through simulation techniques. Unfortunately, the increasing complexity and accuracy of such physical systems require new, more powerful, and flexible tools and techniques for evaluating important system properties and, in particular, the dependability ones such as reliability, safety, and maintainability. In this context, the paper describes some extensions of the Modelica language to support the modeling of system requirements and their relationships. Such extensions enable the requirement verification analysis through native constructs in the Modelica language. Furthermore, they allow exporting a Modelica-based system design as a Bayesian Network in order to analyze its dependability by employing a probabilistic approach. The proposal is exemplified through a case study concerning the dependability analysis of a Tank System.

  19. Fuzzy-Set Based Sentiment Analysis of Big Social Data

    DEFF Research Database (Denmark)

    Mukkamala, Raghava Rao; Hussain, Abid; Vatrapu, Ravi

    Computational approaches to social media analytics are largely limited to graph theoretical approaches such as social network analysis (SNA) informed by the social philosophical approach of relational sociology. There are no other unified modelling approaches to social data that integrate...... from Facebook. Third, we briefly present and discuss the Social Data Analytics Tool (SODATO) that realizes the conceptual model in software and provisions social data analysis based on the conceptual and formal models. Fourth, we use SODATO to fetch social data from the facebook wall of a global brand...... and actors on the facebook page. Sixth and last, we discuss the analytical method and conclude with a discussion of the benefits of set theoretical approaches based on the social philosophical approach of associational sociology....

  20. Fuzzy-Set Based Sentiment Analysis of Big Social Data

    DEFF Research Database (Denmark)

    Mukkamala, Raghava Rao; Hussain, Abid; Vatrapu, Ravi

    2014-01-01

    Abstract—Computational approaches to social media analytics are largely limited to graph theoretical approaches such as social network analysis (SNA) informed by the social philosophical approach of relational sociology. There are no other unified modelling approaches to social data that integrate...... from Facebook. Third, we briefly present and discuss the Social Data Analytics Tool (SODATO) that realizes the conceptual model in software and provisions social data analysis based on the conceptual and formal models. Fourth, we use SODATO to fetch social data from the facebook wall of a global brand...... and actors on the facebook page. Sixth and last, we discuss the analytical method and conclude with a discussion of the benefits of set theoretical approaches based on the social philosophical approach of associational sociology....

  1. Fusing Quantitative Requirements Analysis with Model-based Systems Engineering

    Science.gov (United States)

    Cornford, Steven L.; Feather, Martin S.; Heron, Vance A.; Jenkins, J. Steven

    2006-01-01

    A vision is presented for fusing quantitative requirements analysis with model-based systems engineering. This vision draws upon and combines emergent themes in the engineering milieu. "Requirements engineering" provides means to explicitly represent requirements (both functional and non-functional) as constraints and preferences on acceptable solutions, and emphasizes early-lifecycle review, analysis and verification of design and development plans. "Design by shopping" emphasizes revealing the space of options available from which to choose (without presuming that all selection criteria have previously been elicited), and provides means to make understandable the range of choices and their ramifications. "Model-based engineering" emphasizes the goal of utilizing a formal representation of all aspects of system design, from development through operations, and provides powerful tool suites that support the practical application of these principles. A first step prototype towards this vision is described, embodying the key capabilities. Illustrations, implications, further challenges and opportunities are outlined.

  2. Transmissibility-Based Operational Modal Analysis: Enhanced Stabilisation Diagrams

    Directory of Open Access Journals (Sweden)

    Gert De Sitter

    2012-01-01

    Full Text Available Recently it has been shown that also transmissibilities can be used to identify the modal parameters. This approach has several advantages: because of the deterministic character of the transmissibility functions, the estimated parameters are more accurate than the results obtained with the power spectra based operational modal analysis techniques. Another advantage is that the transmissibility functions do not depend on the colouring of the unknown forces. A disadvantage of the transmissibility based operational modal analysis techniques is that non-physical modes show up in the stabilisation diagrams. In this contribution it will first be shown that those non-physical modes will show up when traditional stabilisation diagrams are used. In a second step, a new approach of selecting the physical modes out of a set of estimated modes will be discussed and the new approach will be validated using data generated with an acoustical Finite Element Model. Finally, the approach will be validated using real acoustical data.

  3. [Ecological security assessment of Tangshan City based on emergy analysis].

    Science.gov (United States)

    Cao, Ming-lan; Li, Ya-dong

    2009-09-01

    Based on 'pressure-state-response' model and by using emergy analysis method, the urban ecological security assessment system and urban ecological security index (EUESI) were constructed, and the variation of ecological security level of Tangshan City in 1995-2005 was evaluated. During this period, the ecological security level of the city increased first and decreased then. The EUESI increased from 0.017 in 1995 to 0.022 in 1996, then dropped yearly, and became unsecure in 2003. The urban ecological security assessment method based on emergy analysis overcame the disadvantages of conventional assessment system, e.g., numerous and repetitive indicators, non-uniform units, and poor comparability, and reflected the urban ecological security state more objectively, being able to provide scientific basis for urban ecological environment management and decision-making.

  4. Image-Analysis Based on Seed Phenomics in Sesame

    Directory of Open Access Journals (Sweden)

    Prasad R.

    2014-10-01

    Full Text Available The seed coat (testa structure of twenty-three cultivated (Sesamum indicum L. and six wild sesame (s. occidentale Regel & Heer., S. mulayanum Nair, S. prostratum Retz., S. radiatum Schumach. & Thonn., S. angustifolium (Oliv. Engl. and S. schinzianum Asch germplasm was analyzed from digital and Scanning Electron Microscopy (SEM images with dedicated software using the descriptors for computer based seed image analysis to understand the diversity of seed morphometric traits, which later on can be extended to screen and evaluate improved genotypes of sesame. Seeds of wild sesame species could conveniently be distinguished from cultivated varieties based on shape and architectural analysis. Results indicated discrete ‘cut off values to identify definite shape and contour of seed for a desirable sesame genotype along with the con-ventional practice of selecting lighter colored testa.

  5. HIV/AIDS counseling: analysis based on Paulo Freire.

    Science.gov (United States)

    Miranda, Karla Corrêa Lima; Barroso, Maria Grasiela Teixeira

    2007-01-01

    The study aimed to investigate the strategies health professionals use in HIV/AIDS counseling. This study is a qualitative research, based on Paulo Freire's theory and practice. Bardin's content analysis was used as the analysis technique. For the studied group, the counseling is focused on cognition, although new concepts permeating this subject are emerging. The main difficulties in counseling are related to the clients and the institution. The main facility is related to the team, which according to the group has a good relationship. Counseling represents a moment of distress, especially because it brings up existential questions to the counselor. It can be inferred that counseling is a special moment, but it does not constitute an educational moment yet. To obtain this goal, a counseling methodology is proposed, based on Paulo Freire's principles and concepts.

  6. CORBA-Based Analysis of Multi Agent Behavior

    Institute of Scientific and Technical Information of China (English)

    Swapan Bhattacharya; Anirban Banerjee; Shibdas Bandyopadhyay

    2005-01-01

    An agent is a computer software that is capable of taking independent action on behalf of its user or owner. It is an entity with goals, actions and domain knowledge, situated in an environment. Multiagent systems comprises of multiple autonomous, interacting computer software, or agents. These systems can successfully emulate the entities active in a distributed environment. The analysis of multiagent behavior has been studied in this paper based on a specific board game problem similar to the famous problem of GO. In this paper a framework is developed to define the states of the multiagent entities and measure the convergence metrics for this problem. An analysis of the changes of states leading to the goal state is also made. We support our study of multiagent behavior by simulations based on a CORBA framework in order to substantiate our findings.

  7. The OASE project: Object-based Analysis and Seamless prediction

    Science.gov (United States)

    Troemel, Silke; Wapler, Kathrin; Bick, Theresa; Diederich, Malte; Deneke, Hartwig; Horvath, Akos; Senf, Fabian; Simmer, Clemens; Simon, Juergen

    2013-04-01

    The research group on Object-based Analysis and SEamless prediction (OASE) is part of the Hans Ertel Centre for Weather Research (HErZ). The group consists of scientists at the Meteorological Institute, University of Bonn, the Leibniz-Institute for Tropospheric Research in Leipzig and the German Weather Service. OASE addresses seamless prediction of convective events from nowcasting to daily predictions by combining radar/satellite compositing and tracking with high-resolution model-based ensemble generation and prediction. While observation-based nowcasting provides good results for lead times between 0-1 hours, numerical weather prediction addresses lead times between 3-21 hours. Especially the discontinuity between 1-3 hours needs to be addressed. Therefore a central goal of the project is a near real-time high-resolved unprecedented data base. A radar and satellite remote sensing-driven 3D observation-microphysics composite covering Germany, currently under development, contains gridded observations and estimated microphysical quantities. Observations and microphysics are intertwined via forward operators and estimated inverse relations, which also provide uncertainties for model ensemble initialisations. The lifetime evolution of dynamics and microphysics in (severe) convective storms is analysed based on 3D scale-space tracking. An object-based analysis condenses the information contained in the dynamic 3D distributions of observables and related microphysics into descriptors, which will allow identifying governing processes leading to the formation and evolution of severe weather events. The object-based approach efficiently characterises and quantifies the process structure and life cycles of severe weather events, and facilitates nowcasting and the generation and initialisation of model prediction ensembles. Observation-based nowcasting will exploit the dual-composite based 3D feature detection and tracking to generate a set of predictions (observation-based

  8. Facial Affect Recognition Using Regularized Discriminant Analysis-Based Algorithms

    Directory of Open Access Journals (Sweden)

    Cheng-Yuan Shih

    2010-01-01

    Full Text Available This paper presents a novel and effective method for facial expression recognition including happiness, disgust, fear, anger, sadness, surprise, and neutral state. The proposed method utilizes a regularized discriminant analysis-based boosting algorithm (RDAB with effective Gabor features to recognize the facial expressions. Entropy criterion is applied to select the effective Gabor feature which is a subset of informative and nonredundant Gabor features. The proposed RDAB algorithm uses RDA as a learner in the boosting algorithm. The RDA combines strengths of linear discriminant analysis (LDA and quadratic discriminant analysis (QDA. It solves the small sample size and ill-posed problems suffered from QDA and LDA through a regularization technique. Additionally, this study uses the particle swarm optimization (PSO algorithm to estimate optimal parameters in RDA. Experiment results demonstrate that our approach can accurately and robustly recognize facial expressions.

  9. [Model-based biofuels system analysis: a review].

    Science.gov (United States)

    Chang, Shiyan; Zhang, Xiliang; Zhao, Lili; Ou, Xunmin

    2011-03-01

    Model-based system analysis is an important tool for evaluating the potential and impacts of biofuels, and for drafting biofuels technology roadmaps and targets. The broad reach of the biofuels supply chain requires that biofuels system analyses span a range of disciplines, including agriculture/forestry, energy, economics, and the environment. Here we reviewed various models developed for or applied to modeling biofuels, and presented a critical analysis of Agriculture/Forestry System Models, Energy System Models, Integrated Assessment Models, Micro-level Cost, Energy and Emission Calculation Models, and Specific Macro-level Biofuel Models. We focused on the models' strengths, weaknesses, and applicability, facilitating the selection of a suitable type of model for specific issues. Such an analysis was a prerequisite for future biofuels system modeling, and represented a valuable resource for researchers and policy makers.

  10. Student Engagement: A Principle-Based Concept Analysis.

    Science.gov (United States)

    Bernard, Jean S

    2015-08-04

    A principle-based concept analysis of student engagement was used to examine the state of the science across disciplines. Four major perspectives of philosophy of science guided analysis and provided a framework for study of interrelationships and integration of conceptual components which then resulted in formulation of a theoretical definition. Findings revealed student engagement as a dynamic reiterative process marked by positive behavioral, cognitive, and affective elements exhibited in pursuit of deep learning. This process is influenced by a broader sociocultural environment bound by contextual preconditions of self-investment, motivation, and a valuing of learning. Outcomes of student engagement include satisfaction, sense of well-being, and personal development. Findings of this analysis prove relevant to nursing education as faculty transition from traditional teaching paradigms, incorporate learner-centered strategies, and adopt innovative pedagogical methodologies. It lends support for curricula reform, development of more accurate evaluative measures, and creation of meaningful teaching-learning environments within the discipline.

  11. SVM-based glioma grading: Optimization by feature reduction analysis.

    Science.gov (United States)

    Zöllner, Frank G; Emblem, Kyrre E; Schad, Lothar R

    2012-09-01

    We investigated the predictive power of feature reduction analysis approaches in support vector machine (SVM)-based classification of glioma grade. In 101 untreated glioma patients, three analytic approaches were evaluated to derive an optimal reduction in features; (i) Pearson's correlation coefficients (PCC), (ii) principal component analysis (PCA) and (iii) independent component analysis (ICA). Tumor grading was performed using a previously reported SVM approach including whole-tumor cerebral blood volume (CBV) histograms and patient age. Best classification accuracy was found using PCA at 85% (sensitivity=89%, specificity=84%) when reducing the feature vector from 101 (100-bins rCBV histogram+age) to 3 principal components. In comparison, classification accuracy by PCC was 82% (89%, 77%, 2 dimensions) and 79% by ICA (87%, 75%, 9 dimensions). For improved speed (up to 30%) and simplicity, feature reduction by all three methods provided similar classification accuracy to literature values (∼87%) while reducing the number of features by up to 98%.

  12. Tariff-based analysis of commercial building electricityprices

    Energy Technology Data Exchange (ETDEWEB)

    Coughlin, Katie M.; Bolduc, Chris A.; Rosenquist, Greg J.; VanBuskirk, Robert D.; McMahon, James E.

    2008-03-28

    This paper presents the results of a survey and analysis ofelectricity tariffs and marginal electricity prices for commercialbuildings. The tariff data come from a survey of 90 utilities and 250tariffs for non-residential customers collected in 2004 as part of theTariff Analysis Project at LBNL. The goals of this analysis are toprovide useful summary data on the marginal electricity prices commercialcustomers actually see, and insight into the factors that are mostimportant in determining prices under different circumstances. We providea new, empirically-based definition of several marginal prices: theeffective marginal price and energy-only anddemand-only prices, andderive a simple formula that expresses the dependence of the effectivemarginal price on the marginal load factor. The latter is a variable thatcan be used to characterize the load impacts of a particular end-use orefficiency measure. We calculate all these prices for eleven regionswithin the continental U.S.

  13. Analysis Of Japans Economy Based On 2014 From Macroeconomics Prospects

    Directory of Open Access Journals (Sweden)

    Dr Mohammad Rafiqul Islam

    2015-02-01

    Full Text Available Abstract Japan is the worlds third largest economy. But currently economic situations of Japan are not stable. It is not increasing as expected. Since 2013 it was world second largest economy but Japan loosed its placed to China in 2014 due to slow growth of important economic indicators. By using the basic Keynesian model we will provide a detailed analysis of the short and long run impacts of the changes for Japans real GDP rate of unemployment and inflation rate. We demonstrated a detailed use of the 45-degree diagram or the AD-IA model and other economic analysis of the macroeconomic principles that underlie the model and concepts. Finally we will recommend the government with a change in fiscal policy what based on the analysis by considering what might be achieved with a fiscal policy response and the extent to which any impact on the stock of public debt might be a consideration

  14. Error Analysis of Robotic Assembly System Based on Screw Theory

    Institute of Scientific and Technical Information of China (English)

    韩卫军; 费燕琼; 赵锡芳

    2003-01-01

    Assembly errors have great influence on assembly quality in robotic assembly systems. Error analysis is directed to the propagations and accumula-tions of various errors and their effect on assembly success.Using the screw coordinates, assembly errors are represented as "error twist", the extremely compact expression. According to the law of screw composition, relative position and orientation errors of mating parts are computed and the necessary condition of assembly success is concluded. A new simple method for measuring assembly errors is also proposed based on the transformation law of a screw.Because of the compact representation of error, the model presented for error analysis can be applied to various part- mating types and especially useful for error analysis of complexity assembly.

  15. GOMA: functional enrichment analysis tool based on GO modules

    Institute of Scientific and Technical Information of China (English)

    Qiang Huang; Ling-Yun Wu; Yong Wang; Xiang-Sun Zhang

    2013-01-01

    Analyzing the function of gene sets is a critical step in interpreting the results of high-throughput experiments in systems biology.A variety of enrichment analysis tools have been developed in recent years,but most output a long list of significantly enriched terms that are often redundant,making it difficult to extract the most meaningful functions.In this paper,we present GOMA,a novel enrichment analysis method based on the new concept of enriched functional Gene Ontology (GO) modules.With this method,we systematically revealed functional GO modules,i.e.,groups of functionally similar GO terms,via an optimization model and then ranked them by enrichment scores.Our new method simplifies enrichment analysis results by reducing redundancy,thereby preventing inconsistent enrichment results among functionally similar terms and providing more biologically meaningful results.

  16. Fractal Analysis Based on Hierarchical Scaling in Complex Systems

    CERN Document Server

    Chen, Yanguang

    2016-01-01

    A fractal is in essence a hierarchy with cascade structure, which can be described with a set of exponential functions. From these exponential functions, a set of power laws indicative of scaling can be derived. Hierarchy structure and spatial network proved to be associated with one another. This paper is devoted to exploring the theory of fractal analysis of complex systems by means of hierarchical scaling. Two research methods are utilized to make this study, including logic analysis method and empirical analysis method. The main results are as follows. First, a fractal system such as Cantor set is described from the hierarchical angle of view; based on hierarchical structure, three approaches are proposed to estimate fractal dimension. Second, the hierarchical scaling can be generalized to describe multifractals, fractal complementary sets, and self-similar curve such as logarithmic spiral. Third, complex systems such as urban system are demonstrated to be a self-similar hierarchy. The human settlements i...

  17. Error Analysis of English Writing Based on Interlanguage Theory

    Institute of Scientific and Technical Information of China (English)

    李玲

    2014-01-01

    Language learning process has been hunted by learner’s errors,which is an unavoidable phenomenon.In the 1950s and 1960s,Contractive Analysis (CA) based on behaviorism and structuralism was generally employed in analyzing learners’ errors. CA soon lost its popularity.Error Analysis (EA),a branch of applied linguistics,has made great contributions to the study of second language learning and throws some light on the process of second language learning.Careful study of the errors reveals the common problems shared by the language learners.Writing is important in language learning process.Under Chinese context,English writing is always a difficult question for Chinese teachers and students,so errors in students’ written works are unavoidable.In this thesis,the author studies on error analysis of English writing with the interlanguage theory as its theoretical guidance.

  18. Frailty phenotypes in the elderly based on cluster analysis

    DEFF Research Database (Denmark)

    Dato, Serena; Montesanto, Alberto; Lagani, Vincenzo

    2012-01-01

    genetic background on the frailty status is still questioned. We investigated the applicability of a cluster analysis approach based on specific geriatric parameters, previously set up and validated in a southern Italian population, to two large longitudinal Danish samples. In both cohorts, we identified...... groups of subjects homogeneous for their frailty status and characterized by different survival patterns. A subsequent survival analysis availing of Accelerated Failure Time models allowed us to formulate an operative index able to correlate classification variables with survival probability. From...... these models, we quantified the differential effect of various parameters on survival, and we estimated the heritability of the frailty phenotype by exploiting the twin pairs in our sample. These data suggest the presence of a genetic influence on the frailty variability and indicate that cluster analysis can...

  19. Computer Vision-Based Image Analysis of Bacteria.

    Science.gov (United States)

    Danielsen, Jonas; Nordenfelt, Pontus

    2017-01-01

    Microscopy is an essential tool for studying bacteria, but is today mostly used in a qualitative or possibly semi-quantitative manner often involving time-consuming manual analysis. It also makes it difficult to assess the importance of individual bacterial phenotypes, especially when there are only subtle differences in features such as shape, size, or signal intensity, which is typically very difficult for the human eye to discern. With computer vision-based image analysis - where computer algorithms interpret image data - it is possible to achieve an objective and reproducible quantification of images in an automated fashion. Besides being a much more efficient and consistent way to analyze images, this can also reveal important information that was previously hard to extract with traditional methods. Here, we present basic concepts of automated image processing, segmentation and analysis that can be relatively easy implemented for use with bacterial research.

  20. Error Analysis of English Writing Based on Interlanguage Theory

    Institute of Scientific and Technical Information of China (English)

    李玲

    2014-01-01

    Language learning process has been hunted by learner’s errors,which is an unavoidable phenomenon.In the 1950 s and 1960 s,Contractive Analysis(CA) based on behaviorism and structuralism was generally employed in analyzing learners’ errors.CA soon lost its popularity.Error Analysis(EA),a branch of applied linguistics,has made great contributions to the study of second language learning and throws some light on the process of second language learning.Careful study of the errors reveals the common problems shared by the language learners.Writing is important in language learning process.Under Chinese context,English writing is always a difficult question for Chinese teachers and students,so errors in students’ written works are unavoidable.In this thesis,the author studies on error analysis of English writing with the interlanguage theory as its theoretical guidance.

  1. INDIVIDUAL COMMUNICATION TRANSMITTER IDENTIFICATION BASED ON MULTIFRACTAL ANALYSIS

    Institute of Scientific and Technical Information of China (English)

    Ren Chunhui; Wei Ping; Lou Zhiyou; Xiao Xianci

    2005-01-01

    In this letter, the communication transmitter transient signals are analyzed based on the time-variant hierarchy exponents of multifractal analysis. The species of optimized sample set is selected as the template of transmitter identification, so that the individual communication transmitter identification can be realized. The turn-on signals of four transmitters are used in the simulation. The experimental results show that the multifractal character of transmitter transient signals is an effective character of individual transmitter identification.

  2. Wavelet Variance Analysis of EEG Based on Window Function

    Institute of Scientific and Technical Information of China (English)

    ZHENG Yuan-zhuang; YOU Rong-yi

    2014-01-01

    A new wavelet variance analysis method based on window function is proposed to investigate the dynamical features of electroencephalogram (EEG).The ex-prienmental results show that the wavelet energy of epileptic EEGs are more discrete than normal EEGs, and the variation of wavelet variance is different between epileptic and normal EEGs with the increase of time-window width. Furthermore, it is found that the wavelet subband entropy (WSE) of the epileptic EEGs are lower than the normal EEGs.

  3. COMMERCIAL VIABILITY ANALYSIS OF LIGNIN BASED CARBON FIBRE

    OpenAIRE

    2014-01-01

    Lignin is a rich renewable source of aromatic compounds. As a potentialpetroleum feedstock replacement, lignin can reduce environmental impacts such ascarbon emission. Due to its complex chemical structure, lignin is currently underutilized.Exploiting lignin as a precursor for carbon fibre adds high economic value to lignin andencourages further development in lignin extraction technology. This report includes apreliminary cost analysis and identifies the key aspects of lignin-based carbon fi...

  4. Study on Segmented Reflector Lamp Design Based on Error Analysis

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    This paper discusses the basic principle and design m ethod for light distribution of car lamp, introduces an important development: h igh efficient and flexible car lamp with reflecting light distribution-segmente d reflector (multi-patch) car lamp, and puts out a design method for segmented reflector based on error analysis. Unlike classical car lamp with refractive lig ht distribution, the method of reflecting light distribution gives car lamp desi gn more flexibility. In the case of guarantying the li...

  5. Contributions to Physics-Based Aeroservoelastic Uncertainty Analysis

    Science.gov (United States)

    Wu, Sang

    The thesis presents the development of a new fully-integrated, MATLAB based simulation capability for aeroservoelastic (ASE) uncertainty analysis that accounts for uncertainties in all disciplines as well as discipline interactions. This new capability allows probabilistic studies of complex configuration at a scope and with depth not known before. Several statistical tools and methods have been integrated into the capability to guide the tasks such as parameter prioritization, uncertainty reduction, and risk mitigation. (Abstract shortened by ProQuest.).

  6. Performance Analysis of STFT Based Timing Approach to OFDM Systems

    Institute of Scientific and Technical Information of China (English)

    KUANG Yu-jun; TENG Yong; YIN Chang-chuan; HAO Jian-jun; YUE Guang-xin

    2003-01-01

    This paper mainly focuses on performance analysis of the previously proposed STFT based 2-D timing approach to OFDM systems and presents simulations results of its performance in AWGN and multipath fading environment and its robustness against the duration of Channel Impulse Response (CIR) and frequency offset. Simulation results suggest that a revised version of Short-Time Fourier Transform (STFT) can be used to greatly reduce computational complexity, especially at higher SNR.

  7. Comparative analysis of some brushless motors based on catalog data

    Directory of Open Access Journals (Sweden)

    Anton Kalapish

    2005-10-01

    Full Text Available Brushless motors (polyphased AC induction, synchronous and brushless DC motors have no alternatives in modern electric drives. They possess highly efficient and very wide range of speeds. The objective of this paper is to represent some relation between the basic parameters and magnitudes of electrical machines. This allows to be made a comparative analysis and a choice of motor concerning each particular case based not only on catalogue data or price for sale.

  8. Dynamic network-based epistasis analysis: Boolean examples

    Directory of Open Access Journals (Sweden)

    Eugenio eAzpeitia

    2011-12-01

    Full Text Available In this review we focus on how the hierarchical and single-path assumptions of epistasis analysis can bias the topologies of gene interactions infered. This has been acknowledged in several previous papers and reviews, but here we emphasize the critical importance of dynamic analyses, and specifically illustrate the use of Boolean network models. Epistasis in a broad sense refers to gene interactions, however, as originally proposed by Bateson (herein, classical epistasis, defined as the blocking of a particular allelic effect due to the effect of another allele at a different locus. Classical epistasis analysis has proven powerful and useful, allowing researchers to infer and assign directionality to gene interactions. As larger data sets are becoming available, the analysis of classical epistasis is being complemented with computer science tools and system biology approaches. We show that when the hierarchical and single-path assumptions are not met in classical epistasis analysis, the access to relevant information and the correct gene interaction topologies are hindered, and it becomes necessary to consider the temporal dynamics of gene interactions. The use of dynamical networks can overcome these limitations. We particularly focus on the use of Boolean networks that, like classical epistasis analysis, relies on logical formalisms, and hence can complement classical epistasis analysis and relax its assumptions. We develop a couple of theoretical examples and analyze them from a dynamic Boolean network model perspective. Boolean networks could help to guide additional experiments and discern among alternative regulatory schemes that would be impossible or difficult to infer without the elimination of these assumption from the classical epistasis analysis. We also use examples from the literature to show how a Boolean network-based approach has resolved ambiguities and guided epistasis analysis. Our review complements previous accounts, not

  9. Moon-Based INSAR Geolocation and Baseline Analysis

    Science.gov (United States)

    Liu, Guang; Ren, Yuanzhen; Ye, Hanlin; Guo, Huadong; Ding, Yixing; Ruan, Zhixing; Lv, Mingyang; Dou, Changyong; Chen, Zhaoning

    2016-07-01

    Earth observation platform is a host, the characteristics of the platform in some extent determines the ability for earth observation. Currently most developing platforms are satellite, in contrast carry out systematic observations with moon based Earth observation platform is still a new concept. The Moon is Earth's only natural satellite and is the only one which human has reached, it will give people different perspectives when observe the earth with sensors from the moon. Moon-based InSAR (SAR Interferometry), one of the important earth observation technology, has all-day, all-weather observation ability, but its uniqueness is still a need for analysis. This article will discuss key issues of geometric positioning and baseline parameters of moon-based InSAR. Based on the ephemeris data, the position, liberation and attitude of earth and moon will be obtained, and the position of the moon-base SAR sensor can be obtained by coordinate transformation from fixed seleno-centric coordinate systems to terrestrial coordinate systems, together with the Distance-Doppler equation, the positioning model will be analyzed; after establish of moon-based InSAR baseline equation, the different baseline error will be analyzed, the influence of the moon-based InSAR baseline to earth observation application will be obtained.

  10. Geographic Object-Based Image Analysis - Towards a new paradigm.

    Science.gov (United States)

    Blaschke, Thomas; Hay, Geoffrey J; Kelly, Maggi; Lang, Stefan; Hofmann, Peter; Addink, Elisabeth; Queiroz Feitosa, Raul; van der Meer, Freek; van der Werff, Harald; van Coillie, Frieke; Tiede, Dirk

    2014-01-01

    The amount of scientific literature on (Geographic) Object-based Image Analysis - GEOBIA has been and still is sharply increasing. These approaches to analysing imagery have antecedents in earlier research on image segmentation and use GIS-like spatial analysis within classification and feature extraction approaches. This article investigates these development and its implications and asks whether or not this is a new paradigm in remote sensing and Geographic Information Science (GIScience). We first discuss several limitations of prevailing per-pixel methods when applied to high resolution images. Then we explore the paradigm concept developed by Kuhn (1962) and discuss whether GEOBIA can be regarded as a paradigm according to this definition. We crystallize core concepts of GEOBIA, including the role of objects, of ontologies and the multiplicity of scales and we discuss how these conceptual developments support important methods in remote sensing such as change detection and accuracy assessment. The ramifications of the different theoretical foundations between the 'per-pixel paradigm' and GEOBIA are analysed, as are some of the challenges along this path from pixels, to objects, to geo-intelligence. Based on several paradigm indications as defined by Kuhn and based on an analysis of peer-reviewed scientific literature we conclude that GEOBIA is a new and evolving paradigm.

  11. Improved nowcasting of precipitation based on convective analysis fields

    Directory of Open Access Journals (Sweden)

    T. Haiden

    2007-04-01

    Full Text Available The high-resolution analysis and nowcasting system INCA (Integrated Nowcasting through Comprehensive Analysis developed at the Austrian national weather service provides three-dimensional fields of temperature, humidity, and wind on an hourly basis, and two-dimensional fields of precipitation rate in 15 min intervals. The system operates on a horizontal resolution of 1 km and a vertical resolution of 100–200 m. It combines surface station data, remote sensing data (radar, satellite, forecast fields of the numerical weather prediction model ALADIN, and high-resolution topographic data. An important application of the INCA system is nowcasting of convective precipitation. Based on fine-scale temperature, humidity, and wind analyses a number of convective analysis fields are routinely generated. These fields include convective boundary layer (CBL flow convergence and specific humidity, lifted condensation level (LCL, convective available potential energy (CAPE, convective inhibition (CIN, and various convective stability indices. Based on the verification of areal precipitation nowcasts it is shown that the pure translational forecast of convective cells can be improved by using a decision algorithm which is based on a subset of the above fields, combined with satellite products.

  12. Weather data analysis based on typical weather sequence analysis. Application: energy building simulation

    CERN Document Server

    David, Mathieu; Garde, Francois; Boyer, Harry

    2014-01-01

    In building studies dealing about energy efficiency and comfort, simulation software need relevant weather files with optimal time steps. Few tools generate extreme and mean values of simultaneous hourly data including correlation between the climatic parameters. This paper presents the C++ Runeole software based on typical weather sequences analysis. It runs an analysis process of a stochastic continuous multivariable phenomenon with frequencies properties applied to a climatic database. The database analysis associates basic statistics, PCA (Principal Component Analysis) and automatic classifications. Different ways of applying these methods will be presented. All the results are stored in the Runeole internal database that allows an easy selection of weather sequences. The extreme sequences are used for system and building sizing and the mean sequences are used for the determination of the annual cooling loads as proposed by Audrier-Cros (Audrier-Cros, 1984). This weather analysis was tested with the datab...

  13. A linear mixture analysis-based compression for hyperspectral image analysis

    Energy Technology Data Exchange (ETDEWEB)

    C. I. Chang; I. W. Ginsberg

    2000-06-30

    In this paper, the authors present a fully constrained least squares linear spectral mixture analysis-based compression technique for hyperspectral image analysis, particularly, target detection and classification. Unlike most compression techniques that directly deal with image gray levels, the proposed compression approach generates the abundance fractional images of potential targets present in an image scene and then encodes these fractional images so as to achieve data compression. Since the vital information used for image analysis is generally preserved and retained in the abundance fractional images, the loss of information may have very little impact on image analysis. In some occasions, it even improves analysis performance. Airborne visible infrared imaging spectrometer (AVIRIS) data experiments demonstrate that it can effectively detect and classify targets while achieving very high compression ratios.

  14. Time-resolved quantitative phosphoproteomics

    DEFF Research Database (Denmark)

    Verano-Braga, Thiago; Schwämmle, Veit; Sylvester, Marc;

    2012-01-01

    identification and phosphorylation site localization. Of these, 121 sites on 79 proteins had their phosphorylation levels significantly changed by Ang-(1-7). Our data suggest that the antiproliferative activity of Ang-(1-7) is due to the activation or inactivation of several target phosphoproteins...

  15. Emergy Analysis and Sustainability Efficiency Analysis of Different Crop-Based Biodiesel in Life Cycle Perspective

    DEFF Research Database (Denmark)

    Ren, Jingzheng; Manzardo, Alessandro; Mazzi, Anna

    2013-01-01

    Biodiesel as a promising alternative energy resource has been a hot spot in chemical engineering nowadays, but there is also an argument about the sustainability of biodiesel. In order to analyze the sustainability of biodiesel production systems and select the most sustainable scenario, various...... kinds of crop-based biodiesel including soybean-, rapeseed-, sunflower-, jatropha- and palm-based biodiesel production options are studied by emergy analysis; soybean-based scenario is recognized as the most sustainable scenario that should be chosen for further study in China. DEA method is used...... to evaluate the sustainability efficiencies of these options, and the biodiesel production systems based on soybean, sunflower, and palm are considered as DEA efficient, whereas rapeseed-based and jatropha-based scenarios are needed to be improved, and the improved methods have also been specified....

  16. Emergy Analysis and Sustainability Efficiency Analysis of Different Crop-Based Biodiesel in Life Cycle Perspective

    Directory of Open Access Journals (Sweden)

    Jingzheng Ren

    2013-01-01

    Full Text Available Biodiesel as a promising alternative energy resource has been a hot spot in chemical engineering nowadays, but there is also an argument about the sustainability of biodiesel. In order to analyze the sustainability of biodiesel production systems and select the most sustainable scenario, various kinds of crop-based biodiesel including soybean-, rapeseed-, sunflower-, jatropha- and palm-based biodiesel production options are studied by emergy analysis; soybean-based scenario is recognized as the most sustainable scenario that should be chosen for further study in China. DEA method is used to evaluate the sustainability efficiencies of these options, and the biodiesel production systems based on soybean, sunflower, and palm are considered as DEA efficient, whereas rapeseed-based and jatropha-based scenarios are needed to be improved, and the improved methods have also been specified.

  17. Emergy analysis and sustainability efficiency analysis of different crop-based biodiesel in life cycle perspective.

    Science.gov (United States)

    Ren, Jingzheng; Manzardo, Alessandro; Mazzi, Anna; Fedele, Andrea; Scipioni, Antonio

    2013-01-01

    Biodiesel as a promising alternative energy resource has been a hot spot in chemical engineering nowadays, but there is also an argument about the sustainability of biodiesel. In order to analyze the sustainability of biodiesel production systems and select the most sustainable scenario, various kinds of crop-based biodiesel including soybean-, rapeseed-, sunflower-, jatropha- and palm-based biodiesel production options are studied by emergy analysis; soybean-based scenario is recognized as the most sustainable scenario that should be chosen for further study in China. DEA method is used to evaluate the sustainability efficiencies of these options, and the biodiesel production systems based on soybean, sunflower, and palm are considered as DEA efficient, whereas rapeseed-based and jatropha-based scenarios are needed to be improved, and the improved methods have also been specified.

  18. Morphometric analysis of the cranial base in Asians.

    Science.gov (United States)

    Chang, Hong-Po; Liu, Pao-Hsin; Tseng, Yu-Chuan; Yang, Yi-Hsin; Pan, Chin-Yun; Chou, Szu-Ting

    2014-01-01

    This study tested the hypothesis that developmental heterogeneity in cranial base morphology increases the prevalence of Class III malocclusion and mandibular prognathism in Asians. Thin-plate spline (TPS) graphical analysis of lateral cephalometric radiographs of the cranial base and the upper midface configuration were compared between a European-American group (24 females and 31 males) and four Asian ethnic groups (100 Chinese, 100 Japanese, 100 Korean and 100 Taiwanese; 50 females and 50 males per group) of young adults with clinically acceptable occlusion and facial profiles. Procrustes analysis was performed to identify statistically significant differences in each configuration of landmarks (P expansion in the anterior portion of the cranial base and upper midface region. The most posterior cranial base region also showed horizontal compression between the basion and Bolton point, with forward displacement of the articulare. Facial flatness and anterior displacement of the temporomandibular joint, resulting from a relative retrusion of the nasomaxillary complex and a relative forward position of the mandible were also noted. These features that tend to cause a prognathic mandible and/or retruded midface indicate a morphologic predisposition of Asian populations for Class III malocclusion.

  19. Direct DNA Analysis with Paper-Based Ion Concentration Polarization.

    Science.gov (United States)

    Gong, Max M; Nosrati, Reza; San Gabriel, Maria C; Zini, Armand; Sinton, David

    2015-11-01

    DNA analysis is essential for diagnosis and monitoring of many diseases. Conventional DNA testing is generally limited to the laboratory. Increasing access to relevant technologies can improve patient care and outcomes in both developed and developing regions. Here, we demonstrate direct DNA analysis in paper-based devices, uniquely enabled by ion concentration polarization at the interface of patterned nanoporous membranes in paper (paper-based ICP). Hepatitis B virus DNA targets in human serum are simultaneously preconcentrated, separated, and detected in a single 10 min operation. A limit of detection of 150 copies/mL is achieved without prior viral load amplification, sufficient for early diagnosis of hepatitis B. We clinically assess the DNA integrity of sperm cells in raw human semen samples. The percent DNA fragmentation results from the paper-based ICP devices strongly correlate (R(2) = 0.98) with the sperm chromatin structure assay. In all cases, agreement was 100% with respect to the clinical decision. Paper-based ICP can provide inexpensive and accessible advanced molecular diagnostics.

  20. Reliability analysis method for slope stability based on sample weight

    Directory of Open Access Journals (Sweden)

    Zhi-gang YANG

    2009-09-01

    Full Text Available The single safety factor criteria for slope stability evaluation, derived from the rigid limit equilibrium method or finite element method (FEM, may not include some important information, especially for steep slopes with complex geological conditions. This paper presents a new reliability method that uses sample weight analysis. Based on the distribution characteristics of random variables, the minimal sample size of every random variable is extracted according to a small sample t-distribution under a certain expected value, and the weight coefficient of each extracted sample is considered to be its contribution to the random variables. Then, the weight coefficients of the random sample combinations are determined using the Bayes formula, and different sample combinations are taken as the input for slope stability analysis. According to one-to-one mapping between the input sample combination and the output safety coefficient, the reliability index of slope stability can be obtained with the multiplication principle. Slope stability analysis of the left bank of the Baihetan Project is used as an example, and the analysis results show that the present method is reasonable and practicable for the reliability analysis of steep slopes with complex geological conditions.

  1. Voxel-based texture analysis of the brain.

    Science.gov (United States)

    Maani, Rouzbeh; Yang, Yee Hong; Kalra, Sanjay

    2015-01-01

    This paper presents a novel voxel-based method for texture analysis of brain images. Texture analysis is a powerful quantitative approach for analyzing voxel intensities and their interrelationships, but has been thus far limited to analyzing regions of interest. The proposed method provides a 3D statistical map comparing texture features on a voxel-by-voxel basis. The validity of the method was examined on artificially generated effects as well as on real MRI data in Alzheimer's Disease (AD). The artificially generated effects included hyperintense and hypointense signals added to T1-weighted brain MRIs from 30 healthy subjects. The AD dataset included 30 patients with AD and 30 age/sex matched healthy control subjects. The proposed method detected artificial effects with high accuracy and revealed statistically significant differences between the AD and control groups. This paper extends the usage of texture analysis beyond the current region of interest analysis to voxel-by-voxel 3D statistical mapping and provides a hypothesis-free analysis tool to study cerebral pathology in neurological diseases.

  2. Validation Database Based Thermal Analysis of an Advanced RPS Concept

    Science.gov (United States)

    Balint, Tibor S.; Emis, Nickolas D.

    2006-01-01

    Advanced RPS concepts can be conceived, designed and assessed using high-end computational analysis tools. These predictions may provide an initial insight into the potential performance of these models, but verification and validation are necessary and required steps to gain confidence in the numerical analysis results. This paper discusses the findings from a numerical validation exercise for a small advanced RPS concept, based on a thermal analysis methodology developed at JPL and on a validation database obtained from experiments performed at Oregon State University. Both the numerical and experimental configurations utilized a single GPHS module enabled design, resembling a Mod-RTG concept. The analysis focused on operating and environmental conditions during the storage phase only. This validation exercise helped to refine key thermal analysis and modeling parameters, such as heat transfer coefficients, and conductivity and radiation heat transfer values. Improved understanding of the Mod-RTG concept through validation of the thermal model allows for future improvements to this power system concept.

  3. Forming Teams for Teaching Programming based on Static Code Analysis

    Directory of Open Access Journals (Sweden)

    Davis Arosemena-Trejos

    2012-03-01

    Full Text Available The use of team for teaching programming can be effective in the classroom because it helps students to generate and acquire new knowledge in less time, but these groups to be formed without taking into account some respects, may cause an adverse effect on the teaching-learning process. This paper proposes a tool for the formation of team based on the semantics of source code (SOFORG. This semantics is based on metrics extracted from the preferences, styles and good programming practices. All this is achieved through a static analysis of code that each student develops. In this way, you will have a record of students with the information extracted; it evaluates the best formation of teams in a given course. The team€™s formations are based on programming styles, skills, pair programming or with leader.

  4. Climate policy decisions require policy-based lifecycle analysis.

    Science.gov (United States)

    Bento, Antonio M; Klotz, Richard

    2014-05-20

    Lifecycle analysis (LCA) metrics of greenhouse gas emissions are increasingly being used to select technologies supported by climate policy. However, LCAs typically evaluate the emissions associated with a technology or product, not the impacts of policies. Here, we show that policies supporting the same technology can lead to dramatically different emissions impacts per unit of technology added, due to multimarket responses to the policy. Using a policy-based consequential LCA, we find that the lifecycle emissions impacts of four US biofuel policies range from a reduction of 16.1 gCO2e to an increase of 24.0 gCO2e per MJ corn ethanol added by the policy. The differences between these results and representative technology-based LCA measures, which do not account for the policy instrument driving the expansion in the technology, illustrate the need for policy-based LCA measures when informing policy decision making.

  5. Forming Teams for Teaching Programming based on Static Code Analysis

    CERN Document Server

    Arosemena-Trejos, Davis; Clunie, Clifton

    2012-01-01

    The use of team for teaching programming can be effective in the classroom because it helps students to generate and acquire new knowledge in less time, but these groups to be formed without taking into account some respects, may cause an adverse effect on the teaching-learning process. This paper proposes a tool for the formation of team based on the semantics of source code (SOFORG). This semantics is based on metrics extracted from the preferences, styles and good programming practices. All this is achieved through a static analysis of code that each student develops. In this way, you will have a record of students with the information extracted; it evaluates the best formation of teams in a given course. The team's formations are based on programming styles, skills, pair programming or with leader.

  6. Least Dependent Component Analysis Based on Mutual Information

    CERN Document Server

    Stögbauer, H; Astakhov, S A; Grassberger, P; St\\"ogbauer, Harald; Kraskov, Alexander; Astakhov, Sergey A.; Grassberger, Peter

    2004-01-01

    We propose to use precise estimators of mutual information (MI) to find least dependent components in a linearly mixed signal. On the one hand this seems to lead to better blind source separation than with any other presently available algorithm. On the other hand it has the advantage, compared to other implementations of `independent' component analysis (ICA) some of which are based on crude approximations for MI, that the numerical values of the MI can be used for: (i) estimating residual dependencies between the output components; (ii) estimating the reliability of the output, by comparing the pairwise MIs with those of re-mixed components; (iii) clustering the output according to the residual interdependencies. For the MI estimator we use a recently proposed k-nearest neighbor based algorithm. For time sequences we combine this with delay embedding, in order to take into account non-trivial time correlations. After several tests with artificial data, we apply the resulting MILCA (Mutual Information based ...

  7. Model-Based Evaluation Of System Scalability: Bandwidth Analysis For Smartphone-Based Biosensing Applications

    DEFF Research Database (Denmark)

    Patou, François; Madsen, Jan; Dimaki, Maria

    2016-01-01

    -engineering efforts for scaling a system specification efficaciously. We demonstrate the value of our methodology by investigating a smartphone-based biosensing instrumentation platform. Specifically, we carry out scalability analysis for the system’s bandwidth specification: the maximum analog voltage waveform...

  8. Average Case Analysis of Some Elimination-Based Data-Flow Analysis Algorithms

    OpenAIRE

    2008-01-01

    The average case of some elimination-based data-flow analysis algorithms is analyzed in a mathematical way. Besides this allows for comparing the timing behavior of the algorithms, it also provides insights into how relevant the underlying statistics are when compared to practical settings.

  9. Ca analysis: an Excel based program for the analysis of intracellular calcium transients including multiple, simultaneous regression analysis.

    Science.gov (United States)

    Greensmith, David J

    2014-01-01

    Here I present an Excel based program for the analysis of intracellular Ca transients recorded using fluorescent indicators. The program can perform all the necessary steps which convert recorded raw voltage changes into meaningful physiological information. The program performs two fundamental processes. (1) It can prepare the raw signal by several methods. (2) It can then be used to analyze the prepared data to provide information such as absolute intracellular Ca levels. Also, the rates of change of Ca can be measured using multiple, simultaneous regression analysis. I demonstrate that this program performs equally well as commercially available software, but has numerous advantages, namely creating a simplified, self-contained analysis workflow.

  10. Techno-Economic Analysis of Biofuels Production Based on Gasification

    Energy Technology Data Exchange (ETDEWEB)

    Swanson, R. M.; Platon, A.; Satrio, J. A.; Brown, R. C.; Hsu, D. D.

    2010-11-01

    This study compares capital and production costs of two biomass-to-liquid production plants based on gasification. The first biorefinery scenario is an oxygen-fed, low-temperature (870?C), non-slagging, fluidized bed gasifier. The second scenario is an oxygen-fed, high-temperature (1,300?C), slagging, entrained flow gasifier. Both are followed by catalytic Fischer-Tropsch synthesis and hydroprocessing to naphtha-range (gasoline blend stock) and distillate-range (diesel blend stock) liquid fractions. Process modeling software (Aspen Plus) is utilized to organize the mass and energy streams and cost estimation software is used to generate equipment costs. Economic analysis is performed to estimate the capital investment and operating costs. Results show that the total capital investment required for nth plant scenarios is $610 million and $500 million for high-temperature and low-temperature scenarios, respectively. Product value (PV) for the high-temperature and low-temperature scenarios is estimated to be $4.30 and $4.80 per gallon of gasoline equivalent (GGE), respectively, based on a feedstock cost of $75 per dry short ton. Sensitivity analysis is also performed on process and economic parameters. This analysis shows that total capital investment and feedstock cost are among the most influential parameters affecting the PV.

  11. Analysis of Host-Based and Network-Based Intrusion Detection System

    Directory of Open Access Journals (Sweden)

    Amrit Pal Singh

    2014-07-01

    Full Text Available Intrusion-detection systems (IDS aim at de-tecting attacks against computer systems and networks or, in general, against information systems. Its basic aim is to protect the system against malwares and unauthorized access of a network or a system. Intrusion Detection is of two types Network-IDS and Host Based- IDS. This paper covers the scope of both the types and their result analysis along with their comparison as stated. OSSEC (HIDS is a free, open source host-base intrusion detection system. It performs log analysis, integrity checking, Windows registry monitoring, rootkit detection, time-based alerting and active response. While Snort (NIDS is a lightweight intrusion detection system that can log packets coming across your network and can alert the user regarding any attack. Both are efficient in their own distinct fields.

  12. Finite element analysis of osteoporosis models based on synchrotron radiation

    Science.gov (United States)

    Xu, W.; Xu, J.; Zhao, J.; Sun, J.

    2016-04-01

    With growing pressure of social aging, China has to face the increasing population of osteoporosis patients as well as the whole world. Recently synchrotron radiation has become an essential tool for biomedical exploration with advantage of high resolution and high stability. In order to study characteristic changes in different stages of primary osteoporosis, this research focused on the different periods of osteoporosis of rats based on synchrotron radiation. Both bone histomorphometry analysis and finite element analysis were then carried on according to the reconstructed three dimensional models. Finally, the changes of bone tissue in different periods were compared quantitatively. Histomorphometry analysis showed that the structure of the trabecular in osteoporosis degraded as the bone volume decreased. For femurs, the bone volume fraction (Bone volume/ Total volume, BV/TV) decreased from 69% to 43%. That led to the increase of the thickness of trabecular separation (from 45.05μ m to 97.09μ m) and the reduction of the number of trabecular (from 7.99 mm-1 to 5.97mm-1). Simulation of various mechanical tests with finite element analysis (FEA) indicated that, with the exacerbation of osteoporosis, the bones' ability of resistance to compression, bending and torsion gradually became weaker. The compression stiffness of femurs decreased from 1770.96 Fμ m-1 to 697.41 Fμ m-1, the bending and torsion stiffness were from 1390.80 Fμ m-1 to 566.11 Fμ m-1 and from 2957.28N.m/o to 691.31 N.m/o respectively, indicated the decrease of bone strength, and it matched the histomorphometry analysis. This study suggested that FEA and synchrotron radiation were excellent methods for analysing bone strength conbined with histomorphometry analysis.

  13. Principle-based concept analysis: Caring in nursing education

    Science.gov (United States)

    Salehian, Maryam; Heydari, Abbas; Aghebati, Nahid; Moonaghi, Hossein Karimi; Mazloom, Seyed Reza

    2016-01-01

    Introduction The aim of this principle-based concept analysis was to analyze caring in nursing education and to explain the current state of the science based on epistemologic, pragmatic, linguistic, and logical philosophical principles. Methods A principle-based concept analysis method was used to analyze the nursing literature. The dataset included 46 English language studies, published from 2005 to 2014, and they were retrieved through PROQUEST, MEDLINE, CINAHL, ERIC, SCOPUS, and SID scientific databases. The key dimensions of the data were collected using a validated data-extraction sheet. The four principles of assessing pragmatic utility were used to analyze the data. The data were managed by using MAXQDA 10 software. Results The scientific literature that deals with caring in nursing education relies on implied meaning. Caring in nursing education refers to student-teacher interactions that are formed on the basis of human values and focused on the unique needs of the students (epistemological principle). The result of student-teacher interactions is the development of both the students and the teachers. Numerous applications of the concept of caring in nursing education are available in the literature (pragmatic principle). There is consistency in the meaning of the concept, as a central value of the faculty-student interaction (linguistic principle). Compared with other related concepts, such as “caring pedagogy,” “value-based education,” and “teaching excellence,” caring in nursing education does not have exact and clear conceptual boundaries (logic principle). Conclusion Caring in nursing education was identified as an approach to teaching and learning, and it is formed based on teacher-student interactions and sustainable human values. A greater understanding of the conceptual basis of caring in nursing education will improve the caring behaviors of teachers, create teaching-learning environments, and help experts in curriculum development

  14. Orbital Energy-Based Reaction Analysis of SN2 Reactions

    Directory of Open Access Journals (Sweden)

    Takao Tsuneda

    2016-07-01

    Full Text Available An orbital energy-based reaction analysis theory is presented as an extension of the orbital-based conceptual density functional theory. In the orbital energy-based theory, the orbitals contributing to reactions are interpreted to be valence orbitals giving the largest orbital energy variation from reactants to products. Reactions are taken to be electron transfer-driven when they provide small variations for the gaps between the contributing occupied and unoccupied orbital energies on the intrinsic reaction coordinates in the initial processes. The orbital energy-based theory is then applied to the calculations of several S N2 reactions. Using a reaction path search method, the Cl− + CH3I → ClCH3 + I− reaction, for which another reaction path called “roundabout path” is proposed, is found to have a precursor process similar to the roundabout path just before this SN2 reaction process. The orbital energy-based theory indicates that this precursor process is obviously driven by structural change, while the successor SN2 reaction proceeds through electron transfer between the contributing orbitals. Comparing the calculated results of the SN2 reactions in gas phase and in aqueous solution shows that the contributing orbitals significantly depend on solvent effects and these orbitals can be correctly determined by this theory.

  15. Relational Analysis based Concurrent Multipath Transfer over Heterogeneous Vehicular Networks

    Directory of Open Access Journals (Sweden)

    Hongke Zhang

    2012-09-01

    Full Text Available In recent years, the growing interest in the Intelligent Transportation Systems (ITS has resulted in variety of peer-reviewed publications. Significant results in this area have enabled many civilian and industry applications. As more and more vehicles are equipped with multiple network interfaces, how to efficient utilize the coexistence of Radio Access Technologies (RAT such as WiFi, UMTS and WiMAX to serve a best Concurrent Multipath Transfer (CMT service is still a challenge in ITS. In this paper, we propose GRA-CMT, a novel Grey Relational Analysis (GRA based Concurrent Multipath Transfer, extension for Stream Control Transport Protocol (SCTP. Depending on the advantages of GRA, a GRA-based Data Distribution algorithm is proposed in GRA-CMT to calculate the Grey Relational Coefficient (GRC value of all candidate paths and offer a more efficient data scheduling algorithm, a further proposed GRA-based CMT Retransmission algorithm devotes to select destination for efficient retransmission. Moreover, the GRA-CMT provides a GRA-based CMT Path Selection scheme to manage candidate paths. Sufficient simulation results obtained by a close realistic simulation topology show how GRA-CMT outperforms existing CMT in heterogeneous SCTP-based vehicular networks.

  16. Weight measurement using image-based pose analysis

    Institute of Scientific and Technical Information of China (English)

    Hong Zhang; Kui Zhang; Ying Mu; Ning Yao; Robert J. Sclabassi; Mingui Sun

    2008-01-01

    Image-based gait analysis as a means of biometric identification has attracted much research attention.Most of the existing methods focus on human identification,posture analysis and movement tracking.There have been few investigations on measuring the carried load based on the carrier's gait characteristics by automatic image processing.Nevertheless,this measurement is very useful in a number of applications,such as the study of the carried load on the postural development of children and adolescence.In this paper,we inves-tigate how to automatically estimate the carried weight from a sequence of images.We present a method to extract human gait silhouette based on an observation that humans tend to minimize the energy during motion.We compute several angles of body leaning and deter-mine the relationship of the carried weight,the leaning angles and the centroid location according to a human kinetic study.Our weight determination method has been verified successfully by experiments.

  17. ALGORITHMS FOR TENNIS RACKET ANALYSIS BASED ON MOTION DATA

    Directory of Open Access Journals (Sweden)

    Maria Skublewska-Paszkowska

    2016-09-01

    Full Text Available Modern technologies, such as motion capture systems (both optical and markerless, are more and more frequently used for athlete performance analysis due to their great precision. Optical systems based on the retro-reflective markers allow for tracking motion of multiple objects of various types. These systems compute human kinetic and kinematic parameters based on biomechanical models. Tracking additional objects like a tennis racket is also a very important aspect for analysing the player’s technique and precision. The motion data gathered by motion capture systems may be used for analysing various aspects that may not be recognised by the human eye or a video camera. This paper presents algorithms for analysis of a tennis racket motion during two of the most important tennis strokes: forehand and backhand. An optical Vicon system was used for obtaining the motion data which was the input for the algorithms. They indicate: the velocity of a tennis racket’s head and the racket’s handle based on the trajectories of attached markers as well as the racket’s orientation. The algorithms were implemented and tested on the data obtained from a professional trainer who participated in the research and performed a series of ten strikes, separately for: 1 forehand without a ball, 2 backhand without a ball, 3 forehand with a ball and 4 backhand with a ball. The computed parameters are gathered in tables and visualised in a graph.

  18. Medical diagnostics by laser-based analysis of exhaled breath

    Science.gov (United States)

    Giubileo, Gianfranco

    2002-08-01

    IMany trace gases can be found in the exhaled breath, some of them giving the possibility of a non invasive diagnosis of related diseases or allowing the monitoring of the disease in the course of its therapy. In the present lecture the principle of medical diagnosis based on the breath analysis will be introduced and the detection of trace gases in exhaled breath by high- resolution molecular spectroscopy in the IR spectral region will be discussed. A number of substrates and the optical systems for their laser detection will be reported. The following laser based experimental systems has been realised in the Molecular Spectroscopy Laboratory in ENEA in Frascati for the analysis of specific substances in the exhaled breath. A tuneable diode laser absorption spectroscopy (TDLAS) appartus for the measurement of 13C/12C isotopic ratio in carbon dioxide, a TDLAS apparatus for the detection of CH4 and a CO2 laser based photoacoustic system to detect trace ethylene at atmospheric pressure. The experimental set-up for each one of the a.m. optical systems will be shown and the related medical applications will be illustrated. The concluding remarks will be focuses on chemical species that are of major interest for medical people today and their diagnostic ability.

  19. ICWorld: An MMOG-Based Approach to Analysis

    Directory of Open Access Journals (Sweden)

    Wyatt Wong

    2008-01-01

    Full Text Available Intelligence analysts routinely work with "wicked" problems—critical,time-sensitive problems where analytical errors can lead to catastrophic consequences for the nation's security. In the analyst's world, important decisions are often made quickly, and are made based on consuming, understanding, and piecing together enormous volumes of data. The data is not only voluminous, but often fragmented, subjective, inaccurate and fluid.Why does multi-player on-line gaming (MMOG technology matter to the IC? Fundamentally, there are two reasons. The first is technological: stripping away the gamelike content, MMOGs are dynamic systems that represent a physical world, where users are presented with (virtual life-and-death challenges that can only be overcome through planning, collaboration and communication. The second is cultural: the emerging generation of analysts is part of what is sometimes called the "Digital Natives" (Prensky 2001 and is fluent with interactive media. MMOGs enable faster visualization, data manipulation, collaboration and analysis than traditional text and imagery.ICWorld is an MMOG approach to intelligence analysis that fuses ideasfrom experts in the fields of gaming and data visualization, with knowledge of current and future intelligence analysis processes and tools. The concept has evolved over the last year as a result of evaluations by allsource analysts from around the IC. When fully developed, the Forterra team believes that ICWorld will fundamentally address major shortcomings of intelligence analysis, and dramatically improve the effectiveness of intelligence products.

  20. SMV model-based safety analysis of software requirements

    Energy Technology Data Exchange (ETDEWEB)

    Koh, Kwang Yong [Department of Nuclear and Quantum Engineering, Korea Advanced Institute of Science and Technology, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701 (Korea, Republic of); Seong, Poong Hyun [Department of Nuclear and Quantum Engineering, Korea Advanced Institute of Science and Technology, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701 (Korea, Republic of)], E-mail: phseong@kaist.ac.kr

    2009-02-15

    Fault tree analysis (FTA) is one of the most frequently applied safety analysis techniques when developing safety-critical industrial systems such as software-based emergency shutdown systems of nuclear power plants and has been used for safety analysis of software requirements in the nuclear industry. However, the conventional method for safety analysis of software requirements has several problems in terms of correctness and efficiency; the fault tree generated from natural language specifications may contain flaws or errors while the manual work of safety verification is very labor-intensive and time-consuming. In this paper, we propose a new approach to resolve problems of the conventional method; we generate a fault tree from a symbolic model verifier (SMV) model, not from natural language specifications, and verify safety properties automatically, not manually, by a model checker SMV. To demonstrate the feasibility of this approach, we applied it to shutdown system 2 (SDS2) of Wolsong nuclear power plant (NPP). In spite of subtle ambiguities present in the approach, the results of this case study demonstrate its overall feasibility and effectiveness.

  1. Psychoacoustic Music Analysis Based on the Discrete Wavelet Packet Transform

    Directory of Open Access Journals (Sweden)

    Xing He

    2008-01-01

    Full Text Available Psychoacoustical computational models are necessary for the perceptual processing of acoustic signals and have contributed significantly in the development of highly efficient audio analysis and coding. In this paper, we present an approach for the psychoacoustic analysis of musical signals based on the discrete wavelet packet transform. The proposed method mimics the multiresolution properties of the human ear closer than other techniques and it includes simultaneous and temporal auditory masking. Experimental results show that this method provides better masking capabilities and it reduces the signal-to-masking ratio substantially more than other approaches, without introducing audible distortion. This model can lead to greater audio compression by permitting further bit rate reduction and more secure watermarking by providing greater signal space for information hiding.

  2. Ground extraction from airborne laser data based on wavelet analysis

    Science.gov (United States)

    Xu, Liang; Yang, Yan; Jiang, Bowen; Li, Jia

    2007-11-01

    With the advantages of high resolution and accuracy, airborne laser scanning data are widely used in topographic mapping. In order to generate a DTM, measurements from object features such as buildings, vehicles and vegetation have to be classified and removed. However, the automatic extraction of bare earth from point clouds acquired by airborne laser scanning equipment remains a problem in LIDAR data filtering nowadays. In this paper, a filter algorithm based on wavelet analysis is proposed. Relying on the capability of detecting discontinuities of continuous wavelet transform and the feature of multi-resolution analysis, the object points can be removed, while ground data are preserved. In order to evaluate the performance of this approach, we applied it to the data set used in the ISPRS filter test in 2003. 15 samples have been tested by the proposed approach. Results showed that it filtered most of the objects like vegetation and buildings, and extracted a well defined ground model.

  3. Carbon nanotube based VLSI interconnects analysis and design

    CERN Document Server

    Kaushik, Brajesh Kumar

    2015-01-01

    The brief primarily focuses on the performance analysis of CNT based interconnects in current research scenario. Different CNT structures are modeled on the basis of transmission line theory. Performance comparison for different CNT structures illustrates that CNTs are more promising than Cu or other materials used in global VLSI interconnects. The brief is organized into five chapters which mainly discuss: (1) an overview of current research scenario and basics of interconnects; (2) unique crystal structures and the basics of physical properties of CNTs, and the production, purification and applications of CNTs; (3) a brief technical review, the geometry and equivalent RLC parameters for different single and bundled CNT structures; (4) a comparative analysis of crosstalk and delay for different single and bundled CNT structures; and (5) various unique mixed CNT bundle structures and their equivalent electrical models.

  4. GIS-based poverty and population distribution analysis in China

    Science.gov (United States)

    Cui, Jing; Wang, Yingjie; Yan, Hong

    2009-07-01

    Geographically, poverty status is not only related with social-economic factors but also strongly affected by geographical environment. In the paper, GIS-based poverty and population distribution analysis method is introduced for revealing their regional differences. More than 100000 poor villages and 592 national key poor counties are chosen for the analysis. The results show that poverty distribution tends to concentrate in most of west China and mountainous rural areas of mid China. Furthermore, the fifth census data are overlaid to those poor areas in order to gain its internal diversity of social-economic characteristics. By overlaying poverty related social-economic parameters, such as sex ratio, illiteracy, education level, percentage of ethnic minorities, family composition, finding shows that poverty distribution is strongly correlated with high illiteracy rate, high percentage minorities, and larger family member.

  5. Multiwave velocity analysis based on Gaussian beam prestack depth migration

    Institute of Scientific and Technical Information of China (English)

    Han Jian-Guang; Wang Yun; Han Ning; Xing Zhan-Tao; Lu Jun

    2014-01-01

    Prestack depth migration of multicomponent seismic data improves the imaging accuracy of subsurface complex geological structures. An accurate velocityfi eld is critical to accurate imaging. Gaussian beam migration was used to perform multicomponent migration velocity analysis of PP- and PS-waves. First, PP- and PS-wave Gaussian beam prestack depth migration algorithms that operate on common-offset gathers are presented to extract offset-domain common-image gathers of PP- and PS-waves. Second, based on the residual moveout equation, the migration velocity fields of P- and S-waves are updated. Depth matching is used to ensure that the depth of the target layers in the PP- and PS-wave migration profi les are consistent, and high-precision P- and S-wave velocities are obtained. Finally, synthetic andfi eld seismic data suggest that the method can be used effectively in multiwave migration velocity analysis.

  6. Bayesian Model Selection with Network Based Diffusion Analysis.

    Science.gov (United States)

    Whalen, Andrew; Hoppitt, William J E

    2016-01-01

    A number of recent studies have used Network Based Diffusion Analysis (NBDA) to detect the role of social transmission in the spread of a novel behavior through a population. In this paper we present a unified framework for performing NBDA in a Bayesian setting, and demonstrate how the Watanabe Akaike Information Criteria (WAIC) can be used for model selection. We present a specific example of applying this method to Time to Acquisition Diffusion Analysis (TADA). To examine the robustness of this technique, we performed a large scale simulation study and found that NBDA using WAIC could recover the correct model of social transmission under a wide range of cases, including under the presence of random effects, individual level variables, and alternative models of social transmission. This work suggests that NBDA is an effective and widely applicable tool for uncovering whether social transmission underpins the spread of a novel behavior, and may still provide accurate results even when key model assumptions are relaxed.

  7. Dynamic analysis of granite rockburst based on the PIV technique

    Institute of Scientific and Technical Information of China (English)

    Wang Hongjian; Liu Da’an; Gong Weili; Li Liyun

    2015-01-01

    This paper describes the deep rockburst simulation system to reproduce the granite instantaneous rock-burst process. Based on the PIV (Particle Image Velocimetry) technique, quantitative analysis of a rock-burst, the images of tracer particle, displacement and strain fields can be obtained, and the debris trajectory described. According to the observation of on-site tests, the dynamic rockburst is actually a gas–solid high speed flow process, which is caused by the interaction of rock fragments and surrounding air. With the help of analysis on high speed video and PIV images, the granite rockburst failure process is composed of six stages of platey fragment spalling and debris ejection. Meanwhile, the elastic energy for these six stages has been calculated to study the energy variation. The results indicate that the rockburst process can be summarized as:an initiating stage, intensive developing stage and gradual decay stage. This research will be helpful for our further understanding of the rockburst mechanism.

  8. Analysis of structural macroeconomic indicators based on harmony approach

    Directory of Open Access Journals (Sweden)

    Knyshenko, Tetyana

    2012-05-01

    Full Text Available In the article the application of fractal theory and proportions of «gold section» is considered to the analysis of macroeconomic indicators, the criteria of optimum and efficiency of structure of economy are selected. An economy is represented as treelike fractal, every level of which is characterized by potential of profits (by the issue and necessity in charges. The types of structural subsystems in an economy are selected: permanent and temporal, the last arise up at violation of systems optimum. An author drew conclusion that on the certain types of economic activity structural subsystems can be effective, but they can not be named optimum, as there are deviations from the «gold section» at the certain type of the technological mode, in the issue, intermediate consumption and GVP. A conclusion about the type of the technological mode based on analysis of investment of spheres and types of economic activity in forming of macroeconomic indexes.

  9. Analysis and design of a smart card based authentication protocol

    Institute of Scientific and Technical Information of China (English)

    Kuo-Hui YEH; Kuo-Yu TSAI; Jia-Li HOU

    2013-01-01

    Numerous smart card based authentication protocols have been proposed to provide strong system security and robust individual privacy for communication between parties these days. Nevertheless, most of them do not provide formal analysis proof, and the security robustness is doubtful. Chang and Cheng (2011) proposed an efficient remote authentication protocol with smart cards and claimed that their proposed protocol could support secure communication in a multi-server environment. Unfortunately, there are opportunities for security enhancement in current schemes. In this paper, we identify the major weakness, i.e., session key disclosure, of a recently published protocol. We consequently propose a novel authentication scheme for a multi-server envi-ronment and give formal analysis proofs for security guarantees.

  10. Seismic Base Isolation Analysis for PASCAR Liquid Metal Reactor

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Kuk Hee; Yoo, Bong; Kim, Yun Jae [Korea Univ., Seoul (Korea, Republic of)

    2008-10-15

    This paper presents a study for developing a seismic isolation system for the PASCAR (Proliferation resistant, Accident-tolerant, Self-supported, Capsular and Assured Reactor) liquid metal reactor design. PASCAR use lead-bismuth eutectic (LBE) as coolant. Because the density (10,000kg/m{sup 3}) of LBE coolant is very heavier than sodium coolant and water, this presents a challenge to designers of the seismic isolation systems that will be used with these heavy liquid metal reactors. Finite element analysis is adapted to determine the characteristics of the isolator device. Results are presented from a study on the use of three-dimensional seismic isolation devices to the full-scale reactor. The seismic analysis responses of the two-dimensional and the three-dimensional isolation systems for the PASCAR are compared with that of the conventional fixed base system.

  11. Kernel-based fisher discriminant analysis for hyperspectral target detection

    Institute of Scientific and Technical Information of China (English)

    GU Yan-feng; ZHANG Ye; YOU Di

    2007-01-01

    A new method based on kernel Fisher discriminant analysis (KFDA) is proposed for target detection of hyperspectral images. The KFDA combines kernel mapping derived from support vector machine and the classical linear Fisher discriminant analysis (LFDA), and it possesses good ability to process nonlinear data such as hyperspectral images. According to the Fisher rule that the ratio of the between-class and within-class scatters is maximized, the KFDA is used to obtain a set of optimal discriminant basis vectors in high dimensional feature space. All pixels in the hyperspectral images are projected onto the discriminant basis vectors and the target detection is performed according to the projection result. The numerical experiments are performed on hyperspectral data with 126 bands collected by Airborne Visible/Infrared Imaging Spectrometer (AVIRIS). The experimental results show the effectiveness of the proposed detection method and prove that this method has good ability to overcome small sample size and spectral variability in the hyperspectral target detection.

  12. Earthquake Analysis of Structure by Base Isolation Technique in SAP

    Directory of Open Access Journals (Sweden)

    T. Subramani

    2014-06-01

    Full Text Available This paper presents an overview of the present state of base isolation techniques with special emphasis and a brief on other techniques developed world over for mitigating earthquake forces on the structures. The dynamic analysis procedure for isolated structures is briefly explained. The provisions of FEMA 450 for base isolated structures are highlighted. The effects of base isolation on structures located on soft soils and near active faults are given in brief. Simple case study on natural base isolation using naturally available soils is presented. Also, the future areas of research are indicated. Earthquakes are one of nature IS greatest hazards; throughout historic time they have caused significant loss offline and severe damage to property, especially to man-made structures. On the other hand, earthquakes provide architects and engineers with a number of important design criteria foreign to the normal design process. From well established procedures reviewed by many researchers, seismic isolation may be used to provide an effective solution for a wide range of seismic design problems. The application of the base isolation techniques to protect structures against damage from earthquake attacks has been considered as one of the most effective approaches and has gained increasing acceptance during the last two decades. This is because base isolation limits the effects of the earthquake attack, a flexible base largely decoupling the structure from the ground motion, and the structural response accelerations are usually less than the ground acceleration. In general, the increase of additional viscous damping in the structure may reduce displacement and acceleration responses of the structure. This study also seeks to evaluate the effects of additional damping on the seismic response when compared with structures without additional damping for the different ground motions.

  13. The calcium-dependent protein kinase 3 of toxoplasma influences basal calcium levels and functions beyond egress as revealed by quantitative phosphoproteome analysis.

    OpenAIRE

    Moritz Treeck; Sanders, John L.; Rajshekhar Y Gaji; Kacie A LaFavers; Child, Matthew A.; Gustavo Arrizabalaga; Elias, Joshua E.; John C Boothroyd

    2014-01-01

    Calcium-dependent protein kinases (CDPKs) are conserved in plants and apicomplexan parasites. In Toxoplasma gondii, TgCDPK3 regulates parasite egress from the host cell in the presence of a calcium-ionophore. The targets and the pathways that the kinase controls, however, are not known. To identify pathways regulated by TgCDPK3, we measured relative phosphorylation site usage in wild type and TgCDPK3 mutant and knock-out parasites by quantitative mass-spectrometry using stable isotope-labelin...

  14. Analysis of manufacturing based on object oriented discrete event simulation

    Directory of Open Access Journals (Sweden)

    Eirik Borgen

    1990-01-01

    Full Text Available This paper describes SIMMEK, a computer-based tool for performing analysis of manufacturing systems, developed at the Production Engineering Laboratory, NTH-SINTEF. Its main use will be in analysis of job shop type of manufacturing. But certain facilities make it suitable for FMS as well as a production line manufacturing. This type of simulation is very useful in analysis of any types of changes that occur in a manufacturing system. These changes may be investments in new machines or equipment, a change in layout, a change in product mix, use of late shifts, etc. The effects these changes have on for instance the throughput, the amount of VIP, the costs or the net profit, can be analysed. And this can be done before the changes are made, and without disturbing the real system. Simulation takes into consideration, unlike other tools for analysis of manufacturing systems, uncertainty in arrival rates, process and operation times, and machine availability. It also shows the interaction effects a job which is late in one machine, has on the remaining machines in its route through the layout. It is these effects that cause every production plan not to be fulfilled completely. SIMMEK is based on discrete event simulation, and the modeling environment is object oriented. The object oriented models are transformed by an object linker into data structures executable by the simulation kernel. The processes of the entity objects, i.e. the products, are broken down to events and put into an event list. The user friendly graphical modeling environment makes it possible for end users to build models in a quick and reliable way, using terms from manufacturing. Various tests and a check of model logic are helpful functions when testing validity of the models. Integration with software packages, with business graphics and statistical functions, is convenient in the result presentation phase.

  15. IMMAN: free software for information theory-based chemometric analysis.

    Science.gov (United States)

    Urias, Ricardo W Pino; Barigye, Stephen J; Marrero-Ponce, Yovani; García-Jacas, César R; Valdes-Martiní, José R; Perez-Gimenez, Facundo

    2015-05-01

    The features and theoretical background of a new and free computational program for chemometric analysis denominated IMMAN (acronym for Information theory-based CheMoMetrics ANalysis) are presented. This is multi-platform software developed in the Java programming language, designed with a remarkably user-friendly graphical interface for the computation of a collection of information-theoretic functions adapted for rank-based unsupervised and supervised feature selection tasks. A total of 20 feature selection parameters are presented, with the unsupervised and supervised frameworks represented by 10 approaches in each case. Several information-theoretic parameters traditionally used as molecular descriptors (MDs) are adapted for use as unsupervised rank-based feature selection methods. On the other hand, a generalization scheme for the previously defined differential Shannon's entropy is discussed, as well as the introduction of Jeffreys information measure for supervised feature selection. Moreover, well-known information-theoretic feature selection parameters, such as information gain, gain ratio, and symmetrical uncertainty are incorporated to the IMMAN software ( http://mobiosd-hub.com/imman-soft/ ), following an equal-interval discretization approach. IMMAN offers data pre-processing functionalities, such as missing values processing, dataset partitioning, and browsing. Moreover, single parameter or ensemble (multi-criteria) ranking options are provided. Consequently, this software is suitable for tasks like dimensionality reduction, feature ranking, as well as comparative diversity analysis of data matrices. Simple examples of applications performed with this program are presented. A comparative study between IMMAN and WEKA feature selection tools using the Arcene dataset was performed, demonstrating similar behavior. In addition, it is revealed that the use of IMMAN unsupervised feature selection methods improves the performance of both IMMAN and WEKA

  16. Pyemu: A Python-Based Framework for Linear-Based Model Uncertainty Analysis

    Science.gov (United States)

    White, J.

    2014-12-01

    pyEMU is an open-source python-based framework for model-independent linear-based parameter and predictive uncertainty analysis. The framework is designed to support the analysis of high-dimensional inverse problems that have thousands of parameters and hundreds of thousands of observations. The code is compatible with the PEST and PEST++ software suite, and implements several forms of linear analysis equations, such as Schur's complement for conditional uncertainty propagation and subspace error variance, including a form of error variance analysis of model structural error. These linear analysis equations are the most common and also the most applicable to large-scale environmental models. Several native python operators (such as multiplication, subtraction, addition, exponentiation) have been overloaded to make equation building more concise as well as to achieve speedup with operations involving diagonal matrices. To help ensure pyEMU is intuitive and easy to use, emphasis was placed on flexibility and concise object instantiation. As a result, several types of arguments can be handled elegantly.

  17. Web-Based Instruction and Learning: Analysis and Needs Assessment

    Science.gov (United States)

    Grabowski, Barbara; McCarthy, Marianne; Koszalka, Tiffany

    1998-01-01

    An analysis and needs assessment was conducted to identify kindergarten through grade 14 (K-14) customer needs with regard to using the World Wide Web (WWW) for instruction and to identify obstacles K-14 teachers face in utilizing NASA Learning Technologies products in the classroom. The needs assessment was conducted as part of the Dryden Learning Technologies Project which is a collaboration between Dryden Flight Research Center (DFRC), Edwards, California and Tne Pennsylvania State University (PSU), University Park, Pennsylvania. The overall project is a multiyear effort to conduct research in the development of teacher training and tools for Web-based science, mathematics and technology instruction and learning.

  18. Stellar Population Analysis of Galaxies based on Genetic Algorithms

    Institute of Scientific and Technical Information of China (English)

    Abdel-Fattah Attia; H.A.Ismail; I.M.Selim; A.M.Osman; I.A.Isaa; M.A.Marie; A.A.Shaker

    2005-01-01

    We present a new method for determining the age and relative contribution of different stellar populations in galaxies based on the genetic algorithm.We apply this method to the barred spiral galaxy NGC 3384, using CCD images in U, B, V, R and I bands. This analysis indicates that the galaxy NGC 3384 is mainly inhabited by old stellar population (age > 109 yr). Some problems were encountered when numerical simulations are used for determining the contribution of different stellar populations in the integrated color of a galaxy. The results show that the proposed genetic algorithm can search efficiently through the very large space of the possible ages.

  19. A Developed Algorithm of Apriori Based on Association Analysis

    Institute of Scientific and Technical Information of China (English)

    LI Pingxiang; CHEN Jiangping; BIAN Fuling

    2004-01-01

    A method for mining frequent itemsets by evaluating their probability of supports based on association analysis is presented. This paper obtains the probability of every 1-itemset by scanning the database, then evaluates the probability of every 2-itemset, every 3-itemset, every k-itemset from the frequent 1-itemsets and gains all the candidate frequent itemsets. This paper also scans the database for verifying the support of the candidate frequent itemsets. Last, the frequent itemsets are mined. The method reduces a lot of time of scanning database and shortens the computation time of the algorithm.

  20. Image edge detection based on multi-fractal spectrum analysis

    Institute of Scientific and Technical Information of China (English)

    WANG Shao-yuan; WANG Yao-nan

    2006-01-01

    In this paper,an image edge detection method based on multi-fractal spectrum analysis is presented.The coarse grain H(o)lder exponent of the image pixels is first computed,then,its multi-fractal spectrum is estimated by the kernel estimation method.Finally,the image edge detection is done by means of different multi-fractal spectrum values.Simulation results show that this method is efficient and has better locality compared with the traditional edge detection methods such as the Sobel method.

  1. Virtual estimator for piecewise linear systems based on observability analysis.

    Science.gov (United States)

    Morales-Morales, Cornelio; Adam-Medina, Manuel; Cervantes, Ilse; Vela-Valdés, Luis G; Beltrán, Carlos Daniel García

    2013-02-27

    This article proposes a virtual sensor for piecewise linear systems based on observability analysis that is in function of a commutation law related with the system's outpu. This virtual sensor is also known as a state estimator. Besides, it presents a detector of active mode when the commutation sequences of each linear subsystem are arbitrary and unknown. For the previous, this article proposes a set of virtual estimators that discern the commutation paths of the system and allow estimating their output. In this work a methodology in order to test the observability for piecewise linear systems with discrete time is proposed. An academic example is presented to show the obtained results.

  2. Cloud for Distributed Data Analysis Based on the Actor Model

    Directory of Open Access Journals (Sweden)

    Ivan Kholod

    2016-01-01

    Full Text Available This paper describes the construction of a Cloud for Distributed Data Analysis (CDDA based on the actor model. The design uses an approach to map the data mining algorithms on decomposed functional blocks, which are assigned to actors. Using actors allows users to move the computation closely towards the stored data. The process does not require loading data sets into the cloud and allows users to analyze confidential information locally. The results of experiments show that the efficiency of the proposed approach outperforms established solutions.

  3. Differentiation-Based Analysis of Environmental Management and Corporate Performance

    Institute of Scientific and Technical Information of China (English)

    SHAN Dong-ming; MU Xin

    2007-01-01

    By building a duopoly model based on product differentiation, both of the clean firm's and the dirty firm's performances are studied under the assumptions that consumers have different preferences for the product environmental attributes, and that the product cost increases with the environmental attribute. The analysis results show that under either the case with no environmental regulation or that with a tariff levied on the dirty product, the clean firm would always get more profit. In addition, the stricter the regulation is, the more profit the clean firm would obtain. This can verify that from the view of product differentiation, a firm could improve its corporate competitiveness with environmental management.

  4. Machine Learning for Vision-Based Motion Analysis

    CERN Document Server

    Wang, Liang; Cheng, Li; Pietikainen, Matti

    2011-01-01

    Techniques of vision-based motion analysis aim to detect, track, identify, and generally understand the behavior of objects in image sequences. With the growth of video data in a wide range of applications from visual surveillance to human-machine interfaces, the ability to automatically analyze and understand object motions from video footage is of increasing importance. Among the latest developments in this field is the application of statistical machine learning algorithms for object tracking, activity modeling, and recognition. Developed from expert contributions to the first and second In

  5. GIS based analysis of future district heating potential in Denmark

    DEFF Research Database (Denmark)

    Nielsen, Steffen; Möller, Bernd

    2013-01-01

    The physical placement of buildings is important when determining the potential for DH (district heating). Good locations for DH are mainly determined by having both a large heat demand within a certain area and having access to local heat resources. In recent years, the locations of buildings...... in Denmark have been mapped in a heat atlas which includes all buildings and their heat demands. This article focuses on developing a method for assessing the costs associated with supplying these buildings with DH. The analysis is based on the existing DH areas in Denmark. By finding the heat production...

  6. Scale-Dependent Representations of Relief Based on Wavelet Analysis

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Automatic generalization of geographic information is the core of multi-scale representation of spatial data,but the scale-dependent generalization methods are far from abundant because of its extreme complicacy.This paper puts forward a new consistency model about scale-dependent representations of relief based on wavelet analysis,and discusses the thresholds in the model so as to acquire the continual representations of relief with different details between scales.The model not only meets the need of automatic generalization but also is scale-dependent completely.Some practical examples are given.

  7. Facility Layout Based on Sequence Analysis: Design of Flowshops

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jin; WU Zhi-ming

    2009-01-01

    A computer-aided method to design a hybrid layout-tree-shape planar flowlines is presented. In new-type flowshop layout, the common machines shared by several flowlines could be located together in functional sections. The approach combines traditional cell formation techniques with sequence alignment algorithms. Firstly, a sequence analysis based cell formation procedure is adopted; then the operation sequences for parts are aligned to maximize machines adjacency in hyperedge representations; finally a tree-shape planar flowline will be obtained for each part family. With the help of a sample of operation sequences obtained from industry, this algorithm is illustrated.

  8. Measurement Uncertainty Analysis of the Strain Gauge Based Stabilographic Platform

    Directory of Open Access Journals (Sweden)

    Walendziuk Wojciech

    2014-08-01

    Full Text Available The present article describes constructing a stabilographic platform which records a standing patient’s deflection from their point of balance. The constructed device is composed of a toughen glass slab propped with 4 force sensors. Power transducers are connected to the measurement system based on a 24-bit ADC transducer which acquires slight body movements of a patient. The data is then transferred to the computer in real time and data analysis is conducted. The article explains the principle of operation as well as the algorithm of measurement uncertainty for the COP (Centre of Pressure surface (x, y.

  9. Virtual Estimator for Piecewise Linear Systems Based on Observability Analysis

    Science.gov (United States)

    Morales-Morales, Cornelio; Adam-Medina, Manuel; Cervantes, Ilse; Vela-Valdés and, Luis G.; García Beltrán, Carlos Daniel

    2013-01-01

    This article proposes a virtual sensor for piecewise linear systems based on observability analysis that is in function of a commutation law related with the system's outpu. This virtual sensor is also known as a state estimator. Besides, it presents a detector of active mode when the commutation sequences of each linear subsystem are arbitrary and unknown. For the previous, this article proposes a set of virtual estimators that discern the commutation paths of the system and allow estimating their output. In this work a methodology in order to test the observability for piecewise linear systems with discrete time is proposed. An academic example is presented to show the obtained results. PMID:23447007

  10. Modal Analysis Based on the Random Decrement Technique

    DEFF Research Database (Denmark)

    Asmussen, J. C.; Brincker, Rune

    1998-01-01

    This article describes the work carried out within the project: Modal Analysis Based on the Random Decrement Technique - Application to Civil Engineering Structures. The project is part of the research programme: Dynamics of Structures sponsored by the Danish Technical Research Counsil. The planned...... contents and the requirement for the project prior to its start are described together with thee results obtained during the 3 year period of the project. The project was mainly carried out as a Ph.D project by the first author from September 1994 to August 1997 in cooperation with associate professor Rune...

  11. Toward a validation process for model based safety analysis

    OpenAIRE

    Adeline, Romain; Cardoso, Janette; Darfeuil, Pierre; Humbert, Sophie; Seguin, Christel

    2010-01-01

    Today, Model Based processes become more and more widespread to achieve the analysis of a system. However, there is no formal testing approach to ensure that the formal model is compliant with the real system. In the paper, we choose to study AltaRica model. We present a general process to well construct and validate an AltaRica formal model. The focus is made on this validation phase, i.e. verifying the compliance between the model and the real system. For it, the proposed process recommends...

  12. Reliability Distribution of Numerical Control Lathe Based on Correlation Analysis

    Institute of Scientific and Technical Information of China (English)

    Xiaoyan Qi; Guixiang Shen; Yingzhi Zhang; Shuguang Sun; Bingkun Chen

    2016-01-01

    Combined Reliability distribution with correlation analysis, a new method has been proposed to make Reliability distribution where considering the elements about structure correlation and failure correlation of subsystems. Firstly, we make a sequence for subsystems by means of TOPSIS which comprehends the considerations of Reliability allocation, and introducing a Copula connecting function to set up a distribution model based on structure correlation, failure correlation and target correlation, and then acquiring reliability target area of all subsystems by Matlab. In this method, not only the traditional distribution considerations are concerned, but also correlation influences are involved, to achieve supplementing information and optimizing distribution.

  13. Operational logs analysis at ALMA observatory based on ELK stack

    Science.gov (United States)

    Gil, Juan P.; Reveco, Johnny; Shen, Tzu-Chiang

    2016-07-01

    During operations, the ALMA observatory generates a huge amount of logs which contain not only valuable information related to specific failures but also for long term performance analysis. We implemented a big data solution based on Elasticsearch, Logstash and Kibana. They are configured as decoupled system which causes zero impact on the existent operations. It is able to keep more than six months of operation logs online. In this paper, we'll describe this infrastructure, applications built on top of it, and the problems that we faced during its implementation.

  14. Sensitivity Analysis of a Bioinspired Refractive Index Based Gas Sensor

    Institute of Scientific and Technical Information of China (English)

    Yang Gao; Qi Xia; Guanglan Liao; Tielin Shi

    2011-01-01

    It was found out that the change of refractive index of ambient gas can lead to obvious change of the color of Morpho butterfly's wing. Such phenomenon has been employed as a sensing principle for detecting gas. In the present study, Rigorous Coupled-Wave Analysis (RCWA) was described briefly, and the partial derivative of optical reflection efficiency with respect to the refractive index of ambient gas, i.e., sensitivity of the sensor, was derived based on RCWA. A bioinspired grating model was constructed by mimicking the nanostructure on the ground scale of Morpho didius butterfly's wing. The analytical sensitivity was verified and the effect of the grating shape on the reflection spectra and its sensitivity were discussed. The results show that by tuning shape parameters of the grating, we can obtain desired reflection spectra and sensitivity, which can be applied to the design of the bioinspired refractive index based gas sensor.

  15. BLAT-based comparative analysis for transposable elements: BLATCAT.

    Science.gov (United States)

    Lee, Sangbum; Oh, Sumin; Kang, Keunsoo; Han, Kyudong

    2014-01-01

    The availability of several whole genome sequences makes comparative analyses possible. In primate genomes, the priority of transposable elements (TEs) is significantly increased because they account for ~45% of the primate genomes, they can regulate the gene expression level, and they are associated with genomic fluidity in their host genomes. Here, we developed the BLAST-like alignment tool (BLAT) based comparative analysis for transposable elements (BLATCAT) program. The BLATCAT program can compare specific regions of six representative primate genome sequences (human, chimpanzee, gorilla, orangutan, gibbon, and rhesus macaque) on the basis of BLAT and simultaneously carry out RepeatMasker and/or Censor functions, which are widely used Windows-based web-server functions to detect TEs. All results can be stored as a HTML file for manual inspection of a specific locus. BLATCAT will be very convenient and efficient for comparative analyses of TEs in various primate genomes.

  16. BLAT-Based Comparative Analysis for Transposable Elements: BLATCAT

    Directory of Open Access Journals (Sweden)

    Sangbum Lee

    2014-01-01

    Full Text Available The availability of several whole genome sequences makes comparative analyses possible. In primate genomes, the priority of transposable elements (TEs is significantly increased because they account for ~45% of the primate genomes, they can regulate the gene expression level, and they are associated with genomic fluidity in their host genomes. Here, we developed the BLAST-like alignment tool (BLAT based comparative analysis for transposable elements (BLATCAT program. The BLATCAT program can compare specific regions of six representative primate genome sequences (human, chimpanzee, gorilla, orangutan, gibbon, and rhesus macaque on the basis of BLAT and simultaneously carry out RepeatMasker and/or Censor functions, which are widely used Windows-based web-server functions to detect TEs. All results can be stored as a HTML file for manual inspection of a specific locus. BLATCAT will be very convenient and efficient for comparative analyses of TEs in various primate genomes.

  17. Discrete directional wavelet bases and frames: analysis and applications

    Science.gov (United States)

    Dragotti, Pier Luigi; Velisavljevic, Vladan; Vetterli, Martin; Beferull-Lozano, Baltasar

    2003-11-01

    The application of the wavelet transform in image processing is most frequently based on a separable construction. Lines and columns in an image are treated independently and the basis functions are simply products of the corresponding one dimensional functions. Such method keeps simplicity in design and computation, but is not capable of capturing properly all the properties of an image. In this paper, a new truly separable discrete multi-directional transform is proposed with a subsampling method based on lattice theory. Alternatively, the subsampling can be omitted and this leads to a multi-directional frame. This transform can be applied in many areas like denoising, non-linear approximation and compression. The results on non-linear approximation and denoising show interesting gains compared to the standard two-dimensional analysis.

  18. Supermarket Analysis Based On Product Discount and Statistics

    Directory of Open Access Journals (Sweden)

    Komal Kumawat

    2014-03-01

    Full Text Available E-commerce has been growing rapidly. Its domain can provide all the right ingredients for successful data mining and it is a significant domain of data mining. E commerce refers to buying and selling of products or services over electronic systems such as internet. Various e commerce systems give discount on product and allow user to buy product online. The basic idea used here is to predict the product sale based on discount applied to the product. Our analysis concentrates on how customer behaves when discount is allotted to him. We have developed a model which finds the customer behaviour when discount is applied to the product. This paper elaborates upon how a different technique like session, click stream is used to collect user data online based on discount applied to the product and how statistics is applied to data set to see the variation in the data.

  19. Single base pair mutation analysis by PNA directed PCR clamping

    DEFF Research Database (Denmark)

    Ørum, H.; Nielsen, P.E.; Egholm, M.;

    1993-01-01

    A novel method that allows direct analysis of single base mutation by the polymerase chain reaction (PCR) is described. The method utilizes the finding that PNAs (peptide nucleic acids) recognize and bind to their complementary nucleic acid sequences with higher thermal stability and specificity...... than the corresponding deoxyribooligonucleotides and that they cannot function as primers for DNA polymerases. We show that a PNA/DNA complex can effectively block the formation of a PCR product when the PNA is targeted against one of the PCR primer sites. Furthermore, we demonstrate that this blockage...... allows selective amplification/suppression of target sequences that differ by only one base pair. Finally we show that PNAs can be designed in such a way that blockage can be accomplished when the PNA target sequence is located between the PCR primers....

  20. Sensitivity analysis of GSI based mechanical characterization of rock mass

    CERN Document Server

    Ván, P

    2012-01-01

    Recently, the rock mechanical and rock engineering designs and calculations are frequently based on Geological Strength Index (GSI) method, because it is the only system that provides a complete set of mechanical properties for design purpose. Both the failure criteria and the deformation moduli of the rock mass can be calculated with GSI based equations, which consists of the disturbance factor, as well. The aim of this paper is the sensitivity analysis of GSI and disturbance factor dependent equations that characterize the mechanical properties of rock masses. The survey of the GSI system is not our purpose. The results show that the rock mass strength calculated by the Hoek-Brown failure criteria and both the Hoek-Diederichs and modified Hoek-Diederichs deformation moduli are highly sensitive to changes of both the GSI and the D factor, hence their exact determination is important for the rock engineering design.