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Sample records for peptide identification repository

  1. Experimental Peptide Identification Repository (EPIR): an integrated peptide-centric platform for validation and mining of tandem mass spectrometry data

    DEFF Research Database (Denmark)

    Kristensen, Dan Bach; Brønd, Jan Christian; Nielsen, Peter Aagaard

    2004-01-01

    LC MS/MS has become an established technology in proteomic studies, and with the maturation of the technology the bottleneck has shifted from data generation to data validation and mining. To address this bottleneck we developed Experimental Peptide Identification Repository (EPIR), which...... is an integrated software platform for storage, validation, and mining of LC MS/MS-derived peptide evidence. EPIR is a cumulative data repository where precursor ions are linked to peptide assignments and protein associations returned by a search engine (e.g. Mascot, Sequest, or PepSea). Any number of datasets can...

  2. ARA-PEPs: a repository of putative sORF-encoded peptides in Arabidopsis thaliana.

    Science.gov (United States)

    Hazarika, Rashmi R; De Coninck, Barbara; Yamamoto, Lidia R; Martin, Laura R; Cammue, Bruno P A; van Noort, Vera

    2017-01-17

    Many eukaryotic RNAs have been considered non-coding as they only contain short open reading frames (sORFs). However, there is increasing evidence for the translation of these sORFs into bioactive peptides with potent signaling, antimicrobial, developmental, antioxidant roles etc. Yet only a few peptides encoded by sORFs are annotated in the model organism Arabidopsis thaliana. To aid the functional annotation of these peptides, we have developed ARA-PEPs (available at http://www.biw.kuleuven.be/CSB/ARA-PEPs ), a repository of putative peptides encoded by sORFs in the A. thaliana genome starting from in-house Tiling arrays, RNA-seq data and other publicly available datasets. ARA-PEPs currently lists 13,748 sORF-encoded peptides with transcriptional evidence. In addition to existing data, we have identified 100 novel transcriptionally active regions (TARs) that might encode 341 novel stress-induced peptides (SIPs). To aid in identification of bioactivity, we add functional annotation and sequence conservation to predicted peptides. To our knowledge, this is the largest repository of plant peptides encoded by sORFs with transcript evidence, publicly available and this resource will help scientists to effortlessly navigate the list of experimentally studied peptides, the experimental and computational evidence supporting the activity of these peptides and gain new perspectives for peptide discovery.

  3. Accelerator assisted repositories, technical issue identification and economic assessment

    Energy Technology Data Exchange (ETDEWEB)

    Myers, T.; Favale, A.; Berwald, D. [Northrop Grumman Corp., Bethpage, NY (United States)

    1995-12-31

    An investigation (technical and economic) is presented to quantify the impact of accelerator assisted repository (AAR) system on the US national nuclear repository strategy. An overview of US spent fuel policy is presented which provides a means to comparatively assess competing spent fuel disposition technologies under investigation. The advantages of an AAR system are highlighted and discussed. A companion economic analysis is presented. The results indicate that although a US geologic waste repository will continue to be required, waste partitioning and accelerator transmutation of selected actinides, and long-lived fission products, can result in substantial benefits. (author) 18 refs.

  4. Optimizing the identification of citrullinated peptides by mass spectrometry

    DEFF Research Database (Denmark)

    Bennike, Tue; Lauridsen, Kasper B.; Olesen, Michael Kruse

    2013-01-01

    Citrullinated proteins have been associated with several diseases and citrullination can most likely function as a target for novel diagnostic agents and unravel disease etiologies. The correct identification of citrullinated proteins is therefore of most importance. Mass spectrometry (MS) driven...... of trypsin, digestion was performed on synthetic peptide sets containing either arginine or citrulline. The peptide sequences originated from disease-associated in vivo citrullinated proteins; some reported as being C-terminal tryptic citrullinated peptides. Furthermore, the proteolytic activity was verified...

  5. Identification of Cyclin A Binders with a Fluorescent Peptide Sensor.

    Science.gov (United States)

    Pazos, Elena; Mascareñas, José L; Vázquez, M Eugenio

    2016-01-01

    A peptide sensor that integrates the 4-dimethylaminophthalimide (4-DMAP) fluorophore in a short cyclin A binding sequence displays a large fluorescence emission increase upon interacting with the cyclin A Binding Groove (CBG). Competitive displacement assays of this probe allow the straightforward identification of peptides that interact with the CBG, which could potentially block the recognition of CDK/cyclin A kinase substrates.

  6. Identification of candidate sites for a near surface repository for radioactive waste

    International Nuclear Information System (INIS)

    Motiejunas, S.

    2004-01-01

    This Report comprises results of the area survey stage, which involves regional screening to define the regions of interest and identification of potential sites within suitable regions. The main goal was to define a few sites potentially suitable for constructing of the near surface repository. It was concluded that a vicinity of Ignalina NPP is among the best suitable regions for the near surface repository. At the present investigation level a ridge in Galilauke village has the most favorable conditions. However, Apvardai site is potentially suitable for the repository too

  7. Protein identification by peptide mass fingerprinting

    DEFF Research Database (Denmark)

    Hjernø, Karin

    2007-01-01

      Peptide mass fingerprinting is an effective way of identifying, e.g., gel-separated proteins, by matching experimentally obtained peptide mass data against large databases. However, several factors are known to influence the quality of the resulting matches, such as proteins contaminating the s...

  8. CPAD, Curated Protein Aggregation Database: A Repository of Manually Curated Experimental Data on Protein and Peptide Aggregation.

    Science.gov (United States)

    Thangakani, A Mary; Nagarajan, R; Kumar, Sandeep; Sakthivel, R; Velmurugan, D; Gromiha, M Michael

    2016-01-01

    Accurate distinction between peptide sequences that can form amyloid-fibrils or amorphous β-aggregates, identification of potential aggregation prone regions in proteins, and prediction of change in aggregation rate of a protein upon mutation(s) are critical to research on protein misfolding diseases, such as Alzheimer's and Parkinson's, as well as biotechnological production of protein based therapeutics. We have developed a Curated Protein Aggregation Database (CPAD), which has collected results from experimental studies performed by scientific community aimed at understanding protein/peptide aggregation. CPAD contains more than 2300 experimentally observed aggregation rates upon mutations in known amyloidogenic proteins. Each entry includes numerical values for the following parameters: change in rate of aggregation as measured by fluorescence intensity or turbidity, name and source of the protein, Uniprot and Protein Data Bank codes, single point as well as multiple mutations, and literature citation. The data in CPAD has been supplemented with five different types of additional information: (i) Amyloid fibril forming hexa-peptides, (ii) Amorphous β-aggregating hexa-peptides, (iii) Amyloid fibril forming peptides of different lengths, (iv) Amyloid fibril forming hexa-peptides whose crystal structures are available in the Protein Data Bank (PDB) and (v) Experimentally validated aggregation prone regions found in amyloidogenic proteins. Furthermore, CPAD is linked to other related databases and resources, such as Uniprot, Protein Data Bank, PUBMED, GAP, TANGO, WALTZ etc. We have set up a web interface with different search and display options so that users have the ability to get the data in multiple ways. CPAD is freely available at http://www.iitm.ac.in/bioinfo/CPAD/. The potential applications of CPAD have also been discussed.

  9. Identification of characteristics which influence repository design domal salt, Task 1. Final report

    International Nuclear Information System (INIS)

    Rawlings, G.; Antonnen, G.; Chamness, M.

    1984-04-01

    The purpose of the complete project is to provide NRC with technical assistance to enable the focused, adequate review by NRC of the aspects related to design and construction of an underground test facility and final geologic repository as presented by the Department of Energy (DOE). The study presented in this report covers the identification of characteristics which influence design and construction of a geologic repository in domal salt. This report has identified five key issues, i.e., constructibility, thermal response, mechanical response, hydrologic response, and geochemical response. This report involves both short-term (up to closure) and long-term (post closure) effects. The characteristics of domal salt and its environment are described under the headings of stratigraphic/structural, tectonic, mechanical, thermal and hydrologic. Characteristics are separated into parameters (quantified and measured) and factors (qualitative). The characteristics are then subjectively ranked by their influence on the key issues. This takes into account the availability and suitability of conservative design/construction techniques, uncertainty in model and model sensitivity to the characteristic

  10. Screening and Identification of Peptides Specifically Targeted to Gastric Cancer Cells from a Phage Display Peptide Library

    Science.gov (United States)

    Sahin, Deniz; Taflan, Sevket Onur; Yartas, Gizem; Ashktorab, Hassan; Smoot, Duane T

    2018-04-25

    Background: Gastric cancer is the second most common cancer among the malign cancer types. Inefficiency of traditional techniques both in diagnosis and therapy of the disease makes the development of alternative and novel techniques indispensable. As an alternative to traditional methods, tumor specific targeting small peptides can be used to increase the efficiency of the treatment and reduce the side effects related to traditional techniques. The aim of this study is screening and identification of individual peptides specifically targeted to human gastric cancer cells using a phage-displayed peptide library and designing specific peptide sequences by using experimentally-eluted peptide sequences. Methods: Here, MKN-45 human gastric cancer cells and HFE-145 human normal gastric epithelial cells were used as the target and control cells, respectively. 5 rounds of biopannning with a phage display 12-peptide library were applied following subtraction biopanning with HFE-145 control cells. The selected phage clones were established by enzyme-linked immunosorbent assay and immunofluorescence detection. We first obtain random phage clones after five biopanning rounds, determine the binding levels of each individual clone. Then, we analyze the frequencies of each amino acid in best binding clones to determine positively overexpressed amino acids for designing novel peptide sequences. Results: DE532 (VETSQYFRGTLS) phage clone was screened positive, showing specific binding on MKN-45 gastric cancer cells. DE-Obs (HNDLFPSWYHNY) peptide, which was designed by using amino acid frequencies of experimentally selected peptides in the 5th round of biopanning, showed specific binding in MKN-45 cells. Conclusion: Selection and characterization of individual clones may give us specifically binding peptides, but more importantly, data extracted from eluted phage clones may be used to design theoretical peptides with better binding properties than even experimentally selected ones

  11. Identification of binding peptides of the ADAM15 disintegrin domain ...

    Indian Academy of Sciences (India)

    Madhsudhan

    ADAM15 disintegrin domain (RADD) that could inhibit melanoma cell adhesion by using Escherichia coli. Second, four specific binding peptides (peptides A, B, C, and D) were selected using a phage display 12-mer peptide library. The screening protocol involved 4 rounds of positive panning on RADD and 2 rounds of ...

  12. Pathogen identification using peptide nanotube biosensors and impedance AFM

    Science.gov (United States)

    Maccuspie, Robert I.

    Pathogen identification at highly sensitive levels is crucial to meet urgent needs in fighting the spread of disease or detecting bioterrorism events. Toward that end, a new method for biosensing utilizing fluorescent antibody nanotubes is proposed. Fundamental studies on the self-assembly of these peptide nanotubes are performed, as are applications of aligning these nanotubes on surfaces. As biosensors, these nanotubes incorporate recognition units with antibodies at their ends and fluorescent signaling units at their sidewalls. When viral pathogens were mixed with these antibody nanotubes in solution, the nanotubes rapidly aggregated around the viruses. The size of the aggregates increased as the concentration of viruses increased, as detected by flow cytometry on the order of attomolar concentrations by changes in fluorescence and light scattering intensities. This enabled determination of the concentrations of viruses at trace levels (102 to 106 pfu/mL) within 30 minutes from the receipt of samples to the final quantitative data analysis, as demonstrated on Adenovirus, Herpes Simplex Virus, Influenza, and Vaccinia virus. As another separate approach, impedance AFM is used to study the electrical properties of individual viruses and nanoparticles used as model systems. The design, development, and implementation of the impedance AFM for an Asylum Research platform is described, as well as its application towards studying the impedance of individual nanoparticles as a model system for understanding the fundamental science of how the life cycle of a virus affects its electrical properties. In combination, these approaches fill a pressing need to quantify viruses both rapidly and sensitively.

  13. Peptomics, identification of novel cationic Arabidopsis peptides with conserved sequence motifs

    DEFF Research Database (Denmark)

    Olsen, Addie Nina; Mundy, John; Skriver, Karen

    2002-01-01

    Arabidopsis family of 34 genes. The predicted peptides are characterized by a conserved C-terminal sequence motif and additional primary structure conservation in a core region. The majority of these genes had not previously been annotated. A subset of the predicted peptides show high overall sequence...... similarity to Rapid Alkalinization Factor (RALF), a peptide isolated from tobacco. We therefore refer to this peptide family as RALFL for RALF-Like. RT-PCR analysis confirmed that several of the Arabidopsis genes are expressed and that their expression patterns vary. The identification of a large gene family...

  14. Classifying Clinical Trial Eligibility Criteria to Facilitate Phased Cohort Identification Using Clinical Data Repositories.

    Science.gov (United States)

    Wang, Amy Y; Lancaster, William J; Wyatt, Matthew C; Rasmussen, Luke V; Fort, Daniel G; Cimino, James J

    2017-01-01

    A major challenge in using electronic health record repositories for research is the difficulty matching subject eligibility criteria to query capabilities of the repositories. We propose categories for study criteria corresponding to the effort needed for querying those criteria: "easy" (supporting automated queries), mixed (initial automated querying with manual review), "hard" (fully manual record review), and "impossible" or "point of enrollment" (not typically in health repositories). We obtained a sample of 292 criteria from 20 studies from ClinicalTrials.gov. Six independent reviewers, three each from two academic research institutions, rated criteria according to our four types. We observed high interrater reliability both within and between institutions. The analysis demonstrated typical features of criteria that map with varying levels of difficulty to repositories. We propose using these features to improve enrollment workflow through more standardized study criteria, self-service repository queries, and analyst-mediated retrievals.

  15. A novel algorithm for validating peptide identification from a shotgun proteomics search engine.

    Science.gov (United States)

    Jian, Ling; Niu, Xinnan; Xia, Zhonghang; Samir, Parimal; Sumanasekera, Chiranthani; Mu, Zheng; Jennings, Jennifer L; Hoek, Kristen L; Allos, Tara; Howard, Leigh M; Edwards, Kathryn M; Weil, P Anthony; Link, Andrew J

    2013-03-01

    Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has revolutionized the proteomics analysis of complexes, cells, and tissues. In a typical proteomic analysis, the tandem mass spectra from a LC-MS/MS experiment are assigned to a peptide by a search engine that compares the experimental MS/MS peptide data to theoretical peptide sequences in a protein database. The peptide spectra matches are then used to infer a list of identified proteins in the original sample. However, the search engines often fail to distinguish between correct and incorrect peptides assignments. In this study, we designed and implemented a novel algorithm called De-Noise to reduce the number of incorrect peptide matches and maximize the number of correct peptides at a fixed false discovery rate using a minimal number of scoring outputs from the SEQUEST search engine. The novel algorithm uses a three-step process: data cleaning, data refining through a SVM-based decision function, and a final data refining step based on proteolytic peptide patterns. Using proteomics data generated on different types of mass spectrometers, we optimized the De-Noise algorithm on the basis of the resolution and mass accuracy of the mass spectrometer employed in the LC-MS/MS experiment. Our results demonstrate De-Noise improves peptide identification compared to other methods used to process the peptide sequence matches assigned by SEQUEST. Because De-Noise uses a limited number of scoring attributes, it can be easily implemented with other search engines.

  16. Identification and screening of potent antimicrobial peptides in arthropod genomes.

    Science.gov (United States)

    Duwadi, Deepesh; Shrestha, Anishma; Yilma, Binyam; Kozlovski, Itamar; Sa-Eed, Munaya; Dahal, Nikesh; Jukosky, James

    2018-05-01

    Using tBLASTn and BLASTp searches, we queried recently sequenced arthropod genomes and expressed sequence tags (ESTs) using a database of known arthropod cecropins, defensins, and attacins. We identified and synthesized 6 potential AMPs and screened them for antimicrobial activity. Using radial diffusion assays and microtiter antimicrobial assays, we assessed the in vitro antimicrobial effects of these peptides against several human pathogens including Gram-positive and Gram-negative bacteria and fungi. We also conducted hemolysis assays to examine the cytotoxicity of these peptides to mammalian cells. Four of the six peptides identified showed antimicrobial effects in these assays. We also created truncated versions of these four peptides to assay their antimicrobial activity. Two cecropins derived from the monarch butterfly genome (Danaus plexippus), DAN1 and DAN2, showed minimum inhibitory concentrations (MICs) in the range of 2-16 μg/ml when screened against Gram-negative bacteria. HOLO1 and LOUDEF1, two defensin-like peptides derived from red flour beetle (Tribolium castaneum) and human body louse (Pediculus humanus humanus), respectively, exhibited MICs in the range of 13-25 μg/ml against Gram-positive bacteria. Furthermore, HOLO1 showed an MIC less than 5 μg/ml against the fungal species Candida albicans. These peptides exhibited no hemolytic activity at concentrations up to 200 μg/ml. The truncated peptides derived from DAN2 and HOLO1 showed very little antimicrobial activity. Our experiments show that the peptides DAN1, DAN2, HOLO1, and LOUDEF1 showed potent antimicrobial activity in vitro against common human pathogens, did not lyse mammalian red blood cells, and indicates their potential as templates for novel therapeutic agents against microbial infection. Copyright © 2018 Elsevier Inc. All rights reserved.

  17. PepArML: A Meta-Search Peptide Identification Platform for Tandem Mass Spectra.

    Science.gov (United States)

    Edwards, Nathan J

    2013-12-01

    The PepArML meta-search peptide identification platform for tandem mass spectra provides a unified search interface to seven search engines; a robust cluster, grid, and cloud computing scheduler for large-scale searches; and an unsupervised, model-free, machine-learning-based result combiner, which selects the best peptide identification for each spectrum, estimates false-discovery rates, and outputs pepXML format identifications. The meta-search platform supports Mascot; Tandem with native, k-score and s-score scoring; OMSSA; MyriMatch; and InsPecT with MS-GF spectral probability scores—reformatting spectral data and constructing search configurations for each search engine on the fly. The combiner selects the best peptide identification for each spectrum based on search engine results and features that model enzymatic digestion, retention time, precursor isotope clusters, mass accuracy, and proteotypic peptide properties, requiring no prior knowledge of feature utility or weighting. The PepArML meta-search peptide identification platform often identifies two to three times more spectra than individual search engines at 10% FDR.

  18. Identification and characterization of potential discharge areas for radionuclide transport by groundwater from a nuclear waste repository in Sweden.

    Science.gov (United States)

    Berglund, Sten; Bosson, Emma; Selroos, Jan-Olof; Sassner, Mona

    2013-05-01

    This paper describes solute transport modeling carried out as a part of an assessment of the long-term radiological safety of a planned deep rock repository for spent nuclear fuel in Forsmark, Sweden. Specifically, it presents transport modeling performed to locate and describe discharge areas for groundwater potentially carrying radionuclides from the repository to the surface where man and the environment could be affected by the contamination. The modeling results show that topography to large extent determines the discharge locations. Present and future lake and wetland objects are central for the radionuclide transport and dose calculations in the safety assessment. Results of detailed transport modeling focusing on the regolith and the upper part of the rock indicate that the identification of discharge areas and objects considered in the safety assessment is robust in the sense that it does not change when a more detailed model representation is used.

  19. Probabilistic consensus scoring improves tandem mass spectrometry peptide identification.

    Science.gov (United States)

    Nahnsen, Sven; Bertsch, Andreas; Rahnenführer, Jörg; Nordheim, Alfred; Kohlbacher, Oliver

    2011-08-05

    Database search is a standard technique for identifying peptides from their tandem mass spectra. To increase the number of correctly identified peptides, we suggest a probabilistic framework that allows the combination of scores from different search engines into a joint consensus score. Central to the approach is a novel method to estimate scores for peptides not found by an individual search engine. This approach allows the estimation of p-values for each candidate peptide and their combination across all search engines. The consensus approach works better than any single search engine across all different instrument types considered in this study. Improvements vary strongly from platform to platform and from search engine to search engine. Compared to the industry standard MASCOT, our approach can identify up to 60% more peptides. The software for consensus predictions is implemented in C++ as part of OpenMS, a software framework for mass spectrometry. The source code is available in the current development version of OpenMS and can easily be used as a command line application or via a graphical pipeline designer TOPPAS.

  20. Peptidomic Identification of Cysteine-Rich Peptides from Plants.

    Science.gov (United States)

    Hemu, Xinya; Serra, Aida; Darwis, Dina A; Cornvik, Tobias; Sze, Siu Kwan; Tam, James P

    2018-01-01

    Plant cysteine-rich peptides (CRPs) constitute a majority of plant-derived peptides with high molecular diversity. This protocol describes a rapid and efficient peptidomic approach to identify a whole spectrum of CRPs in a plant extract and decipher their molecular diversity and bioprocessing mechanism. Cyclotides from C. ternatea are used as the model CRPs to demonstrate our methodology. Cyclotides exist naturally in both cyclic and linear forms, although the linear forms (acyclotide) are generally present at much lower concentrations. Both cyclotides and acyclotides require linearization of their backbone prior to fragmentation and sequencing. A novel and practical three-step chemoenzymatic treatment was developed to linearize and distinguish both forms: (1) N-terminal acetylation that pre-labels the acyclotides; (2) conversion of Cys into pseudo-Lys through aziridine-mediated S-alkylation to reduce disulfide bonds and to increase the net charge of peptides; and (3) opening of cyclic backbones by the novel asparaginyl endopeptidase butelase 2 that cleaves at the native bioprocessing site. The treated peptides are subsequently analyzed by liquid chromatography coupled to mass spectrometry using electron transfer dissociation fragmentation and sequences are identified by matching the MS/MS spectra directly with the transcriptomic database.

  1. MSblender: A probabilistic approach for integrating peptide identifications from multiple database search engines.

    Science.gov (United States)

    Kwon, Taejoon; Choi, Hyungwon; Vogel, Christine; Nesvizhskii, Alexey I; Marcotte, Edward M

    2011-07-01

    Shotgun proteomics using mass spectrometry is a powerful method for protein identification but suffers limited sensitivity in complex samples. Integrating peptide identifications from multiple database search engines is a promising strategy to increase the number of peptide identifications and reduce the volume of unassigned tandem mass spectra. Existing methods pool statistical significance scores such as p-values or posterior probabilities of peptide-spectrum matches (PSMs) from multiple search engines after high scoring peptides have been assigned to spectra, but these methods lack reliable control of identification error rates as data are integrated from different search engines. We developed a statistically coherent method for integrative analysis, termed MSblender. MSblender converts raw search scores from search engines into a probability score for every possible PSM and properly accounts for the correlation between search scores. The method reliably estimates false discovery rates and identifies more PSMs than any single search engine at the same false discovery rate. Increased identifications increment spectral counts for most proteins and allow quantification of proteins that would not have been quantified by individual search engines. We also demonstrate that enhanced quantification contributes to improve sensitivity in differential expression analyses.

  2. Enhancing bioactive peptide release and identification using targeted enzymatic hydrolysis of milk proteins.

    Science.gov (United States)

    Nongonierma, Alice B; FitzGerald, Richard J

    2018-06-01

    Milk proteins have been extensively studied for their ability to yield a range of bioactive peptides following enzymatic hydrolysis/digestion. However, many hurdles still exist regarding the widespread utilization of milk protein-derived bioactive peptides as health enhancing agents for humans. These mostly arise from the fact that most milk protein-derived bioactive peptides are not highly potent. In addition, they may be degraded during gastrointestinal digestion and/or have a low intestinal permeability. The targeted release of bioactive peptides during the enzymatic hydrolysis of milk proteins may allow the generation of particularly potent bioactive hydrolysates and peptides. Therefore, the development of milk protein hydrolysates capable of improving human health requires, in the first instance, optimized targeted release of specific bioactive peptides. The targeted hydrolysis of milk proteins has been aided by a range of in silico tools. These include peptide cutters and predictive modeling linking bioactivity to peptide structure [i.e., molecular docking, quantitative structure activity relationship (QSAR)], or hydrolysis parameters [design of experiments (DOE)]. Different targeted enzymatic release strategies employed during the generation of milk protein hydrolysates are reviewed herein and their limitations are outlined. In addition, specific examples are provided to demonstrate how in silico tools may help in the identification and discovery of potent milk protein-derived peptides. It is anticipated that the development of novel strategies employing a range of in silico tools may help in the generation of milk protein hydrolysates containing potent and bioavailable peptides, which in turn may be used to validate their health promoting effects in humans. Graphical abstract The targeted enzymatic hydrolysis of milk proteins may allow the generation of highly potent and bioavailable bioactive peptides.

  3. Charge State Coalescence During Electrospray Ionization Improves Peptide Identification by Tandem Mass Spectrometry

    Science.gov (United States)

    Meyer, Jesse G.; A. Komives, Elizabeth

    2012-08-01

    We report the effects of supercharging reagents dimethyl sulphoxide (DMSO) and m-nitrobenzyl alcohol ( m-NBA) applied to untargeted peptide identification, with special emphasis on non-tryptic peptides. Peptides generated from a mixture of five standard proteins digested with trypsin, elastase, or pepsin were separated with nanoflow liquid chromatography using mobile phases modified with either 5 % DMSO or 0.1 % m-NBA. Eluting peptides were ionized by online electrospray and sequenced by both CID and ETD using data-dependent MS/MS. Statistically significant improvements in peptide identifications were observed with DMSO co-solvent. In order to understand this observation, we assessed the effects of supercharging reagents on the chromatographic separation and the electrospray quality. The increase in identifications was not due to supercharging, which was greater for the 0.1 % m-NBA co-solvent and not observed for the 5.0 % DMSO co-solvent. The improved MS/MS efficiency using the DMSO modified mobile phase appeared to result from charge state coalescence.

  4. Identification of Alternative Splice Variants Using Unique Tryptic Peptide Sequences for Database Searches.

    Science.gov (United States)

    Tran, Trung T; Bollineni, Ravi C; Strozynski, Margarita; Koehler, Christian J; Thiede, Bernd

    2017-07-07

    Alternative splicing is a mechanism in eukaryotes by which different forms of mRNAs are generated from the same gene. Identification of alternative splice variants requires the identification of peptides specific for alternative splice forms. For this purpose, we generated a human database that contains only unique tryptic peptides specific for alternative splice forms from Swiss-Prot entries. Using this database allows an easy access to splice variant-specific peptide sequences that match to MS data. Furthermore, we combined this database without alternative splice variant-1-specific peptides with human Swiss-Prot. This combined database can be used as a general database for searching of LC-MS data. LC-MS data derived from in-solution digests of two different cell lines (LNCaP, HeLa) and phosphoproteomics studies were analyzed using these two databases. Several nonalternative splice variant-1-specific peptides were found in both cell lines, and some of them seemed to be cell-line-specific. Control and apoptotic phosphoproteomes from Jurkat T cells revealed several nonalternative splice variant-1-specific peptides, and some of them showed clear quantitative differences between the two states.

  5. Identification of structures, systems, and components important to safety at the potential repository at Yucca Mountain

    International Nuclear Information System (INIS)

    Hartman, D.J.; Miller, D.D.; Klamerus, L.J.

    1991-10-01

    This study recommends which structures, systems, and components of the potential repository at Yucca Mountain are important to safety. The assessment was completed in April 1990 and uses the reference repository configuration in the Site Characterization Plan Conceptual Design Report and follows the methodology required at that time by DOE Procedure AP6.10-Q. Failures of repository items during the preclosure period are evaluated to determine the potential offsite radiation doses and associated probabilities. Items are important to safety if, in the event they fail to perform their intended function, an accident could result which causes a dose commitment greater than 0.5 rem to the whole body or any organ of an individual in an unrestricted area. This study recommends that these repository items include the structures that house spent fuel and high-level waste, the associated filtered ventilation exhaust systems, certain waste- handling equipment, the waste containers, the waste treatment building structure, the underground waste transporters, and other items listed in this report. This work was completed April 1990. 27 refs., 7 figs., 9 tabs

  6. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites

    DEFF Research Database (Denmark)

    Nielsen, Henrik; Engelbrecht, Jacob; Brunak, Søren

    1997-01-01

    We have developed a new method for the identification of signal peptides and their cleavage based on neural networks trained on separate sets of prokaryotic and eukaryotic sequence. The method performs significantly better than previous prediction schemes and can easily be applied on genome...

  7. Identification and characterization of peptide: N- glycanase from Dictyostelium discoideum

    Directory of Open Access Journals (Sweden)

    Gosain Anuradha

    2012-06-01

    Full Text Available Abstract Background Peptide: N- glycanase (PNGase enzyme cleaves oligosaccharides from the misfolded glycoproteins and prepares them for degradation. This enzyme plays a role in the endoplasmic reticulum associated degradation (ERAD pathway in yeast and mice but its biological importance and role in multicellular development remain largely unknown. Results In this study, the PNGase from the cellular slime mold, Dictyostelium discoideum (DdPNGase was identified based on the presence of a common TG (transglutaminase core domain and its sequence homology with the known PNGases. The domain architecture and the sequence comparison validated the presence of probable functional domains in DdPNGase. The tertiary structure matched with the mouse PNGase. Here we show that DdPNGase is an essential protein, required for aggregation during multicellular development and a knockout strain of it results in small sized aggregates, all of which did not form fruiting bodies. The in situ hybridization and RT-PCR results show higher level of expression during the aggregate stage. The expression gets restricted to the prestalk region during later developmental stages. DdPNGase is a functional peptide:N-glycanase enzyme possessing deglycosylation activity, but does not possess any significant transamidation activity. Conclusions We have identified and characterized a novel PNGase from D. discoideum and confirmed its deglycosylation activity. The results emphasize the importance of PNGase in aggregation during multicellular development of this organism.

  8. High-Throughput Identification of Antimicrobial Peptides from Amphibious Mudskippers

    Directory of Open Access Journals (Sweden)

    Yunhai Yi

    2017-11-01

    Full Text Available Widespread existence of antimicrobial peptides (AMPs has been reported in various animals with comprehensive biological activities, which is consistent with the important roles of AMPs as the first line of host defense system. However, no big-data-based analysis on AMPs from any fish species is available. In this study, we identified 507 AMP transcripts on the basis of our previously reported genomes and transcriptomes of two representative amphibious mudskippers, Boleophthalmus pectinirostris (BP and Periophthalmus magnuspinnatus (PM. The former is predominantly aquatic with less time out of water, while the latter is primarily terrestrial with extended periods of time on land. Within these identified AMPs, 449 sequences are novel; 15 were reported in BP previously; 48 are identically overlapped between BP and PM; 94 were validated by mass spectrometry. Moreover, most AMPs presented differential tissue transcription patterns in the two mudskippers. Interestingly, we discovered two AMPs, hemoglobin β1 and amylin, with high inhibitions on Micrococcus luteus. In conclusion, our high-throughput screening strategy based on genomic and transcriptomic data opens an efficient pathway to discover new antimicrobial peptides for ongoing development of marine drugs.

  9. High-Throughput Identification of Antimicrobial Peptides from Amphibious Mudskippers.

    Science.gov (United States)

    Yi, Yunhai; You, Xinxin; Bian, Chao; Chen, Shixi; Lv, Zhao; Qiu, Limei; Shi, Qiong

    2017-11-22

    Widespread existence of antimicrobial peptides (AMPs) has been reported in various animals with comprehensive biological activities, which is consistent with the important roles of AMPs as the first line of host defense system. However, no big-data-based analysis on AMPs from any fish species is available. In this study, we identified 507 AMP transcripts on the basis of our previously reported genomes and transcriptomes of two representative amphibious mudskippers, Boleophthalmus pectinirostris (BP) and Periophthalmus magnuspinnatus (PM). The former is predominantly aquatic with less time out of water, while the latter is primarily terrestrial with extended periods of time on land. Within these identified AMPs, 449 sequences are novel; 15 were reported in BP previously; 48 are identically overlapped between BP and PM; 94 were validated by mass spectrometry. Moreover, most AMPs presented differential tissue transcription patterns in the two mudskippers. Interestingly, we discovered two AMPs, hemoglobin β1 and amylin, with high inhibitions on Micrococcus luteus . In conclusion, our high-throughput screening strategy based on genomic and transcriptomic data opens an efficient pathway to discover new antimicrobial peptides for ongoing development of marine drugs.

  10. Identification and accurate quantification of structurally related peptide impurities in synthetic human C-peptide by liquid chromatography-high resolution mass spectrometry.

    Science.gov (United States)

    Li, Ming; Josephs, Ralf D; Daireaux, Adeline; Choteau, Tiphaine; Westwood, Steven; Wielgosz, Robert I; Li, Hongmei

    2018-06-04

    Peptides are an increasingly important group of biomarkers and pharmaceuticals. The accurate purity characterization of peptide calibrators is critical for the development of reference measurement systems for laboratory medicine and quality control of pharmaceuticals. The peptides used for these purposes are increasingly produced through peptide synthesis. Various approaches (for example mass balance, amino acid analysis, qNMR, and nitrogen determination) can be applied to accurately value assign the purity of peptide calibrators. However, all purity assessment approaches require a correction for structurally related peptide impurities in order to avoid biases. Liquid chromatography coupled to high resolution mass spectrometry (LC-hrMS) has become the key technique for the identification and accurate quantification of structurally related peptide impurities in intact peptide calibrator materials. In this study, LC-hrMS-based methods were developed and validated in-house for the identification and quantification of structurally related peptide impurities in a synthetic human C-peptide (hCP) material, which served as a study material for an international comparison looking at the competencies of laboratories to perform peptide purity mass fraction assignments. More than 65 impurities were identified, confirmed, and accurately quantified by using LC-hrMS. The total mass fraction of all structurally related peptide impurities in the hCP study material was estimated to be 83.3 mg/g with an associated expanded uncertainty of 3.0 mg/g (k = 2). The calibration hierarchy concept used for the quantification of individual impurities is described in detail. Graphical abstract ᅟ.

  11. Identification of coeliac disease triggering glutenin peptides in adults.

    Science.gov (United States)

    Donnelly, Suzanne C; Šuligoj, Tanja; Ellis, H Julia; Ciclitira, Paul J

    2016-07-01

    Coeliac disease affects approximately 1% of Northern American and European populations. It is caused by an inappropriate immune response to dietary gluten. Gluten comprises of two major protein fractions: gliadins and glutenins. Glutenins have recently been found to be toxic to coeliac individuals. Proliferation assays suggest in some but not all paediatric coeliac individuals there may be immunological stimulation with high molecular weight (HMW) glutenins. Less evidence pertains to low molecular weight (LMW) glutenins. The aim is to assess adaptive, T-cell driven, and innate immune response in adult coeliac individuals towards HMW glutenin peptide, glut04, and LMW glutenin peptide, glt156. Coeliac patients were recruited attending endoscopy for routine monitoring. Adaptive immune response towards glut04 and glt156 was measured by proliferation assays and measurement of interferon-γ secretion in 28 T-cell lines. The innate immune response was assessed by measurement of enterocyte cell height (ECH) in coeliac small intestinal biopsies following overnight incubation in organ culture chambers in a further nine individuals. There were 3/28 and 2/28 positive proliferation results using gluten-sensitive T-cells with glut04 and glt156, respectively. All coeliac biopsies tested in organ culture chambers demonstrated clear reduction in ECH with peptic-tryptic digest of whole industrial gluten, glut04 and glt156 when compared to negative control ovalbumin (p < 0.005). Three individuals had both T-cell and organ culture study data. Their proliferation assays showed no stimulation of the T-cells. This study demonstrates glutenin epitopes glut04 and glt156, while minor T-cell epitopes, are important in their ability to trigger the innate immune response.

  12. In-Silico identification of peptides for the diagnostics of paratuberculosis

    DEFF Research Database (Denmark)

    Tang, Sheila Tuyet; Lund, Ole; Jungersen, Gregers

    Identification of bovine MHC class II reactive peptides that are specific/unique to paratuberculosis and conserved across pathogenic variations of the paratuberculosis proteome will be of high value for development of new vaccines and immune based diagnostics. Here, we present an in silico screen...... by statistical significance. BMC Bioinformatics, 2003. 4: p. 21. 2. Nielsen, M., et al., Quantitative predictions of peptide binding to any HLA-DR molecule of known sequence: NetMHCIIpan. PLoS Comput Biol, 2008. 4(7): p. e1000107....

  13. A peptide affinity column for the identification of integrin alpha IIb-binding proteins.

    Science.gov (United States)

    Daxecker, Heide; Raab, Markus; Bernard, Elise; Devocelle, Marc; Treumann, Achim; Moran, Niamh

    2008-03-01

    To understand the regulation of integrin alpha(IIb)beta(3), a critical platelet adhesion molecule, we have developed a peptide affinity chromatography method using the known integrin regulatory motif, LAMWKVGFFKR. Using standard Fmoc chemistry, this peptide was synthesized onto a Toyopearl AF-Amino-650 M resin on a 6-aminohexanoic acid (Ahx) linker. Peptide density was controlled by acetylation of 83% of the Ahx amino groups. Four recombinant human proteins (CIB1, PP1, ICln and RN181), previously identified as binding to this integrin regulatory motif, were specifically retained by the column containing the integrin peptide but not by a column presenting an irrelevant peptide. Hemoglobin, creatine kinase, bovine serum albumin, fibrinogen and alpha-tubulin failed to bind under the chosen conditions. Immunodetection methods confirmed the binding of endogenous platelet proteins, including CIB1, PP1, ICln RN181, AUP-1 and beta3-integrin, from a detergent-free platelet lysate. Thus, we describe a reproducible method that facilitates the reliable extraction of specific integrin-binding proteins from complex biological matrices. This methodology may enable the sensitive and specific identification of proteins that interact with linear, membrane-proximal peptide motifs such as the integrin regulatory motif LAMWKVGFFKR.

  14. Identification of key radionuclides in a nuclear waste repository in basalt

    International Nuclear Information System (INIS)

    Barney, G.S.; Wood, B.J.

    1980-05-01

    Radionuclides were identified which appear to pose the greatest potential hazard to man during long-term storage of nuclear waste in a repository mined in the Columbia Plateau basalt formation. The criteria used to select key radionuclides were as follows: quantity of radionuclide in stored waste; biological toxicity; leach rate of the wastes into groundwater; and transport rate via groundwater flow. The waste forms were assumed to be either unreprocessed spent fuel or borosilicate glass containing reprocessed high-level waste. The nuclear waste composition was assumed to be that from a light water reactor. Radionuclides were ranked according to quantity, toxicity, and release rate from the repository. These rankings were combined to obtain a single list of key radionuclides. The ten most important radionuclides in order of decreasing hazard are: 99 Tc, 129 I, 237 Np, 226 Ra, 107 Pd, 230 Th, 210 Pb, 126 Sn, 79 Se, and 242 Pu. Safety assessment studies and the design of engineered barriers should concentrate on containment of radionuclides in this list

  15. A Genetic Programming Method for the Identification of Signal Peptides and Prediction of Their Cleavage Sites

    Directory of Open Access Journals (Sweden)

    David Lennartsson

    2004-01-01

    Full Text Available A novel approach to signal peptide identification is presented. We use an evolutionary algorithm for automatic evolution of classification programs, so-called programmatic motifs. The variant of evolutionary algorithm used is called genetic programming where a population of solution candidates in the form of full computer programs is evolved, based on training examples consisting of signal peptide sequences. The method is compared with a previous work using artificial neural network (ANN approaches. Some advantages compared to ANNs are noted. The programmatic motif can perform computational tasks beyond that of feed-forward neural networks and has also other advantages such as readability. The best motif evolved was analyzed and shown to detect the h-region of the signal peptide. A powerful parallel computer cluster was used for the experiment.

  16. Identification of small peptides arising from hydrolysis of meat proteins in dry fermented sausages.

    Science.gov (United States)

    López, Constanza M; Bru, Elena; Vignolo, Graciela M; Fadda, Silvina G

    2015-06-01

    In this study, proteolysis and low molecular weight (LMW) peptides (<3kDa) from commercial Argentinean fermented sausages were characterized by applying a peptidomic approach. Protein profiles and peptides obtained by Tricine-SDS-PAGE and RP-HPLC-MS, respectively, allowed distinguishing two different types of fermented sausages, although no specific biomarkers relating to commercial brands or quality were recognized. From electrophoresis, α-actin, myoglobin, creatine kinase M-type and L-lactate dehydrogenase were degraded at different intensities. In addition, a partial characterization of fermented sausage peptidome through the identification of 36 peptides, in the range of 1000-2100 Da, arising from sarcoplasmic (28) and myofibrillar (8) proteins was achieved. These peptides had been originated from α-actin, myoglobin, and creatine kinase M-type, but also from the hydrolysis of other proteins not previously reported. Although muscle enzymes exerted a major role on peptidogenesis, microbial contribution cannot be excluded as it was postulated herein. This work represents a first peptidomic approach for fermented sausages, thereby providing a baseline to define key peptides acting as potential biomarkers. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Identification of ligand-selective peptidic ActRIIB-antagonists using phage display technology

    Directory of Open Access Journals (Sweden)

    Kotaro Sakamoto

    2017-09-01

    Full Text Available ActRIIB (activin receptor type-2B is an activin receptor subtype constitutively expressed in the whole body, playing a role in cellular proliferation, differentiation, and metabolism. For its various physiological activities, ActRIIB interacts with activin and multiple other ligands including myostatin (MSTN, growth differentiation factor 11 (GDF11, and bone morphogenetic protein 9 (BMP9. Notably, the protein-protein interaction (PPI between ActRIIB and MSTN negatively controls muscular development. Therefore, this PPI has been targeted for effective treatment of muscle degenerative diseases such as muscular dystrophy and sarcopenia. Here, we report the identification of ligand-selective peptidic ActRIIB-antagonists by phage display technology. Our peptides bound to the extracellular domain of ActRIIB, inhibited PPIs between ActRIIB expressed on the cell surface and its ligands, and subsequently suppressed activation of Smad that serves as the downstream signal of the ActRIIB pathway. Interestingly, these peptidic antagonists displayed different ligand selectivities; the AR2mini peptide inhibited multiple ligands (activin A, MSTN, GDF11, and BMP9, AR9 inhibited MSTN and GDF11, while AR8 selectively inhibited MSTN. This is the first report of artificial peptidic ActRIIB-antagonists possessing ligand-selectivity.

  18. Identification of human embryonic progenitor cell targeting peptides using phage display.

    Directory of Open Access Journals (Sweden)

    Paola A Bignone

    Full Text Available Human pluripotent stem (hPS cells are capable of differentiation into derivatives of all three primary embryonic germ layers and can self-renew indefinitely. They therefore offer a potentially scalable source of replacement cells to treat a variety of degenerative diseases. The ability to reprogram adult cells to induced pluripotent stem (iPS cells has now enabled the possibility of patient-specific hPS cells as a source of cells for disease modeling, drug discovery, and potentially, cell replacement therapies. While reprogramming technology has dramatically increased the availability of normal and diseased hPS cell lines for basic research, a major bottleneck is the critical unmet need for more efficient methods of deriving well-defined cell populations from hPS cells. Phage display is a powerful method for selecting affinity ligands that could be used for identifying and potentially purifying a variety of cell types derived from hPS cells. However, identification of specific progenitor cell-binding peptides using phage display may be hindered by the large cellular heterogeneity present in differentiating hPS cell populations. We therefore tested the hypothesis that peptides selected for their ability to bind a clonal cell line derived from hPS cells would bind early progenitor cell types emerging from differentiating hPS cells. The human embryonic stem (hES cell-derived embryonic progenitor cell line, W10, was used and cell-targeting peptides were identified. Competition studies demonstrated specificity of peptide binding to the target cell surface. Efficient peptide targeted cell labeling was accomplished using multivalent peptide-quantum dot complexes as detected by fluorescence microscopy and flow cytometry. The cell-binding peptides were selective for differentiated hPS cells, had little or no binding on pluripotent cells, but preferential binding to certain embryonic progenitor cell lines and early endodermal hPS cell derivatives. Taken

  19. Identification of a novel Plasmopara halstedii elicitor protein combining de novo peptide sequencing algorithms and RACE-PCR

    Directory of Open Access Journals (Sweden)

    Madlung Johannes

    2010-05-01

    Full Text Available Abstract Background Often high-quality MS/MS spectra of tryptic peptides do not match to any database entry because of only partially sequenced genomes and therefore, protein identification requires de novo peptide sequencing. To achieve protein identification of the economically important but still unsequenced plant pathogenic oomycete Plasmopara halstedii, we first evaluated the performance of three different de novo peptide sequencing algorithms applied to a protein digests of standard proteins using a quadrupole TOF (QStar Pulsar i. Results The performance order of the algorithms was PEAKS online > PepNovo > CompNovo. In summary, PEAKS online correctly predicted 45% of measured peptides for a protein test data set. All three de novo peptide sequencing algorithms were used to identify MS/MS spectra of tryptic peptides of an unknown 57 kDa protein of P. halstedii. We found ten de novo sequenced peptides that showed homology to a Phytophthora infestans protein, a closely related organism of P. halstedii. Employing a second complementary approach, verification of peptide prediction and protein identification was performed by creation of degenerate primers for RACE-PCR and led to an ORF of 1,589 bp for a hypothetical phosphoenolpyruvate carboxykinase. Conclusions Our study demonstrated that identification of proteins within minute amounts of sample material improved significantly by combining sensitive LC-MS methods with different de novo peptide sequencing algorithms. In addition, this is the first study that verified protein prediction from MS data by also employing a second complementary approach, in which RACE-PCR led to identification of a novel elicitor protein in P. halstedii.

  20. Data for identification of GPI-anchored peptides and ω-sites in cancer cell lines

    Directory of Open Access Journals (Sweden)

    Yusuke Masuishi

    2016-06-01

    Full Text Available We present data obtained using a focused proteomics approach to identify the glycosylphosphatidylinositol (GPI-anchored peptides in 19 human cancer cell lines. GPI-anchored proteins (GPI-APs, which localize to the outer leaflet of the membrane microdomains commonly referred to as lipid rafts play important roles in diverse biological processes. Due to the complex structure of the GPI-anchor moiety, it has been difficult to identify GPI-anchored peptide sequences on the proteomic scale by database searches using tools such as MASCOT. Here we provide data from 73 ω-sites derived from 49 GPI-APs in 19 human cancer cell lines. This article contains data related to the research article entitled “Identification of glycosylphosphatidylinositol-anchored proteins and ω-sites using TiO2-based affinity purification followed by hydrogen fluoride treatment” (Masuishi et al., 2016 [1].

  1. Classification of Clinical Research Study Eligibility Criteria to Support Multi-Stage Cohort Identification Using Clinical Data Repositories.

    Science.gov (United States)

    Cimino, James J; Lancaster, William J; Wyatt, Mathew C

    2017-01-01

    One of the challenges to using electronic health record (EHR) repositories for research is the difficulty mapping study subject eligibility criteria to the query capabilities of the repository. We sought to characterize criteria as "easy" (searchable in a typical repository), "hard" (requiring manual review of the record data), and "impossible" (not typically available in EHR repositories). We obtained 292 criteria from 20 studies available from Clinical Trials.gov and rated them according to our three types, plus a fourth "mixed" type. We had good agreement among three independent reviewers and chose 274 criteria that were characterized by single types for further analysis. The resulting analysis showed typical features of criteria that do and don't map to repositories. We propose that these features be used to guide researchers in specifying eligibility criteria to improve development of enrollment workflow, including the definition of EHR repository queries for self-service or analyst-mediated retrievals.

  2. Brute-Force Approach for Mass Spectrometry-Based Variant Peptide Identification in Proteogenomics without Personalized Genomic Data

    Science.gov (United States)

    Ivanov, Mark V.; Lobas, Anna A.; Levitsky, Lev I.; Moshkovskii, Sergei A.; Gorshkov, Mikhail V.

    2018-02-01

    In a proteogenomic approach based on tandem mass spectrometry analysis of proteolytic peptide mixtures, customized exome or RNA-seq databases are employed for identifying protein sequence variants. However, the problem of variant peptide identification without personalized genomic data is important for a variety of applications. Following the recent proposal by Chick et al. (Nat. Biotechnol. 33, 743-749, 2015) on the feasibility of such variant peptide search, we evaluated two available approaches based on the previously suggested "open" search and the "brute-force" strategy. To improve the efficiency of these approaches, we propose an algorithm for exclusion of false variant identifications from the search results involving analysis of modifications mimicking single amino acid substitutions. Also, we propose a de novo based scoring scheme for assessment of identified point mutations. In the scheme, the search engine analyzes y-type fragment ions in MS/MS spectra to confirm the location of the mutation in the variant peptide sequence.

  3. Processed Meat Protein and Heat-Stable Peptide Marker Identification Using Microwave-Assisted Tryptic Digestion

    Directory of Open Access Journals (Sweden)

    Magdalena Montowska

    2016-01-01

    Full Text Available New approaches to rapid examination of proteins and peptides in complex food matrices are of great interest to the community of food scientists. The aim of the study is to examine the influence of microwave irradiation on the acceleration of enzymatic cleavage and enzymatic digestion of denatured proteins in cooked meat of five species (cattle, horse, pig, chicken and turkey and processed meat products (coarsely minced, smoked, cooked and semi-dried sausages. Severe protein aggregation occurred not only in heated meat under harsh treatment at 190 °C but also in processed meat products. All the protein aggregates were thoroughly hydrolyzed aft er 1 h of trypsin treatment with short exposure times of 40 and 20 s to microwave irradiation at 138 and 303 W. There were much more missed cleavage sites observed in all microwave-assisted digestions. Despite the incompleteness of microwave-assisted digestion, six unique peptide markers were detected, which allowed unambiguous identification of processed meat derived from the examined species. Although the microwave-assisted tryptic digestion can serve as a tool for rapid and high-throughput protein identification, great caution and pre-evaluation of individual samples is recommended in protein quantitation.

  4. Maximizing the sensitivity and reliability of peptide identification in large-scale proteomic experiments by harnessing multiple search engines.

    Science.gov (United States)

    Yu, Wen; Taylor, J Alex; Davis, Michael T; Bonilla, Leo E; Lee, Kimberly A; Auger, Paul L; Farnsworth, Chris C; Welcher, Andrew A; Patterson, Scott D

    2010-03-01

    Despite recent advances in qualitative proteomics, the automatic identification of peptides with optimal sensitivity and accuracy remains a difficult goal. To address this deficiency, a novel algorithm, Multiple Search Engines, Normalization and Consensus is described. The method employs six search engines and a re-scoring engine to search MS/MS spectra against protein and decoy sequences. After the peptide hits from each engine are normalized to error rates estimated from the decoy hits, peptide assignments are then deduced using a minimum consensus model. These assignments are produced in a series of progressively relaxed false-discovery rates, thus enabling a comprehensive interpretation of the data set. Additionally, the estimated false-discovery rate was found to have good concordance with the observed false-positive rate calculated from known identities. Benchmarking against standard proteins data sets (ISBv1, sPRG2006) and their published analysis, demonstrated that the Multiple Search Engines, Normalization and Consensus algorithm consistently achieved significantly higher sensitivity in peptide identifications, which led to increased or more robust protein identifications in all data sets compared with prior methods. The sensitivity and the false-positive rate of peptide identification exhibit an inverse-proportional and linear relationship with the number of participating search engines.

  5. Endogenous Plasma Peptide Detection and Identification in the Rat by a Combination of Fractionation Methods and Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Fabrice Bertile

    2007-01-01

    Full Text Available Mass spectrometry-based analyses are essential tools in the field of biomarker research. However, detection and characterization of plasma low abundance and/or low molecular weight peptides is challenged by the presence of highly abundant proteins, salts and lipids. Numerous strategies have already been tested to reduce the complexity of plasma samples. The aim of this study was to enrich the low molecular weight fraction of rat plasma. To this end, we developed and compared simple protocols based on membrane filtration, solid phase extraction, and a combination of both. As assessed by UV absorbance, an albumin depletion 99% was obtained. The multistep fractionation strategy (including reverse phase HPLC allowed detection, in a reproducible manner (CV [1] 30%–35%, of more than 450 peaks below 3000 Da by MALDI-TOF/MS. A MALDI-TOF/MS-determined LOD as low as 1 fmol/μL was obtained, thus allowing nanoLC-Chip/ MS/MS identification of spiked peptides representing ∼10–6% of total proteins, by weight. Signal peptide recovery ranged between 5%–100% according to the spiked peptide considered. Tens of peptide sequence tags from endogenous plasma peptides were also obtained and high confidence identifications of low abundance fibrinopeptide A and B are reported here to show the efficiency of the protocol. It is concluded that the fractionation protocol presented would be of particular interest for future differential (high throughput analyses of the plasma low molecular weight fraction.

  6. Isolation and identification of a new intracellular antimicrobial peptide produced by Paenibacillus alvei AN5.

    Science.gov (United States)

    Alkotaini, Bassam; Anuar, Nurina; Kadhum, Abdul Amir Hassan; Sani, Asmahani Azira Abdu

    2014-04-01

    A wild-type, Gram-positive, rod-shaped, endospore-forming and motile bacteria has been isolated from palm oil mill sludge in Malaysia. Molecular identification using 16S rRNA gene sequence analysis indicated that the bacteria belonged to genus Paenibacillus. With 97 % similarity to P. alvei (AUG6), the isolate was designated as P. alvei AN5. An antimicrobial compound was extracted from P. alvei AN5-pelleted cells using 95 % methanol and was then lyophilized. Precipitates were re-suspended in phosphate buffered saline (PBS), producing an antimicrobial crude extract (ACE). The ACE showed antimicrobial activity against Salmonella enteritidis ATCC 13076, Escherichia coli ATCC 29522, Bacillus cereus ATCC 14579 and Lactobacillus plantarum ATCC 8014. By using SP-Sepharose cation exchange chromatography, Sephadex G-25 gel filtration and Tricine SDS-PAGE, the ACE was purified, which produced a ~2-kDa active band. SDS-PAGE and infrared (IR) spectroscopy indicated the proteinaceous nature of the antimicrobial compound in the ACE, and liquid chromatography electrospray ionization mass spectroscopy and de novo sequencing using an automatic, Q-TOF premier system detected a peptide with the amino acid sequence F-C-K-S-L-P-L-P-L-S-V-K (1,330.7789 Da). This novel peptide was designated as AN5-2. The antimicrobial peptide exhibited stability from pH 3 to 12 and maintained its activity after being heated to 90 °C. It also remained active after incubation with denaturants (urea, SDS and EDTA).

  7. Identification of MHC class II restricted T‐cell‐mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides

    DEFF Research Database (Denmark)

    Wang, Mingjun; Tang, Sheila Tuyet; Stryhn, Anette

    2011-01-01

    Major histocompatibility complex (MHC) class I restricted cytotoxic T lymphocytes (CTL) are known to play an important role in the control of Mycobacterium tuberculosis infection so identification of CTL epitopes from M. tuberculosis is of importance for the development of effective peptide...

  8. IL17eScan: A Tool for the Identification of Peptides Inducing IL-17 Response

    Directory of Open Access Journals (Sweden)

    Sudheer Gupta

    2017-10-01

    Full Text Available IL-17 cytokines are pro-inflammatory cytokines and are crucial in host defense against various microbes. Induction of these cytokines by microbial antigens has been investigated in the case of ischemic brain injury, gingivitis, candidiasis, autoimmune myocarditis, etc. In this study, we have investigated the ability of amino acid sequence of antigens to induce IL-17 response using machine-learning approaches. A total of 338 IL-17-inducing and 984 IL-17 non-inducing peptides were retrieved from Immune Epitope Database. 80% of the data were randomly selected as training dataset and rest 20% as validation dataset. To predict the IL-17-inducing ability of peptides/protein antigens, different sequence-based machine-learning models were developed. The performance of support vector machine (SVM and random forest (RF was compared with different parameters to predict IL-17-inducing epitopes (IIEs. The dipeptide composition-based SVM-model displayed an accuracy of 82.4% with Matthews correlation coefficient = 0.62 at polynomial (t = 1 kernel on 10-fold cross-validation and outperformed RF. Amino acid residues Leu, Ser, Arg, Asn, and Phe and dipeptides LL, SL, LK, IL, LI, NL, LR, FK, SF, and LE are abundant in IIEs. The present tool helps in the identification of IIEs using machine-learning approaches. The induction of IL-17 plays an important role in several inflammatory diseases, and identification of such epitopes would be of great help to the immunologists. It is freely available at http://metagenomics.iiserb.ac.in/IL17eScan/ and http://metabiosys.iiserb.ac.in/IL17eScan/.

  9. From GenBank to GBIF: Phylogeny-Based Predictive Niche Modeling Tests Accuracy of Taxonomic Identifications in Large Occurrence Data Repositories.

    Science.gov (United States)

    Smith, B Eugene; Johnston, Mark K; Lücking, Robert

    2016-01-01

    Accuracy of taxonomic identifications is crucial to data quality in online repositories of species occurrence data, such as the Global Biodiversity Information Facility (GBIF), which have accumulated several hundred million records over the past 15 years. These data serve as basis for large scale analyses of macroecological and biogeographic patterns and to document environmental changes over time. However, taxonomic identifications are often unreliable, especially for non-vascular plants and fungi including lichens, which may lack critical revisions of voucher specimens. Due to the scale of the problem, restudy of millions of collections is unrealistic and other strategies are needed. Here we propose to use verified, georeferenced occurrence data of a given species to apply predictive niche modeling that can then be used to evaluate unverified occurrences of that species. Selecting the charismatic lichen fungus, Usnea longissima, as a case study, we used georeferenced occurrence records based on sequenced specimens to model its predicted niche. Our results suggest that the target species is largely restricted to a narrow range of boreal and temperate forest in the Northern Hemisphere and that occurrence records in GBIF from tropical regions and the Southern Hemisphere do not represent this taxon, a prediction tested by comparison with taxonomic revisions of Usnea for these regions. As a novel approach, we employed Principal Component Analysis on the environmental grid data used for predictive modeling to visualize potential ecogeographical barriers for the target species; we found that tropical regions conform a strong barrier, explaining why potential niches in the Southern Hemisphere were not colonized by Usnea longissima and instead by morphologically similar species. This approach is an example of how data from two of the most important biodiversity repositories, GenBank and GBIF, can be effectively combined to remotely address the problem of inaccuracy of

  10. The preparation and identification of peptide imaging agent of lung cancer

    International Nuclear Information System (INIS)

    Chu Liping; Wang Yan; Wang Yueying; Liu Jinjian; Wu Hongying; Liu Jianfeng

    2010-01-01

    Objective: To screen in vivo lung cancer specific binding 7-peptide from T7 phage display random peptide library and prepare peptide imaging agent in early in early diagnosis of lung cancer. Methods: Used phage display in vivo technology to get the 7-peptide phage that can bind the lung cancer specifically, then sequenced and synthesized 7-peptide. After being labeled by 125 I, this 7-peptide was injected into mice via vein and the distribution in the mice tumor mold was observed. Results: One 7-peptide was obtained after four rounds of screening, and the peptide could bind lung cancer tissue specifically. Metabolism of this peptide in mice was fast and imaging of lung cancer was best two hours later after injection. The distribution in vivo decreased and almost disappeared after six hours. Conclusion: This 7-peptide could be used to image and diagnose of lung cancer effectively. (authors)

  11. STEPS: a grid search methodology for optimized peptide identification filtering of MS/MS database search results.

    Science.gov (United States)

    Piehowski, Paul D; Petyuk, Vladislav A; Sandoval, John D; Burnum, Kristin E; Kiebel, Gary R; Monroe, Matthew E; Anderson, Gordon A; Camp, David G; Smith, Richard D

    2013-03-01

    For bottom-up proteomics, there are wide variety of database-searching algorithms in use for matching peptide sequences to tandem MS spectra. Likewise, there are numerous strategies being employed to produce a confident list of peptide identifications from the different search algorithm outputs. Here we introduce a grid-search approach for determining optimal database filtering criteria in shotgun proteomics data analyses that is easily adaptable to any search. Systematic Trial and Error Parameter Selection--referred to as STEPS--utilizes user-defined parameter ranges to test a wide array of parameter combinations to arrive at an optimal "parameter set" for data filtering, thus maximizing confident identifications. The benefits of this approach in terms of numbers of true-positive identifications are demonstrated using datasets derived from immunoaffinity-depleted blood serum and a bacterial cell lysate, two common proteomics sample types. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. RAId_DbS: mass-spectrometry based peptide identification web server with knowledge integration

    Directory of Open Access Journals (Sweden)

    Ogurtsov Aleksey Y

    2008-10-01

    Full Text Available Abstract Background Existing scientific literature is a rich source of biological information such as disease markers. Integration of this information with data analysis may help researchers to identify possible controversies and to form useful hypotheses for further validations. In the context of proteomics studies, individualized proteomics era may be approached through consideration of amino acid substitutions/modifications as well as information from disease studies. Integration of such information with peptide searches facilitates speedy, dynamic information retrieval that may significantly benefit clinical laboratory studies. Description We have integrated from various sources annotated single amino acid polymorphisms, post-translational modifications, and their documented disease associations (if they exist into one enhanced database per organism. We have also augmented our peptide identification software RAId_DbS to take into account this information while analyzing a tandem mass spectrum. In principle, one may choose to respect or ignore the correlation of amino acid polymorphisms/modifications within each protein. The former leads to targeted searches and avoids scoring of unnecessary polymorphism/modification combinations; the latter explores possible polymorphisms in a controlled fashion. To facilitate new discoveries, RAId_DbS also allows users to conduct searches permitting novel polymorphisms as well as to search a knowledge database created by the users. Conclusion We have finished constructing enhanced databases for 17 organisms. The web link to RAId_DbS and the enhanced databases is http://www.ncbi.nlm.nih.gov/CBBResearch/qmbp/RAId_DbS/index.html. The relevant databases and binaries of RAId_DbS for Linux, Windows, and Mac OS X are available for download from the same web page.

  13. Optimization of hydrolysis conditions, isolation, and identification of neuroprotective peptides derived from seahorse Hippocampus trimaculatus.

    Science.gov (United States)

    Pangestuti, Ratih; Ryu, Bomi; Himaya, Swa; Kim, Se-Kwon

    2013-08-01

    Hippocampus trimaculatus is one of the most heavily traded seahorse species for traditional medicine purposes in many countries. In the present study, we showed neuroprotective effects of peptide derived from H. trimaculatus against amyloid-β42 (Aβ42) toxicity which are central to the pathogenesis of Alzheimer's diseases (AD). Firstly, H. trimaculatus was separately hydrolyzed by four different enzymes and tested for their protective effect on Aβ42-induced neurotoxicity in differentiated PC12 cells. Pronase E hydrolysate exerted highest protection with cell viability value of 88.33 ± 3.33 %. Furthermore, we used response surface methodology to optimize pronase E hydrolysis conditions and found that temperature at 36.69 °C with the hydrolysis time 20.01 h, enzyme to substrate (E/S) ratio of 2.02 % and pH 7.34 were the most optimum conditions. Following several purification steps, H. trimaculatus-derived neuroprotective peptides (HTP-1) sequence was identified as Gly-Thr-Glu-Asp-Glu-Leu-Asp-Lys (906.4 Da). HTP-1 protected PC12 cells from Aβ42-induced neuronal death with the cell viability value of 85.52 ± 2.22 % and up-regulated pro-survival gene (Bcl-2) expressions. These results suggest that HTP-1 has the potential to be used in treatment of neurodegenerative diseases, particularly AD. Identification, characterization, and synthesis of bioactive components derived from H. trimaculatus have the potential to replace or at least complement the use of seahorse as traditional medicine, which further may become an approach to minimize seahorse exploitation in traditional medicine.

  14. Identification and mapping of linear antibody epitopes in human serum albumin using high-density Peptide arrays.

    Directory of Open Access Journals (Sweden)

    Lajla Bruntse Hansen

    Full Text Available We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm(2. Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA and from rabbit serum albumin (RSA were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level.

  15. Identification and mapping of linear antibody epitopes in human serum albumin using high-density Peptide arrays.

    Science.gov (United States)

    Hansen, Lajla Bruntse; Buus, Soren; Schafer-Nielsen, Claus

    2013-01-01

    We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm(2). Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA) as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids) at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level.

  16. Identification of Cellular Binding Sites for a Novel Human Anti-Breast Cancer Peptide

    National Research Council Canada - National Science Library

    DeFreest, Lori

    2004-01-01

    ... breast cancer growth. We have developed and optimized an affinity chromatography procedure to identify the receptor for AFPep by using the peptide as "bait" to isolate proteins from solublized cells which have an affinity for the peptide...

  17. Fungal Secretome Analysis via PepSAVI-MS: Identification of the Bioactive Peptide KP4 from Ustilago maydis

    Science.gov (United States)

    Kirkpatrick, Christine L.; Parsley, Nicole C.; Bartges, Tessa E.; Cooke, Madeline E.; Evans, Wilaysha S.; Heil, Lilian R.; Smith, Thomas J.; Hicks, Leslie M.

    2018-05-01

    Fungal secondary metabolites represent a rich and largely untapped source for bioactive molecules, including peptides with substantial structural diversity and pharmacological potential. As methods proceed to take a deep dive into fungal genomes, complimentary methods to identify bioactive components are required to keep pace with the expanding fungal repertoire. We developed PepSAVI-MS to expedite the search for natural product bioactive peptides and herein demonstrate proof-of-principle applicability of the pipeline for the discovery of bioactive peptides from fungal secretomes via identification of the antifungal killer toxin KP4 from Ustilago maydis P4. This work opens the door to investigating microbial secretomes with a new lens, and could have broad applications across human health, agriculture, and food safety. [Figure not available: see fulltext.

  18. Signal peptide discrimination and cleavage site identification using SVM and NN.

    Science.gov (United States)

    Kazemian, H B; Yusuf, S A; White, K

    2014-02-01

    About 15% of all proteins in a genome contain a signal peptide (SP) sequence, at the N-terminus, that targets the protein to intracellular secretory pathways. Once the protein is targeted correctly in the cell, the SP is cleaved, releasing the mature protein. Accurate prediction of the presence of these short amino-acid SP chains is crucial for modelling the topology of membrane proteins, since SP sequences can be confused with transmembrane domains due to similar composition of hydrophobic amino acids. This paper presents a cascaded Support Vector Machine (SVM)-Neural Network (NN) classification methodology for SP discrimination and cleavage site identification. The proposed method utilises a dual phase classification approach using SVM as a primary classifier to discriminate SP sequences from Non-SP. The methodology further employs NNs to predict the most suitable cleavage site candidates. In phase one, a SVM classification utilises hydrophobic propensities as a primary feature vector extraction using symmetric sliding window amino-acid sequence analysis for discrimination of SP and Non-SP. In phase two, a NN classification uses asymmetric sliding window sequence analysis for prediction of cleavage site identification. The proposed SVM-NN method was tested using Uni-Prot non-redundant datasets of eukaryotic and prokaryotic proteins with SP and Non-SP N-termini. Computer simulation results demonstrate an overall accuracy of 0.90 for SP and Non-SP discrimination based on Matthews Correlation Coefficient (MCC) tests using SVM. For SP cleavage site prediction, the overall accuracy is 91.5% based on cross-validation tests using the novel SVM-NN model. © 2013 Published by Elsevier Ltd.

  19. Identification of release scenarios for a repository of radioactive waste in a salt dome in the Netherlands

    International Nuclear Information System (INIS)

    Glasbergen, P.; Hamstra, J.

    1981-01-01

    A review is presented of the long-term scenarios used in the safety analysis which was carried out for the disposal of radioactive waste in salt domes in the Netherlands. The long-term analysis involved the following natural processes or events: climatological and sea-level changes, glacial erosion, diapirism, subsidence, faulting and dissolution. The model calculations which were carried out showed the dominant parameters: the rate of diapirism and the rate of subsurface dissolution of rock salt. During the operational period the intrusion of water in the repository was considered to be the most hazardous event. Because the layout of the disposal mine, the disposal geometry and the disposal mining procedures were still under consideration, the first approach of a release scenario was made on a generic basis. A generic scenario is presented for the events during the flooding of the repository. The transport ways of water through the repository and its surroundings are indicated. It is concluded that release scenario analysis for long-term periods and for the operational period provides essential information to optimize the overall disposal system in an iterative process

  20. Complex mixture analysis of peptides using LC/LC-MS/MS and data-dependent protein identification

    International Nuclear Information System (INIS)

    Wasinger, V.; Corthals, G.

    2001-01-01

    The comprehensive identification of proteins within complex solutions by mass-spectrometry largely depends on the sensitivity, resolving power and sampling efficiency of the technology. An integrated orthogonal approach using Strong Cation Exchange-Reverse Phase-MS/MS (SCX-RP-MS/MS) was used to evaluate the data-dependent Collision Induced Dissociation (CID) of yeast peptides. Reverse phase gradient times of 4, 10. 30, 90, and 180 minutes allowed the identification of hundreds of proteins in a nearly automated fashion from nuclear, membrane, and cytosolic distributions. Many proteins from typically difficult to resolve regions of two-dimensional gels, such as >100kDa, > pI 9.0 and Codon Adaptation Index < 0.2, were also identified using this multi-dimensional separation technology. Few low mass proteins (<10kDa) were identified. The impact of scan-range and duty-cycle on CID of peptides will be discussed

  1. MULTIPRED2: A computational system for large-scale identification of peptides predicted to bind to HLA supertypes and alleles

    DEFF Research Database (Denmark)

    Zhang, Guang Lan; DeLuca, David S.; Keskin, Derin B.

    2011-01-01

    binding peptides and immunological hotspots in an intuitive manner and also to provide a global view of results as heat maps. Another function of MULTIPRED2, which has direct relevance to vaccine design, is the calculation of population coverage. Currently it calculates population coverage in five major...... groups in North America. MULTIPRED2 is an important tool to complement wet-lab experimental methods for identification of T-cell epitopes. It is available at http://cvc.dfci.harvard.edu/multipred2/....

  2. Use of synthetic peptide libraries for the H-2Kd binding motif identification.

    Science.gov (United States)

    Quesnel, A; Casrouge, A; Kourilsky, P; Abastado, J P; Trudelle, Y

    1995-01-01

    To identify Kd-binding peptides, an approach based on small peptide libraries has been developed. These peptide libraries correspond to all possible single-amino acid variants of a particular Kd-binding peptide, SYIPSAEYI, an analog of the Plasmodium berghei 252-260 antigenic peptide SYIPSAEKI. In the parent sequence, each position is replaced by all the genetically encoded amino acids (except cysteine). The multiple analog syntheses are performed either by the Divide Couple and Recombine method or by the Single Resin method and generate mixtures containing 19 peptides. The present report deals with the synthesis, the purification, the chemical characterization by amino acid analysis and electrospray mass spectrometry (ES-MS), and the application of such mixtures in binding tests with a soluble, functionally empty, single-chain H-2Kd molecule denoted SC-Kd. For each mixture, bound peptides were eluted and analyzed by sequencing. Since the binding tests were realized in noncompetitive conditions, our results show that a much broader set of peptides bind to Kd than expected from previous studies. This may be of practical importance when looking for low affinity peptides such as tumor peptides capable of eliciting protective immune response.

  3. Repositories; Repositorios

    Energy Technology Data Exchange (ETDEWEB)

    Freire, Carolina Braccini; Tello, Cledola Cassia Oliveira de [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)]. E-mails: cbf@cdtn.br; tellocc@cdtn.br

    2007-11-15

    The use of the nuclear energy is increasing in all areas. Then the radioactive waste management is in continuous development to comply the national and international established requirements. The final objective is to assure that it will not have any contamination of the public or the environmental, and that the exposition doses will be lower than the radiological protection limits. The multi barrier concept for the repository is internationally recognized. Among the repository types, the most used are: near surface, geological formations and of deposition in rock cavities. This article explains the concept and the types of repository and gives some examples of them. (author)

  4. Mass-spectrometric identification of a novel angiotensin peptide in human plasma

    DEFF Research Database (Denmark)

    Jankowski, Vera; Vanholder, Raymond; van der Giet, Markus

    2007-01-01

    Angiotensin peptides play a central role in cardiovascular physiology and pathology. Among these peptides, angiotensin II (Ang II) has been investigated most intensively. However, further angiotensin peptides such as Ang 1-7, Ang III, and Ang IV also contribute to vascular regulation, and may eli...... elicit additional, different, or even opposite effects to Ang II. Here, we describe a novel Ang II-related, strong vasoconstrictive substance in plasma from healthy humans and end-stage renal failure patients....

  5. Peptidomic Analysis of Fetal Heart Tissue for Identification of Endogenous Peptides Involved in Tetralogy of Fallot.

    Science.gov (United States)

    Zhang, Jingjing; Liang, Dong; Cheng, Qing; Cao, Li; Wu, Yun; Wang, Yan; Han, Shuping; Yu, Zhangbin; Cui, Xianwei; Xu, Tianhui; Ma, Dingyuan; Hu, Ping; Xu, Zhengfeng

    2017-06-01

    Tetralogy of fallot (TOF) is one of the most prevalent types of congenital heart diseases. As a category of bioactive molecules, peptides have been proved to participate in various biological processes. However, the role of endogenous peptides in the pathogenesis of TOF has not been studied. In this study, we performed a comparative peptidomic profile in the fetal heart of TOF and the control group for the first time by liquid chromatography-tandem mass spectrometry. Our data demonstrated that a total of 201 peptides derived from 176 precursor proteins were differentially expressed in the heart tissues of TOF fetuses compared with normal controls, including 41 upregulated peptides and 160 downregulated peptides. After analyzing the characteristics of these differentially expressed peptides and their precursor proteins, we found that these peptides were potentially involved in different biological processes, especially cardiogenesis and congenital anomaly of the cardiovascular system. Interestingly, we detected several extracellular matrix-derived peptides involved in our differentially expressed peptidomic profile. In summary, our study constructed a comparative peptidomic profile from the heart tissues of TOF fetuses and normal controls, and it identified a series of peptides that could potentially participate in heart development and TOF formation. The emergence of our peptidomics study indicated a new perspective to explore the pathogenesis of abnormal heart morphology, especially TOF.

  6. Identification of Angiotensin I-Converting Enzyme Inhibitory Peptides Derived from Enzymatic Hydrolysates of Razor Clam Sinonovacula constricta

    Directory of Open Access Journals (Sweden)

    Yun Li

    2016-06-01

    Full Text Available Angiotensin I-converting enzyme (ACE inhibitory activity of razor clam hydrolysates produced using five proteases, namely, pepsin, trypsin, alcalase, flavourzyme and proteases from Actinomucor elegans T3 was investigated. Flavourzyme hydrolysate showed the highest level of degree of hydrolysis (DH (45.87% followed by A. elegans T3 proteases hydrolysate (37.84% and alcalase (30.55%. The A. elegans T3 proteases was observed to be more effective in generating small peptides with ACE-inhibitory activity. The 3 kDa membrane permeate of A. elegans T3 proteases hydrolysate showed the highest ACE-inhibitory activity with an IC50 of 0.79 mg/mL. After chromatographic separation by Sephadex G-15 gel filtration and reverse phase-high performance liquid chromatography, the potent fraction was subjected to MALDI/TOF-TOF MS/MS for identification. A novel ACE-inhibitory peptide (VQY was identified exhibiting an IC50 of 9.8 μM. The inhibitory kinetics investigation by Lineweaver-Burk plots demonstrated that the peptide acts as a competitive ACE inhibitor. The razor clam hydrolysate obtained by A. elegans T3 proteases could serve as a source of functional peptides with ACE-inhibitory activity for physiological benefits.

  7. Design and application of a data-independent precursor and product ion repository.

    Science.gov (United States)

    Thalassinos, Konstantinos; Vissers, Johannes P C; Tenzer, Stefan; Levin, Yishai; Thompson, J Will; Daniel, David; Mann, Darrin; DeLong, Mark R; Moseley, M Arthur; America, Antoine H; Ottens, Andrew K; Cavey, Greg S; Efstathiou, Georgios; Scrivens, James H; Langridge, James I; Geromanos, Scott J

    2012-10-01

    The functional design and application of a data-independent LC-MS precursor and product ion repository for protein identification, quantification, and validation is conceptually described. The ion repository was constructed from the sequence search results of a broad range of discovery experiments investigating various tissue types of two closely related mammalian species. The relative high degree of similarity in protein complement, ion detection, and peptide and protein identification allows for the analysis of normalized precursor and product ion intensity values, as well as standardized retention times, creating a multidimensional/orthogonal queryable, qualitative, and quantitative space. Peptide ion map selection for identification and quantification is primarily based on replication and limited variation. The information is stored in a relational database and is used to create peptide- and protein-specific fragment ion maps that can be queried in a targeted fashion against the raw or time aligned ion detections. These queries can be conducted either individually or as groups, where the latter affords pathway and molecular machinery analysis of the protein complement. The presented results also suggest that peptide ionization and fragmentation efficiencies are highly conserved between experiments and practically independent of the analyzed biological sample when using similar instrumentation. Moreover, the data illustrate only minor variation in ionization efficiency with amino acid sequence substitutions occurring between species. Finally, the data and the presented results illustrate how LC-MS performance metrics can be extracted and utilized to ensure optimal performance of the employed analytical workflows.

  8. Characterization and identification of novel antidiabetic and anti-obesity peptides from camel milk protein hydrolysates.

    Science.gov (United States)

    Mudgil, Priti; Kamal, Hina; Yuen, Gan Chee; Maqsood, Sajid

    2018-09-01

    In-vitro inhibitory properties of peptides released from camel milk proteins against dipeptidyl peptidase-IV (DPP-IV), porcine pancreatic α-amylase (PPA), and porcine pancreatic lipase (PPL) were studied. Results revealed that upon hydrolysis by different enzymes, camel milk proteins displayed dramatic increase in inhibition of DPP-IV and PPL, but slight improvement in PPA inhibition was noticed. Peptide sequencing revealed a total of 20 and 3 peptides for A9 and B9 hydrolysates respectively, obtained the score of 0.8 or more on peptide ranker and were categorized as potential DPP-IV inhibitory peptides. KDLWDDFKGL in A9 and MPSKPPLL in B9 were identified as most potent PPA inhibitory peptide. For PPL inhibition only 7 and 2 peptides qualified as PPL inhibitory peptides from hydrolysates A9 and B9, respectively. The present study report for the first time PPA and PPL inhibitory and only second for DPP-IV inhibitory potential of protein hydrolysates from camel milk. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. Automated multi-dimensional liquid chromatography : sample preparation and identification of peptides from human blood filtrate

    NARCIS (Netherlands)

    Machtejevas, Egidijus; John, Harald; Wagner, Knut; Standker, Ludger; Marko-Varga, Gyorgy; Georg Forssmann, Wolf; Bischoff, Rainer; K. Unger, Klaus

    2004-01-01

    A comprehensive on-line sample clean-up with an integrated two-dimensional HPLC system was developed for the analysis of natural peptides. Samples comprised of endogenous peptides with molecular weights up to 20 kDa were generated from human hemofiltrate (HF) obtained from patients with chronic

  10. Hydrolysis of Whey Protein Isolate with Bacillus licheniformis Protease: Fractionation and Identification of Aggregating Peptides

    NARCIS (Netherlands)

    Creusot, N.P.; Gruppen, H.

    2007-01-01

    The objective of this work was to identify the dominant aggregating peptides from a whey protein hydrolysate (degree of hydrolysis of 6.8%) obtained with Bacillus licheniformis protease. The aggregating peptides were fractionated with preparative reversed-phase chromatography and identified with

  11. Interactions of photoactive DNAs with terminal deoxynucleotidyl transferase: Identification of peptides in the DNA binding domain

    International Nuclear Information System (INIS)

    Farrar, Y.J.K.; Evans, R.K.; Beach, C.M.; Coleman, M.S.

    1991-01-01

    Terminal deoxynucleotidyl transferase (terminal transferase) was specifically modified in the DNA binding site by a photoactive DNA substrate (hetero-40-mer duplex containing eight 5-azido-dUMP residues at one 3' end). Under optimal photolabeling conditions, 27-40% of the DNA was covalently cross-linked to terminal transferase. The specificity of the DNA and protein interaction was demonstrated by protection of photolabeling at the DNA binding domain with natural DNA substrates. In order to recover high yields of modified peptides from limited amounts of starting material, protein modified with 32 P-labeled photoactive DNA and digested with trypsin was extracted 4 times with phenol followed by gel filtration chromatography. All peptides not cross-linked to DNA were extracted into the phenol phase while the photolyzed DNA and the covalently cross-linked peptides remained in the aqueous phase. The 32 P-containing peptide-DNA fraction was subjected to amino acid sequence analysis. Two sequences, Asp 221 -Lys 231 (peptide B8) and Cys 234 -Lys 249 (peptide B10), present in similar yield, were identified. Structure predictions placed the two peptides in an α-helical array of 39 angstrom which would accommodate a DNA helix span of 11 nucleotides. These peptides share sequence similarity with a region in DNA polymerase β that has been implicated in the binding of DNA template

  12. Identification of diverse archaeal proteins with class III signal peptides cleaved by distinct archaeal prepilin peptidases

    NARCIS (Netherlands)

    Szabó, Zalán; Oliveira Stahl, Adriana; Albers, Sonja-V.; Kissinger, Jessica C.; Driessen, Arnold J.M.; Pohlschröder, Mechthild; Pohlschroder, M.

    2007-01-01

    Most secreted archaeal proteins are targeted to the membrane via a tripartite signal composed of a charged N terminus and a hydrophobic domain, followed by a signal peptidase-processing site. Signal peptides of archaeal flagellins, similar to class III signal peptides of bacterial type IV pilins,

  13. Isolation and identification of a cardioactive peptide from Tenebrio molitor and Spodoptera eridania.

    Science.gov (United States)

    Furuya, K; Liao, S; Reynolds, S E; Ota, R B; Hackett, M; Schooley, D A

    1993-12-01

    We isolated several cardioactive peptides from extracts of whole heads of the mealworm, Tenebrio molitor, and the southern armyworm, Spodoptera eridania, using a semi-isolated heart of Manduca sexta for bioassay. We have now isolated from each species the peptide with the strongest effect on rate of contraction of the heart. The peptides were identified using micro Edman sequencing and mass spectrometric methods. This cardioactive peptide has the same primary structure from both species: Pro-Phe-Cys-Asn-Ala-Phe-Thr-Gly-Cys-NH2, a cyclic nonapeptide which is identical to crustacean cardioactive peptide (CCAP) originally isolated from the shore crab, Carcinus maenas, and subsequently isolated from Locusta migratoria and Manduca sexta. This is additional evidence that CCAP has widespread occurrence in arthropoda.

  14. Fractionation and identification of novel antioxidant peptides from buffalo and bovine casein hydrolysates.

    Science.gov (United States)

    Shazly, Ahmed Behdal; He, Zhiyong; El-Aziz, Mahmoud Abd; Zeng, Maomao; Zhang, Shuang; Qin, Fang; Chen, Jie

    2017-10-01

    Buffalo and bovine caseins were hydrolysed by alcalase and trypsin to produce novel antioxidant peptides. The casein hydrolysates were purified using ultrafiltration (UF) and further characterized by RP-HPLC. The fractions produced higher antioxidant activities were identified for their peptides using LC MS/MS. All UF-VI (MWcasein (UF-VI with 54.84-fold purification) showed higher antioxidant activity than that obtained by trypsin. Trypsin hydrolysate contained high amount of hydrophobic amino acids while alcalase hydrolysate consisted mainly of Ser, Arg, Ala and Leu. The antioxidant peptides identified by LC MS/MS were RELEE, MEDNKQ and TVA, EQL in buffalo casein hydrolysates produced by trypsin and alcalase, respectively. Mechanism and reaction pathways of selected antioxidant peptides with ABTS were proposed. Conclusively, buffalo casein provided antioxidant peptides similar to bovine, suggesting that buffalo casein is a novel source of antioxidant. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Identification of Peptides in Flowers of Sambucus nigra with Antimicrobial Activity against Aquaculture Pathogens.

    Science.gov (United States)

    Álvarez, Claudio Andrés; Barriga, Andrés; Albericio, Fernando; Romero, María Soledad; Guzmán, Fanny

    2018-04-27

    The elder ( Sambucus spp.) tree has a number of uses in traditional medicine. Previous studies have demonstrated the antimicrobial properties of elderberry liquid extract against human pathogenic bacteria and also influenza viruses. These properties have been mainly attributed to phenolic compounds. However, other plant defense molecules, such as antimicrobial peptides (AMPs), may be present. Here, we studied peptide extracts from flowers of Sambucus nigra L. The mass spectrometry analyses determined peptides of 3 to 3.6 kDa, among them, cysteine-rich peptides were identified with antimicrobial activity against various Gram-negative bacteria, including recurrent pathogens of Chilean aquaculture. In addition, membrane blebbing on the bacterial surface after exposure to the cyclotide was visualized by SEM microscopy and SYTOX Green permeabilization assay showed the ability to disrupt the bacterial membrane. We postulate that these peptides exert their action by destroying the bacterial membrane.

  16. Identification and applicability of analogues for a safety case for a HLW repository in evaporites: results from a NEA workshop

    Energy Technology Data Exchange (ETDEWEB)

    Noseck, U.; Wolf, J. [Gesellschaft für Anlagen und Reaktorsicherheit (GRS) mbH, Brunswick (Germany); Steininger, W. [Project Management Agency Karslruhe Water Technology and Waste Management, PTKA-WTE, Karlsruhe Institute of Technology, KIT, Eggenstein-Leopoldshafen (Germany); Miller, B. [AMEC, The Renaissance Center, Warrington (United Kingdom)

    2015-06-15

    A workshop was held in September 2012 in Braunschweig, Germany, to discuss the potential for natural and anthropogenic analogue studies to contribute to safety cases for radioactive waste repositories constructed in salt formations. Presentations were given on many analogue sites and systems from different countries. Discussions at the workshop then addressed the following aspects that are particularly relevant to the safety concept for radioactive waste disposal in salt: (1) the long-term integrity of rock salt formations, (2) the integrity of technical barriers, and (3) microbial, chemical and transport processes. A diverse range of natural systems were discussed as potential analogues for the integrity of rock salt. These included the deformation of anhydrite layers in rock salt; the response of rock salt to mechanical and thermal loads; and the isotopic signatures of syngenetic waters contained in fluid inclusions. Some anthropogenic examples drawn from the oil and gas industries, and from hazardous waste disposal, were proposed as analogues for the integrity of (geo)technical barriers. A broad range of studies on natural and anthropogenic salt-brine systems were identified as potential analogues for the radionuclide sorption and (co)precipitation process that may take place in the repository near and far fields, as well as for understanding the significance of hydrocarbons and microbial processes. It was evident from discussions at the workshop that there are some specific technical issues that may benefit from further analogue study, particularly the compaction of crushed salt backfill, the viability of microbes in the near-field, the stability of plugs and seals, the deformation of anhydrite, and isotope signatures in fluid inclusions. (authors)

  17. Identification of Four-Jointed Box 1 (FJX1-Specific Peptides for Immunotherapy of Nasopharyngeal Carcinoma.

    Directory of Open Access Journals (Sweden)

    San Jiun Chai

    Full Text Available Nasopharyngeal carcinoma (NPC is highly prevalent in South East Asia and China. The poor outcome is due to late presentation, recurrence, distant metastasis and limited therapeutic options. For improved treatment outcome, immunotherapeutic approaches focusing on dendritic and autologous cytotoxic T-cell based therapies have been developed, but cost and infrastructure remain barriers for implementing these in low-resource settings. As our prior observations had found that four-jointed box 1 (FJX1, a tumor antigen, is overexpressed in NPCs, we investigated if short 9-20 amino acid sequence specific peptides matching to FJX1 requiring only intramuscular immunization to train host immune systems would be a better treatment option for this disease. Thus, we designed 8 FJX1-specific peptides and implemented an assay system to first, assess the binding of these peptides to HLA-A2 molecules on T2 cells. After, ELISPOT assays were used to determine the peptides immunogenicity and ability to induce potential cytotoxicity activity towards cancer cells. Also, T-cell proliferation assay was used to evaluate the potential of MHC class II peptides to stimulate the expansion of isolated T-cells. Our results demonstrate that these peptides are immunogenic and peptide stimulated T-cells were able to induce peptide-specific cytolytic activity specifically against FJX1-expressing cancer cells. In addition, we demonstrated that the MHC class II peptides were capable of inducing T-cell proliferation. Our results suggest that these peptides are capable of inducing specific cytotoxic cytokines secretion against FJX1-expressing cancer cells and serve as a potential vaccine-based therapy for NPC patients.

  18. Comparison of trapping profiles between d-peptides and glutathione in the identification of reactive metabolites

    Directory of Open Access Journals (Sweden)

    Jaana E. Laine

    2015-01-01

    Full Text Available Qualitative trapping profile of reactive metabolites arising from six structurally different compounds was tested with three different d-peptide isomers (Peptide 1, gly–tyr–pro–cys–pro–his-pro; Peptide 2, gly–tyr–pro–ala–pro–his–pro; Peptide 3, gly–tyr–arg–pro–cys–pro–his–lys–pro and glutathione (GSH using mouse and human liver microsomes as the biocatalyst. The test compounds were classified either as clinically “safe” (amlodipine, caffeine, ibuprofen, or clinically as “risky” (clozapine, nimesulide, ticlopidine; i.e., associated with severe clinical toxicity outcomes. Our working hypothesis was as follows: could the use of short different amino acid sequence containing d-peptides in adduct detection confer any add-on value to that obtained with GSH? All “risky” agents’ resulted in the formation of several GSH adducts in the incubation mixture and with at least one peptide adduct with both microsomal preparations. Amlodipine did not form any adducts with any of the trapping agents. No GSH and peptide 2 and 3 adducts were found with caffeine, but with peptide 1 one adduct with human liver microsomes was detected. Ibuprofen produced one Peptide 1-adduct with human and mouse liver microsomes but not with GSH. In conclusion, GSH still remains the gold trapping standard for reactive metabolites. However, targeted d-peptides could provide additional information about protein binding potential of electrophilic agents, but their clinical significance needs to be clarified using a wider spectrum of chemicals together with other safety estimates.

  19. Identification of a peptide binding protein that plays a role in antigen presentation

    International Nuclear Information System (INIS)

    Lakey, E.K.; Margoliash, E.; Pierce, S.K.

    1987-01-01

    The helper T-cell response to globular proteins appears, in general, to require intracellular processing of the antigen, such that a peptide fragment containing the T-cell antigenic determinant is released and transported to and held on the surface of an Ia-expressing, antigen-presenting cell. However, the molecular details underlying these phenomena are largely unknown. The means by which antigenic peptides are anchored on the antigen-presenting cell surface was investigated. A cell surface protein is identified that was isolated by it ability to bind to a 24-amino acid peptide fragment of pigeon cytochrome c, residues 81-104, containing the major antigenic determinant for B10.A mouse T cells. This peptide binding protein, purified from [ 35 S]methionine-labeled cells, appears as two discrete bands of ≅72 and 74 kDa after NaDodSO 4 /PAGE. The protein can be eluted from the peptide affinity column with equivalent concentrations of either the antigenic pigeon cytochrome c peptide or the corresponding nonantigenic peptide of mouse cytochrome c. However, it does not bind to the native cytochromes c, either of pigeon or mouse, and thus the protein appears to recognize some structure available only in the free peptides. This protein plays a role in antigen presentation. Its expression is not major histocompatibility complex-restricted in that the blocking activity of the antisera can be absorbed on spleen cells from mice of different haplotypes. This peptide binding protein can be isolated from a variety of cell types, including B cells, T cells, and fibroblasts. The anchoring of processed peptides on the cell surface by such a protein may play a role in antigen presentation

  20. Identification of Four-Jointed Box 1 (FJX1)-Specific Peptides for Immunotherapy of Nasopharyngeal Carcinoma

    Science.gov (United States)

    Chai, San Jiun; Yap, Yoke Yeow; Foo, Yoke Ching; Yap, Lee Fah; Ponniah, Sathibalan; Teo, Soo Hwang; Cheong, Sok Ching; Patel, Vyomesh; Lim, Kue Peng

    2015-01-01

    Nasopharyngeal carcinoma (NPC) is highly prevalent in South East Asia and China. The poor outcome is due to late presentation, recurrence, distant metastasis and limited therapeutic options. For improved treatment outcome, immunotherapeutic approaches focusing on dendritic and autologous cytotoxic T-cell based therapies have been developed, but cost and infrastructure remain barriers for implementing these in low-resource settings. As our prior observations had found that four-jointed box 1 (FJX1), a tumor antigen, is overexpressed in NPCs, we investigated if short 9–20 amino acid sequence specific peptides matching to FJX1 requiring only intramuscular immunization to train host immune systems would be a better treatment option for this disease. Thus, we designed 8 FJX1-specific peptides and implemented an assay system to first, assess the binding of these peptides to HLA-A2 molecules on T2 cells. After, ELISPOT assays were used to determine the peptides immunogenicity and ability to induce potential cytotoxicity activity towards cancer cells. Also, T-cell proliferation assay was used to evaluate the potential of MHC class II peptides to stimulate the expansion of isolated T-cells. Our results demonstrate that these peptides are immunogenic and peptide stimulated T-cells were able to induce peptide-specific cytolytic activity specifically against FJX1-expressing cancer cells. In addition, we demonstrated that the MHC class II peptides were capable of inducing T-cell proliferation. Our results suggest that these peptides are capable of inducing specific cytotoxic cytokines secretion against FJX1-expressing cancer cells and serve as a potential vaccine-based therapy for NPC patients. PMID:26536470

  1. Comprehensive Identification of Glycated Peptides and Their Glycation Motifs in Plasma and Erythrocytes of Control and Diabetic Subjects

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Qibin; Monroe, Matthew E.; Schepmoes, Athena A.; Clauss, Therese RW; Gritsenko, Marina A.; Meng, Da; Petyuk, Vladislav A.; Smith, Richard D.; Metz, Thomas O.

    2011-07-01

    Non-enzymatic glycation of proteins is implicated in diabetes mellitus and its related complications. In this report, we extend our previous development and refinement of proteomics-based methods for the analysis of non-enzymatically glycated proteins to comprehensively identify glycated proteins in normal and diabetic human plasma and erythrocytes. Using immunodepletion, enrichment, and fractionation strategies, we identified 7749 unique glycated peptides, corresponding to 3742 unique glycated proteins. Semi-quantitative comparisons revealed a number of proteins with glycation levels significantly increased in diabetes relative to control samples and that erythrocyte proteins are more extensively glycated than plasma proteins. A glycation motif analysis revealed amino acids that are favored more than others in the protein primary structures in the vicinity of the glycation sites in both sample types. The glycated peptides and corresponding proteins reported here provide a foundation for the potential identification of novel markers for diabetes, glycemia, or diabetic complications.

  2. Preliminary screening and identification of the hepatocarcinoma cell-binding peptide

    International Nuclear Information System (INIS)

    Zhu Xiaohua; Wu Hua

    2004-01-01

    Objective: To explore the feasibility of screening and isolating homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display random peptide library and to develop a new peptide which may be potentially used as targeting delivery carrier in the biological targeted diagnosis or therapy for liver cancer. Methods: A 12-mer peptide phage display library was used to screen and isolate peptides that bind to human hepatocarcinoma cells, and four rounds of subtractive panning were carried out with the human hepatocarcinoma cell line HepG2 as the target. The affinities of selected phage clones for human hepatocarcinoma cells were determined with enzyme-linked immunosorbent assay (ELISA) and compared with that to human liver cell and other tumor cells of different tissue origins, respectively. In addition, the binding site in the tumor cells was observed with immunofluorescence analysis under confocal light microscopy. The amino acid sequences of phages that bind HepG2 specifically were deduced through DNA sequencing. Based on the results of DNA sequence, a 16-mer peptide (WH16) was designed and synthesized. Binding ability of the new peptide, WH16, was determined with competitive inhibition test. Results: After four rounds of panning, the phages that were bound to and internalized in human hepatocarcinoma cells were isolated. ELISA and immunofluorescence analysis confirmed the affinity of these phages for hepatocarcinoma cells. 56.67%(17/30) of the isolated phages displayed repeated sequence FLLEPHLMDTSM, and FLEP was defined as conservative motif . Binding of the selected phage to HepG2 cells was inhibited by synthesized peptide WH16, that strongly support that cellular binding of the phage is mediated through its displayed peptide, and WH16 can also bind to HepG2. Conclusions: It is feasible to screen and isolate homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display random peptide

  3. Preliminary screening and identification of the peptide binding to hepatocarcinoma cell

    International Nuclear Information System (INIS)

    Zhu Xiaohua; Wu Ha

    2004-01-01

    Objective: The present study was performed to screen and isolate homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display of random peptide library with the purpose of developing a new peptide which may be potentially used as target delivery carrier in the biological target diagnosis or therapy for liver cancer. Methods: A peptide 12-mer phage display library was used to screen and isolate peptide that bind to human hepatocarcinoma cell, and four rounds subtractive panning were carried out with the human hepatocarcinoma cell line HepG2 as the target. The affinities of selected phage clones to human hepatocarcinoma cell were determined with ELISA and compared with human liver cell and other tumor cells of different tissue origins respectively. In addition, the binding site in the tumor cells was observed with immunofluorescence analysis under confocal light microscopy. The amino acid sequences of phages that bind HepG2 specifically were deduced though DNA sequencing. Based on the results of DNA sequence, a 16-mer peptide (WH16) was designed and synthesized. Binding ability of the new peptide WH16 was determined with competitive inhibition test. Results: After four rounds panning, the phages that bound to and internalized in human hepatocarcinoma cell were isolated. ELISA and immunofluorescence analysis confirmed the affinity of these phages to hepatpcarcinoma cells 56.57%(17/30) of the isolated phages displayed repeated sequence FLLEPHLMDTSM, and FLEP was defined as conservative motif. Binding of the selected phage to HepG2 cells was inhibited by synthesized peptide WH16, which strongly support that cellular binding of phage is mediated though its displayed peptide and WH16 can also bind to HepG2. Conclusion: It is feasible to screen and isolate homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display of random peptide libraries. The sequence of peptide that can bind to

  4. Preliminary screening and identification of the hepatocarcinoma cell-binding peptide

    Energy Technology Data Exchange (ETDEWEB)

    Xiaohua, Zhu; Hua, Wu [Department of Nuclear Medicine, Tongji Hospital, Tongji Medical College, Huazhong Univ. of Science and Technology, Wuhan (China)

    2004-12-15

    Objective: To explore the feasibility of screening and isolating homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display random peptide library and to develop a new peptide which may be potentially used as targeting delivery carrier in the biological targeted diagnosis or therapy for liver cancer. Methods: A 12-mer peptide phage display library was used to screen and isolate peptides that bind to human hepatocarcinoma cells, and four rounds of subtractive panning were carried out with the human hepatocarcinoma cell line HepG2 as the target. The affinities of selected phage clones for human hepatocarcinoma cells were determined with enzyme-linked immunosorbent assay (ELISA) and compared with that to human liver cell and other tumor cells of different tissue origins, respectively. In addition, the binding site in the tumor cells was observed with immunofluorescence analysis under confocal light microscopy. The amino acid sequences of phages that bind HepG2 specifically were deduced through DNA sequencing. Based on the results of DNA sequence, a 16-mer peptide (WH16) was designed and synthesized. Binding ability of the new peptide, WH16, was determined with competitive inhibition test. Results: After four rounds of panning, the phages that were bound to and internalized in human hepatocarcinoma cells were isolated. ELISA and immunofluorescence analysis confirmed the affinity of these phages for hepatocarcinoma cells. 56.67%(17/30) of the isolated phages displayed repeated sequence FLLEPHLMDTSM, and FLEP was defined as conservative motif . Binding of the selected phage to HepG2 cells was inhibited by synthesized peptide WH16, that strongly support that cellular binding of the phage is mediated through its displayed peptide, and WH16 can also bind to HepG2. Conclusions: It is feasible to screen and isolate homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display random peptide

  5. Preliminary screening and identification of the peptide binding to hepatocarcinoma cell

    Energy Technology Data Exchange (ETDEWEB)

    Xiaohua, Zhu; Ha, Wu [Department of Nuclear Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan (China)

    2004-07-01

    Objective: The present study was performed to screen and isolate homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display of random peptide library with the purpose of developing a new peptide which may be potentially used as target delivery carrier in the biological target diagnosis or therapy for liver cancer. Methods: A peptide 12-mer phage display library was used to screen and isolate peptide that bind to human hepatocarcinoma cell, and four rounds subtractive panning were carried out with the human hepatocarcinoma cell line HepG2 as the target. The affinities of selected phage clones to human hepatocarcinoma cell were determined with ELISA and compared with human liver cell and other tumor cells of different tissue origins respectively. In addition, the binding site in the tumor cells was observed with immunofluorescence analysis under confocal light microscopy. The amino acid sequences of phages that bind HepG2 specifically were deduced though DNA sequencing. Based on the results of DNA sequence, a 16-mer peptide (WH16) was designed and synthesized. Binding ability of the new peptide WH16 was determined with competitive inhibition test. Results: After four rounds panning, the phages that bound to and internalized in human hepatocarcinoma cell were isolated. ELISA and immunofluorescence analysis confirmed the affinity of these phages to hepatpcarcinoma cells 56.57%(17/30) of the isolated phages displayed repeated sequence FLLEPHLMDTSM, and FLEP was defined as conservative motif. Binding of the selected phage to HepG2 cells was inhibited by synthesized peptide WH16, which strongly support that cellular binding of phage is mediated though its displayed peptide and WH16 can also bind to HepG2. Conclusion: It is feasible to screen and isolate homing peptides that bind specifically, or preferentially, to hepatocarcinoma cells using phage display of random peptide libraries. The sequence of peptide that can bind to

  6. Identification and Relative Quantification of Bioactive Peptides Sequentially Released during Simulated Gastrointestinal Digestion of Commercial Kefir.

    Science.gov (United States)

    Liu, Yufang; Pischetsrieder, Monika

    2017-03-08

    Health-promoting effects of kefir may be partially caused by bioactive peptides. To evaluate their formation or degradation during gastrointestinal digestion, we monitored changes of the peptide profile in a model of (1) oral, (2) gastric, and (3) small intestinal digestion of kefir. Matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy analyses revealed clearly different profiles between digests 2/3 and kefir/digest 1. Subsequent ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry identified 92 peptides in total (25, 25, 43, and 30, partly overlapping in kefir and digests 1, 2, and 3, respectively), including 16 peptides with ascribed bioactivity. Relative quantification in scheduled multiple reaction monitoring mode showed that many bioactive peptides were released by simulated digestion. Most prominently, the concentration of angiotensin-converting enzyme inhibitor β-casein 203-209 increased approximately 10 000-fold after combined oral, gastric, and intestinal digestion. Thus, physiological digestive processes may promote bioactive peptide formation from proteins and oligopeptides in kefir. Furthermore, bioactive peptides present in certain compartments of the gastrointestinal tract may exert local physiological effects.

  7. Identification of novel peptides for horse meat speciation in highly processed foodstuffs.

    Science.gov (United States)

    Claydon, Amy J; Grundy, Helen H; Charlton, Adrian J; Romero, M Rosario

    2015-01-01

    There is a need for robust analytical methods to support enforcement of food labelling legislation. Proteomics is emerging as a complementary methodology to existing tools such as DNA and antibody-based techniques. Here we describe the development of a proteomics strategy for the determination of meat species in highly processed foods. A database of specific peptides for nine relevant animal species was used to enable semi-targeted species determination. This principle was tested for horse meat speciation, and a range of horse-specific peptides were identified as heat stable marker peptides for the detection of low levels of horse meat in mixtures with other species.

  8. Repository design

    Energy Technology Data Exchange (ETDEWEB)

    John, C M

    1982-01-01

    Various technical issues of radioactive waste design are addressed in this paper. Two approaches to repository design considered herein are: (1) design to minimize the disturbance of the hot rock; and (2) designs that intentionally modify the hot rock to insure better containment of the wastes. The latter designs range from construction of a highly impermeable barrier around a spherical cavern to creating a matrix of tunnels and boreholes to form a cage within which the hydraulic pressure is nearly constant. Examples of these design alternatives are described in some detail. It is concluded that proposed designs for repositories illustrate that performance criteria considered acceptable for such facilities can be met by appropriate site selection and repository engineering. With these technically feasible design concepts, it is also felt that socioeconomic and institutional issues can be better resolved. (BLM)

  9. Lactoferrin-derived Peptides Active towards Influenza: Identification of Three Potent Tetrapeptide Inhibitors.

    Science.gov (United States)

    Scala, Maria Carmina; Sala, Marina; Pietrantoni, Agostina; Spensiero, Antonia; Di Micco, Simone; Agamennone, Mariangela; Bertamino, Alessia; Novellino, Ettore; Bifulco, Giuseppe; Gomez-Monterrey, Isabel M; Superti, Fabiana; Campiglia, Pietro

    2017-09-06

    Bovine lactoferrin is a biglobular multifunctional iron binding glycoprotein that plays an important role in innate immunity against infections. We have previously demonstrated that selected peptides from bovine lactoferrin C-lobe are able to prevent both Influenza virus hemagglutination and cell infection. To deeper investigate the ability of lactoferrin derived peptides to inhibit Influenza virus infection, in this study we identified new bovine lactoferrin C-lobe derived sequences and corresponding synthetic peptides were synthesized and assayed to check their ability to prevent viral hemagglutination and infection. We identified three tetrapeptides endowed with broad anti-Influenza activity and able to inhibit viral infection in a concentration range femto- to picomolar. Our data indicate that these peptides may constitute a non-toxic tool for potential applications as anti-Influenza therapeutics.

  10. LC-MS/MS Identification of Species-Specific Muscle Peptides in Processed Animal Proteins.

    Science.gov (United States)

    Marchis, Daniela; Altomare, Alessandra; Gili, Marilena; Ostorero, Federica; Khadjavi, Amina; Corona, Cristiano; Ru, Giuseppe; Cappelletti, Benedetta; Gianelli, Silvia; Amadeo, Francesca; Rumio, Cristiano; Carini, Marina; Aldini, Giancarlo; Casalone, Cristina

    2017-12-06

    An innovative analytical strategy has been applied to identify signature peptides able to distinguish among processed animal proteins (PAPs) derived from bovine, pig, fish, and milk products. Proteomics was first used to elucidate the proteome of each source. Starting from the identified proteins and using a funnel based approach, a set of abundant and well characterized peptides with suitable physical-chemical properties (signature peptides) and specific for each source was selected. An on-target LC-ESI-MS/MS method (MRM mode) was set up using standard peptides and was then applied to selectively identify the PAP source and also to distinguish proteins from bovine carcass and milk proteins. We believe that the method described meets the request of the European Commission which has developed a strategy for gradually lifting the "total ban" toward "species to species ban", therefore requiring official methods for species-specific discrimination in feed.

  11. Identification of pro-opiomelanocortin and secretion of its peptide fragments in bovine adrenals

    Energy Technology Data Exchange (ETDEWEB)

    Tennov, A.V.; Dmitriev, A.D.; Kizim, E.A.; Ustinova, E.E.

    1986-01-01

    This paper describes the results of an investigation to show that biosynthesis of POMC, its proteolytic processing, an secretion of the peptide products of that processing take place in the bovine adrenals. Rabbit antisera against endorphins were obtained and used for radioimmunoassay of peptides. I 125-labeled peptides were obtained by the chloramine method and purified from free I 125 on Sephadex G-10 (0.7 x 5 cm, centrifugation for 10 min at 1500 g). To detect secretion of peptide fragments of POMC in the adrenals experiments were undertaken to determine the beta-endorphin content in perfusates obtained during retrograde perfusion of the bovine adrenals. It was found that immunoreactive compounds, indistinguishable in their immunochemical properties from beta-endorphin, are present in the perfusates, just as in the tissue extracts.

  12. Identification of novel dipeptidyl peptidase IV (DPP-IV) inhibitory peptides in camel milk protein hydrolysates.

    Science.gov (United States)

    Nongonierma, Alice B; Paolella, Sara; Mudgil, Priti; Maqsood, Sajid; FitzGerald, Richard J

    2018-04-01

    Nine novel dipeptidyl peptidase IV (DPP-IV) inhibitory peptides (FLQY, FQLGASPY, ILDKEGIDY, ILELA, LLQLEAIR, LPVP, LQALHQGQIV, MPVQA and SPVVPF) were identified in camel milk proteins hydrolysed with trypsin. This was achieved using a sequential approach combining liquid chromatography tandem mass spectrometry (LC-MS/MS), qualitative/quantitative structure activity relationship (QSAR) and confirmatory studies with synthetic peptides. The most potent camel milk protein-derived DPP-IV inhibitory peptides, LPVP and MPVQA, had DPP-IV half maximal inhibitory concentrations (IC 50 ) of 87.0 ± 3.2 and 93.3 ± 8.0 µM, respectively. DPP-IV inhibitory peptide sequences identified within camel and bovine milk protein hydrolysates generated under the same hydrolysis conditions differ. This was linked to differences in enzyme selectivity for peptide bond cleavage of camel and bovine milk proteins as well as dissimilarities in their amino acid sequences. Camel milk proteins contain novel DPP-IV inhibitory peptides which may play a role in the regulation of glycaemia in humans. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Identification of ageing-associated naturally occurring peptides in human urine

    Science.gov (United States)

    Nkuipou-Kenfack, Esther; Bhat, Akshay; Klein, Julie; Jankowski, Vera; Mullen, William; Vlahou, Antonia; Dakna, Mohammed; Koeck, Thomas; Schanstra, Joost P.; Zürbig, Petra; Rudolph, Karl L.; Schumacher, Björn; Pich, Andreas; Mischak, Harald

    2015-01-01

    To assess normal and pathological peptidomic changes that may lead to an improved understanding of molecular mechanisms underlying ageing, urinary peptidomes of 1227 healthy and 10333 diseased individuals between 20 and 86 years of age were investigated. The diseases thereby comprised diabetes mellitus, renal and cardiovascular diseases. Using age as a continuous variable, 116 peptides were identified that significantly (p age in the healthy cohort. The same approach was applied to the diseased cohort. Upon comparison of the peptide patterns of the two cohorts 112 common age-correlated peptides were identified. These 112 peptides predominantly originated from collagen, uromodulin and fibrinogen. While most fibrillar and basement membrane collagen fragments showed a decreased age-related excretion, uromodulin, beta-2-microglobulin and fibrinogen fragments showed an increase. Peptide-based in silico protease analysis was performed and 32 proteases, including matrix metalloproteinases and cathepsins, were predicted to be involved in ageing. Identified peptides, predicted proteases and patient information were combined in a systems biology pathway analysis to identify molecular pathways associated with normal and/or pathological ageing. While perturbations in collagen homeostasis, trafficking of toll-like receptors and endosomal pathways were commonly identified, degradation of insulin-like growth factor-binding proteins was uniquely identified in pathological ageing. PMID:26431327

  14. Repository exploration

    International Nuclear Information System (INIS)

    Pentz, D.L.

    1984-01-01

    This paper discusses exploration objectives and requirements for a nuclear repository in the U.S.A. The importance of designing the exploration program to meet the system performance objectives is emphasized and some examples of the extent of exploration required before the License Application for Construction Authorization is granted are also discussed

  15. Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysate of Spirulina platensis.

    Science.gov (United States)

    Yu, Jie; Hu, Yuanliang; Xue, Mingxiong; Dun, Yaohao; Li, Shenao; Peng, Nan; Liang, Yunxiang; Zhao, Shumao

    2016-07-28

    The aim of this study was to isolate antioxidant peptides from an enzymatic hydrolysate of Spirulina platensis. A novel antioxidant peptide was obtained by ultrafiltration, gel filtration chromatography, and reverse-phase high-performance liquid chromatography, with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay used to measure the antioxidant activity, and the sequence was determined to be Pro-Asn-Asn (343.15 Da) by electrospray ionization tandem mass spectrometry. This peptide was synthesized to confirm its antioxidant properties, and it exhibited 81.44 ± 0.43% DPPH scavenging activity at 100 µg/ml, which was similar to that of glutathione (82.63 ± 0.56%). Furthermore, the superoxide anion and hydroxyl free-radical scavenging activities and the SOD activity of the peptide were 47.84 ± 0.49%, 54.01 ± 0.82%, and 12.55 ± 0.75%, respectively, at 10 mg/ml. These results indicate that S. platensis is a good source of antioxidant peptides, and that its hydrolysate may have important applications in the pharmaceutical and food industries.

  16. Identification of dipeptidyl peptidase-IV inhibitory peptides from mare whey protein hydrolysates.

    Science.gov (United States)

    Song, J J; Wang, Q; Du, M; Ji, X M; Mao, X Y

    2017-09-01

    Inhibition of dipeptidyl peptidase-IV (DPP-IV) activity is a promising strategy for treatment of type 2 diabetes. In the current study, DPP-IV inhibitory peptides were identified from mare whey protein hydrolysates obtained by papain. The results showed that all the mare whey protein hydrolysates obtained at various hydrolysis durations possessed more potent DPP-IV inhibitory activity compared with intact whey protein. The 4-h hydrolysates showed the greatest DPP-IV inhibitory activity with half-maximal inhibitory concentration of 0.18 mg/mL. The 2 novel peptides from 4-h hydrolysate fractions separated by successive chromatographic steps were characterized by liquid chromatography-electrospray ionization tandem mass spectrometry. The novel peptides Asn-Leu-Glu-Ile-Ile-Leu-Arg and Thr-Gln-Met-Val-Asp-Glu-Glu-Ile-Met-Glu-Lys-Phe-Arg, which corresponded to β-lactoglobulin 1 f(71-77) and β-lactoglobulin 1 f(143-155), demonstrated DPP-IV inhibitory activity with half-maximal inhibitory concentrations of 86.34 and 69.84 μM, respectively. The DPP-IV inhibitory activity of the 2 peptides was retained or even improved after simulated gastrointestinal digestion in vitro. Our findings indicate that mare whey protein-derived peptides may possess potential as functional food ingredients in the management of type 2 diabetes. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  17. On Comparison of SimTandem with State-of-the-Art Peptide Identification Tools, Efficiency of Precursor Mass Filter and Dealing with Variable Modifications

    Directory of Open Access Journals (Sweden)

    Novák Jiří

    2013-12-01

    Full Text Available The similarity search in theoretical mass spectra generated from protein sequence databases is a widely accepted approach for identification of peptides from query mass spectra produced by shotgun proteomics. Growing protein sequence databases and noisy query spectra demand database indexing techniques and better similarity measures for the comparison of theoretical spectra against query spectra. We employ a modification of previously proposed parameterized Hausdorff distance for comparisons of mass spectra. The new distance outperforms the original distance, the angle distance and state-of-the-art peptide identification tools OMSSA and X!Tandem in the number of identified peptides even though the q-value is only 0.001. When a precursor mass filter is used as a database indexing technique, our method outperforms OMSSA in the speed of search. When variable modifications are not searched, the search time is similar to X!Tandem. We show that the precursor mass filter is an efficient database indexing technique for high-accuracy data even though many variable modifications are being searched. We demonstrate that the number of identified peptides is bigger when variable modifications are searched separately by more search runs of a peptide identification engine. Otherwise, the false discovery rates are affected by mixing unmodified and modified spectra together resulting in a lower number of identified peptides. Our method is implemented in the freely available application SimTandem which can be used in the framework TOPP based on OpenMS.

  18. Identification of a peptide inhibitor for the histone methyltransferase WHSC1.

    Directory of Open Access Journals (Sweden)

    Michael J Morrison

    Full Text Available WHSC1 is a histone methyltransferase that is responsible for mono- and dimethylation of lysine 36 on histone H3 and has been implicated as a driver in a variety of hematological and solid tumors. Currently, there is a complete lack of validated chemical matter for this important drug discovery target. Herein we report on the first fully validated WHSC1 inhibitor, PTD2, a norleucine-containing peptide derived from the histone H4 sequence. This peptide exhibits micromolar affinity towards WHSC1 in biochemical and biophysical assays. Furthermore, a crystal structure was solved with the peptide in complex with SAM and the SET domain of WHSC1L1. This inhibitor is an important first step in creating potent, selective WHSC1 tool compounds for the purposes of understanding the complex biology in relation to human disease.

  19. Role of peptide processing predictions in T cell epitope identification : contribution of different prediction programs

    NARCIS (Netherlands)

    Calis, Jorg J A; Reinink, Peter; Keller, Christin; Kloetzel, Peter M; Kesmir, Can

    2015-01-01

    Proteolysis is the general term to describe the process of protein degradation into peptides. Proteasomes are the main actors in cellular proteolysis, and their activity can be measured in in vitro digestion experiments. However, in vivo proteolysis can be different than what is measured in these

  20. MALDI-based identification of stable hazelnut protein derived tryptic marker peptides.

    Science.gov (United States)

    Cucu, T; De Meulenaer, B; Devreese, B

    2012-01-01

    Food allergy is an important health problem especially in industrialised countries. Tree nuts, among which are hazelnuts (Corylus avellana), are typically causing serious and life-threatening symptoms in sensitive subjects. Hazelnut is used as a food ingredient in pastry, confectionary products, ice cream and meat products, therefore undeclared hazelnut can be often present as a cross-contaminant representing a threat for allergic consumers. Mass spectrometric techniques are used for the detection of food allergens in processed foods, but limited information regarding stable tryptic peptide markers for hazelnut is available. The aim of this study was to detect stable peptide markers from modified hazelnut protein through the Maillard reaction and oxidation in a buffered solution. Peptides ³⁹⁵Gly-Arg⁴⁰³ from Cor a 11 and ²⁰⁹Gln-Arg²¹⁷, ³⁵¹Ile-Arg³⁶³, ⁴⁶⁴Ala-Arg⁴⁷⁸ and ⁴⁰¹Val-Arg⁴¹⁷ from Cor a 9 hazelnut allergens proved to be the most stable and could be detected and confirmed with high scores in most of the modified samples. The identified peptides can be further used as analytical targets for the development of more robust quantitative methods for hazelnut detection in processed foods.

  1. CrossWork: Software-assisted identification of cross-linked peptides

    DEFF Research Database (Denmark)

    Rasmussen, Morten; Refsgaard, Jan; Peng, Li

    2011-01-01

    Work searches batches of tandem mass-spectrometric data, and identifies cross-linked and non-cross-linked peptides using a standard PC. We tested CrossWork by searching mass-spectrometric datasets of cross-linked complement factor C3 against small (1 protein) and large (1000 proteins) search spaces, and show...

  2. Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition

    DEFF Research Database (Denmark)

    Yuhong, Huang; Busk, Peter Kamp; Grell, Morten Nedergaard

    2014-01-01

    Mucor circinelloides produces plant cell wall degrading enzymes that allow it to grow on complex polysaccharides. Although the genome of M. circinelloides has been sequenced, only few plant cell wall degrading enzymes are annotated in this species. We applied peptide pattern recognition, which...

  3. Identification of CJC-1295, a growth-hormone-releasing peptide, in an unknown pharmaceutical preparation.

    Science.gov (United States)

    Henninge, John; Pepaj, Milaim; Hullstein, Ingunn; Hemmersbach, Peter

    2010-01-01

    Several peptide drugs are being manufactured illicitly, and in some cases they are being made available to the public before entering or completing clinical trials. At the request of Norwegian police and customs authorities, unknown pharmaceutical preparations suspected to contain peptide drugs are regularly subjected to analysis. In 2009, an unknown pharmaceutical preparation was submitted for analysis by liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS). The preparation was found to contain a 29 amino acid peptide with a C-terminal amide function. Based on the interpretation of mass spectrometric data, an amino acid sequence was proposed. The sequence is consistent with a peptide currently marketed under the name CJC-1295. CJC-1295 is a releasing factor for growth hormone and is therefore considered a Prohibited Substance under Section S2 of the WADA Prohibited List. This substance has potential performance-enhancing effects, it is readily available, and there is reason to believe that it is being used within the bodybuilding community. Copyright © 2010 John Wiley & Sons, Ltd.

  4. Identification of 11-amino acid peptides that disrupt Notch-mediated processes in Drosophila

    Directory of Open Access Journals (Sweden)

    Yeh Hsiao-Fong

    2011-06-01

    Full Text Available Abstract Background The conserved Notch signaling pathway regulates cell fate decisions and maintains stem cells in multicellular organisms. Up-regulation of Notch signaling is observed in several types of cancer and is causally involved in proliferation and survival of cancer cells. Thus, it is of great interest to look for anti-Notch reagents for therapeutic purposes. In model animal Drosophila, Notch signaling restricts selection of sensory organ precursors (SOPs during external sensory (ES organ development. To look for novel genes that can suppress Notch signaling, we performed a gain-of-function modifier screen to look for genes that enhance the phenotype of ectopic ES organs induced by overexpression of phyllopod, a gene required for SOP specification. Results From the gain-of-function screen, we discovered that overexpression of polished rice/tarsal-less (pri/tal increases the numbers of ES organs as well as SOPs. pri/tal is a polycistronic gene that contains four short open reading frames encoding three 11-amino acid and one 32-amino acid peptides. Ectopic expression of the 11 amino-acid peptides recapitulates the pri/tal misexpression phenotype in ectopic ES organ formation. In situ hybridization experiment reveals that pri/tal mRNA is expressed in the SOPs of the chemosensory organs and the stretch-sensing chordotonal organs. In Drosophila wing development, the Notch signaling pathway mediates the formation of the dorsal-ventral (DV compartmental boundary and the restriction of the vein width from the primordial veins, the proveins. We also found that pri/tal mRNA is expressed in the DV boundary and the longitudinal proveins, and overexpression of Pri/Tal peptides disrupts the DV boundary formation and helps to expand the width of the wing vein. Genetic analyses further show that a Notch loss-of-function allele strongly enhances these two phenotypes. Cut and E(splmβ are target genes of the Notch pathway in DV boundary formation and

  5. Identification and characterization of pyrokinin and CAPA peptides, and corresponding GPCRs from spotted wing drosophila, Drosophila suzukii

    Science.gov (United States)

    To date, some biological activities have been confirmed as different named peptides, however, most FXPRLamide peptides are still poorly understood although these peptides are found in all insects. So, the study of receptors for these peptides is particularly important. Receptors of FXPRLamide peptid...

  6. Optimization of Search Engines and Postprocessing Approaches to Maximize Peptide and Protein Identification for High-Resolution Mass Data.

    Science.gov (United States)

    Tu, Chengjian; Sheng, Quanhu; Li, Jun; Ma, Danjun; Shen, Xiaomeng; Wang, Xue; Shyr, Yu; Yi, Zhengping; Qu, Jun

    2015-11-06

    The two key steps for analyzing proteomic data generated by high-resolution MS are database searching and postprocessing. While the two steps are interrelated, studies on their combinatory effects and the optimization of these procedures have not been adequately conducted. Here, we investigated the performance of three popular search engines (SEQUEST, Mascot, and MS Amanda) in conjunction with five filtering approaches, including respective score-based filtering, a group-based approach, local false discovery rate (LFDR), PeptideProphet, and Percolator. A total of eight data sets from various proteomes (e.g., E. coli, yeast, and human) produced by various instruments with high-accuracy survey scan (MS1) and high- or low-accuracy fragment ion scan (MS2) (LTQ-Orbitrap, Orbitrap-Velos, Orbitrap-Elite, Q-Exactive, Orbitrap-Fusion, and Q-TOF) were analyzed. It was found combinations involving Percolator achieved markedly more peptide and protein identifications at the same FDR level than the other 12 combinations for all data sets. Among these, combinations of SEQUEST-Percolator and MS Amanda-Percolator provided slightly better performances for data sets with low-accuracy MS2 (ion trap or IT) and high accuracy MS2 (Orbitrap or TOF), respectively, than did other methods. For approaches without Percolator, SEQUEST-group performs the best for data sets with MS2 produced by collision-induced dissociation (CID) and IT analysis; Mascot-LFDR gives more identifications for data sets generated by higher-energy collisional dissociation (HCD) and analyzed in Orbitrap (HCD-OT) and in Orbitrap Fusion (HCD-IT); MS Amanda-Group excels for the Q-TOF data set and the Orbitrap Velos HCD-OT data set. Therefore, if Percolator was not used, a specific combination should be applied for each type of data set. Moreover, a higher percentage of multiple-peptide proteins and lower variation of protein spectral counts were observed when analyzing technical replicates using Percolator

  7. Identification and characterization of histidine-rich peptides from hard ticks Ixodes ricinus and Ixodes scapularis.

    OpenAIRE

    DORŇÁKOVÁ, Veronika

    2011-01-01

    Antimicrobial (cationic) proteins play an important role in innate imunity. Such proteins can possess antibacterial, antiendotoxic or fungicidal abilities. The rising resistence of microbes to common antibiotics evokes acute need of studying more endogenous proteins to reveal new potential antibiotics. Ticks, the blood-feeding ectoparasites with effectual defense system, present an endless source of newly described and unknown antimicrobial peptides/proteins with significant theurapeutic pote...

  8. The use of interaction matrices for identification, structuring and ranking of FEPs in a repository system. Application on the far-field of a deep geological repository for spent fuel

    Energy Technology Data Exchange (ETDEWEB)

    Skagius, K; Wiborgh, M [Kemakta, Stockholm (Sweden); Stroem, A [Swedish Nuclear Fuel and Waste Management Co., Stockholm (Sweden)

    1995-11-01

    The basic device in the Rock Engineering Systems approach, the interaction matrix, has been used to identify, structure and rank Features, Events, and Processes (FEPs) describing barrier performance and radionuclide behaviour in the far-field of a deep geologic repository for spent fuel. The result is a first version of the Process System (PS), for the far-field of a deep repository, structured in an interaction matrix with supporting documentation. The documentation is compiled in databases, one containing matrix specific information and one containing general FEP descriptions. The study has shown that an interaction matrix is feasible to use both for the structuring of the PS and for visualisation of the PS. The developed documentation system increases the transparency of the system description and makes it possible to trace back the judgements made during the construction of the matrix. This will facilitate review work and future revisions as well as consistent treatment of different issues in the system. This study is a first step in the application of a systematic method to establish a structured description of the PS for a deep repository for spent fuel. The work could be seen as a part of the preparation for the forthcoming performance and safety analysis. The next step would be to develop the PS for the remaining parts of the repository system to the same level as has been done for the far-field system. Before the PS is evaluated for different selected system premises, a scientific review of the contents of the PS for the whole repository system would be beneficial. 5 refs.

  9. The use of interaction matrices for identification, structuring and ranking of FEPs in a repository system. Application on the far-field of a deep geological repository for spent fuel

    International Nuclear Information System (INIS)

    Skagius, K.; Wiborgh, M.; Stroem, A.

    1995-11-01

    The basic device in the Rock Engineering Systems approach, the interaction matrix, has been used to identify, structure and rank Features, Events, and Processes (FEPs) describing barrier performance and radionuclide behaviour in the far-field of a deep geologic repository for spent fuel. The result is a first version of the Process System (PS), for the far-field of a deep repository, structured in an interaction matrix with supporting documentation. The documentation is compiled in databases, one containing matrix specific information and one containing general FEP descriptions. The study has shown that an interaction matrix is feasible to use both for the structuring of the PS and for visualisation of the PS. The developed documentation system increases the transparency of the system description and makes it possible to trace back the judgements made during the construction of the matrix. This will facilitate review work and future revisions as well as consistent treatment of different issues in the system. This study is a first step in the application of a systematic method to establish a structured description of the PS for a deep repository for spent fuel. The work could be seen as a part of the preparation for the forthcoming performance and safety analysis. The next step would be to develop the PS for the remaining parts of the repository system to the same level as has been done for the far-field system. Before the PS is evaluated for different selected system premises, a scientific review of the contents of the PS for the whole repository system would be beneficial. 5 refs

  10. Identification of a Peptide-Pheromone that Enhances Listeria monocytogenes Escape from Host Cell Vacuoles

    Science.gov (United States)

    Xayarath, Bobbi; Alonzo, Francis; Freitag, Nancy E.

    2015-01-01

    Listeria monocytogenes is a Gram-positive facultative intracellular bacterial pathogen that invades mammalian cells and escapes from membrane-bound vacuoles to replicate within the host cell cytosol. Gene products required for intracellular bacterial growth and bacterial spread to adjacent cells are regulated by a transcriptional activator known as PrfA. PrfA becomes activated following L. monocytogenes entry into host cells, however the signal that stimulates PrfA activation has not yet been defined. Here we provide evidence for L. monocytogenes secretion of a small peptide pheromone, pPplA, which enhances the escape of L. monocytogenes from host cell vacuoles and may facilitate PrfA activation. The pPplA pheromone is generated via the proteolytic processing of the PplA lipoprotein secretion signal peptide. While the PplA lipoprotein is dispensable for pathogenesis, bacteria lacking the pPplA pheromone are significantly attenuated for virulence in mice and have a reduced efficiency of bacterial escape from the vacuoles of nonprofessional phagocytic cells. Mutational activation of PrfA restores virulence and eliminates the need for pPplA-dependent signaling. Experimental evidence suggests that the pPplA peptide may help signal to L. monocytogenes its presence within the confines of the host cell vacuole, stimulating the expression of gene products that contribute to vacuole escape and facilitating PrfA activation to promote bacterial growth within the cytosol. PMID:25822753

  11. Identification of the bioactive and consensus peptide motif from Momordica charantia insulin receptor-binding protein.

    Science.gov (United States)

    Lo, Hsin-Yi; Li, Chia-Cheng; Ho, Tin-Yun; Hsiang, Chien-Yun

    2016-08-01

    Many food bioactive peptides with diverse functions have been discovered by studying plant proteins. We have previously identified a 68-residue insulin receptor (IR)-binding protein (mcIRBP) from Momordica charantia that exhibits hypoglycemic effects in mice via interaction with IR. By in vitro digestion, we found that mcIRBP-19, spanning residues 50-68 of mcIRBP, enhanced the binding of insulin to IR, stimulated the phosphorylation of PDK1 and Akt, induced the expression of glucose transporter 4, and stimulated both the uptake of glucose in cells and the clearance of glucose in diabetic mice. Furthermore, mcIRBP-19 homologs were present in various plants and shared similar β-hairpin structures and IR kinase-activating abilities to mcIRBP-19. In conclusion, our findings suggested that mcIRBP-19 is a blood glucose-lowering bioactive peptide that exhibits IR-binding potentials. Moreover, we newly identified novel IR-binding bioactive peptides in various plants which belonged to different taxonomic families. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Verification of Single-Peptide Protein Identifications by the Application of Complementary Database Search Algorithms

    National Research Council Canada - National Science Library

    Rohrbough, James G; Breci, Linda; Merchant, Nirav; Miller, Susan; Haynes, Paul A

    2005-01-01

    .... One such technique, known as the Multi-Dimensional Protein Identification Technique, or MudPIT, involves the use of computer search algorithms that automate the process of identifying proteins...

  13. Identification of novel peptide ligands for the cancer-specific receptor mutation EFGRvIII using a mixture-based synthetic combinatorial library

    DEFF Research Database (Denmark)

    Denholt, Charlotte Lund; Hansen, Paul Robert; Pedersen, Nina

    2009-01-01

    We report here, the design and synthesis of a positional scanning synthetic combinatorial library for the identification of novel peptide ligands targeted against the cancer-specific epidermal growth factor tyrosine kinase receptor mutation variant III (EGFRvIII). This receptor is expressed in se...

  14. Deconvolution of mixture spectra and increased throughput of Peptide identification by utilization of intensified complementary ions formed in tandem mass spectrometry

    DEFF Research Database (Denmark)

    Kryuchkov, Fedor; Verano-Braga, Thiago; Hansen, Thomas Aarup

    2013-01-01

    -resolution orbitrap mass spectrometry, an increase in the number of peptide identifications was obtained relative to the original CAD MS/MS spectra when intensified golden complementary (+18.6%) and CAD complementary pairs (+17.2%) were submitted to the Mascot search engine. This also exceeded the results obtained...

  15. Screening and identification of RhD antigen mimic epitopes from a phage display random peptide library for the serodiagnosis of haemolytic disease of the foetus and newborn.

    Science.gov (United States)

    Wang, Jiao; Song, Jingjing; Zhou, Shuimei; Fu, Yourong; Bailey, Jeffrey A; Shen, Changxin

    2018-01-16

    Identification of RhD antigen epitopes is a key component in understanding the pathogenesis of haemolytic disease of the foetus and newborn. Research has indicated that phage display libraries are useful tools for identifying novel mimic epitopes (mimotopes) which may help to determine antigen specificity. We selected the mimotopes of blood group RhD antigen by affinity panning a phage display library using monoclonal anti-D. After three rounds of biopanning, positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and then sent for sequencing and peptides synthesis. Next, competitive ELISA and erythrocyte haemagglutination inhibition tests were carried out to confirm the inhibitory activity of the synthetic peptide. To evaluate the diagnostic performance of the synthetic peptide, a diagnostic ELISA was examined. Fourteen of 35 phage clones that were chosen randomly from the titering plate were considered to be positive. Following DNA sequencing and translation, 11 phage clones were found to represent the same peptide - RMKMLMMLMRRK (P4) - whereas each of the other three clones represented a unique peptide. Through the competitive ELISA and erythrocyte haemagglutination inhibition tests, the peptide (P4) was verified to have the ability to mimic the RhD antigen. The diagnostic ELISA for P4 proved to be sensitive (82.61%) and specific (88.57%). This study reveals that the P4 peptide can mimic RhD antigen and paves the way for the development of promising targeted diagnostic and therapeutic platforms for haemolytic disease of the foetus and newborn.

  16. Identification of novel dipeptidyl peptidase-IV and angiotensin-I-converting enzyme inhibitory peptides from meat proteins using in silico analysis.

    Science.gov (United States)

    Lafarga, Tomas; O'Connor, Paula; Hayes, Maria

    2014-09-01

    Angiotensin-I-converting enzyme (ACE-I, EC 3.4.15.1), renin (EC 3.4.23.15), and dipeptidyl peptidase-IV (DPP-IV, EC 3.4.14.5) play key roles in the control of hypertension and the development of type-2 diabetes and other diseases associated with metabolic syndrome. The aim of this work was to utilize known in silico methodologies, peptide databases and software including ProtParam (http://web.expasy.org/protparam/), Basic Local Alignment Tool (BLAST), ExPASy PeptideCutter (http://web.expasy.org/peptide_cutter/) and BIOPEP (http://www.uwm.edu.pl/biochemia/index.php/pl/biopep) to assess the release of potentially bioactive DPP-IV, renin and ACE-I inhibitory peptides from bovine and porcine meat proteins including hemoglobin, collagen and serum albumin. These proteins were chosen as they are found commonly in meat by-products such as bone, blood and low-value meat cuts. In addition, the bioactivities of identified peptides were confirmed using chemical synthesis and in vitro bioassays. The concentration of peptide required to inhibit the activity of ACE-I and DPP-IV by 50% was determined for selected, active peptides. Novel ACE-I and DPP-IV inhibitory peptides were identified in this study using both in silico analysis and a literature search to streamline enzyme selection for peptide production. These novel peptides included the ACE-I inhibitory tri-peptide Ile-Ile-Tyr and the DPP-IV inhibitory tri-peptide Pro-Pro-Leu corresponding to sequences f (182-184) and f (326-328) of both porcine and bovine serum albumin which can be released following hydrolysis with the enzymes papain and pepsin, respectively. This work demonstrates that meat proteins are a suitable resource for the generation of bioactive peptides and further demonstrates the usefulness of in silico methodologies to streamline identification and generation of bioactive peptides. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Identification of diagnostic peptide regions that distinguish Zika virus from related mosquito-borne Flaviviruses.

    Directory of Open Access Journals (Sweden)

    Alexandra J Lee

    Full Text Available Zika virus (ZIKV is a member of the Flavivirus genus of positive-sense single-stranded RNA viruses, which includes Dengue, West Nile, Yellow Fever, and other mosquito-borne arboviruses. Infection by ZIKV can be difficult to distinguish from infection by other mosquito-borne Flaviviruses due to high sequence similarity, serum antibody cross-reactivity, and virus co-circulation in endemic areas. Indeed, existing serological methods are not able to consistently differentiate ZIKV from other Flaviviruses, which makes it extremely difficult to accurately calculate the incidence rate of Zika-associated Guillain-Barre in adults, microcephaly in newborns, or asymptomatic infections within a geographical area. In order to identify Zika-specific peptide regions that could be used as serology reagents, we have applied comparative genomics and protein structure analyses to identify amino acid residues that distinguish each of 10 Flavivirus species and subtypes from each other by calculating the specificity, sensitivity, and surface exposure of each residue in relevant target proteins. For ZIKV we identified 104 and 116 15-mer peptides in the E glycoprotein and NS1 non-structural protein, respectively, that contain multiple diagnostic sites and are located in surface-exposed regions in the tertiary protein structure. These sensitive, specific, and surface-exposed peptide regions should serve as useful reagents for seroprevalence studies to better distinguish between prior infections with any of these mosquito-borne Flaviviruses. The development of better detection methods and diagnostic tools will enable clinicians and public health workers to more accurately estimate the true incidence rate of asymptomatic infections, neurological syndromes, and birth defects associated with ZIKV infection.

  18. Frog secretions and hunting magic in the upper Amazon: identification of a peptide that interacts with an adenosine receptor.

    Science.gov (United States)

    Daly, J W; Caceres, J; Moni, R W; Gusovsky, F; Moos, M; Seamon, K B; Milton, K; Myers, C W

    1992-11-15

    A frog used for "hunting magic" by several groups of Panoan-speaking Indians in the borderline between Brazil and Peru is identified as Phyllomedusa bicolor. This frog's skin secretion, which the Indians introduce into the body through fresh burns, is rich in peptides. These include vasoactive peptides, opioid peptides, and a peptide that we have named adenoregulin, with the sequence GLWSKIKEVGKEAAKAAAKAAGKAALGAVSEAV as determined from mass spectrometry and Edman degradation. The natural peptide may contain a D amino acid residue, since it is not identical in chromatographic properties to the synthetic peptide. Adenoregulin enhances binding of agonists to A1 adenosine receptors; it is accompanied in the skin secretion by peptides that inhibit binding. The vasoactive peptide sauvagine, the opioid peptides, and adenoregulin and related peptides affect behavior in mice and presumably contribute to the behavioral sequelae observed in humans.

  19. Identification of MHC class I H-2 Kb/Db-restricted immunogenic peptides derived from retinal proteins

    DEFF Research Database (Denmark)

    Wang, Mingjun; Bai, Fang; Pries, Mette

    2006-01-01

    PURPOSE: To identify H-2 Kb/Db-binding immunogenic peptides derived from retinal proteins. METHODS: Computer-based prediction was used to identify potentially H-2 Kb/Db-binding peptides derived from the interphotoreceptor retinol-binding protein (IRBP), soluble retinal antigen (S...... on day 21 after immunization with IRBP or IRBP and the immunogenic peptides. RESULTS: All the 21 predicted peptides were found to upregulate expression of H-2 Kb/Db on RMA-S cells. Five peptides, the two IRBP-derived peptides IRBP89-96 and IRBP(101-108), and the three PEDF-derived peptides, PEDF389....... The immunogenic peptides alone did not induce inflammation in the eyes, but they could enhance severity of uveitis induced by IRBP. CONCLUSIONS: Five of 21 H-2 Kb/Db-binding retinal protein-derived peptides were found to be immunogenic, suggesting that these peptides could function as autoantigenic epitopes...

  20. Identification of novel selective V2 receptor non-peptide agonists.

    Science.gov (United States)

    Del Tredici, Andria L; Vanover, Kim E; Knapp, Anne E; Bertozzi, Sine M; Nash, Norman R; Burstein, Ethan S; Lameh, Jelveh; Currier, Erika A; Davis, Robert E; Brann, Mark R; Mohell, Nina; Olsson, Roger; Piu, Fabrice

    2008-10-30

    Peptides with agonist activity at the vasopressin V(2) receptor are used clinically to treat fluid homeostasis disorders such as polyuria and central diabetes insipidus. Of these peptides, the most commonly used is desmopressin, which displays poor bioavailability as well as potent activity at the V(1b) receptor, with possible stress-related adverse effects. Thus, there is a strong need for the development of small molecule chemistries with selective V(2) receptor agonist activity. Using the functional cell-based assay Receptor Selection and Amplification Technology (R-SAT((R))), a screening effort identified three small molecule chemotypes (AC-94544, AC-88324, and AC-110484) with selective agonist activity at the V(2) receptor. One of these compounds, AC-94544, displayed over 180-fold selectivity at the V(2) receptor compared to related vasopressin and oxytocin receptors and no activity at 28 other G protein-coupled receptors (GPCRs). All three compounds also showed partial agonist activity at the V(2) receptor in a cAMP accumulation assay. In addition, in a rat model of central diabetes insipidus, AC-94544 was able to significantly reduce urine output in a dose-dependent manner. Thus, AC-94544, AC-88324, and AC-110484 represent novel opportunities for the treatment of disorders associated with V(2) receptor agonist deficiency.

  1. Pepsin Digested Oat Bran Proteins: Separation, Antioxidant Activity, and Identification of New Peptides

    Directory of Open Access Journals (Sweden)

    Ariane Vanvi

    2016-01-01

    Full Text Available The aim of this study was to determine pepsin hydrolysis conditions to produce digested oat bran proteins with higher radical scavenging activities and separate and identify peptides. Isolated proteins were then digested with different concentrations of pepsin and incubation times. Hydrolysates produced with 1 : 30 enzyme substrate (E/S ratio and 2 h possessed the highest peroxyl radical scavenging activity, 608 ± 17 µM TE/g (compared to 456–474 µM TE/g for other digests, and was therefore subsequently fractionated into eight fractions (F1–F8 by high performance liquid chromatography (HPLC. F1 and F2 had little activity because of their low protein contents. Activities of F3–F8 were 447–874 µM TE/g, 20–36%, and 10–14% in the peroxyl, superoxide anion, and hydroxyl radical tests, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS was used to identify a total of fifty peptides that may have contributed to the activity of F3, a fraction that better scavenged radicals.

  2. Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes for Rapid Identification of Candida albicans Directly from Blood Culture Bottles

    Science.gov (United States)

    Rigby, Susan; Procop, Gary W.; Haase, Gerhard; Wilson, Deborah; Hall, Geraldine; Kurtzman, Cletus; Oliveira, Kenneth; Von Oy, Sabina; Hyldig-Nielsen, Jens J.; Coull, James; Stender, Henrik

    2002-01-01

    A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described. The test (the C. albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C. albicans 26S rRNA. The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55°C. Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy. The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C. albicans (n = 72), C. dubliniensis (n = 58), C. glabrata (n = 5), C. krusei (n = 2), C. parapsilosis (n = 4), and C. tropicalis (n = 3). The performance of the C. albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity. It is concluded that this 2.5-h method for the definitive identification of C. albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management. PMID:12037084

  3. Rapid Identification of Staphylococcus aureus Directly from Blood Cultures by Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes

    Science.gov (United States)

    Oliveira, Kenneth; Procop, Gary W.; Wilson, Deborah; Coull, James; Stender, Henrik

    2002-01-01

    A new fluorescence in situ hybridization (FISH) method with peptide nucleic acid (PNA) probes for identification of Staphylococcus aureus directly from positive blood culture bottles that contain gram-positive cocci in clusters (GPCC) is described. The test (the S. aureus PNA FISH assay) is based on a fluorescein-labeled PNA probe that targets a species-specific sequence of the 16S rRNA of S. aureus. Evaluations with 17 reference strains and 48 clinical isolates, including methicillin-resistant and methicillin-susceptible S. aureus species, coagulase-negative Staphylococcus species, and other clinically relevant and phylogenetically related bacteria and yeast species, showed that the assay had 100% sensitivity and 96% specificity. Clinical trials with 87 blood cultures positive for GPCC correctly identified 36 of 37 (97%) of the S. aureus-positive cultures identified by standard microbiological methods. The positive and negative predictive values were 100 and 98%, respectively. It is concluded that this rapid method (2.5 h) for identification of S. aureus directly from blood culture bottles that contain GPCC offers important information for optimal antibiotic therapy. PMID:11773123

  4. Learning Object Repositories

    Science.gov (United States)

    Lehman, Rosemary

    2007-01-01

    This chapter looks at the development and nature of learning objects, meta-tagging standards and taxonomies, learning object repositories, learning object repository characteristics, and types of learning object repositories, with type examples. (Contains 1 table.)

  5. A novel strategy for global mapping of O-GlcNAc proteins and peptides using selective enzymatic deglycosylation, HILIC enrichment and mass spectrometry identification.

    Science.gov (United States)

    Shen, Bingquan; Zhang, Wanjun; Shi, Zhaomei; Tian, Fang; Deng, Yulin; Sun, Changqing; Wang, Guangshun; Qin, Weijie; Qian, Xiaohong

    2017-07-01

    O-GlcNAcylation is a kind of dynamic O-linked glycosylation of nucleocytoplasmic and mitochondrial proteins. It serves as a major nutrient sensor to regulate numerous biological processes including transcriptional regulation, cell metabolism, cellular signaling, and protein degradation. Dysregulation of cellular O-GlcNAcylated levels contributes to the etiologies of many diseases such as diabetes, neurodegenerative disease and cancer. However, deeper insight into the biological mechanism of O-GlcNAcylation is hampered by its extremely low stoichiometry and the lack of efficient enrichment approaches for large-scale identification by mass spectrometry. Herein, we developed a novel strategy for the global identification of O-GlcNAc proteins and peptides using selective enzymatic deglycosylation, HILIC enrichment and mass spectrometry analysis. Standard O-GlcNAc peptides can be efficiently enriched even in the presence of 500-fold more abundant non-O-GlcNAc peptides and identified by mass spectrometry with a low nanogram detection sensitivity. This strategy successfully achieved the first large-scale enrichment and characterization of O-GlcNAc proteins and peptides in human urine. A total of 474 O-GlcNAc peptides corresponding to 457 O-GlcNAc proteins were identified by mass spectrometry analysis, which is at least three times more than that obtained by commonly used enrichment methods. A large number of unreported O-GlcNAc proteins related to cell cycle, biological regulation, metabolic and developmental process were found in our data. The above results demonstrated that this novel strategy is highly efficient in the global enrichment and identification of O-GlcNAc peptides. These data provide new insights into the biological function of O-GlcNAcylation in human urine, which is correlated with the physiological states and pathological changes of human body and therefore indicate the potential of this strategy for biomarker discovery from human urine. Copyright

  6. Molecular identification of a Drosophila G protein-coupled receptor specific for crustacean cardioactive peptide

    DEFF Research Database (Denmark)

    Cazzamali, Giuseppe; Hauser, Frank; Kobberup, Sune

    2003-01-01

    The Drosophila Genome Project website (www.flybase.org) contains the sequence of an annotated gene (CG6111) expected to code for a G protein-coupled receptor. We have cloned this receptor and found that its gene was not correctly predicted, because an annotated neighbouring gene (CG14547) was also...... part of the receptor gene. DNA corresponding to the corrected gene CG6111 was expressed in Chinese hamster ovary cells, where it was found to code for a receptor that could be activated by low concentrations of crustacean cardioactive peptide, which is a neuropeptide also known to occur in Drosophila...... and other insects (EC(50), 5.4 x 10(-10)M). Other known Drosophila neuropeptides, such as adipokinetic hormone, did not activate the receptor. The receptor is expressed in all developmental stages from Drosophila, but only very weakly in larvae. In adult flies, the receptor is mainly expressed in the head...

  7. Identification of Symptomatic Fetuses Infected with Cytomegalovirus Using Amniotic Fluid Peptide Biomarkers.

    Directory of Open Access Journals (Sweden)

    Cyrille Desveaux

    2016-01-01

    Full Text Available Cytomegalovirus (CMV is the most common cause of congenital infection, and is a major cause of sensorineural hearing loss and neurological disabilities. Evaluating the risk for a CMV infected fetus to develop severe clinical symptoms after birth is crucial to provide appropriate guidance to pregnant women who might have to consider termination of pregnancy or experimental prenatal medical therapies. However, establishing the prognosis before birth remains a challenge. This evaluation is currently based upon fetal imaging and fetal biological parameters, but the positive and negative predictive values of these parameters are not optimal, leaving room for the development of new prognostic factors. Here, we compared the amniotic fluid peptidome between asymptomatic fetuses who were born as asymptomatic neonates and symptomatic fetuses who were either terminated in view of severe cerebral lesions or born as severely symptomatic neonates. This comparison allowed us to identify a 34-peptide classifier in a discovery cohort of 13 symptomatic and 13 asymptomatic neonates. This classifier further yielded 89% sensitivity, 75% specificity and an area under the curve of 0.90 to segregate 9 severely symptomatic from 12 asymptomatic neonates in a validation cohort, showing an overall better performance than that of classical fetal laboratory parameters. Pathway analysis of the 34 peptides underlined the role of viral entry in fetuses with severe brain disease as well as the potential importance of both beta-2-microglobulin and adiponectin to protect the injured fetal brain infected with CMV. The results also suggested the mechanistic implication of the T calcium channel alpha-1G (CACNA1G protein in the development of seizures in severely CMV infected children. These results open a new field for potential therapeutic options. In conclusion, this study demonstrates that amniotic fluid peptidome analysis can effectively predict the severity of congenital CMV

  8. Identification of Small Peptides in Human Cerebrospinal Fluid upon Amyloid-β Degradation.

    Science.gov (United States)

    Mizuta, Naoki; Yanagida, Kanta; Kodama, Takashi; Tomonaga, Takeshi; Takami, Mako; Oyama, Hiroshi; Kudo, Takashi; Ikeda, Manabu; Takeda, Masatoshi; Tagami, Shinji; Okochi, Masayasu

    2017-01-01

    Amyloid-β (Aβ) degradation in brains of Alzheimer disease patients is a crucial focus for the clarification of disease pathogenesis. Nevertheless, the mechanisms underlying Aβ degradation in the human brain remain unclear. This study aimed to quantify the levels of small C-terminal Aβ fragments generated upon Aβ degradation in human cerebrospinal fluid (CSF). A fraction containing small peptides was isolated and purified from human CSF by high-pressure liquid chromatography. Degradation products of Aβ C termini were identified and measured by liquid chromatography-tandem mass spectrometry. The C-terminal fragments of Aβ in the conditioned medium of cultured cells transfected with the Swedish variant of βAPP (sw βAPP) were analyzed. These fragments in brains of PS1 I213T knock-in transgenic mice, overexpressing sw βAPP, were also analyzed. The peptide fragments GGVV and GVV, produced by the cleavage of Aβ40, were identified in human CSF as well as in the brains of the transgenic mice and in the conditioned medium of the cultured cells. Relative to Aβ40 levels, GGVV and GVV levels were 7.6 ± 0.81 and 1.5 ± 0.18%, respectively, in human CSF. Levels of the GGVV fragment did not increase by the introduction of genes encoding neprilysin and insulin-degrading enzyme to the cultured cells. Our results indicate that a substantial amount of Aβ40 in human brains is degraded via a neprilysin- or insulin-degrading enzyme-independent pathway. © 2017 S. Karger AG, Basel.

  9. Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum.

    Science.gov (United States)

    Cui, Pengfei; Dong, Yuan; Li, Zhijian; Zhang, Yubo; Zhang, Shicui

    2016-07-01

    The global ever-growing concerns about multi-drug resistant (MDR) microbes leads to urgent demands for exploration of new antibiotics including antimicrobial peptides (AMPs). Here we demonstrated that a cDNA from Ciliata Paramecium caudatum, designated Pcamp1, coded for a protein with features characteristic of AMPs, which is not homologous to any AMPs currently known. Both the C-terminal 91 amino acid residues of PcAMP1, cPcAMP1, expressed in Escherichia coli and the C-terminal 26 amino acid residues (predicted mature AMP), cPcAMP1/26, synthesized, underwent a coil-to-helix transition in the presence of TFE, SDS or DPC. Functional assays revealed that cPcAMP1 and cPcAMP1/26 were both able to kill Aeromonas hydrophila and Staphylococcus aureus. ELISA showed that cPcAMP1 and cPcAMP1/26 were able to bind to microbe-associated molecular pattern molecules LPS and LTA, which was further corroborated by the observations that cPcAMP1 could deposit onto the bacterial membranes. Importantly, both cPcAMP1 and cPcAMP1/26 were able to induce bacterial membrane permeabilization and depolarization, and to increase intracellular ROS levels. Additionally, cPcAMP1 and cPcAMP1/26 were not cytotoxic to mammalian cells. Taken together, our results show that PcAMP1 is a potential AMP with a membrane selectivity towards bacterial cells, which renders it a promising template for the design of novel peptide antibiotics against MDR microbes. It also shows that use of signal conserved sequence of AMPs can be an effective tool to identify potential AMPs across different animal classes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Expression of recombinant Arabian camel lactoferricin-related peptide in Pichia pastoris and its antimicrobial identification.

    Science.gov (United States)

    Chahardooli, Mahmood; Niazi, Ali; Aram, Farzaneh; Sohrabi, Seyyed Mohsen

    2016-01-30

    Lactoferricin (LFcin) is a strong cationic peptide released from the N-terminus of lactoferrin by gastric pepsin digestion. LFcin has some important properties, including high antimicrobial activity. To date, lactoferricins have been isolated and characterised from various animal species, but not from camel. The aim of this study was to characterise and express recombinant camel lactoferricin (LFcinC) in Pichia pastoris and investigate its antimicrobial activity. After methanol induction, LFcinC was expressed and secreted into a culture broth medium and the results determined by concentrated supernatant culture medium showed high antimicrobial activity against the following microorganisms: Escherichia coli PTCC 1330 (ATCC 8739), Staphylococcus aureus PTCC 1112 (ATCC 6538), Pseudomonas aeruginosa PTCC 1074 (ATCC 9027), Bacillus subtilis PTCC 1023 (ATCC 6633), and Candida albicans PTCC 5027 (ATCC 10231). Thermal stability was clarified with antibacterial activity against Escherichia coli PTCC 1330 (ATCC 8739). Results confirmed that camel lactoferricin had suitable antimicrobial activity and its production by Pichia pastoris can be used for recombinant production. © 2015 Society of Chemical Industry.

  11. Identification of a Minimal Peptide Tag for in Vivo and in Vitro Loading of Encapsulin.

    Science.gov (United States)

    Cassidy-Amstutz, Caleb; Oltrogge, Luke; Going, Catherine C; Lee, Antony; Teng, Poh; Quintanilla, David; East-Seletsky, Alexandra; Williams, Evan R; Savage, David F

    2016-06-21

    The encapsulation of enzymes and other proteins within a proteinaceous shell has been observed in many bacteria and archaea, but the function and utility of many such compartments are enigmatic. Efforts to study these functions have been complicated by the size and complexity of traditional protein compartments. One potential system for investigating the effect of compartmentalization is encapsulin, a large and newly discovered class of protein shells that are typically composed of two proteins: a protomer that assembles into the icosahedral shell and a cargo protein packaged inside. Encapsulins are some of the simplest known protein shell systems and readily self-assemble in vivo. Systematic characterization of the effects of compartmentalization requires the ability to load a wide range of cargo proteins. Here, we demonstrate that foreign cargo can be loaded into the encapsulin from Thermotoga maritima both in vivo and in vitro by fusion of the cargo protein with a short C-terminal peptide present in the native cargo. To facilitate biochemical characterization, we also develop a simple and rapid purification protocol and demonstrate the thermal and pH stability of the shell. Efforts to study the biophysical effects of protein encapsulation have been problematic in complex compartments, but the simplicity of assembling and loading encapsulin makes it an ideal system for future experiments exploring the effects of encapsulation on proteins.

  12. [Construction and identification of Nogo extra cellular peptide residues 1-40 gene lentiviral vector].

    Science.gov (United States)

    Yuan, Haifeng; Song, Yueming; Liu, Hao; Zhou, Chunguang; Kong, Qingquan; Liu, Liming; Gong, Quan

    2012-02-01

    To construct a lentiviral expression vector carrying Nogo extra cellular peptide residues 1-40 (NEP1-40) and to obtain NEP1-40 efficient and stable expression in mammalian cells. The DNA fragment of NEP1-40 coding sequence was amplified by PCR with designed primer from the cDNA library including NEP1-40 gene, and then subcloned into pGC-FU vector with in-fusion technique to generate the lentiviral expression vector, pGC-FU-NEP1-40. The positive clones were screened by PCR and the correct NEP1-40 was confirmed by sequencing. Recombinant lentiviruses were produced in 293T cells after the cotransfection of pGC-FU-NEP1-40, and packaging plasmids of pHelper 1.0 and pHelper 2.0. Green fluorescent protein (GFP) expression of infected 293T cells was observed to evaluate gene delivery efficiency. NEP1-40 protein expression in 293T cells was detected by Western blot. The lentiviral expression vector carrying NEP1-40 was successfully constructed by GFP observation, and NEP1-40 protein expression was detected in 293T cells by Western blot. The recombinant lentivirus pGC-FU-NEP1-40 is successfully constructed and it lays a foundation for further molecular function study of NEP 1-40.

  13. Ligand and Structure-Based Approaches for the Identification of Peptide Deformylase Inhibitors as Antibacterial Drugs.

    Science.gov (United States)

    Gao, Jian; Liang, Li; Zhu, Yasheng; Qiu, Shengzhi; Wang, Tao; Zhang, Ling

    2016-07-15

    Peptide deformylase (PDF) is a metalloprotease catalyzing the removal of a formyl group from newly synthesized proteins, which makes it an important antibacterial drug target. Given the importance of PDF inhibitors like actinonin in antibacterial drug discovery, several reported potent PDF inhibitors were used to develop pharmacophore models using the Galahad module of Sybyl 7.1 software. Generated pharmacophore models were composed of two donor atom centers, four acceptor atom centers and two hydrophobic groups. Model-1 was screened against the Zinc database and several compounds were retrieved as hits. Compounds with Qfit values of more than 60 were employed to perform a molecular docking study with the receptor Escherichia coli PDF, then compounds with docking score values of more than 6 were used to predict the in silico pharmacokinetic and toxicity risk via OSIRIS property explorer. Two known PDF inhibitors were also used to perform a molecular docking study with E. coli PDF as reference molecules. The results of the molecular docking study were validated by reproducing the crystal structure of actinonin. Molecular docking and in silico pharmacokinetic and toxicity prediction studies suggested that ZINC08740166 has a relatively high docking score of 7.44 and a drug score of 0.78.

  14. The Pig PeptideAtlas

    DEFF Research Database (Denmark)

    Hesselager, Marianne Overgaard; Codrea, Marius; Sun, Zhi

    2016-01-01

    Biological research of Sus scrofa, the domestic pig, is of immediate relevance for food production sciences, and for developing pig as a model organism for human biomedical research. Publicly available data repositories play a fundamental role for all biological sciences, and protein data...... repositories are in particular essential for the successful development of new proteomic methods. Cumulative proteome data repositories, including the PeptideAtlas, provide the means for targeted proteomics, system-wide observations, and cross-species observational studies, but pigs have so far been...... underrepresented in existing repositories. We here present a significantly improved build of the Pig PeptideAtlas, which includes pig proteome data from 25 tissues and three body fluid types mapped to 7139 canonical proteins. The content of the Pig PeptideAtlas reflects actively ongoing research within...

  15. Repository performance confirmation

    International Nuclear Information System (INIS)

    Hansen, Francis D.

    2011-01-01

    Yucca Mountain license application identified a broad suite of monitoring activities. A revision of the plan was expected to winnow the number of activities down to a manageable size. As a result, an objective process for the next stage of performance confirmation planning was developed as an integral part of an overarching long-term testing and monitoring strategy. The Waste Isolation Pilot Plant compliance monitoring program at once reflects its importance to stakeholders while demonstrating adequate understanding of relevant monitoring parameters. The compliance criteria were stated by regulation and are currently monitored as part of the regulatory rule for disposal. At the outset, the screening practice and parameter selection were not predicated on a direct or indirect correlation to system performance metrics, as was the case for Yucca Mountain. Later on, correlation to performance was established, and the Waste Isolation Pilot Plant continues to monitor ten parameters originally identified in the compliance certification documentation. The monitoring program has proven to be effective for the technical intentions and societal or public assurance. The experience with performance confirmation in the license application process for Yucca Mountain helped identify an objective, quantitative methodology for this purpose. Revision of the existing plan would be based on findings of the total system performance assessment. Identification and prioritization of confirmation activities would then derive from performance metrics associated with performance assessment. Given the understanding of repository performance confirmation, as reviewed in this paper, it is evident that the performance confirmation program for the Yucca Mountain project could be readily re-engaged if licensing activities resumed.

  16. Identification of genetic loci required for Campylobacter resistance to fowlicidin-1, a chicken host defense peptide

    Directory of Open Access Journals (Sweden)

    Ky Van Hoang

    2012-03-01

    Full Text Available Antimicrobial peptides (AMPs are critical components of host defense limiting bacterial infections at the gastrointestinal mucosal surface. Bacterial pathogens have co-evolved with host innate immunity and developed means to counteract the effect of endogenous AMPs. However, molecular mechanisms of AMP resistance in Campylobacter, an important human food borne pathogen with poultry as a major reservoir, are still largely unknown. In this study, random transposon mutagenesis and targeted site-directed mutagenesis approaches were used to identify genetic loci contributing Campylobacter resistance to fowlicidin-1, a chicken AMP belonging to cathelicidin family. An efficient transposon mutagenesis approach (EZ::TNTM Transposome in conjunction with a microtiter plate screening identified three mutants whose susceptibilities to fowlicidin-1 were significantly increased. Backcrossing of the transposon mutations into parent strain confirmed that the AMP-sensitive phenotype in each mutant was linked to the specific transposon insertion. Direct sequencing showed that these mutants have transposon inserted in the genes encoding two-component regulator CbrR, transporter CjaB, and putative trigger factor Tig. Genomic analysis also revealed an operon (Cj1580c-1584c that is homologous to sapABCDF, an operon conferring resistance to AMP in other pathogens. Insertional inactivation of Cj1583c (sapB significantly increased susceptibility of Campylobacter to fowlicidin-1. The sapB as well as tig and cjaB mutants were significantly impaired in their ability to compete with their wild-type strain 81-176 to colonize the chicken cecum. Together, this study identified four genetic loci in Campylobacter that will be useful for characterizing molecular basis of Campylobacter resistance to AMPs, a significant knowledge gap in Campylobacter pathogenesis.

  17. Identification and expression analysis of a novel stylicin antimicrobial peptide from Kuruma shrimp (Marsupenaeus japonicus).

    Science.gov (United States)

    Liu, Hong-tao; Wang, Jun; Mao, Yong; Liu, Min; Niu, Su-fang; Qiao, Ying; Su, Yong-quan; Wang, Chun-zhong; Zheng, Zhi-peng

    2015-12-01

    Antimicrobial peptides (AMPs) are important components of the innate immune system and function as the first line of defense against invading pathogens. In current study we identified, cloned and characterized a novel stylicin AMP from Kuruma shrimp Marsupenaeus japonicus (Mj-sty). The full-length cDNA of Mj-sty was 428 bp with an open reading frame of 315 bp that encoded 104 amino acids. The theoretical molecular mass of mature Mj-sty was 8.693 kDa with an isoelectric point (pI) of 4.79. A proline-rich N-terminal region and a C-terminal region contained 13 cysteine residues were identified. Genomic sequence analysis with respect to its cDNA showed that Mj-sty was organized into two exons interrupted by one intron. Tissue-specific expression revealed that Mj-sty was mainly transcribed in gills and hemocytes. Expression of Mj-sty in early developmental stages demonstrated that Mj-sty mRNA were present from fertilized eggs to post-larvae of 17 days (PL17), and the expression levels showed a significant variation in different developmental stages. After challenge of white spot syndrome virus (WSSV), the time-dependent expression pattern of Mj-sty in both gills and hepatopancrease showed down-regulation at the early hours of infection, subsequently up-regulation and down-regulation, and then up-regulation at the end hours to almost the half of the controls. The results indicate that Mj-sty is potentially involved in the ontogenesis and immune responses against WSSV. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition.

    Science.gov (United States)

    Huang, Yuhong; Busk, Peter Kamp; Grell, Morten Nedergaard; Zhao, Hai; Lange, Lene

    2014-12-01

    Mucor circinelloides produces plant cell wall degrading enzymes that allow it to grow on complex polysaccharides. Although the genome of M. circinelloides has been sequenced, only few plant cell wall degrading enzymes are annotated in this species. We applied peptide pattern recognition, which is a non-alignment based method for sequence analysis to map conserved sequences in glycoside hydrolase families. The conserved sequences were used to identify similar genes in the M. circinelloides genome. We found 12 different novel genes encoding members of the GH3, GH5, GH9, GH16, GH38, GH47 and GH125 families in M. circinelloides. One of the two GH3-encoding genes was predicted to encode a β-glucosidase (EC 3.2.1.21). We expressed this gene in Pichia pastoris KM71H and found that the purified recombinant protein had relative high β-glucosidase activity (1.73U/mg) at pH5 and 50°C. The Km and Vmax with p-nitrophenyl-β-d-glucopyranoside as substrate was 0.20mM and 2.41U/mg, respectively. The enzyme was not inhibited by glucose and retained 84% activity at glucose concentrations up to 140mM. Although zygomycetes are not considered to be important degraders of lignocellulosic biomass in nature, the present finding of an active β-glucosidase in M. circinelloides demonstrates that enzymes from this group of fungi have a potential for cellulose degradation. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Multivalent peptidic linker enables identification of preferred sites of conjugation for a potent thialanstatin antibody drug conjugate.

    Directory of Open Access Journals (Sweden)

    Sujiet Puthenveetil

    Full Text Available Antibody drug conjugates (ADCs are no longer an unknown entity in the field of cancer therapy with the success of marketed ADCs like ADCETRIS and KADCYLA and numerous others advancing through clinical trials. The pursuit of novel cytotoxic payloads beyond the mictotubule inhibitors and DNA damaging agents has led us to the recent discovery of an mRNA splicing inhibitor, thailanstatin, as a potent ADC payload. In our previous work, we observed that the potency of this payload was uniquely tied to the method of conjugation, with lysine conjugates showing much superior potency as compared to cysteine conjugates. However, the ADC field is rapidly shifting towards site-specific ADCs due to their advantages in manufacturability, characterization and safety. In this work we report the identification of a highly efficacious site-specific thailanstatin ADC. The site of conjugation played a critical role on both the in vitro and in vivo potency of these ADCs. During the course of this study, we developed a novel methodology of loading a single site with multiple payloads using an in situ generated multi-drug carrying peptidic linker that allowed us to rapidly screen for optimal conjugation sites. Using this methodology, we were able to identify a double-cysteine mutant ADC delivering four-loaded thailanstatin that was very efficacious in a gastric cancer xenograft model at 3mg/kg and was also shown to be efficacious against T-DM1 resistant and MDR1 overexpressing tumor cell lines.

  20. Multivalent peptidic linker enables identification of preferred sites of conjugation for a potent thialanstatin antibody drug conjugate.

    Science.gov (United States)

    Puthenveetil, Sujiet; He, Haiyin; Loganzo, Frank; Musto, Sylvia; Teske, Jesse; Green, Michael; Tan, Xingzhi; Hosselet, Christine; Lucas, Judy; Tumey, L Nathan; Sapra, Puja; Subramanyam, Chakrapani; O'Donnell, Christopher J; Graziani, Edmund I

    2017-01-01

    Antibody drug conjugates (ADCs) are no longer an unknown entity in the field of cancer therapy with the success of marketed ADCs like ADCETRIS and KADCYLA and numerous others advancing through clinical trials. The pursuit of novel cytotoxic payloads beyond the mictotubule inhibitors and DNA damaging agents has led us to the recent discovery of an mRNA splicing inhibitor, thailanstatin, as a potent ADC payload. In our previous work, we observed that the potency of this payload was uniquely tied to the method of conjugation, with lysine conjugates showing much superior potency as compared to cysteine conjugates. However, the ADC field is rapidly shifting towards site-specific ADCs due to their advantages in manufacturability, characterization and safety. In this work we report the identification of a highly efficacious site-specific thailanstatin ADC. The site of conjugation played a critical role on both the in vitro and in vivo potency of these ADCs. During the course of this study, we developed a novel methodology of loading a single site with multiple payloads using an in situ generated multi-drug carrying peptidic linker that allowed us to rapidly screen for optimal conjugation sites. Using this methodology, we were able to identify a double-cysteine mutant ADC delivering four-loaded thailanstatin that was very efficacious in a gastric cancer xenograft model at 3mg/kg and was also shown to be efficacious against T-DM1 resistant and MDR1 overexpressing tumor cell lines.

  1. P19-S Managing Proteomics Data from Data Generation and Data Warehousing to Central Data Repository and Journal Reviewing Processes

    Science.gov (United States)

    Thiele, H.; Glandorf, J.; Koerting, G.; Reidegeld, K.; Blüggel, M.; Meyer, H.; Stephan, C.

    2007-01-01

    In today’s proteomics research, various techniques and instrumentation bioinformatics tools are necessary to manage the large amount of heterogeneous data with an automatic quality control to produce reliable and comparable results. Therefore a data-processing pipeline is mandatory for data validation and comparison in a data-warehousing system. The proteome bioinformatics platform ProteinScape has been proven to cover these needs. The reprocessing of HUPO BPP participants’ MS data was done within ProteinScape. The reprocessed information was transferred into the global data repository PRIDE. ProteinScape as a data-warehousing system covers two main aspects: archiving relevant data of the proteomics workflow and information extraction functionality (protein identification, quantification and generation of biological knowledge). As a strategy for automatic data validation, different protein search engines are integrated. Result analysis is performed using a decoy database search strategy, which allows the measurement of the false-positive identification rate. Peptide identifications across different workflows, different MS techniques, and different search engines are merged to obtain a quality-controlled protein list. The proteomics identifications database (PRIDE), as a public data repository, is an archiving system where data are finally stored and no longer changed by further processing steps. Data submission to PRIDE is open to proteomics laboratories generating protein and peptide identifications. An export tool has been developed for transferring all relevant HUPO BPP data from ProteinScape into PRIDE using the PRIDE.xml format. The EU-funded ProDac project will coordinate the development of software tools covering international standards for the representation of proteomics data. The implementation of data submission pipelines and systematic data collection in public standards–compliant repositories will cover all aspects, from the generation of MS data

  2. SWATH Mass Spectrometry Performance Using Extended Peptide MS/MS Assay Libraries*

    Science.gov (United States)

    Wu, Jemma X.; Song, Xiaomin; Pascovici, Dana; Zaw, Thiri; Care, Natasha; Krisp, Christoph; Molloy, Mark P.

    2016-01-01

    The use of data-independent acquisition methods such as SWATH for mass spectrometry based proteomics is usually performed with peptide MS/MS assay libraries which enable identification and quantitation of peptide peak areas. Reference assay libraries can be generated locally through information dependent acquisition, or obtained from community data repositories for commonly studied organisms. However, there have been no studies performed to systematically evaluate how locally generated or repository-based assay libraries affect SWATH performance for proteomic studies. To undertake this analysis, we developed a software workflow, SwathXtend, which generates extended peptide assay libraries by integration with a local seed library and delivers statistical analysis of SWATH-quantitative comparisons. We designed test samples using peptides from a yeast extract spiked into peptides from human K562 cell lysates at three different ratios to simulate protein abundance change comparisons. SWATH-MS performance was assessed using local and external assay libraries of varying complexities and proteome compositions. These experiments demonstrated that local seed libraries integrated with external assay libraries achieve better performance than local assay libraries alone, in terms of the number of identified peptides and proteins and the specificity to detect differentially abundant proteins. Our findings show that the performance of extended assay libraries is influenced by the MS/MS feature similarity of the seed and external libraries, while statistical analysis using multiple testing corrections increases the statistical rigor needed when searching against large extended assay libraries. PMID:27161445

  3. Identification of sixteen peptides reflecting heat and/or storage induced processes by profiling of commercial milk samples.

    Science.gov (United States)

    Ebner, Jennifer; Baum, Florian; Pischetsrieder, Monika

    2016-09-16

    Peptide profiles of different drinking milk samples were examined to study how the peptide fingerprint of milk reflects processing conditions. The combination of a simple and fast method for peptide extraction using stage tips and MALDI-TOF-MS enabled the fast and easy generation and relative quantification of peptide fingerprints for high-temperature short-time (HTST), extended shelf life (ESL) and ultra-high temperature (UHT) milk of the same dairies. The relative quantity of 16 peptides changed as a function of increasing heat load. Additional heating experiments showed that among those, the intensity of peptide β-casein 196-209 (m/z 1460.9Da) was most heavily influenced by heat treatment indicating a putative marker peptide for milk processing conditions. Storage experiments with HTST- and UHT milk revealed that the differences between different types of milk samples were not only caused by the heating process. Relevant was also the proteolytic activity of enzymes during storage, which were differently influenced by the heat treatment. These results indicate that the peptide profile may be suitable to monitor processing as well as storage conditions of milk. In the present study, peptide profiling of different types of milk was carried out by MALDI-TOF-MS after stage-tip extraction and relative quantification using an internal reference peptide. Although MALDI-TOF-MS covers only part of the peptidome, the method is easy and quick and is, therefore, suited for routine analysis to address several aspects of food authenticity. Using this method, 16 native peptides were detected in milk that could be modulated by different industrial processes. Subsequent heating and storage experiments with pasteurized and UHT milk confirmed that these peptides are indeed related to the production or storage conditions of the respective products. Furthermore, the heating experiments revealed one peptide, namely the β-casein-derived sequence β-casein 196-209, which underwent

  4. Isolation and identification of calcium-chelating peptides from Pacific cod skin gelatin and their binding properties with calcium.

    Science.gov (United States)

    Wu, Wenfei; Li, Bafang; Hou, Hu; Zhang, Hongwei; Zhao, Xue

    2017-12-13

    A calcium-chelating peptide is considered to have the ability to improve calcium absorption. In this study, Pacific cod skin gelatin hydrolysates treated with trypsin for 120 min exhibited higher calcium-chelating activity. Sequential chromatography, involving hydroxyapatite affinity chromatography and reversed phase high performance liquid chromatography, was used for the purification of calcium-chelating peptides. Two novel peptides with the typical characteristics of collagen were sequenced as GDKGESGEAGER and GEKGEGGHR based on LC-HRMS/MS, which showed a high affinity to calcium. Calcium-peptide complexation was further characterized by ESI-MS (MS and MS/MS) and FTIR spectroscopy. The results showed that the complexation of the two peptides with calcium was conducted mainly at the ratio of 1 : 1. The amino terminal group and the peptide bond of the peptide backbone as well as the amino group of the lysine side chain and the carboxylate of the glutamate side chain were the possible calcium binding sites for the two peptides. Meanwhile, several amino acid side chain groups, including the hydroxyl (Ser) and carboxylate (Asp) of GDKGESGEAGER and the imine (His) of GEKGEGGHR, were crucial in the complexation. The arginine residue in GEKGEGGHR also participated in the calcium coordination. Additionally, several active fragments with calcium-chelating activity were obtained using MS/MS spectra, including GDKGESGEAGE, GEAGER, GEK, EKG and KGE. This study suggests that gelatin-derived peptides have the potential to be used as a calcium-chelating ingredient to combat calcium deficiency.

  5. Identification of structural traits that increase the antimicrobial activity of a chimeric peptide of human β-defensins 2 and 3.

    Science.gov (United States)

    Spudy, Björn; Sönnichsen, Frank D; Waetzig, Georg H; Grötzinger, Joachim; Jung, Sascha

    2012-10-12

    Antimicrobial peptides participate in the first line of defence of many organisms against pathogens. In humans, the family of β-defensins plays a pivotal role in innate immunity. Two human β-defensins, β-defensin-2 and -3 (HBD2 and HBD3), show substantial sequence identity and structural similarity. However, HBD3 kills Staphylococcus (S.) aureus with a 4- to 8-fold higher efficiency compared to HBD2, whereas their activities against Escherichia (E.) coli are very similar. The generation of six HBD2/HBD3-chimeric molecules led to the identification of distinct molecular regions which mediate their divergent killing properties. One of the chimeras (chimera C3) killed both E. coli and S. aureus with an even higher efficacy compared to the wild-type molecules. Due to the broad spectrum of its antimicrobial activity against many human multidrug-resistant pathogens, this HBD2/HBD3-chimeric peptide represents a promising candidate for a new class of antibiotics. In order to investigate the structural basis of its exceptional antimicrobial activity, the peptide's tertiary structure was determined by NMR spectroscopy, which allowed its direct comparison to the published structures of HBD2 and HBD3 and the identification of the activity-increasing molecular features. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Process mining software repositories

    NARCIS (Netherlands)

    Poncin, W.; Serebrenik, A.; Brand, van den M.G.J.

    2011-01-01

    Software developers' activities are in general recorded in software repositories such as version control systems, bug trackers and mail archives. While abundant information is usually present in such repositories, successful information extraction is often challenged by the necessity to

  7. Breast Cancer Tissue Repository

    National Research Council Canada - National Science Library

    Iglehart, J

    1997-01-01

    The Breast Tissue Repository at Duke enters its fourth year of finding. The purpose of the Repository at Duke is to provide substantial quantities of frozen tissue for explorative molecular studies...

  8. Identification and Characterization of a Small Inhibitory Peptide That Can Target DNA-PKcs Autophosphorylation and Increase Tumor Radiosensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Sun Xiaonan [Department of Radiation Oncology, Sir Run Run Shaw Hospital, Sir Run Run Shaw Institute of Clinical Medicine of Zhejiang University, Hangzhou (China); Yang Chunying [Department of Radiation Oncology, Methodist Hospital Research Institute, Weill Cornell Medical College, Houston, TX (United States); Liu Hai; Wang Qi [Department of Radiation Oncology, Sir Run Run Shaw Hospital, Sir Run Run Shaw Institute of Clinical Medicine of Zhejiang University, Hangzhou (China); Wu Shixiu [Department of Radiation Oncology, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou (China); Li Xia; Xie Tian [Research Center of Biomedicine and Health, Hangzhou Normal University, Hangzhou (China); Brinkman, Kathryn L.; Teh, Bin S.; Butler, E. Brian [Department of Radiation Oncology, Methodist Hospital Research Institute, Weill Cornell Medical College, Houston, TX (United States); Xu Bo, E-mail: bxu@tmhs.org [Department of Radiation Oncology, Methodist Hospital Research Institute, Weill Cornell Medical College, Houston, TX (United States); Zheng, Shu, E-mail: zhengshu@zju.edu.cn [Cancer Institute, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou (China)

    2012-12-01

    Purpose: The DNA protein kinase catalytic subunit (DNA-PKcs) is one of the critical elements involved in the DNA damage repair process. Inhibition of DNA-PKcs results in hypersensitivity to ionizing radiation (IR); therefore, this approach has been explored to develop molecular targeted radiosensitizers. Here, we aimed to develop small inhibitory peptides that could specifically target DNA-PKcs autophosphorylation, a critical step for the enzymatic activation of the kinase in response to IR. Methods and Materials: We generated several small fusion peptides consisting of 2 functional domains, 1 an internalization domain and the other a DNA-PKcs autophosphorylation inhibitory domain. We characterized the internalization, toxicity, and radiosensitization activities of the fusion peptides. Furthermore, we studied the mechanisms of the inhibitory peptides on DNA-PKcs autophosphorylation and DNA repair. Results: We found that among several peptides, the biotin-labeled peptide 3 (BTW3) peptide, which targets DNA-PKcs threonine 2647 autophosphorylation, can abrogate IR-induced DNA-PKcs activation and cause prolonged {gamma}-H2AX focus formation. We demonstrated that BTW3 exposure led to hypersensitivity to IR in DNA-PKcs-proficient cells but not in DNA-PKcs-deficient cells. Conclusions: The small inhibitory peptide BTW3 can specifically target DNA-PKcs autophosphorylation and enhance radiosensitivity; therefore, it can be further developed as a novel class of radiosensitizer.

  9. Identification and Characterization of a Small Inhibitory Peptide That Can Target DNA-PKcs Autophosphorylation and Increase Tumor Radiosensitivity

    International Nuclear Information System (INIS)

    Sun Xiaonan; Yang Chunying; Liu Hai; Wang Qi; Wu Shixiu; Li Xia; Xie Tian; Brinkman, Kathryn L.; Teh, Bin S.; Butler, E. Brian; Xu Bo; Zheng, Shu

    2012-01-01

    Purpose: The DNA protein kinase catalytic subunit (DNA-PKcs) is one of the critical elements involved in the DNA damage repair process. Inhibition of DNA-PKcs results in hypersensitivity to ionizing radiation (IR); therefore, this approach has been explored to develop molecular targeted radiosensitizers. Here, we aimed to develop small inhibitory peptides that could specifically target DNA-PKcs autophosphorylation, a critical step for the enzymatic activation of the kinase in response to IR. Methods and Materials: We generated several small fusion peptides consisting of 2 functional domains, 1 an internalization domain and the other a DNA-PKcs autophosphorylation inhibitory domain. We characterized the internalization, toxicity, and radiosensitization activities of the fusion peptides. Furthermore, we studied the mechanisms of the inhibitory peptides on DNA-PKcs autophosphorylation and DNA repair. Results: We found that among several peptides, the biotin-labeled peptide 3 (BTW3) peptide, which targets DNA-PKcs threonine 2647 autophosphorylation, can abrogate IR-induced DNA-PKcs activation and cause prolonged γ-H2AX focus formation. We demonstrated that BTW3 exposure led to hypersensitivity to IR in DNA-PKcs-proficient cells but not in DNA-PKcs-deficient cells. Conclusions: The small inhibitory peptide BTW3 can specifically target DNA-PKcs autophosphorylation and enhance radiosensitivity; therefore, it can be further developed as a novel class of radiosensitizer.

  10. Identification and functional analysis of a novel bradykinin inhibitory peptide in the venoms of New World Crotalinae pit vipers

    International Nuclear Information System (INIS)

    James Graham, Robert Leslie; Graham, Ciaren; McClean, Stephen; Chen, Tianbao; O'Rourke, Martin; Hirst, David; Theakston, David; Shaw, Chris

    2005-01-01

    A novel undecapeptide has been isolated and structurally characterized from the venoms of three species of New World pit vipers from the subfamily, Crotalinae. These include the Mexican moccasin (Agkistrodon bilineatus), the prairie rattlesnake (Crotalus viridis viridis), and the South American bushmaster (Lachesis muta). The peptide was purified from all three venoms using a combination of gel permeation chromatography and reverse-phase HPLC. Automated Edman degradation sequencing and MALDI-TOF mass spectrometry established its peptide primary structure as: Thr-Pro-Pro-Ala-Gly-Pro-Asp-Val-Gly-Pro-Arg-OH, with a non-protonated molecular mass of 1063.18 Da. A synthetic replicate of the peptide was found to be an antagonist of bradykinin action at the rat vascular B2 receptor. This is the first bradykinin inhibitory peptide isolated from snake venom. Database searching revealed the peptide to be highly structurally related (10/11 residues) with a domain residing between the bradykinin-potentiating peptide and C-type natriuretic peptide domains of a recently cloned precursor from tropical rattlesnake (Crotalus durissus terrificus) venom gland. BIP thus represents a novel biological entity from snake venom

  11. Identification and molecular docking study of novel angiotensin-converting enzyme inhibitory peptides from Salmo salar using in silico methods.

    Science.gov (United States)

    Yu, Zhipeng; Chen, Yang; Zhao, Wenzhu; Li, Jianrong; Liu, Jingbo; Chen, Feng

    2018-01-25

    In order to circumvent some challenges of the classical approach, the in silico method has been applied to the discovery of angiotensin-converting enzyme (ACE) inhibitory peptides from food proteins. In this study, some convenient and efficient in silico tools were utilized to identify novel ACE inhibitory peptides from Salmo salar. Collagen from Salmo salar was digested in silico into hundreds of peptides. Results revealed that tetrapeptides PGAR and IGPR showed potent ACE inhibitory activity, with IC 50 values of 0.598 ± 0.12 and 0.43 ± 0.09 mmol L -1 , respectively. The molecular docking result showed that PGAR and IGPR interact with ACE mostly via hydrogen bonds and attractive charge. Peptide IGPR interacts with Zn + at the ACE active site, showing high inhibitory activity. Interaction with Zn + in ACE may lead to higher inhibitory activity of peptides, and Pi interactions may promote the effect of peptides on ACE. The in silico method can be an effective method to predict potent ACE inhibitory peptides from food proteins. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  12. Application of Ga-Al discrimination plots in identification of high strength granitic host rocks for deep geological repository of high level radioactive waste

    International Nuclear Information System (INIS)

    Bajpai, R.K.; Narayan, P.K.; Trivedi, R.K.; Purohit, M.K.

    2010-01-01

    The permanent disposal of vitrified high level wastes and in some cases even spent fuel, is being planned in specifically designed and built deep geological repository located in the depth range of 500-600m in appropriate host rock at carefully selected sites. Such facilities are expected to provide very long term isolation and confinement to the disposed waste by means of long term mechanical stability of such structures that results from very high strength and homogeneity of the chosen rock, geochemical compatible environment around the disposed waste and general lack of groundwater. In Indian geological repository development programme, granites have been selected as target host rock and large scale characterization studies have been undertaken to develop database of mineralogy, petrology, geochemistry and rock mechanical characteristics. The paper proposes a new approach for demarcation of high strength homogeneous granite rocks from within an area of about 100 square kilometres wherein a cocktail of granites of different origins with varying rock mass characteristics co exists. The study area is characterised by the presence of A, S and I type granites toughly intermixed. The S type granites are derived from sedimentary parent material and therefore carry relics of parent fabric and at times undigested material with resultant reduction in their strength and increased inhomogeneity. On the other hand I type varieties are derived from igneous parents and are more homogeneous with sufficient strength. The A type granites are emplaced as molten mass in a complete non-tectonic setting with resultant homogeneous compositions, absence of tectonic fabric and very high strength. Besides they are silica rich with less vulnerability to alterations with time. Thus A type granites are most suited for construction of Deep Geological Repository. For developing a geochemical approach for establishing relation between chemical compositions and rock strength parameters, a

  13. Peptide immunisation of HLA-DR-transgenic mice permits the identification of a novel HLA-DRbeta1*0101- and HLA-DRbeta1*0401-restricted epitope from p53.

    Science.gov (United States)

    Rojas, José Manuel; McArdle, Stephanie E B; Horton, Roger B V; Bell, Matthew; Mian, Shahid; Li, Geng; Ali, Selman A; Rees, Robert C

    2005-03-01

    Because of the central role of CD4(+) T cells in antitumour immunity, the identification of the MHC class II-restricted peptides to which CD4(+) T cells respond has become a priority of tumour immunologists. Here, we describe a strategy permitting us to rapidly determine the immunogenicity of candidate HLA-DR-restricted peptides using peptide immunisation of HLA-DR-transgenic mice, followed by assessment of the response in vitro. This strategy was successfully applied to the reported haemaglutinin influenza peptide HA(307-319), and then extended to three candidate HLA-DR-restricted p53 peptides predicted by the evidence-based algorithm SYFPEITHI to bind to HLA-DRbeta1*0101 (HLA-DR1) and HLA-DRbeta1*0401 (HLA-DR4) molecules. One of these peptides, p53(108-122), consistently induced responses in HLA-DR1- and in HLA-DR4-transgenic mice. Moreover, this peptide was naturally processed by dendritic cells (DCs), and induced specific proliferation in the splenocytes of mice immunised with p53 cDNA, demonstrating that immune responses could be naturally mounted to the peptide. Furthermore, p53(108-122) peptide was also immunogenic in HLA-DR1 and HLA-DR4 healthy donors. Thus, the use of this transgenic model permitted the identification of a novel HLA-DR-restricted epitope from p53 and constitutes an attractive approach for the rapid identification of novel immunogenic MHC class II-restricted peptides from tumour antigens, which can ultimately be incorporated in immunotherapeutic protocols.

  14. Proteomics data repositories: Providing a safe haven for your data and acting as a springboard for further research

    Science.gov (United States)

    Vizcaíno, Juan Antonio; Foster, Joseph M.; Martens, Lennart

    2010-01-01

    Despite the fact that data deposition is not a generalised fact yet in the field of proteomics, several mass spectrometry (MS) based proteomics repositories are publicly available for the scientific community. The main existing resources are: the Global Proteome Machine Database (GPMDB), PeptideAtlas, the PRoteomics IDEntifications database (PRIDE), Tranche, and NCBI Peptidome. In this review the capabilities of each of these will be described, paying special attention to four key properties: data types stored, applicable data submission strategies, supported formats, and available data mining and visualization tools. Additionally, the data contents from model organisms will be enumerated for each resource. There are other valuable smaller and/or more specialized repositories but they will not be covered in this review. Finally, the concept behind the ProteomeXchange consortium, a collaborative effort among the main resources in the field, will be introduced. PMID:20615486

  15. Analysis of Swine Leukocyte Antigen Peptide Binding Profiles and the Identification of T cell Epitopes by Tetramer Staining

    DEFF Research Database (Denmark)

    Pedersen, Lasse Eggers

    class I peptide binding characteristics in relation to immune responses to vaccination or infection. Applying proven technologies to newly produced, recombinant swine leukocyte antigen (SLA) class I proteins yielded a body of data for peptide:SLA:β2m (pSLA) complex affinity and stability. Mapping...... system to specifically identify and react upon non-self peptide fragments unique only to the foreign intruder. The polymorphism of the MHC molecule effectively individualizes the immune response of each member of any given species. Moreover, responding T cells recognize antigen ligands, only...... in the context of peptide:MHC:β2m (pMHC) complex. The gene encoding the MHC is one of the most polymorphic regions of the genome known. Despite thousands of different human leukocyte antigen (HLA) variants identified, each member of a species only inherits and expresses a few of these MHC alleles. The “MHC...

  16. Identification of a novel antimicrobial peptide from human hepatitis B virus core protein arginine-rich domain (ARD.

    Directory of Open Access Journals (Sweden)

    Heng-Li Chen

    Full Text Available The rise of multidrug-resistant (MDR pathogens causes an increasing challenge to public health. Antimicrobial peptides are considered a possible solution to this problem. HBV core protein (HBc contains an arginine-rich domain (ARD at its C-terminus, which consists of 16 arginine residues separated into four clusters (ARD I to IV. In this study, we demonstrated that the peptide containing the full-length ARD I-IV (HBc147-183 has a broad-spectrum antimicrobial activity at micro-molar concentrations, including some MDR and colistin (polymyxin E-resistant Acinetobacter baumannii. Furthermore, confocal fluorescence microscopy and SYTOX Green uptake assay indicated that this peptide killed Gram-negative and Gram-positive bacteria by membrane permeabilization or DNA binding. In addition, peptide ARD II-IV (HBc153-176 and ARD I-III (HBc147-167 were found to be necessary and sufficient for the activity against P. aeruginosa and K. peumoniae. The antimicrobial activity of HBc ARD peptides can be attenuated by the addition of LPS. HBc ARD peptide was shown to be capable of direct binding to the Lipid A of lipopolysaccharide (LPS in several in vitro binding assays. Peptide ARD I-IV (HBc147-183 had no detectable cytotoxicity in various tissue culture systems and a mouse animal model. In the mouse model by intraperitoneal (i.p. inoculation with Staphylococcus aureus, timely treatment by i.p. injection with ARD peptide resulted in 100-fold reduction of bacteria load in blood, liver and spleen, as well as 100% protection of inoculated animals from death. If peptide was injected when bacterial load in the blood reached its peak, the protection rate dropped to 40%. Similar results were observed in K. peumoniae using an IVIS imaging system. The finding of anti-microbial HBc ARD is discussed in the context of commensal gut microbiota, development of intrahepatic anti-viral immunity and establishment of chronic infection with HBV. Our current results suggested that

  17. Identification of novel inhibitors of Pseudomonas aeruginosa MurC enzyme derived from phage-displayed peptide libraries.

    Science.gov (United States)

    El Zoeiby, Ahmed; Sanschagrin, François; Darveau, André; Brisson, Jean-Robert; Levesque, Roger C

    2003-03-01

    The machinery of peptidoglycan biosynthesis is an ideal site at which to look for novel antimicrobial targets. Phage display was used to develop novel peptide inhibitors for MurC, an essential enzyme involved in the early steps of biosynthesis of peptidoglycan monomer. We cloned and overexpressed the murA, -B and -C genes from Pseudomonas aeruginosa in the pET expression vector, adding a His-tag to their C termini. The three proteins were overproduced in Escherichia coli and purified to homogeneity in milligram quantities. MurA and -B were combinatorially used to synthesize the MurC substrate UDP-N-acetylmuramate, the identity of which was confirmed by mass spectrometry and nuclear magnetic resonance analysis. Two phage-display libraries were screened against MurC in order to identify peptide ligands to the enzyme. Three rounds of biopanning were carried out, successively increasing elution specificity from round 1 to 3. The third round was accomplished with both non-specific elution and competitive elution with each of the three MurC substrates, UDP-N-acetylmuramic acid (UNAM), ATP and L-alanine. The DNA of 10 phage, selected randomly from each group, was extracted and sequenced, and consensus peptide sequences were elucidated. Peptides were synthesized and tested for inhibition of the MurC-catalysed reaction, and two peptides were shown to be inhibitors of MurC activity with IC(50)s of 1.5 and 0.9 mM, respectively. The powerful selection technique of phage display allowed us to identify two peptide inhibitors of the essential bacterial enzyme MurC. The peptide sequences represent the basis for the synthesis of inhibitory peptidomimetic molecules.

  18. Identification of a haemolysin-like peptide with antibacterial activity using the draft genome sequence of Staphylococcus epidermidis strain A487.

    Science.gov (United States)

    Al-Mahrous, Mohammed M; Jack, Ralph W; Sandiford, Stephanie K; Tagg, John R; Beatson, Scott A; Upton, Mathew

    2011-08-01

    Our interest in Staphylococcus epidermidis strain A487 was prompted by the unusual nature of its inhibitory activity in screening tests against methicillin-resistant Staphylococcus aureus isolates. The inhibitory activity was detected in deferred antagonism tests only if the agar plate was preheated for at least 35 min at ≥ 55 °C before inoculation of the indicator bacteria, this phenomenon indicating possible involvement of a heat-labile immunity agent or protease. The inhibitor was purified to homogeneity by ammonium sulphate precipitation, followed by cation-exchange and reversed-phase chromatography. Tandem MS revealed a novel peptide of molecular weight 2588.4 Da. The draft genome sequence of strain A487 was determined using 454 GS FLX technology, allowing the identification of the structural gene (hlp) encoding the mature peptide MQFITDLIKKAVDFFKGLFGNK. The deduced amino acid sequence of peptide 487 exhibited 70.8% similarity to that of a putative haemolysin from Staphylococcus cohnii. Analysis of the genome of strain A487 showed several additional inhibitor-encoding genes, including hld, the determinant for staphylococcal δ-lysin. This work indicates that potentially useful inhibitors could be overlooked in agar-based inhibitor screening programmes lacking a heat pretreatment step and also highlights the utility of draft genome sequence examination in antibacterial agent discovery. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  19. Repository Rodeo Redux

    CERN Document Server

    Anez, Melissa; Donohue, Tim; Fyson, Will; Simko, Tibor; Wilcox, David

    2017-01-01

    You’ve got more repository questions and we’ve got more answers! Last year’s Repository Rodeo panel was a huge success, so we’re taking the show on the road to Brisbane for OR2017. Join representatives from the DSpace, Eprints, Fedora, Hydra, and Islandora communities as we (briefly) explain what each of our repositories actually does. We'll also talk about the directions of our respective technical and community developments, and related to the conference theme of Open: Innovation Knowledge Repositories, offer brief observations about the latest, most promising and/or most surprising innovations in our space. This panel will be a great opportunity for newcomers to Open Repositories to get a crash course on the major repository options and meet representatives from each of their communities. After a brief presentation from each representative, we'll open the session up for questions from the audience.

  20. The fusion protein signal-peptide-coding region of canine distemper virus: a useful tool for phylogenetic reconstruction and lineage identification.

    Directory of Open Access Journals (Sweden)

    Nicolás Sarute

    Full Text Available Canine distemper virus (CDV; Paramyxoviridae, Morbillivirus is the etiologic agent of a multisystemic infectious disease affecting all terrestrial carnivore families with high incidence and mortality in domestic dogs. Sequence analysis of the hemagglutinin (H gene has been widely employed to characterize field strains, permitting the identification of nine CDV lineages worldwide. Recently, it has been established that the sequences of the fusion protein signal-peptide (Fsp coding region are extremely variable, suggesting that analysis of its sequence might be useful for strain characterization studies. However, the divergence of Fsp sequences among worldwide strains and its phylogenetic resolution has not yet been evaluated. We constructed datasets containing the Fsp-coding region and H gene sequences of the same strains belonging to eight CDV lineages. Both datasets were used to evaluate their phylogenetic resolution. The phylogenetic analysis revealed that both datasets clustered the same strains into eight different branches, corresponding to CDV lineages. The inter-lineage amino acid divergence was fourfold greater for the Fsp peptide than for the H protein. The likelihood mapping revealed that both datasets display strong phylogenetic signals in the region of well-resolved topologies. These features indicate that Fsp-coding region sequence analysis is suitable for evolutionary studies as it allows for straightforward identification of CDV lineages.

  1. The fusion protein signal-peptide-coding region of canine distemper virus: a useful tool for phylogenetic reconstruction and lineage identification.

    Science.gov (United States)

    Sarute, Nicolás; Calderón, Marina Gallo; Pérez, Ruben; La Torre, José; Hernández, Martín; Francia, Lourdes; Panzera, Yanina

    2013-01-01

    Canine distemper virus (CDV; Paramyxoviridae, Morbillivirus) is the etiologic agent of a multisystemic infectious disease affecting all terrestrial carnivore families with high incidence and mortality in domestic dogs. Sequence analysis of the hemagglutinin (H) gene has been widely employed to characterize field strains, permitting the identification of nine CDV lineages worldwide. Recently, it has been established that the sequences of the fusion protein signal-peptide (Fsp) coding region are extremely variable, suggesting that analysis of its sequence might be useful for strain characterization studies. However, the divergence of Fsp sequences among worldwide strains and its phylogenetic resolution has not yet been evaluated. We constructed datasets containing the Fsp-coding region and H gene sequences of the same strains belonging to eight CDV lineages. Both datasets were used to evaluate their phylogenetic resolution. The phylogenetic analysis revealed that both datasets clustered the same strains into eight different branches, corresponding to CDV lineages. The inter-lineage amino acid divergence was fourfold greater for the Fsp peptide than for the H protein. The likelihood mapping revealed that both datasets display strong phylogenetic signals in the region of well-resolved topologies. These features indicate that Fsp-coding region sequence analysis is suitable for evolutionary studies as it allows for straightforward identification of CDV lineages.

  2. Identification of cyst nematode B-type CLE peptides and modulation of the vascular stem cell pathway for feeding cell formation.

    Directory of Open Access Journals (Sweden)

    Xiaoli Guo

    2017-02-01

    Full Text Available Stem cell pools in the SAM (shoot apical meristem, RAM (root apical meristem and vascular procambium/cambium are regulated by CLE-receptor kinase-WOX signaling modules. Previous data showed that cyst nematode CLE-like effector proteins delivered into host cells through a stylet, act as ligand mimics of plant A-type CLE peptides and are pivotal for successful parasitism. Here we report the identification of a new class of CLE peptides from cyst nematodes with functional similarity to the B-type CLE peptide TDIF (tracheary element differentiation inhibitory factor encoded by the CLE41 and CLE44 genes in Arabidopsis. We further demonstrate that the TDIF-TDR (TDIF receptor-WOX4 pathway, which promotes procambial meristem cell proliferation, is involved in beet cyst nematode Heterodera schachtii parasitism. We observed activation of the TDIF pathway in developing feeding sites, reduced nematode infection in cle41 and tdr-1 wox4-1 mutants, and compromised syncytium size in cle41, tdr-1, wox4-1 and tdr-1 wox4-1 mutants. By qRT-PCR and promoter:GUS analyses, we showed that the expression of WOX4 is decreased in a clv1-101 clv2-101 rpk2-5 mutant, suggesting that WOX4 is a potential downstream target of nematode CLEs. Exogenous treatment with both nematode A-type and B-type CLE peptides induced massive cell proliferation in wild type roots, suggesting that the two types of CLEs may regulate cell proliferation during feeding site formation. These findings highlight an important role of the procambial cell proliferation pathway in cyst nematode feeding site formation.

  3. CAED Document Repository

    Data.gov (United States)

    U.S. Environmental Protection Agency — Compliance Assurance and Enforcement Division Document Repository (CAEDDOCRESP) provides internal and external access of Inspection Records, Enforcement Actions, and...

  4. Administrative Data Repository (ADR)

    Data.gov (United States)

    Department of Veterans Affairs — The Administrative Data Repository (ADR) was established to provide support for the administrative data elements relative to multiple categories of a person entity...

  5. Olfactory identification in patients with schizophrenia - the influence of β-endorphin and calcitonin gene-related peptide concentrations.

    Science.gov (United States)

    Urban-Kowalczyk, M; Śmigielski, J; Strzelecki, D

    2017-03-01

    The relationship between the olfactory system and emotional processing is an area of growing interest in schizophrenia research. Both the orbitofrontal cortex and amygdala are involved in the processing of olfactory information, and olfactory deficits may be also influenced by endogenous opioids and calcitonin gene-related peptide (CGRP), which is probably involved in dopaminergic transmission. However, the relationship between endorphins and dopaminergic transmission has not been fully explored. Odor identification performance and valence interaction was evaluated among 50 schizophrenic patients and 50 controls. Schizophrenia symptoms were assessed using the Positive and Negative Syndrome Scale (PANSS). All study participants were subjected to the University of Pennsylvania Smell Identification Test (UPSIT), blood β-endorphin (BE) and CGRP measurement. Insignificantly higher BE concentrations were observed in the patient group, while significantly higher UPSIT scores were seen in controls (mean UPSIT 32.48 vs 26.82). The patients demonstrated significantly more identification errors for pleasant (P=0.000) and neutral (P=0.055) odors than for unpleasant odors. Patients with higher BE concentrations made more identification errors concerning pleasant (R s =-0.292; P=0.04) and neutral odors (R s =-0.331; P=0.019). Although the concentration of CGRP was significantly higher in the patient sample (Pidentification by valence for pleasant and neutral odors (UPSIT n/16: R s =-0.450, P=0.001; UPSIT n/15: R s =-0.586, P=0.000), and a weak negative correlation between PANSS N score and identification of unpleasant odors (UPSIT n/9: R s =-0.325, P=0.021). Schizophrenic patients present a unique pattern of smell identification characterized by aberrant hedonic ratings for pleasant odors but not unpleasant ones. Individuals with predominant negative symptoms and higher BE concentrations are most able to identify negative odors. Copyright © 2016 Elsevier Masson SAS. All rights

  6. Identification of a preferred substrate peptide for transglutaminase 3 and detection of in situ activity in skin and hair follicles.

    Science.gov (United States)

    Yamane, Asaka; Fukui, Mina; Sugimura, Yoshiaki; Itoh, Miho; Alea, Mileidys Perez; Thomas, Vincent; El Alaoui, Said; Akiyama, Masashi; Hitomi, Kiyotaka

    2010-09-01

    Transglutaminases (TGases) are a family of enzymes that catalyze cross-linking reactions between proteins. During epidermal differentiation, these enzymatic reactions are essential for formation of the cornified envelope, which consists of cross-linked structural proteins. Two main transglutaminases isoforms, epidermal-type (TGase 3) and keratinocyte-type (TGase 1), are cooperatively involved in this process of differentiating keratinocytes. Information regarding their substrate preference is of great importance to determine the functional role of these isozymes and clarify their possible co-operative action. Thus far, we have identified highly reactive peptide sequences specifically recognized by TGases isozymes such as TGase 1, TGase 2 (tissue-type isozyme) and the blood coagulation isozyme, Factor XIII. In this study, several substrate peptide sequences for human TGase 3 were screened from a phage-displayed peptide library. The preferred substrate sequences for TGase 3 were selected and evaluated as fusion proteins with mutated glutathione S-transferase. From these studies, a highly reactive and isozyme-specific sequence (E51) was identified. Furthermore, this sequence was found to be a prominent substrate in the peptide form and was suitable for detection of in situ TGase 3 activity in the mouse epidermis. TGase 3 enzymatic activity was detected in the layers of differentiating keratinocytes and hair follicles with patterns distinct from those of TGase 1. Our findings provide new information on the specific distribution of TGase 3 and constitute a useful tool to clarify its functional role in the epidermis.

  7. Identification of novel bacteriophage peptides using a combination of gene sequence LC-MS-MS analysis and BLASTP

    Science.gov (United States)

    Introduction: In an effort to characterize novel bacteriophage with lytic activity against pathogenic E.coli associated with foodborne illness, gene sequencing and mass spectrometry have been used to identify expressed peptides which differentiate isolated bacteriophage from other known phage. Here,...

  8. Identification of three novel peptides isolated from the venom of the neotropical social wasp Polistes major major

    Czech Academy of Sciences Publication Activity Database

    Čeřovský, Václav; Pohl, J.; Yang, Z.; Alam, N.; Attygalle, A. B.

    2007-01-01

    Roč. 13, č. 7 (2007), s. 445-450 ISSN 1075-2617 Institutional research plan: CEZ:AV0Z40550506 Keywords : mastoparans * wasp kinins * chemotactic peptides Subject RIV: CC - Organic Chemistry Impact factor: 1.768, year: 2007

  9. Identification of New Anti-inflammatory Peptides from Zein Hydrolysate after Simulated Gastrointestinal Digestion and Transport in Caco-2 Cells.

    Science.gov (United States)

    Liang, Qiufang; Chalamaiah, Meram; Ren, Xiaofeng; Ma, Haile; Wu, Jianping

    2018-02-07

    Chronic inflammation is an underlying contributor to various chronic diseases. The objectives of this study were to investigate the anti-inflammatory activity of zein hydrolysate after simulated gastrointestinal digestion and Caco-2 cell absorption and to identify novel anti-inflammatory peptides after transport across Caco-2 cells. Three zein hydrolysates were prepared and further digested using gastrointestinal proteases; their transports were studied in Caco-2 cells. Anti-inflammatory activity was studied in endothelial EA.hy926 cells. Three zein hydrolysates and their digests significantly decreased the expression of tumor necrosis factor-α (TNF-α) induced pro-inflammatory vascular cell adhesion molecule-1 (VCAM-1) by 37.3-66.0%. Eleven novel peptides with 5-9 amino acid residues were sequenced; three peptides showed strong anti-inflammatory activity by inhibiting the VCAM-1 by 54-38.9% and intercellular cell adhesion molecule-1 (ICAM-1) by 36.5-28.6% at 0.2 mM. A new approach to identify novel anti-inflammatory peptides that could survive gastrointestinal digestion and absorption was developed.

  10. A neural network method for identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites

    DEFF Research Database (Denmark)

    Nielsen, Henrik; Engelbrecht, Jacob; Brunak, Søren

    1997-01-01

    We have developed a new method for the identication of signal peptides and their cleavage sites based on neural networks trained on separate sets of prokaryotic and eukaryotic sequences. The method performs signicantly better than previous prediction schemes, and can easily be applied to genome...

  11. Identification of antihyperuricemic peptides in the proteolytic digest of shark cartilage water extract using in vivo activity-guided fractionation.

    Science.gov (United States)

    Murota, Itsuki; Taguchi, Satoko; Sato, Nobuyuki; Park, Eun Young; Nakamura, Yasushi; Sato, Kenji

    2014-03-19

    A peptide that exerts antihyperuricemic activity after oral administration was identified from a microbial protease (alcalase) digest of the water extract of shark cartilage by in vivo activity-guided fractionation, using oxonate-induced hyperuricemic rats. Water extract of shark cartilage was first fractionated by preparative ampholine-free isoelectric focusing, followed by preparative reversed-phase liquid chromatography. The antihyperuricemic activity of the alcalse digests of the obtained fractions was evaluated using an animal model. Alcalase digests of the basic and hydrophobic fractions exerted antihyperuricemic activity. A total of 18 peptides were identified in the alcalase digest of the final active fraction. These peptides were chemically synthesized and evaluated for antihyperuricemic activity. Tyr-Leu-Asp-Asn-Tyr and Ser-Pro-Pro-Tyr-Trp-Pro-Tyr lowered the serum uric acid level via intravenous injection at 5 mg/kg of body weight. Furthermore, orally administered Tyr-Leu-Asp-Asn-Tyr showed antihyperuricemic activity. Therefore, these peptides are at least partially responsible for the antihyperuricemic activity of the alcalase digest of shark cartilage.

  12. Identification of a Peptide for Systemic Brain Delivery of a Morpholino Oligonucleotide in Mouse Models of Spinal Muscular Atrophy

    Science.gov (United States)

    Shabanpoor, Fazel; Hammond, Suzan M; Abendroth, Frank; Hazell, Gareth; Wood, Matthew J.A.

    2017-01-01

    Splice-switching antisense oligonucleotides are emerging treatments for neuromuscular diseases, with several splice-switching oligonucleotides (SSOs) currently undergoing clinical trials such as for Duchenne muscular dystrophy (DMD) and spinal muscular atrophy (SMA). However, the development of systemically delivered antisense therapeutics has been hampered by poor tissue penetration and cellular uptake, including crossing of the blood–brain barrier (BBB) to reach targets in the central nervous system (CNS). For SMA application, we have investigated the ability of various BBB-crossing peptides for CNS delivery of a splice-switching phosphorodiamidate morpholino oligonucleotide (PMO) targeting survival motor neuron 2 (SMN2) exon 7 inclusion. We identified a branched derivative of the well-known ApoE (141–150) peptide, which as a PMO conjugate was capable of exon inclusion in the CNS following systemic administration, leading to an increase in the level of full-length SMN2 transcript. Treatment of newborn SMA mice with this peptide-PMO (P-PMO) conjugate resulted in a significant increase in the average lifespan and gains in weight, muscle strength, and righting reflexes. Systemic treatment of adult SMA mice with this newly identified P-PMO also resulted in small but significant increases in the levels of SMN2 pre-messenger RNA (mRNA) exon inclusion in the CNS and peripheral tissues. This work provides proof of principle for the ability to select new peptide paradigms to enhance CNS delivery and activity of a PMO SSO through use of a peptide-based delivery platform for the treatment of SMA potentially extending to other neuromuscular and neurodegenerative diseases. PMID:28118087

  13. Nuclear waste repository siting

    International Nuclear Information System (INIS)

    Soloman, B.D.; Cameron, D.M.

    1987-01-01

    This paper discusses the geopolitics of nuclear waste disposal in the USA. Constitutional choice and social equity perspectives are used to argue for a more open and just repository siting program. The authors assert that every potential repository site inevitably contains geologic, environmental or other imperfections and that the political process is the correct one for determining sites selected

  14. Radioactive waste repository study

    International Nuclear Information System (INIS)

    1978-11-01

    This is the second part of a report of a preliminary study for AECL. It considers the requirements for an underground waste repository for the disposal of wastes produced by the Canadian Nuclear Fuel Program. The following topics are discussed with reference to the repository: 1) geotechnical assessment, 2) hydrogeology and waste containment, 3) thermal loading and 4) rock mechanics. (author)

  15. Detachable strong cation exchange monolith, integrated with capillary zone electrophoresis and coupled with pH gradient elution, produces improved sensitivity and numbers of peptide identifications during bottom-up analysis of complex proteomes.

    Science.gov (United States)

    Zhang, Zhenbin; Yan, Xiaojing; Sun, Liangliang; Zhu, Guijie; Dovichi, Norman J

    2015-04-21

    A detachable sulfonate-silica hybrid strong cation-exchange monolith was synthesized in a fused silica capillary, and used for solid phase extraction with online pH gradient elution during capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) proteomic analysis. Tryptic digests were prepared in 50 mM formic acid and loaded onto the strong cation-exchange monolith. Fractions were eluted using a series of buffers with lower concentration but higher pH values than the 50 mM formic acid background electrolyte. This combination of elution and background electrolytes results in both sample stacking and formation of a dynamic pH junction and allows use of relatively large elution buffer volumes while maintaining reasonable peak efficiency and resolution. A series of five pH bumps were applied to elute E. coli tryptic peptides from the monolith, followed by analysis using CZE coupled to an LTQ-Orbitrap Velos mass spectrometer; 799 protein groups and 3381 peptides were identified from 50 ng of the digest in a 2.5 h analysis, which approaches the identification rate for this organism that was obtained with an Orbitrap Fusion. We attribute the improved numbers of peptide and protein identifications to the efficient fractionation by the online pH gradient elution, which decreased the complexity of the sample in each elution step and improved the signal intensity of low abundance peptides. We also performed a comparative analysis using a nanoACQUITY UltraPerformance LCH system. Similar numbers of protein and peptide identifications were produced by the two methods. Protein identifications showed significant overlap between the two methods, whereas peptide identifications were complementary.

  16. Identification of natural antimicrobial agents to treat dengue infection: In vitro analysis of latarcin peptide activity against dengue virus.

    Science.gov (United States)

    Rothan, Hussin A; Bahrani, Hirbod; Rahman, Noorsaadah Abd; Yusof, Rohana

    2014-05-31

    Although there have been considerable advances in the study of dengue virus, no vaccines or anti-dengue drugs are currently available for humans. Therefore, new approaches are necessary for the development of potent anti-dengue drugs. Natural antimicrobial peptides (AMPs) with potent antiviral activities are potential hits-to-leads for antiviral drug discovery. We performed this study to identify and characterise the inhibitory potential of the latarcin peptide (Ltc 1, SMWSGMWRRKLKKLRNALKKKLKGE) against dengue virus replication in infected cells. The Ltc 1 peptide showed a significantly inhibitory effect against the dengue protease NS2B-NS3pro at 37°C, a physiological human temperature, (IC50, 12.68 ± 3.2 μM), and greater inhibitory effect was observed at 40°C, a temperature similar to a high fever (IC50, 6.58 ± 4.1 μM). A greater reduction in viral load (p.f.u./ml) was observed at simultaneous (0.7 ± 0.3 vs. 7.2 ± 0.5 control) and post-treatment (1.8 ± 0.7 vs. 6.8 ± 0.6 control) compared to the pre-treatment (4.5 ± 0.6 vs. 6.9 ± 0.5 control). Treatment with the Ltc 1 peptide reduced the viral RNA in a dose-dependent manner with EC50 values of 8.3 ± 1.2, 7.6 ± 2.7 and 6.8 ± 2.5 μM at 24, 48 and 72 h, respectively. The Ltc 1 peptide exhibited significant inhibitory effects against dengue NS2B-NS3pro and virus replication in the infected cells. Therefore, further investigation is necessary to develop the Ltc 1 peptide as a new anti-dengue therapeutic.

  17. Identification and characterization of pyrokinin and CAPA peptides, and corresponding GPCRs from spotted wing drosophila, Drosophila suzukii.

    Science.gov (United States)

    Choi, Man-Yeon; Ahn, Seung-Joon; Kim, A Young; Koh, Youngho

    2017-05-15

    The family of FXPRLamide peptides serves as a major insect hormone. It is characterized by a core active amino acid sequence conserved at the C-terminal ends, and provides various physiological roles across the Insecta. In this study we identified and characterized pyrokinin (PK) and CAPA cDNAs encoding two FXPRLamide peptides, pyrokinin and CAPA-DH (diapause hormone), and two corresponding G protein-coupled receptors (GPCRs) from spotted wing drosophila (SWD), Drosophila suzukii. Expressions of PK and CAPA mRNAs were differentially observed during all life stages except the embryo, and the detection of CAPA transcription was relatively strong compared with the PK gene in SWD. Both D. suzukii pyrokinin receptor (DrosuPKr) and CAPA-DH receptor (DrosuCAPA-DHr) were functionally expressed and confirmed through binding to PK and DH peptides. Differential expression of two GPCRs occurred during all life stages; a strong transcription of DrosuPKr was observed in the 3rd instar. DrosuCAPA-DHr was clearly expressed from the embryo to the larva, but not detected in the adult. Gene regulation during the life stages was not synchronized between ligand and receptor. For example, SWD CAPA mRNA has been up-regulated in the adult while CAPA-DHr was down-regulated. The difference could be from the CAPA mRNA translating multiple peptides including CAPA-DH and two CAPA-PVK (periviscerokinin) peptides to act on different receptors. Comparing the genes of SWD PK, CAPA, PKr and CAPA-DHr to four corresponding genes of D. melanogaster, SWD CAPA and the receptor are more similar to D. melanogaster than PK and the receptor. These data suggest that the CAPA gene could be evolutionally more conserved to have a common biological role in insects. In addition, the effect of Kozak sequences was investigated by the expression of the GPCRs with or without Kozak sequences in Sf9 insect cells. The Kozak sequenced PK receptor was significantly less active than the original (= no Kozak sequenced

  18. Rapid identification of Staphylococcus aureus and methicillin resistance by flow cytometry using a peptide nucleic acid probe.

    Science.gov (United States)

    Shrestha, Nabin K; Scalera, Nikole M; Wilson, Deborah A; Brehm-Stecher, Byron; Procop, Gary W

    2011-09-01

    A total of 56 Staphylococcus aureus isolates incubated for 2 h in the presence or absence of oxacillin were analyzed by flow cytometry after labeling with an S. aureus-specific peptide nucleic acid (PNA) probe. Two defined ratios, the paired signal count ratio (PSCR) and the gate signal count ratio (GSCR), differentiated methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) with sensitivities of 100% each and specificities of 96% and 100%, respectively.

  19. Rapid Identification of Staphylococcus aureus and Methicillin Resistance by Flow Cytometry Using a Peptide Nucleic Acid Probe ▿

    Science.gov (United States)

    Shrestha, Nabin K.; Scalera, Nikole M.; Wilson, Deborah A.; Brehm-Stecher, Byron; Procop, Gary W.

    2011-01-01

    A total of 56 Staphylococcus aureus isolates incubated for 2 h in the presence or absence of oxacillin were analyzed by flow cytometry after labeling with an S. aureus-specific peptide nucleic acid (PNA) probe. Two defined ratios, the paired signal count ratio (PSCR) and the gate signal count ratio (GSCR), differentiated methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) with sensitivities of 100% each and specificities of 96% and 100%, respectively. PMID:21795508

  20. Identification of small secreted peptides (SSPs) in maize and expression analysis of partial SSP genes in reproductive tissues.

    Science.gov (United States)

    Li, Ye Long; Dai, Xin Ren; Yue, Xun; Gao, Xin-Qi; Zhang, Xian Sheng

    2014-10-01

    Maize 1,491 small secreted peptides were identified, which were classified according to the character of peptide sequences. Partial SSP gene expressions in reproductive tissues were determined by qRT-PCR. Small secreted peptides (SSPs) are important cell-cell communication messengers in plants. Most information on plant SSPs come from Arabidopsis thaliana and Oryza sativa, while little is known about the SSPs of other grass species such as maize (Zea mays). In this study, we identified 1,491 SSP genes from maize genomic sequences. These putative SSP genes were distributed throughout the ten maize chromosomes. Among them, 611 SSPs were classified into 198 superfamilies according to their conserved domains, and 725 SSPs with four or more cysteines at their C-termini shared similar cysteine arrangements with their counterparts in other plant species. Moreover, the SSPs requiring post-translational modification, as well as defensin-like (DEFL) proteins, were identified. Further, the expression levels of 110 SSP genes were analyzed in reproductive tissues, including male flower, pollen, silk, and ovary. Most of the genes encoding basal-layer antifungal peptide-like, small coat proteins-like, thioredoxin-like proteins, γ-thionins-like, and DEFL proteins showed high expression levels in the ovary and male flower compared with their levels in silk and mature pollen. The rapid alkalinization factor-like genes were highly expressed only in the mature ovary and mature pollen, and pollen Ole e 1-like genes showed low expression in silk. The results of this study provide basic information for further analysis of SSP functions in the reproductive process of maize.

  1. Identification and Characterization of Bioactive Peptides of Fermented Goat Milk as a Sources of Antioxidant as a Therapeutic Natural Product

    Science.gov (United States)

    Mahdi, Chanif; Untari, Handayu; Cendrakasih Padaga, Masdiana

    2018-01-01

    The increasing of functional food is rising in line with public awareness for healthy food consumption. Provision of functional food source is developed through enhanced bioactive that has a regulatory function for body. Bioactive peptides in milk is known have variety of beneficial function of the body such as immunomodulator, immunostimulatory, anti-hypertension, anti-hyper cholesterol, as well as a variety of other beneficial function. The aim of this study is to obtain fermentation methods to product functional dairy product contain bioactive peptides and beneficial of fermented goat milk. The result of this study showed that goat milk fermented using 3 % commercial starter able to produce the best yoghurt than using local yoghurt starter. Analysis of protein content showed that the fermentation processing increased the amount of protein in goat milk sample. Using SDS-PAGE showed that the breakdown of protein into fraction of fermented goat milk greater than unfermented goat milk. The result of fractional protein was analyzed by LC MS/MS and showed that there were three kind bioactive sequences of bioactive peptides. Each of which consist of 16 amino acids that safely protected from gastrointestinal animal model that fed by dietary treatment of hypercholesterolemia.

  2. Identification of the cognate peptide-MHC target of T cell receptors using molecular modeling and force field scoring

    DEFF Research Database (Denmark)

    Lanzarotti, Esteban; Marcatili, Paolo; Nielsen, Morten

    2018-01-01

    Interactions of T cell receptors (TCR) to peptides in complex with MHC (p:MHC) are key features that mediate cellular immune responses. While MHC binding is required for a peptide to be presented to T cells, not all MHC binders are immunogenic. The interaction of a TCR to the p:MHC complex holds...... terms. Building a benchmark of TCR:p:MHC complexes where epitopes and non-epitopes are modelled using state-of-the-art molecular modelling tools, scoring p:MHC to a given TCR using force-fields, optimized in a cross-validation setup to evaluate TCR inter atomic interactions involved with each p:MHC, we...... and model the TCR:p:MHC complex structure. In summary, we conclude that it is possible to identify the TCR cognate target among different candidate peptides by using a force-field based model, and believe this works could lay the foundation for future work within prediction of TCR:p:MHC interactions....

  3. Identification of a Bacteria Using Phylogenetic Relationships Revealed by MS/MS Sequencing of Tryptic Peptides Derived From Cellular Proteins

    National Research Council Canada - National Science Library

    Dworzanski, J. P; Wick, H; Snyder, A. P; Deshpande, S. V; Chen, R; Li, L

    2004-01-01

    .... In addition four hundred bacterial genome-sequencing projects are in progress. These achievements provide new possibilities for reliable identification of bacteria on a molecular level by retrieving their genomic information...

  4. Gas generation in repositories

    International Nuclear Information System (INIS)

    Biddle, P.; Rees, J.H.; McGahan, D.; Rushbrook, P.E.

    1987-09-01

    The nature and quantities of gases likely to be produced by various processes in repositories for low level and intermediate level radioactive wastes are examined in this preliminary study. Many simplifying assumptions are made where published or experimental data is unavailable. The corrosion of the canisters and metallic components in wastes is likely to be the major gas production process in both types of repository. A significant contribution from microbiological activity is expected to occur in low level repositories, predominantly where no cement grouting of the cans has been carried out. A number of areas for further research, required before a more comprehensive study could be carried out, have been identified. (author)

  5. Centralized mouse repositories.

    Science.gov (United States)

    Donahue, Leah Rae; Hrabe de Angelis, Martin; Hagn, Michael; Franklin, Craig; Lloyd, K C Kent; Magnuson, Terry; McKerlie, Colin; Nakagata, Naomi; Obata, Yuichi; Read, Stuart; Wurst, Wolfgang; Hörlein, Andreas; Davisson, Muriel T

    2012-10-01

    Because the mouse is used so widely for biomedical research and the number of mouse models being generated is increasing rapidly, centralized repositories are essential if the valuable mouse strains and models that have been developed are to be securely preserved and fully exploited. Ensuring the ongoing availability of these mouse strains preserves the investment made in creating and characterizing them and creates a global resource of enormous value. The establishment of centralized mouse repositories around the world for distributing and archiving these resources has provided critical access to and preservation of these strains. This article describes the common and specialized activities provided by major mouse repositories around the world.

  6. Stepwise identification of HLA-A*0201-restricted CD8+ T-cell epitope peptides from herpes simplex virus type 1 genome boosted by a StepRank scheme.

    Science.gov (United States)

    Bi, Jianjun; Song, Rengang; Yang, Huilan; Li, Bingling; Fan, Jianyong; Liu, Zhongrong; Long, Chaoqin

    2011-01-01

    Identification of immunodominant epitopes is the first step in the rational design of peptide vaccines aimed at T-cell immunity. To date, however, it is yet a great challenge for accurately predicting the potent epitope peptides from a pool of large-scale candidates with an efficient manner. In this study, a method that we named StepRank has been developed for the reliable and rapid prediction of binding capabilities/affinities between proteins and genome-wide peptides. In this procedure, instead of single strategy used in most traditional epitope identification algorithms, four steps with different purposes and thus different computational demands are employed in turn to screen the large-scale peptide candidates that are normally generated from, for example, pathogenic genome. The steps 1 and 2 aim at qualitative exclusion of typical nonbinders by using empirical rule and linear statistical approach, while the steps 3 and 4 focus on quantitative examination and prediction of the interaction energy profile and binding affinity of peptide to target protein via quantitative structure-activity relationship (QSAR) and structure-based free energy analysis. We exemplify this method through its application to binding predictions of the peptide segments derived from the 76 known open-reading frames (ORFs) of herpes simplex virus type 1 (HSV-1) genome with or without affinity to human major histocompatibility complex class I (MHC I) molecule HLA-A*0201, and find that the predictive results are well compatible with the classical anchor residue theory and perfectly match for the extended motif pattern of MHC I-binding peptides. The putative epitopes are further confirmed by comparisons with 11 experimentally measured HLA-A*0201-restrcited peptides from the HSV-1 glycoproteins D and K. We expect that this well-designed scheme can be applied in the computational screening of other viral genomes as well.

  7. Identification, characterization, and synthesis of peptide epitopes and a recombinant six-epitope protein for Trichomonas vaginalis serodiagnosis

    Directory of Open Access Journals (Sweden)

    Alderete JF

    2013-08-01

    Full Text Available J F Alderete, Calvin J NeaceSchool of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, WA, USAAbstract: There is a need for a rapid, accurate serodiagnostic test useful for both women and men infected by Trichomonas vaginalis, which causes the number one sexually transmitted infection (STI. Women and men exposed to T. vaginalis make serum antibody to fructose-1,6-bisphosphate aldolase (ALD, α-enolase (ENO, and glyceraldehyde-3-phosphate dehydrogenase (GAP. We identified, by epitope mapping, the common and distinct epitopes of each protein detected by the sera of women patients with trichomonosis and by the sera of men highly seropositive to the immunogenic protein α-actinin (positive control sera. We analyzed the amino acid sequences to determine the extent of identity of the epitopes of each protein with other proteins in the databanks. This approach identified epitopes unique to T. vaginalis, indicating these peptide-epitopes as possible targets for a serodiagnostic test. Individual or combinations of 15-mer peptide epitopes with low to no identity with other proteins were reactive with positive control sera from both women and men but were unreactive with negative control sera. These analyses permitted the synthesis of a recombinant His6 fusion protein of 111 amino acids with an Mr of ~13.4 kDa, which consisted of 15-mer peptides of two distinct epitopes each for ALD, ENO, and GAP. This recombinant protein was purified by affinity chromatography. This composite protein was detected by enzyme-linked immunosorbent assay (ELISA, dot blots, and immunoblots, using positive control sera from women and men. These data indicate that it is possible to identify epitopes and that either singly, in combination, or as a composite protein represent targets for a point-of-care serodiagnostic test for T. vaginalis.Keywords: diagnostics, point-of-care, targets, trichomonosis

  8. Identification of Poly(ethylene glycol) and Poly(ethylene glycol)-Based Detergents Using Peptide Search Engines.

    Science.gov (United States)

    Ahmadi, Shiva; Winter, Dominic

    2018-06-05

    Poly(ethylene glycol) (PEG) is one of the most common polymer contaminations in mass spectrometry (MS) samples. At present, the detection of PEG and other polymers relies largely on manual inspection of raw data, which is laborious and frequently difficult due to sample complexity and retention characteristics of polymer species in reversed-phase chromatography. We developed a new strategy for the automated identification of PEG molecules from tandem mass spectrometry (MS/MS) data using protein identification algorithms in combination with a database containing "PEG-proteins". Through definition of variable modifications, we extend the approach for the identification of commonly used PEG-based detergents. We exemplify the identification of different types of polymers by static nanoelectrospray tandem mass spectrometry (nanoESI-MS/MS) analysis of pure detergent solutions and data analysis using Mascot. Analysis of liquid chromatography-tandem mass spectrometry (LC-MS/MS) runs of a PEG-contaminated sample by Mascot identified 806 PEG spectra originating from four PEG species using a defined set of modifications covering PEG and common PEG-based detergents. Further characterization of the sample for unidentified PEG species using error-tolerant and mass-tolerant searches resulted in identification of 3409 and 3187 PEG-related MS/MS spectra, respectively. We further demonstrate the applicability of the strategy for Protein Pilot and MaxQuant.

  9. National Radwaste Repository Mochovce

    International Nuclear Information System (INIS)

    2000-01-01

    In this leaflet the National Radioactive Waste Repository in Mochovce (Repository) is described. The Mochovce National Radioactive Waste Repository is a surface multi-barrier type storage facility for solid and treated solidified radioactive wastes generated from the Slovak Republic nuclear power plants operation and decommissioning, research institutes, laboratories and hospitals. The Repository comprises a system of single- and double-row storage boxes. The first double-row is enclosed by a steel-structure building. The 18 x 6 x 5.5 m storage boxes are made of reinforced concrete. The wall thickness is 600 mm. Two-double-rows, i.e. 80 storage boxes were built as part of Stage I (1 row = 20 storage boxes). Each storage box has a storage capacity of 90 fibre concrete containers of 3.1 m 3 volume. The total storage capacity is 7200 containers with the overall storage volume of 22320 m 3

  10. NIA Aging Cell Repository

    Data.gov (United States)

    Federal Laboratory Consortium — To facilitate aging research on cells in culture, the NIA provides support for the NIA Aging Cell Repository, located at the Coriell Institute for Medical Research...

  11. NIDDK Central Repository

    Data.gov (United States)

    U.S. Department of Health & Human Services — The NIDDK Central Repository stores biosamples, genetic and other data collected in designated NIDDK-funded clinical studies. The purpose of the NIDDK Central...

  12. Managing and Evaluating Digital Repositories

    Science.gov (United States)

    Zuccala, Alesia; Oppenheim, Charles; Dhiensa, Rajveen

    2008-01-01

    Introduction: We examine the role of the digital repository manager, discuss the future of repository management and evaluation and suggest that library and information science schools develop new repository management curricula. Method: Face-to-face interviews were carried out with managers of five different types of repositories and a Web-based…

  13. Radioactive waste repository study

    International Nuclear Information System (INIS)

    1978-11-01

    This is the first part of a report of a preliminary study for Atomic Energy of Canada Limited. It considers the requirements for an underground waste repository for the disposal of wastes produced by the Canadian Nuclear Fuel Program. The following topics are discussed with reference to the repository: 1) underground layout, 2) cost estimates, 3) waste handling, 4) retrievability, decommissioning, sealing and monitoring, and 5) research and design engineering requirements. (author)

  14. MAJOR REPOSITORY DESIGN ISSUES

    International Nuclear Information System (INIS)

    JACK N. BAILEY, DWAYNE CHESTNUT, JAMES COMPTON AND RICHARD D. SNELL

    1997-01-01

    The Yucca Mountain Project is focused on producing a four-part viability assessment in late FY98. Its four components (design, performance assessment, cost estimate, and licensing development plan) must be consistent. As a tool to compare design and performance assessment options, a series of repository pictures were developed for the sequential time phases of a repository. The boundaries of the time phases correspond to evolution in the engineered barrier system (EBS)

  15. Repository simulation tests

    International Nuclear Information System (INIS)

    Wicks, G.G.; Bibler, N.E.; Jantzen, C.M.; Plodinec, M.J.

    1984-01-01

    The repository simulation experiments described in this paper are designed to assess the performance of SRP waste glass under the most realistic repository conditions that can be obtained in the laboratory. These tests simulate the repository environment as closely as possible and introduce systematically the variability of the geology, groundwater chemistry, and waste package components during the leaching of the waste glass. The tests evaluate waste form performance under site-specific conditions, which differ for each of the geologic repositories under consideration. Data from these experiments will aid in the development of a realistic source term that can describe the release of radionuclides from SRP waste glass as a component of proposed waste packages. Hence, this information can be useful to optimize waste package design for SRP waste glass and to provide data for predicting long-term performance and subsequent conformance to regulations. The repository simulation tests also help to bridge the gap in interpreting results derived from tests performed under the control of the laboratory to the uncertainity and variability of field tests. In these experiments, site-specific repository components and conditions are emphasized and only the site specific materials contact the waste forms. An important feature of these tests is that both actual and simulated waste glasses are tested identically. 7 figures, 2 tables

  16. Repository operational criteria analysis

    International Nuclear Information System (INIS)

    Hageman, J.P.; Chowdhury, A.H.

    1992-08-01

    The objective of the ''Repository Operational Criteria (ROC) Feasibility Studies'' (or ROC task) was to conduct comprehensive and integrated analyses of repository design, construction, and operations criteria in 10 CFR Part 60 regulations, considering the interfaces and impacts of any potential changes to those regulations. The study addresses regulatory criteria related to the preclosure aspects of the geologic repository. The study task developed regulatory concepts or potential repository operational criteria (PROC) based on analysis of a repository's safety functions and other regulations for similar facilities. These regulatory concepts or PROC were used as a basis to assess the sufficiency and adequacy of the current criteria in 10 CFR Part 60. Where the regulatory concepts were same as current operational criteria, these criteria were referenced. The operations criteria referenced or the PROC developed are given in this report. Detailed analyses used to develop the regulatory concepts and any necessary PROC for those regulations that may require a minor change are also presented. The results of the ROC task showed a need for further analysis and possible major rule change related to the design bases of a geologic repository operations area, siting, and radiological emergency planning

  17. The Equine PeptideAtlas

    DEFF Research Database (Denmark)

    Bundgaard, Louise; Jacobsen, Stine; Sørensen, Mette Aamand

    2014-01-01

    Progress in MS-based methods for veterinary research and diagnostics is lagging behind compared to the human research, and proteome data of domestic animals is still not well represented in open source data repositories. This is particularly true for the equine species. Here we present a first...... Equine PeptideAtlas encompassing high-resolution tandem MS analyses of 51 samples representing a selection of equine tissues and body fluids from healthy and diseased animals. The raw data were processed through the Trans-Proteomic Pipeline to yield high quality identification of proteins and peptides....... The current release comprises 24 131 distinct peptides representing 2636 canonical proteins observed at false discovery rates of 0.2% at the peptide level and 1.4% at the protein level. Data from the Equine PeptideAtlas are available for experimental planning, validation of new datasets, and as a proteomic...

  18. Identification and characterization of Cd-induced peptides in Egeria densa (water weed): Putative role in Cd detoxification

    International Nuclear Information System (INIS)

    Malec, Przemyslaw; Maleva, Maria G.; Prasad, M.N.V.; Strzalka, Kazimierz

    2009-01-01

    Egeria densa has ability to grow in heavy metal contaminated and polluted bodies of water. Shoots exposed to Cd at concentrations up to 300 μM for 7 days showed a pronounced decrease in chlorophyll a and in total protein concentration. Thiol-containing compounds and low-molecular-weight polypeptides were detected in Cd-treated plant extracts by gel filtration chromatography. Two Cd-binding fractions, a thiol-enriched fraction and a non-thiol fraction with a lower molecular weight were identified in extracts by gel filtration. The main fraction of thiol-containing polypeptide, purified by gel filtration and anion-exchange chromatography had a molecular weight of ∼10 kDa. This peptide was characterized by a broad absorption band specific to mercaptide bonds and Cd-sensitive fluorescence emission of aromatic amino acid residues. Our results indicate that cadmium exposure of plants resulted in both a formation of thiol-enriched cadmium complexing peptides and a synthesis of low-molecular-weight metal chelators. The putative role of these compounds in Cd detoxification is discussed.

  19. Identification of avocado (Persea americana) pulp proteins by nano-LC-MS/MS via combinatorial peptide ligand libraries.

    Science.gov (United States)

    Esteve, Clara; D'Amato, Alfonsina; Marina, María Luisa; García, María Concepción; Righetti, Pier Giorgio

    2012-09-01

    Avocado (Persea americana) proteins have been scarcely studied despite their importance, especially in food related allergies. The proteome of avocado pulp was explored in depth by extracting proteins with capture by combinatorial peptide ligand libraries at pH 7.4 and under conditions mimicking reverse-phase capture at pH 2.2. The total number of unique gene products identified amounts to 1012 proteins, of which 174 are in common with the control, untreated sample, 190 are present only in the control and 648 represent the new species detected via combinatorial peptide ligand libraries of all combined eluates and likely represent low-abundance proteins. Among the 1012 proteins, it was possible to identify the already known avocado allergen Pers a 1 and different proteins susceptible to be allergens such as a profilin, a polygalacturonase, a thaumatin-like protein, a glucanase, and an isoflavone reductase like protein. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Accurate pan-specific prediction of peptide-MHC class II binding affinity with improved binding core identification

    DEFF Research Database (Denmark)

    Andreatta, Massimo; Karosiene, Edita; Rasmussen, Michael

    2015-01-01

    with known binding registers, the new method NetMHCIIpan-3.1 significantly outperformed the earlier 3.0 version. We illustrate the impact of accurate binding core identification for the interpretation of T cell cross-reactivity using tetramer double staining with a CMV epitope and its variants mapped...

  1. Direct identification of on-bead peptides using surface-enhanced Raman spectroscopic barcoding system for high-throughput bioanalysis.

    Science.gov (United States)

    Kang, Homan; Jeong, Sinyoung; Koh, Yul; Geun Cha, Myeong; Yang, Jin-Kyoung; Kyeong, San; Kim, Jaehi; Kwak, Seon-Yeong; Chang, Hye-Jin; Lee, Hyunmi; Jeong, Cheolhwan; Kim, Jong-Ho; Jun, Bong-Hyun; Kim, Yong-Kweon; Hong Jeong, Dae; Lee, Yoon-Sik

    2015-05-28

    Recently, preparation and screening of compound libraries remain one of the most challenging tasks in drug discovery, biomarker detection, and biomolecular profiling processes. So far, several distinct encoding/decoding methods such as chemical encoding, graphical encoding, and optical encoding have been reported to identify those libraries. In this paper, a simple and efficient surface-enhanced Raman spectroscopic (SERS) barcoding method using highly sensitive SERS nanoparticles (SERS ID) is presented. The 44 kinds of SERS IDs were able to generate simple codes and could possibly generate more than one million kinds of codes by incorporating combinations of different SERS IDs. The barcoding method exhibited high stability and reliability under bioassay conditions. The SERS ID encoding based screening platform can identify the peptide ligand on the bead and also quantify its binding affinity for specific protein. We believe that our SERS barcoding technology is a promising method in the screening of one-bead-one-compound (OBOC) libraries for drug discovery.

  2. Ghrelin receptor inverse agonists: identification of an active peptide core and its interaction epitopes on the receptor

    DEFF Research Database (Denmark)

    Holst, Birgitte; Lang, Manja; Brandt, Erik

    2006-01-01

    [D-Arg1,D-Phe5,D-Trp7,9,Leu11]Substance P functions as a low-potency antagonist but a high-potency full inverse agonist on the ghrelin receptor. Through a systematic deletion and substitution analysis of this peptide, the C-terminal carboxyamidated pentapeptide wFwLX was identified as the core...... structure, which itself displayed relatively low inverse agonist potency. Mutational analysis at 17 selected positions in the main ligand-binding crevice of the ghrelin receptor demonstrated that ghrelin apparently interacts only with residues in the middle part of the pocket [i.e., between transmembrane...... upon both AspII:20 and GluIII:09. The identified pharmacophore can possibly serve as the basis for targeted discovery of also nonpeptide inverse agonists for the ghrelin receptor....

  3. Antiplatelet Aggregation and Antithrombosis Efficiency of Peptides in the Snake Venom of Deinagkistrodon acutus: Isolation, Identification, and Evaluation

    Directory of Open Access Journals (Sweden)

    Bin Ding

    2015-01-01

    Full Text Available Two peptides of Pt-A (Glu-Asn-Trp 429 Da and Pt-B (Glu-Gln-Trp 443 Da were isolated from venom liquor of Deinagkistrodon acutus. Their antiplatelet aggregation effects were evaluated with platelet-rich human plasma in vitro; the respective IC50 of Pt-A and Pt-B was 66 μM and 203 μM. Both peptides exhibited protection effects on ADP-induced paralysis in mice. After ADP administration, the paralysis time of different concentration of Pt-A and Pt-B lasted as the following: 80 mg/kg Pt-B (152.8 ± 57.8 s < 40 mg/kg Pt-A (163.5 ± 59.8 s < 20 mg/kg Pt-A (253.5 ± 74.5 s < 4 mg/kg clopidogrel (a positive control, 254.5 ± 41.97 s < 40 mg/kg Pt-B (400.8 ± 35.9 s < 10 mg/kg Pt-A (422.8 ± 55.4 s, all of which were statistically shorter than the saline treatment (666 ± 28 s. Pulmonary tissue biopsy confirmed that Pt-A and Pt-B prevented the formation of thrombi in the lung. Unlike ADP injection alone, which caused significant reduction of peripheral platelet count, Pt-A treatment prevented the drop of peripheral platelet counts; interestingly, Pt-B could not, even though the same amount of Pt-B also showed protection effects on ADP-induced paralysis and thrombosis. More importantly, intravenous injection of Pt-A and Pt-B did not significantly increase the hemorrhage risks as clopidogrel.

  4. Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures.

    Science.gov (United States)

    Lamer, S; Jungblut, P R

    2001-03-10

    In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.

  5. Trust in Digital Repositories

    Directory of Open Access Journals (Sweden)

    Elizabeth Yakel

    2013-06-01

    Full Text Available ISO 16363:2012, Space Data and Information Transfer Systems - Audit and Certification of Trustworthy Digital Repositories (ISO TRAC, outlines actions a repository can take to be considered trustworthy, but research examining whether the repository’s designated community of users associates such actions with trustworthiness has been limited. Drawing from this ISO document and the management and information systems literatures, this paper discusses findings from interviews with 66 archaeologists and quantitative social scientists. We found similarities and differences across the disciplines and among the social scientists. Both disciplinary communities associated trust with a repository’s transparency. However, archaeologists mentioned guarantees of preservation and sustainability more frequently than the social scientists, who talked about institutional reputation. Repository processes were also linked to trust, with archaeologists more frequently citing metadata issues and social scientists discussing data selection and cleaning processes. Among the social scientists, novices mentioned the influence of colleagues on their trust in repositories almost twice as much as the experts. We discuss the implications our findings have for identifying trustworthy repositories and how they extend the models presented in the management and information systems literatures.

  6. Sellafield repository design concept

    International Nuclear Information System (INIS)

    1998-01-01

    Between 1989 and 1997, UK Nirex Ltd carried out a programme of investigations to evaluate the potential of a site adjacent to the BNFL Sellafield works to host a deep repository for the United Kingdom's intermediate-level and certain low-level radioactive waste. The programme of investigations was wound down following the decision in March 1997 to uphold the rejection of the Company's planning application for the Rock Characterisation Facility (RCF), an underground laboratory which would have allowed further investigations to confirm whether or not the site would be suitable. Since that time, the Company's efforts in relation to the Sellafield site have been directed towards documenting and publishing the work carried out. The design concept for a repository at Sellafield was developed in parallel with the site investigations through an iterative process as knowledge of the site and understanding of the repository system performance increased. This report documents the Sellafield repository design concept as it had been developed, from initial design considerations in 1991 up to the point when the RCF planning application was rejected. It shows, from the context of a project at that particular site, how much information and experience has been gained that will be applicable to the development of a deep waste repository at other potential sites

  7. Socioeconomic impacts of repositories

    International Nuclear Information System (INIS)

    Thomas, J.K.; Hamm, R.R.; Murdock, S.H.

    1983-01-01

    Federal and state decision makers, community leaders, and residents must know how communities will be changed by the impacts of a high-level nuclear waste repository. This chapter identifies the factors affecting an assessment of socioeconomic impacts and the types of impacts (economic, demographic, fiscal, community service, and social) likely to occur as a result of repository development. Each of these types can be divided into standard (those which typically results from any large-scale development) and special impact categories (those which result from the fact that radioactive materials will be handled). 3 tables

  8. Safeguards for geological repositories

    International Nuclear Information System (INIS)

    Fattah, A.

    2000-01-01

    Direct disposal of spent nuclear fuel in geological repositories is a recognised option for closing nuclear fuel cycles. Geological repositories are at present in stages of development in a number of countries and are expected to be built and operated early next century. A State usually has an obligation to safely store any nuclear material, which is considered unsuitable to re-enter the nuclear fuel cycle, isolated from the biosphere. In conjunction with this, physical protection has to be accounted for to prevent inadvertent access to such material. In addition to these two criteria - which are fully under the State's jurisdiction - a third criterion reflecting international non-proliferation commitments needs to be addressed. Under comprehensive safeguards agreements a State concedes verification of nuclear material for safeguards purposes to the IAEA. The Agency can thus provide assurance to the international community that such nuclear material has been used for peaceful purposes only as declared by the State. It must be emphasised that all three criteria mentioned constitute a 'unit'. None can be sacrificed for the sake of the other, but compromises may have to be sought in order to make their combination as effective as possible. Based on comprehensive safeguards agreements signed and ratified by the State, safeguards can be terminated only when the material has been consumed or diluted in such a way that it can no longer be utilised for any nuclear activities or has become practicably irrecoverable. As such safeguards for nuclear material in geological repositories have to be continued even after the repository has been back-filled and sealed. The effective application of safeguards must assure continuity-of-knowledge that the nuclear material in the repository has not been diverted for an unknown purpose. The nuclear material disposed in a geological repository may eventually have a higher and long term proliferation risk because the inventory is

  9. Magnetic microparticle-based SELEX process for the identification of highly specific aptamers of heart marker--brain natriuretic peptide

    International Nuclear Information System (INIS)

    Wang, Ying; Cao, Jinxuan; Wu, Jingjing; Xue, Feng; Teng, Jun; Chen, Wei; Chen, Yinji; Lu, Chunxia

    2015-01-01

    The brain natriuretic peptide (BNP) is known to be an effective indicator of heart failure. It has been widely adopted as a parameter for the evaluation of heart function of cardiovascular and cerebrovascular diseases (CVDs). Current immune-recognition based methods for the detection of BNP are limited, to a certain extent, by the poor stability of the antibody and by high costs. The availability of an aptamer specific for BNP would greatly assist in the rapid and early diagnosis of CVDs. In order to screen for such an aptamer by the SELEX method, we have used magnetic microparticles (m-MPs) as the separation substrate for immobilization of target BNP. The use of m-MPs for rapid separation of combined aptamers enables bound oligonucleotides to be separated directly, quickly, and with high efficiency. After 14 rounds of selection, a panel of six aptamers against BNP was identified. Their dissociation constants range from 12.5 to 139 nM. The classical technique for conjugation of a target to m-MPs is known to be applicable to various fields, and we conclude that this m-MP-based SELEX process provides a general strategy for screening of specific aptamers against various analytes. (author)

  10. Identification of a boron nitride nanosphere-binding peptide for the intracellular delivery of CpG oligodeoxynucleotides

    Science.gov (United States)

    Zhang, Huijie; Yamazaki, Tomohiko; Zhi, Chunyi; Hanagata, Nobutaka

    2012-09-01

    CpG oligonucleotides (CpG ODNs) interact with Toll-like receptor 9 (TLR9), which results in the induction of immunostimulatory cytokines. We delivered CpG ODNs intracellularly using boron nitride nanospheres (BNNS). To enhance the loading capacity of CpG ODNs on BNNS, we used a phage display technique to identify a 12-amino acid peptide designated as BP7, with specific affinity for BNNS, and used it as a linker to load CpG ODNs on BNNS. The tyrosine residue (Y) at the eighth position from the N-terminus played a crucial role in the affinity of BP7 to BNNS. BNNS that bound BP7 (BNNS-BP7) were taken up by cells and showed no cytotoxicity, and CpG ODNs were successfully crosslinked with BP7 to create BP7-CpG ODN conjugates. Using BP7 as a linker, the loading efficiency of CpG ODNs on BNNS increased 5-fold compared to the direct binding of CpG ODNs to BNNS. Furthermore, the BP7-CpG ODN conjugate-loaded BNNS had a greater capacity to induce interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) production from peripheral blood mononuclear cells (PBMCs) than that of CpG ODNs directly loaded on BNNS. The higher amount of cytokine induction by BP7-CpG ODN conjugate-loaded BNNS may be attributed to a higher loading capacity and stronger binding to BNNS of the linker BP7. The greater functionality of BP7-conjugated CpG ODNs on BNNS expands the potential of BNNS for drug delivery applications.CpG oligonucleotides (CpG ODNs) interact with Toll-like receptor 9 (TLR9), which results in the induction of immunostimulatory cytokines. We delivered CpG ODNs intracellularly using boron nitride nanospheres (BNNS). To enhance the loading capacity of CpG ODNs on BNNS, we used a phage display technique to identify a 12-amino acid peptide designated as BP7, with specific affinity for BNNS, and used it as a linker to load CpG ODNs on BNNS. The tyrosine residue (Y) at the eighth position from the N-terminus played a crucial role in the affinity of BP7 to BNNS. BNNS that bound BP7

  11. Process model repositories and PNML

    NARCIS (Netherlands)

    Hee, van K.M.; Post, R.D.J.; Somers, L.J.A.M.; Werf, van der J.M.E.M.; Kindler, E.

    2004-01-01

    Bringing system and process models together in repositories facilitates the interchange of model information between modelling tools, and allows the combination and interlinking of complementary models. Petriweb is a web application for managing such repositories. It supports hierarchical process

  12. Low level waste repositories

    International Nuclear Information System (INIS)

    Hill, P.R.H.; Wilson, M.A.

    1983-11-01

    Factors in selecting a site for low-level radioactive waste disposal are discussed. South Australia has used a former tailings dam in a remote, arid location as a llw repository. There are also low-level waste disposal procedures at the Olympic Dam copper/uranium project

  13. CRIS and Institutional Repositories

    Directory of Open Access Journals (Sweden)

    A Asserson

    2010-04-01

    Full Text Available CRIS (Current Research Information Systems provide researchers, research managers, innovators, and others with a view over the research activity of a domain. IRs (institutional repositories provide a mechanism for an organisation to showcase through OA (open access its intellectual property. Increasingly, organizations are mandating that their employed researchers deposit peer-reviewed published material in the IR. Research funders are increasingly mandating that publications be deposited in an open access repository: some mandate a central (or subject-based repository, some an IR. In parallel, publishers are offering OA but replacing subscription-based access with author (or author institution payment for publishing. However, many OA repositories have metadata based on DC (Dublin Core which is inadequate; a CERIF (Common-European Research Information Format CRIS provides metadata describing publications with formal syntax and declared semantics thus facilitating interoperation or homogeneous access over heterogeneous sources. The formality is essential for research output metrics, which are increasingly being used to determine future funding for research organizations.

  14. Salt repository design approach

    International Nuclear Information System (INIS)

    Matthews, S.C.

    1983-01-01

    This paper presents a summary discussion of the approaches that have been and will be taken in design of repository facilities for use with disposal of radioactive wastes in salt formations. Since specific sites have yet to be identified, the discussion is at a general level, supplemented with illustrative examples where appropriate. 5 references, 1 figure

  15. Repository site characterization

    International Nuclear Information System (INIS)

    Voss, J.W.; Pentz, D.L.

    1987-01-01

    The characterization of candidate repository sites has a number of programmatic objectives. Principal among these is the acquisition of data: a) to determine the suitability of a site relative to the DOE repository siting guidelines, b) to support model development and calculations to determine the suitability of a site relative to the post closure criteria of the NRC and EPA, c) to support the design of a disposal system, including the waste package and the engineered barrier system, as well as the shafts and underground openings of the repository. In meeting the gaols of site characterization, the authors have an obligation to conduct their investigations within an appropriate budget and schedule. This mandates that a well-constructed and systematic plan for field investigations be developed. Such a plan must fully account for the mechanisms which will control the radiologic performance in the repository. The plan must also flexibly and dynamically respond to the results of each step of field investigation, responding to the spatial variability of earth as well as to enhanced understandings of the performance of the disposal system. Such a plan must ensure that sufficient data are available to support the necessary probabilistic calculations of performance. This paper explores the planning for field data acquisition with specific reference to requirements for demonstrations of the acceptable performance for disposal systems

  16. Radioactive waste repository study

    International Nuclear Information System (INIS)

    1978-11-01

    This is the third part of a report of a preliminary study for AECL. It summarizes the topics considered in reports AECL-6188-1 and AECL-6188-2 as requirements for an undergpound repository for disposal of wastes produced by the Canadian Nuclear Fuel Program. (author)

  17. Computational Materials Repository

    DEFF Research Database (Denmark)

    Landis, David

    , different abstraction levels and enables users to analyze their own results, and allows to share data with collaborators. The approach of the Computational Materials Repository (CMR) is to convert data to an internal format that maintains the original variable names without insisting on any semantics...

  18. The Computational Materials Repository

    DEFF Research Database (Denmark)

    Landis, David D.; Hummelshøj, Jens S.; Nestorov, Svetlozar

    2012-01-01

    The possibilities for designing new materials based on quantum physics calculations are rapidly growing, but these design efforts lead to a significant increase in the amount of computational data created. The Computational Materials Repository (CMR) addresses this data challenge and provides...

  19. Peptide Nucleic Acid Based Molecular Authentication for Identification of Four Medicinal Paeonia Species Using Melting Array Analysis of the Internal Transcribed Spacer 2 Region

    Directory of Open Access Journals (Sweden)

    Wook Jin Kim

    2017-11-01

    Full Text Available Accurate taxonomic identification of plant materials in herbal medicines is important for product quality control. The genus Paeonia (Saxifragales is the source of the herbal preparations Paeoniae Radix (Paeoniae Radix Alba and Paeoniae Radix Rubra and Moutan Radicis Cotex. However, confusion has arisen regarding their contents due to linguistic and taxonomic ambiguities, similar morphologies and different definitions of Paeoniae Radix in the Korean and Chinese national pharmacopoeias, leading to the distribution of adulterated products. To develop a method for identifying the four Paeonia species used in these medicines, three fluorescently-labeled peptide nucleic acid (PNA probes were designed against ITS2 sequences containing single nucleotide polymorphisms (SNPs and used in a real-time PCR melting curve assay. Each of the four Paeonia species was accurately identified using this analysis. The accuracy and analytical stability of the PNA melting curve assay was confirmed using commercially available samples of the four Paeonia species. This assay is a reliable genetic tool to distinguish between different Paeonia-derived herbal medicines and identify the botanical origins of Paeoniae Radix and Moutan Radicis Cortex. This technique may also contribute to quality control and standardization of herbal medicines by providing a reliable authentication tool and preventing the distribution of inauthentic adulterants.

  20. Peptide Nucleic Acid Based Molecular Authentication for Identification of Four Medicinal Paeonia Species Using Melting Array Analysis of the Internal Transcribed Spacer 2 Region.

    Science.gov (United States)

    Kim, Wook Jin; Yang, Sungyu; Choi, Goya; Moon, Byeong Cheol

    2017-11-07

    Accurate taxonomic identification of plant materials in herbal medicines is important for product quality control. The genus Paeonia (Saxifragales) is the source of the herbal preparations Paeoniae Radix (Paeoniae Radix Alba and Paeoniae Radix Rubra) and Moutan Radicis Cotex. However, confusion has arisen regarding their contents due to linguistic and taxonomic ambiguities, similar morphologies and different definitions of Paeoniae Radix in the Korean and Chinese national pharmacopoeias, leading to the distribution of adulterated products. To develop a method for identifying the four Paeonia species used in these medicines, three fluorescently-labeled peptide nucleic acid (PNA) probes were designed against ITS2 sequences containing single nucleotide polymorphisms (SNPs) and used in a real-time PCR melting curve assay. Each of the four Paeonia species was accurately identified using this analysis. The accuracy and analytical stability of the PNA melting curve assay was confirmed using commercially available samples of the four Paeonia species. This assay is a reliable genetic tool to distinguish between different Paeonia -derived herbal medicines and identify the botanical origins of Paeoniae Radix and Moutan Radicis Cortex. This technique may also contribute to quality control and standardization of herbal medicines by providing a reliable authentication tool and preventing the distribution of inauthentic adulterants.

  1. jmzIdentML API: A Java interface to the mzIdentML standard for peptide and protein identification data.

    Science.gov (United States)

    Reisinger, Florian; Krishna, Ritesh; Ghali, Fawaz; Ríos, Daniel; Hermjakob, Henning; Vizcaíno, Juan Antonio; Jones, Andrew R

    2012-03-01

    We present a Java application programming interface (API), jmzIdentML, for the Human Proteome Organisation (HUPO) Proteomics Standards Initiative (PSI) mzIdentML standard for peptide and protein identification data. The API combines the power of Java Architecture of XML Binding (JAXB) and an XPath-based random-access indexer to allow a fast and efficient mapping of extensible markup language (XML) elements to Java objects. The internal references in the mzIdentML files are resolved in an on-demand manner, where the whole file is accessed as a random-access swap file, and only the relevant piece of XMLis selected for mapping to its corresponding Java object. The APIis highly efficient in its memory usage and can handle files of arbitrary sizes. The APIfollows the official release of the mzIdentML (version 1.1) specifications and is available in the public domain under a permissive licence at http://www.code.google.com/p/jmzidentml/. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Identification of putative agouti-related protein(87-132)-melanocortin-4 receptor interactions by homology molecular modeling and validation using chimeric peptide ligands.

    Science.gov (United States)

    Wilczynski, Andrzej; Wang, Xiang S; Joseph, Christine G; Xiang, Zhimin; Bauzo, Rayna M; Scott, Joseph W; Sorensen, Nicholas B; Shaw, Amanda M; Millard, William J; Richards, Nigel G; Haskell-Luevano, Carrie

    2004-04-22

    Agouti-related protein (AGRP) is one of only two naturally known antagonists of G-protein-coupled receptors (GPCRs) identified to date. Specifically, AGRP antagonizes the brain melanocortin-3 and -4 receptors involved in energy homeostasis. Alpha-melanocyte stimulating hormone (alpha-MSH) is one of the known endogenous agonists for these melanocortin receptors. Insight into putative interactions between the antagonist AGRP amino acids with the melanocortin-4 receptor (MC4R) may be important for the design of unique ligands for the treatment of obesity related diseases and is currently lacking in the literature. A three-dimensional homology molecular model of the mouse MC4 receptor complex with the hAGRP(87-132) ligand docked into the receptor has been developed to identify putative antagonist ligand-receptor interactions. Key putative AGRP-MC4R interactions include the Arg111 of hAGRP(87-132) interacting in a negatively charged pocket located in a cavity formed by transmembrane spanning (TM) helices 1, 2, 3, and 7, capped by the acidic first extracellular loop (EL1) and specifically with the conserved melanocortin receptor residues mMC4R Glu92 (TM2), mMC4R Asp114 (TM3), and mMC4R Asp118 (TM3). Additionally, Phe112 and Phe113 of hAGRP(87-132) putatively interact with an aromatic hydrophobic pocket formed by the mMC4 receptor residues Phe176 (TM4), Phe193 (TM5), Phe253 (TM6), and Phe254 (TM6). To validate the AGRP-mMC4R model complex presented herein from a ligand perspective, we generated nine chimeric peptide ligands based on a modified antagonist template of the hAGRP(109-118) (Tyr-c[Asp-Arg-Phe-Phe-Asn-Ala-Phe-Dpr]-Tyr-NH(2)). In these chimeric ligands, the antagonist AGRP Arg-Phe-Phe residues were replaced by the melanocortin agonist His/D-Phe-Arg-Trp amino acids. These peptides resulted in agonist activity at the mouse melanocortin receptors (mMC1R and mMC3-5Rs). The most notable results include the identification of a novel subnanomolar melanocortin peptide

  3. Use of 99mTc-HYNIC-βAla-Bombesina(7-14) peptide for the identification of prostate tumor, LNCaP line, in an experimental model

    International Nuclear Information System (INIS)

    Fuscaldi, Leonardo Lima

    2012-01-01

    Prostate cancer is one of the most prevalent tumors in men, showing high mortality rates. Current diagnostic methods are not able to identify early prostate carcinoma, often resulting in a late diagnosis with established metastasis. Thus, there is by the scientific community an incessant search for diagnostic methods for early assessment of prostate cancer, facilitating the treatment and increasing the chances of cure. In this context, nuclear medicine provides a diagnostic method which can detect tumors at an early stage, because it is based on biochemical and physiological changes of the tissue, such as overexpression of gastrin releasing peptide receptors (GRPr's) by prostate cancer cells. Bombesin, a tetradecapeptide isolated from the frog Bombina bombina, has a high affinity for the GRPr's, since it is analogous to gastrin releasing peptide. Therefore, this study aims to prepare the complex 99m Tc-HYNIC-βAla-Bombesina (7-14) and use it for the identification of prostate tumor, LNCaP line, in an experimental model. For in vitro assays, aliquots of 0.026 MBq of the radiopeptide were incubated with 2x10 6 LNCaP cells in a water bath at 37 deg C, for 1 and 4 hours, with and without prior addition of cold peptide (n=3). Prostate tumors were induced into the upper right flank of male BALB/c nude mice by subcutaneous injection of 5x106 LNCaP cells resuspended in 150 μL of Matrigel:RPMI-1640 medium (1:1). Biodistribution profile (n=5) and scintigraphic images (n=3) were obtained at 1 and 4 hours after intravenous injection of 7.4 MBq of 99m Tc-HYNIC-βAla-Bombesina (7-14) . To assess this, healthy male BALB/c mice and tumor-bearing male BALB/c nude mice with 15, 20 and 25 days of tumor development were used. In vitro study results showed that the fraction of the radiopeptide which bound to LNCaP cells was 2.08 +- 0.30% (1 hour) and 2.44 +- 0.18% (4 hours). From the percentage which was bound, the internalized fractions were 25.64 +- 3.14% (1 hour) and 25.27 +- 2

  4. Consortial routes to effective repositories

    OpenAIRE

    Moyle, M.; Proudfoot, R.

    2009-01-01

    A consortial approach to the establishment of repository services can help a group of Higher Education Institutions (HEIs) to share costs, share technology and share expertise. Consortial repository work can tap into existing structures, or it can involve new groupings of institutions with a common interest in exploring repository development. This Briefing Paper outlines some of the potential benefits of collaborative repository activity, and highlights some of the technical and organisation...

  5. A modern approach for epitope prediction: identification of foot-and-mouth disease virus peptides binding bovine leukocyte antigen (BoLA) class I molecules

    DEFF Research Database (Denmark)

    Pandya, Mital; Rasmussen, Michael; Hansen, Andreas

    2015-01-01

    Major histocompatibility complex (MHC) class I molecules regulate adaptive immune responses through the presentation of antigenic peptides to CD8+ T cells. Polymorphisms in the peptide binding region of class I molecules determine peptide binding affinity and stability during antigen presentation......, and different antigen peptide motifs are associated with specific genetic sequences of class I molecules. Understanding bovine leukocyte antigen (BoLA), peptide-MHC class I binding specificities may facilitate development of vaccines or reagents for quantifying the adaptive immune response to intracellular...... pathogens, such as foot-and-mouth disease virus (FMDV). Six synthetic BoLA class I (BoLA-I) molecules were produced, and the peptide binding motif was generated for five of the six molecules using a combined approach of positional scanning combinatorial peptide libraries (PSCPLs) and neural network...

  6. Autoimmunity to Tropomyosin-Specific Peptides Induced by Mycobacterium leprae in Leprosy Patients: Identification of Mimicking Proteins.

    Science.gov (United States)

    Singh, Itu; Yadav, Asha Ram; Mohanty, Keshar Kunja; Katoch, Kiran; Sharma, Prashant; Pathak, Vinay Kumar; Bisht, Deepa; Gupta, Umesh D; Sengupta, Utpal

    2018-01-01

    It has been shown earlier that there is a rise in the levels of autoantibodies and T cell response to cytoskeletal proteins in leprosy. Our group recently demonstrated a rise in both T and B cell responses to keratin and myelin basic protein in all types of leprosy patients and their associations in type 1 reaction (T1R) group of leprosy. In this study, we investigated the association of levels of autoantibodies and lymphoproliferation against myosin in leprosy patients across the spectrum and tried to find out the mimicking proteins or epitopes between host protein and protein/s of Mycobacterium leprae . One hundred and sixty-nine leprosy patients and 55 healthy controls (HC) were enrolled in the present study. Levels of anti-myosin antibodies and T-cell responses against myosin were measured by ELISA and lymphoproliferation assay, respectively. Using 2-D gel electrophoresis, western blot and MALDI-TOF/TOF antibody-reactive spots were identified. Three-dimensional structure of mimicking proteins was modeled by online server. B cell epitopes of the proteins were predicted by BCPREDS server 1.0 followed by identification of mimicking epitopes. Mice of inbred BALB/c strain were hyperimmunized with M. leprae soluble antigen (MLSA) and splenocytes and lymph node cells of these animals were adoptively transferred to naïve mice. Highest level of anti-myosin antibodies was noted in sera of T1R leprosy patients. We observed significantly higher levels of lymphoproliferative response ( p  leprae . We found four mimicking epitopes between these sequences. These data suggest that these mimicking proteins tropomyosin and ATP-dependent Clp protease ATP-binding subunit of M. leprae or more precisely mimicking epitopes (four B cell epitopes) might be responsible for extensive tissue damage during type1 reaction in leprosy.

  7. Publishers and repositories

    CERN Multimedia

    CERN. Geneva

    2007-01-01

    The impact of self-archiving on journals and publishers is an important topic for all those involved in scholarly communication. There is some evidence that the physics arXiv has had no impact on physics journals, while 'economic common sense' suggests that some impact is inevitable. I shall review recent studies of librarian attitudes towards repositories and journals, and place this in the context of IOP Publishing's experiences with arXiv. I shall offer some possible reasons for the mis-match between these perspectives and then discuss how IOP has linked with arXiv and experimented with OA publishing. As well as launching OA journals we have co-operated with Cornell and the arXiv on Eprintweb.org, a platform that offers new features to repository users. View Andrew Wray's biography

  8. Distributed Web Service Repository

    Directory of Open Access Journals (Sweden)

    Piotr Nawrocki

    2015-01-01

    Full Text Available The increasing availability and popularity of computer systems has resulted in a demand for new, language- and platform-independent ways of data exchange. That demand has in turn led to a significant growth in the importance of systems based on Web services. Alongside the growing number of systems accessible via Web services came the need for specialized data repositories that could offer effective means of searching of available services. The development of mobile systems and wireless data transmission technologies has allowed the use of distributed devices and computer systems on a greater scale. The accelerating growth of distributed systems might be a good reason to consider the development of distributed Web service repositories with built-in mechanisms for data migration and synchronization.

  9. Shared Medical Imaging Repositories.

    Science.gov (United States)

    Lebre, Rui; Bastião, Luís; Costa, Carlos

    2018-01-01

    This article describes the implementation of a solution for the integration of ownership concept and access control over medical imaging resources, making possible the centralization of multiple instances of repositories. The proposed architecture allows the association of permissions to repository resources and delegation of rights to third entities. It includes a programmatic interface for management of proposed services, made available through web services, with the ability to create, read, update and remove all components resulting from the architecture. The resulting work is a role-based access control mechanism that was integrated with Dicoogle Open-Source Project. The solution has several application scenarios like, for instance, collaborative platforms for research and tele-radiology services deployed at Cloud.

  10. 1-(3-aminopropyl)-3-butylimidazolium bromide for carboxyl group derivatization: potential applications in high sensitivity peptide identification by mass spectrometry.

    Science.gov (United States)

    Qiao, Xiaoqiang; Zhou, Yuan; Hou, Chunyan; Zhang, Xiaodan; Yang, Kaiguang; Zhang, Lihua; Zhang, Yukui

    2013-03-01

    The cationic reagent 1-(3-aminopropyl)-3-butylimidazolium bromide (BAPI) was exploited for the derivatization of carboxyl groups on peptides. Nearly 100% derivatization efficiency was achieved with the synthetic peptide RVYVHPI (RI-7). Furthermore, the peptide derivative was stable in a 0.1% TFA/water solution or a 0.1% (v/v) TFA/acetonitrile/water solution for at least one week. The effect of BAPI derivatization on the ionization of the peptide RI-7 was further investigated, and the detection sensitivity was improved >42-fold via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), thus outperforming the commercial piperazine derivatization approach. Moreover, the charge states of the peptide were largely increased via BAPI derivatization by electrospray ionization (ESI) MS. The results indicate the potential merits of BAPI derivatization for high sensitivity peptide analysis by MS.

  11. Identification of the bacteria-binding peptide domain on salivary agglutinin (gp-340/DMBT1), a member of the scavenger receptor cysteine-rich superfamily

    DEFF Research Database (Denmark)

    Bikker, Floris J; Ligtenberg, Antoon J M; Nazmi, Kamran

    2002-01-01

    Salivary agglutinin is encoded by DMBT1 and identical to gp-340, a member of the scavenger receptor cysteine-rich (SRCR) superfamily. Salivary agglutinin/DMBT1 is known for its Streptococcus mutans agglutinating properties. This 300-400 kDa glycoprotein is composed of conserved peptide motifs: 14...... containing exclusively SRCR and SID domains that binds to S. mutans. To define more closely the S. mutans-binding domain, consensus-based peptides of the SRCR domains and SIDs were designed and synthesized. Only one of the SRCR peptides, designated SRCRP2, and none of the SID peptides bound to S. mutans....... Strikingly, this peptide was also able to induce agglutination of S. mutans and a number of other bacteria. The repeated presence of this peptide in the native molecule endows agglutinin/DMBT1 with a general bacterial binding feature with a multivalent character. Moreover, our studies demonstrate...

  12. Czech Republic. Dukovany repository

    International Nuclear Information System (INIS)

    2001-01-01

    Full text: The repository at the Dukovany site is a structure located above the land surface. It consists of two double-rows of reinforced concrete vaults. Each double-row has dimensions 38x160x6 meters and contains 2x28 vaults. The internal dimensions of each vault are 18x6x5.4 meters. The repository serves for reactor wastes from the Dukovany and Temelin nuclear power plants (NPPs). Its capacity is 55,000 m 3 or 130,000 drums. The repository is a fully engineered facility with multiple barriers. The first engineered barrier is the waste form (in the case of waste from the Dukovany NPP, the waste form is mainly bitumen, but concrete and glass are also considered as suitable solidification products). The second barrier is the container (a 200 litre steel drum or a HIC container), whereas the third consists of cut-off reinforced concrete walls with asphalt-based hydro-insulation. The fourth barrier is a cap which should protect the vaults against infiltration of rainwater and should serve also as an intrusion and erosion barrier. The fifth barrier is a drainage system around the repository which is composed of layers of gravel and sand. The void space in drums around the waste is filled with specially composed grout. Such waste packages are emplaced into the disposal vault, which is covered by pre-fabricated panels. Thereafter, joints between the panels are sealed and a provisional coverage added; the final cover, however, will be constructed only over the whole row of 28 vaults, until all vaults are filled with waste. The final cover will encompass the following components: reinforced concrete pre-fabricated panels (500 mm); cement overcoat (30 mm); insulation foil; concrete layer for cap levelling (5-150 mm); layer of asphalto-propylene concrete (150 mm); soil (450 mm); geotextile foil with topsoil (top surface vegetation). (author)

  13. Autoimmunity to Tropomyosin-Specific Peptides Induced by Mycobacterium leprae in Leprosy Patients: Identification of Mimicking Proteins

    Directory of Open Access Journals (Sweden)

    Itu Singh

    2018-04-01

    Full Text Available BackgroundIt has been shown earlier that there is a rise in the levels of autoantibodies and T cell response to cytoskeletal proteins in leprosy. Our group recently demonstrated a rise in both T and B cell responses to keratin and myelin basic protein in all types of leprosy patients and their associations in type 1 reaction (T1R group of leprosy.ObjectivesIn this study, we investigated the association of levels of autoantibodies and lymphoproliferation against myosin in leprosy patients across the spectrum and tried to find out the mimicking proteins or epitopes between host protein and protein/s of Mycobacterium leprae.MethodologyOne hundred and sixty-nine leprosy patients and 55 healthy controls (HC were enrolled in the present study. Levels of anti-myosin antibodies and T-cell responses against myosin were measured by ELISA and lymphoproliferation assay, respectively. Using 2-D gel electrophoresis, western blot and MALDI-TOF/TOF antibody-reactive spots were identified. Three-dimensional structure of mimicking proteins was modeled by online server. B cell epitopes of the proteins were predicted by BCPREDS server 1.0 followed by identification of mimicking epitopes. Mice of inbred BALB/c strain were hyperimmunized with M. leprae soluble antigen (MLSA and splenocytes and lymph node cells of these animals were adoptively transferred to naïve mice.ResultsHighest level of anti-myosin antibodies was noted in sera of T1R leprosy patients. We observed significantly higher levels of lymphoproliferative response (p < 0.05 with myosin in all types of leprosy patients compared to HC. Further, hyperimmunization of inbred BALB/c strain of female mice and rabbit with MLSA revealed that both hyperimmunized rabbit and mice evoked heightened levels of antibodies against myosin and this autoimmune response could be adoptively transferred from hyperimmunized to naïve mice. Tropomyosin was found to be mimicking with ATP-dependent Clp protease ATP

  14. Staged Repository Development Programmes

    International Nuclear Information System (INIS)

    Isaacs, T

    2003-01-01

    Programs to manage and ultimately dispose of high-level radioactive wastes are unique from scientific and technological as well as socio-political aspects. From a scientific and technological perspective, high-level radioactive wastes remain potentially hazardous for geological time periods-many millennia-and scientific and technological programs must be put in place that result in a system that provides high confidence that the wastes will be isolated from the accessible environment for these many thousands of years. Of course, ''proof'' in the classical sense is not possible at the outset, since the performance of the system can only be known with assurance, if ever, after the waste has been emplaced for those geological time periods. Adding to this challenge, many uncertainties exist in both the natural and engineered systems that are intended to isolate the wastes, and some of the uncertainties will remain regardless of the time and expense in attempting to characterize the system and assess its performance. What was perhaps underappreciated in the early days of waste management and repository program development were the unique and intense reactions that the institutional, political, and public bodies would have to repository program development, particularly in programs attempting to identify and then select sites for characterization, design, licensing, and ultimate development. Reactions in most nations were strong, focused, unrelenting, and often successful in hindering, derailing, and even stopping national repository programs. The reasons for such reactions and the measures to successfully respond to them are still evolving and continue to be the focus of many national program and political leaders. Adaptive Staging suggests an approach to repository program development that reflects the unique challenges associated with the disposal of high-level radioactive waste. The step-wise, incremental, learn-as-you-go approach is intended to maximize the

  15. Peptide dendrimers

    Czech Academy of Sciences Publication Activity Database

    Niederhafner, Petr; Šebestík, Jaroslav; Ježek, Jan

    2005-01-01

    Roč. 11, - (2005), 757-788 ISSN 1075-2617 R&D Projects: GA ČR(CZ) GA203/03/1362 Institutional research plan: CEZ:AV0Z40550506 Keywords : multiple antigen peptides * peptide dendrimers * synthetic vaccine * multipleantigenic peptides Subject RIV: CC - Organic Chemistry Impact factor: 1.803, year: 2005

  16. New screening strategy and analysis for identification of allosteric modulators for glucagon-like peptide-1 receptor using GLP-1 (9-36) amide.

    Science.gov (United States)

    Nakane, Atsushi; Gotoh, Yusuke; Ichihara, Junji; Nagata, Hidetaka

    2015-12-15

    The glucagon-like peptide-1 receptor (GLP-1R) is an important physiologic regulator of insulin secretion and a major therapeutic target for diabetes mellitus. GLP-1 (7-36) amide (active form of GLP-1) is truncated to GLP-1 (9-36) amide, which has been described as a weak agonist of GLP-1R and the major form of GLP-1 in the circulation. New classes of positive allosteric modulators (PAMs) for GLP-1R may offer improved therapeutic profiles. To identify these new classes, we developed novel and robust primary and secondary high-throughput screening (HTS) systems in which PAMs were identified to enhance the GLP-1R signaling induced by GLP-1 (9-36) amide. Screening enabled identification of two compounds, HIT-465 and HIT-736, which possessed new patterns of modulation of GLP-1R. We investigated the ability of these compounds to modify GLP-1R signaling enhanced GLP-1 (9-36) amide- and/or GLP-1 (7-36) amide-mediated cyclic adenosine monophosphate (cAMP) accumulation. These compounds also had unique profiles with regard to allosteric modulation of multiple downstream signaling (PathHunter β-arrestin signaling, PathHunter internalization signaling, microscopy-based internalization assay). We found allosteric modulation patterns to be obviously different among HIT-465, HIT-736, and Novo Nordisk compound 2. This work may enable the design of new classes of drug candidates by targeting modulation of GLP-1 (7-36) amide and GLP-1 (9-36) amide. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Evaluation of repository safety

    Energy Technology Data Exchange (ETDEWEB)

    Sagar, B.; Patrick, W.; Dasgupta, B.; Mohanty, S. [Center for Nuclear Waste Regulatory Analyses, San Antonio (United States)

    2002-07-01

    The United States high-level waste program requires evaluation of radiological safety during two distinct time intervals. The first interval, commonly referred to as the preclosure period, deals with receipt of waste at the site, transfer into disposal containers, if needed, emplacement in the underground openings, monitoring and maintenance activities, backfill and closure of the underground openings, and decontamination and decommissioning of the surface facilities of the geologic repository. The preclosure period may extend from a few tens of years to as long as a few hundred of years, depending on repository design and societal norms regarding a final decision to permanently seal the repository. During the preclosure or operational period, performance confirmation studies are conducted to provide a basis for updating and reevaluating estimates of postclosure performance and, finally, to provide a basis for a closure decision. The postclosure period during which expected repository performance must meet certain standards may range from ten thousands years, as it does in the United States, to millions of years, as it does in some European nations. Waste handling operations in the preclosure period are to be evaluated in relation to their potential effect on workers, members of general public, and the general environment. During this period, releases of radioactivity are to be monitored and appropriate actions taken whenever established limits are approached or exceeded. Preclosure safety is highly dependent on facility design, operational hardware and automated systems, operational sequences, and reliability of humans involved in operations. Preclosure safety analyses conducted before operations begin play a major role in the design process, selection of equipment, and development of operational procedures. Because of the complexity, duration, and spatial scales of the operations, analyses are conducted using mathematical models implemented in computer codes

  18. Evaluation of repository safety

    International Nuclear Information System (INIS)

    Sagar, B.; Patrick, W.; Dasgupta, B.; Mohanty, S.

    2002-01-01

    The United States high-level waste program requires evaluation of radiological safety during two distinct time intervals. The first interval, commonly referred to as the preclosure period, deals with receipt of waste at the site, transfer into disposal containers, if needed, emplacement in the underground openings, monitoring and maintenance activities, backfill and closure of the underground openings, and decontamination and decommissioning of the surface facilities of the geologic repository. The preclosure period may extend from a few tens of years to as long as a few hundred of years, depending on repository design and societal norms regarding a final decision to permanently seal the repository. During the preclosure or operational period, performance confirmation studies are conducted to provide a basis for updating and reevaluating estimates of postclosure performance and, finally, to provide a basis for a closure decision. The postclosure period during which expected repository performance must meet certain standards may range from ten thousands years, as it does in the United States, to millions of years, as it does in some European nations. Waste handling operations in the preclosure period are to be evaluated in relation to their potential effect on workers, members of general public, and the general environment. During this period, releases of radioactivity are to be monitored and appropriate actions taken whenever established limits are approached or exceeded. Preclosure safety is highly dependent on facility design, operational hardware and automated systems, operational sequences, and reliability of humans involved in operations. Preclosure safety analyses conducted before operations begin play a major role in the design process, selection of equipment, and development of operational procedures. Because of the complexity, duration, and spatial scales of the operations, analyses are conducted using mathematical models implemented in computer codes

  19. "Design and application of a data-independent precursor and product ion repository."

    NARCIS (Netherlands)

    Thalassinos, K.; Vissers, J.P.; Tenzer, S.; Levin, Y.; Thompson, J.W.; Daniel, D.; Mann, D.; Delong, M.R.; Moseley, M.A.; America, A.H.P.; Ottens, A.K.; Cavey, G.S.; Efstathiou, G.; Scrivens, J.H.; Langridge, J.I.; Geromanos, S.J.

    2012-01-01

    The functional design and application of a data-independent LC-MS precursor and product ion repository for protein identification, quantification, and validation is conceptually described. The ion repository was constructed from the sequence search results of a broad range of discovery experiments

  20. Identification of constrained peptides that bind to and preferentially inhibit the activity of the hepatitis C viral RNA-dependent RNA polymerase

    International Nuclear Information System (INIS)

    Amin, Anthony; Zaccardi, Joe; Mullen, Stanley; Olland, Stephane; Orlowski, Mark; Feld, Boris; Labonte, Patrick; Mak, Paul

    2003-01-01

    A class of disulfide constrained peptides containing a core motif FPWG was identified from a screen of phage displayed library using the HCV RNA-dependent RNA polymerase (NS5B) as a bait. Surface plasmon resonance studies showed that three highly purified synthetic constrained peptides bound to immobilized NS5B with estimated K d values ranging from 30 to 60 μM. In addition, these peptides inhibited the NS5B activity in vitro with IC 50 ranging from 6 to 48 μM, whereas in contrast they had no inhibitory effect on the enzymatic activities of calf thymus polymerase α, human polymerase β, RSV polymerase, and HIV reverse transcriptase in vitro. Two peptides demonstrated conformation-dependent inhibition since their synthetic linear versions were not inhibitory in the NS5B assay. A constrained peptide with the minimum core motif FPWG retained selective inhibition of NS5B activity with an IC 50 of 50 μM. Alanine scan analyses of a representative constrained peptide, FPWGNTW, indicated that residues F1 and W7 were critical for the inhibitory effect of this peptide, although residues P2 and N5 had some measurable inhibitory effect as well. Further analyses of the mechanism of inhibition indicated that these peptides inhibited the formation of preelongation complexes required for the elongation reaction. However, once the preelongation complex was formed, its activity was refractory to peptide inhibition. Furthermore, the constrained peptide FPWGNTW inhibited de novo initiated RNA synthesis by NS5B from a poly(rC) template. These data indicate that the peptides confer selective inhibition of NS5B activity by binding to the enzyme and perturbing an early step preceding the processive elongation step of RNA synthesis

  1. Repository simulation model: Final report

    International Nuclear Information System (INIS)

    1988-03-01

    This report documents the application of computer simulation for the design analysis of the nuclear waste repository's waste handling and packaging operations. The Salt Repository Simulation Model was used to evaluate design alternatives during the conceptual design phase of the Salt Repository Project. Code development and verification was performed by the Office of Nuclear Waste Isolation (ONWL). The focus of this report is to relate the experience gained during the development and application of the Salt Repository Simulation Model to future repository design phases. Design of the repository's waste handling and packaging systems will require sophisticated analysis tools to evaluate complex operational and logistical design alternatives. Selection of these design alternatives in the Advanced Conceptual Design (ACD) and License Application Design (LAD) phases must be supported by analysis to demonstrate that the repository design will cost effectively meet DOE's mandated emplacement schedule and that uncertainties in the performance of the repository's systems have been objectively evaluated. Computer simulation of repository operations will provide future repository designers with data and insights that no other analytical form of analysis can provide. 6 refs., 10 figs

  2. Towards Interoperable Preservation Repositories: TIPR

    Directory of Open Access Journals (Sweden)

    Priscilla Caplan

    2010-07-01

    Full Text Available Towards Interoperable Preservation Repositories (TIPR is a project funded by the Institute of Museum and Library Services to create and test a Repository eXchange Package (RXP. The package will make it possible to transfer complex digital objects between dissimilar preservation repositories.  For reasons of redundancy, succession planning and software migration, repositories must be able to exchange copies of archival information packages with each other. Every different repository application, however, describes and structures its archival packages differently. Therefore each system produces dissemination packages that are rarely understandable or usable as submission packages by other repositories. The RXP is an answer to that mismatch. Other solutions for transferring packages between repositories focus either on transfers between repositories of the same type, such as DSpace-to-DSpace transfers, or on processes that rely on central translation services.  Rather than build translators between many dissimilar repository types, the TIPR project has defined a standards-based package of metadata files that can act as an intermediary information package, the RXP, a lingua franca all repositories can read and write.

  3. From Viral genome to specific peptide epitopes - Methods for identifying porcine T cell epitopes based on in silico predictions, in vitro identification and ex vivo verification

    DEFF Research Database (Denmark)

    Pedersen, Lasse Eggers; Rasmussen, Michael; Harndahl, Mikkel

    The affinity for and stability of peptides bound by major histocompatibility complex (MHC) class I molecules are instrumental factors in presentation of viral epitopes to cytotoxic T lymphocytes (CTLs). In swine, such peptide presentations by swine leukocyte antigens (SLA) are crucial for swine i...

  4. Purification and identification of O-GlcNAc-modified peptides using phosphate-based alkyne CLICK chemistry in combination with titanium dioxide chromatography and mass spectrometry

    DEFF Research Database (Denmark)

    Parker, Benjamin L; Gupta, Pankaj; Cordwell, Stuart

    2011-01-01

    A selective method for the enrichment of O-GlcNAcylated peptides using a novel CLICK chemistry reagent is described. Peptides modified by O-GlcNAc were enzymatically labeled with N-azidoacetylgalactosamine. The azide was then reacted with a phospho-alkyne using CLICK chemistry and O-GlcNAcGalNAzPO4...

  5. From viral genome to specific peptide epitopes: methods for identifying porcine T cell epitopes based on in silico predictions, in vitro identification and ex vivo verification

    DEFF Research Database (Denmark)

    Pedersen, Lasse Eggers; Rasmussen, Michael; Harndah, Mikkel

    2013-01-01

    to predict likely candidates for peptide-SLA binding. These results were combined with binding predictions generated by the algorithm, NetMHCpan (http://www.cbs.dtu.dk/services/NetMHCpan/) in order to select peptide candidates for in vitro analysis. The correlation between high affinity and high stability.......000 peptides. T cell epitopes were identified using peptide-SLA complexes assembled into fluorescent tetramers to stain swine influenza specific CTLs derived from immunized animals and MHC-defined pigs vaccinated against foot-and-mouth disease virus. These results demonstrate the broad applicability of methods...... originally developed for analysis of human leukocyte antigen (HLA) presentation of peptides. The methods presented provide a timely and cost-effective approach to CTL epitope discovery that can be applied to diseases of swine and of other mammalian species of interest....

  6. NASA Biological Specimen Repository

    Science.gov (United States)

    McMonigal, K. A.; Pietrzyk, R. A.; Sams, C. F.; Johnson, M. A.

    2010-01-01

    The NASA Biological Specimen Repository (NBSR) was established in 2006 to collect, process, preserve and distribute spaceflight-related biological specimens from long duration ISS astronauts. This repository provides unique opportunities to study longitudinal changes in human physiology spanning may missions. The NBSR collects blood and urine samples from all participating ISS crewmembers who have provided informed consent. These biological samples are collected once before flight, during flight scheduled on flight days 15, 30, 60, 120 and within 2 weeks of landing. Postflight sessions are conducted 3 and 30 days after landing. The number of in-flight sessions is dependent on the duration of the mission. Specimens are maintained under optimal storage conditions in a manner that will maximize their integrity and viability for future research The repository operates under the authority of the NASA/JSC Committee for the Protection of Human Subjects to support scientific discovery that contributes to our fundamental knowledge in the area of human physiological changes and adaptation to a microgravity environment. The NBSR will institute guidelines for the solicitation, review and sample distribution process through establishment of the NBSR Advisory Board. The Advisory Board will be composed of representatives of all participating space agencies to evaluate each request from investigators for use of the samples. This process will be consistent with ethical principles, protection of crewmember confidentiality, prevailing laws and regulations, intellectual property policies, and consent form language. Operations supporting the NBSR are scheduled to continue until the end of U.S. presence on the ISS. Sample distribution is proposed to begin with selections on investigations beginning in 2017. The availability of the NBSR will contribute to the body of knowledge about the diverse factors of spaceflight on human physiology.

  7. INSTITUTIONAL REPOSITORY: EMPLOYMENT IN EDUCATION

    Directory of Open Access Journals (Sweden)

    Vasyl P. Oleksyuk

    2012-11-01

    Full Text Available The article investigated the concept of «institutional repository» and determined the aspects of institutional repositories in higher education. Institutional Repositories are information systems that allow preserving, storing and disseminating scientific knowledge produced in higher education and scientific research institutions. This study presented the main aspects using institutional repositories in educational process (such as storage of scientific and educational information, means of organization activity of students, object of studying. This article produced the structure of communities and collections of the institutional. It is described the experience of implementing of DSpace in the learning process.

  8. Object linking in repositories

    Science.gov (United States)

    Eichmann, David (Editor); Beck, Jon; Atkins, John; Bailey, Bill

    1992-01-01

    This topic is covered in three sections. The first section explores some of the architectural ramifications of extending the Eichmann/Atkins lattice-based classification scheme to encompass the assets of the full life cycle of software development. A model is considered that provides explicit links between objects in addition to the edges connecting classification vertices in the standard lattice. The second section gives a description of the efforts to implement the repository architecture using a commercially available object-oriented database management system. Some of the features of this implementation are described, and some of the next steps to be taken to produce a working prototype of the repository are pointed out. In the final section, it is argued that design and instantiation of reusable components have competing criteria (design-for-reuse strives for generality, design-with-reuse strives for specificity) and that providing mechanisms for each can be complementary rather than antagonistic. In particular, it is demonstrated how program slicing techniques can be applied to customization of reusable components.

  9. Identification of a new androgen receptor (AR) co-regulator BUD31 and related peptides to suppress wild-type and mutated AR-mediated prostate cancer growth via peptide screening and X-ray structure analysis.

    Science.gov (United States)

    Hsu, Cheng-Lung; Liu, Jai-Shin; Wu, Po-Long; Guan, Hong-Hsiang; Chen, Yuh-Ling; Lin, An-Chi; Ting, Huei-Ju; Pang, See-Tong; Yeh, Shauh-Der; Ma, Wen-Lung; Chen, Chung-Jung; Wu, Wen-Guey; Chang, Chawnshang

    2014-12-01

    Treatment with individual anti-androgens is associated with the development of hot-spot mutations in the androgen receptor (AR). Here, we found that anti-androgens-mt-ARs have similar binary structure to the 5α-dihydrotestosterone-wt-AR. Phage display revealed that these ARs bound to similar peptides, including BUD31, containing an Fxx(F/H/L/W/Y)Y motif cluster with Tyr in the +5 position. Structural analyses of the AR-LBD-BUD31 complex revealed formation of an extra hydrogen bond between the Tyr+5 residue of the peptide and the AR. Functional studies showed that BUD31-related peptides suppressed AR transactivation, interrupted AR N-C interaction, and suppressed AR-mediated cell growth. Combination of peptide screening and X-ray structure analysis may serve as a new strategy for developing anti-ARs that simultaneously suppress both wt and mutated AR function. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  10. Accelerator Physics Code Web Repository

    CERN Document Server

    Zimmermann, Frank; Bellodi, G; Benedetto, E; Dorda, U; Giovannozzi, Massimo; Papaphilippou, Y; Pieloni, T; Ruggiero, F; Rumolo, G; Schmidt, F; Todesco, E; Zotter, Bruno W; Payet, J; Bartolini, R; Farvacque, L; Sen, T; Chin, Y H; Ohmi, K; Oide, K; Furman, M; Qiang, J; Sabbi, G L; Seidl, P A; Vay, J L; Friedman, A; Grote, D P; Cousineau, S M; Danilov, V; Holmes, J A; Shishlo, A; Kim, E S; Cai, Y; Pivi, M; Kaltchev, D I; Abell, D T; Katsouleas, Thomas C; Boine-Frankenheim, O; Franchetti, G; Hofmann, I; Machida, S; Wei, J

    2006-01-01

    In the framework of the CARE HHH European Network, we have developed a web-based dynamic acceleratorphysics code repository. We describe the design, structure and contents of this repository, illustrate its usage, and discuss our future plans, with emphasis on code benchmarking.

  11. Safety analysis in subsurface repositories

    International Nuclear Information System (INIS)

    1985-06-01

    The development of mathematical models to represent the repository-geosphere-biosphere system, and the development of a structure for data acquisition, processing, and use to analyse the safety of subsurface repositories, are presented. To study the behavior of radionuclides in geosphere a laboratory to determine the hydrodynamic dispersion coefficient was constructed. (M.C.K.) [pt

  12. Granite-repository - geochemical environment

    International Nuclear Information System (INIS)

    1979-04-01

    Some geochemical data of importance for a radioactive waste repository in hard rock are reviewed. The ground water composition at depth is assessed. The ground water chemistry in the vicinity of uranium ores is discussed. The redox system in Swedish bedrock is described. Influences of extreme climatic changes and of repository mining and construction are also evaluated

  13. ACCELERATION PHYSICS CODE WEB REPOSITORY.

    Energy Technology Data Exchange (ETDEWEB)

    WEI, J.

    2006-06-26

    In the framework of the CARE HHH European Network, we have developed a web-based dynamic accelerator-physics code repository. We describe the design, structure and contents of this repository, illustrate its usage, and discuss our future plans, with emphasis on code benchmarking.

  14. Technology overview of mined repositories

    International Nuclear Information System (INIS)

    Gimera, R.; Thirumalai, K.

    1982-01-01

    Mined repositories present an environmentally viable option for permanent disposal of nuclear waste. This paper reviews the state-of-the-art mining technologies and identifies technological issues and developments necessary to mine a repository in basalt. The thermal loading, isolation, and retrieval requirements of a repository present unique technological challenges unknown to conventional mining practice. The technology issues and developments required in the areas of excavation, roof and ground support, equipment development, instrumentation development, and sealing are presented. Performance assessment methods must be developed to evaluate the adequacies of technologies developed to design, construct, operate, and decommission a repository. A stepwise test-and-development approach is used in the Basalt Waste Isolation Project to develop cost-effective technologies for a repository

  15. Influence analysis of Github repositories.

    Science.gov (United States)

    Hu, Yan; Zhang, Jun; Bai, Xiaomei; Yu, Shuo; Yang, Zhuo

    2016-01-01

    With the support of cloud computing techniques, social coding platforms have changed the style of software development. Github is now the most popular social coding platform and project hosting service. Software developers of various levels keep entering Github, and use Github to save their public and private software projects. The large amounts of software developers and software repositories on Github are posing new challenges to the world of software engineering. This paper tries to tackle one of the important problems: analyzing the importance and influence of Github repositories. We proposed a HITS based influence analysis on graphs that represent the star relationship between Github users and repositories. A weighted version of HITS is applied to the overall star graph, and generates a different set of top influential repositories other than the results from standard version of HITS algorithm. We also conduct the influential analysis on per-month star graph, and study the monthly influence ranking of top repositories.

  16. Identification of the peptide derived from S1 domain that inhibits type I and type II feline infectious peritonitis virus infection.

    Science.gov (United States)

    Doki, Tomoyoshi; Takano, Tomomi; Koyama, Yusuke; Hohdatsu, Tsutomu

    2015-06-02

    Feline infectious peritonitis virus (FIPV) can cause a lethal disease in cats, feline infectious peritonitis (FIP). A therapeutic drug that is effective against FIP has not yet been developed. Peptides based on viral protein amino acid sequences have recently been attracting attention as new antiviral drugs. In the present study, we synthesized 30 overlapping peptides based on the amino acid sequence of the S1 domain of the type I FIPV strain KU-2 S protein, and investigated their inhibitory effects on FIPV infection. To evaluate the inhibitory effects on type I FIPV infection of these peptides, we investigated a method to increase the infection efficiency of poorly replicative type I FIPV. The efficiency of type I FIPV infection was increased by diluting the virus with medium containing a polycation. Of the 30 peptides, I-S1-8 (S461-S480), I-S1-9 (S471-S490), I-S1-10 (S481-S500), I-S1-16 (S541-S560), and I-S1-22 (S601-S620) significantly decreased the infectivity of FIPV strain KU-2 while I-S1-9 and I-S1-16 exhibited marked inhibitory effects on FIPV infection. The inhibitory effects on FIPV infection of these 2 peptides on other type I and type II FIPV strains, feline herpesvirus (FHV), and feline calicivirus (FCV) were also examined. These 2 peptides specifically inhibited type I and type II FIPV, but did FHV or FCV infection. In conclusion, the possibility of peptides derived from the S protein of type I FIPV strain KU-2 as anti-FIPV agents effective not only for type I, but also type II FIPV was demonstrated in vitro. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. LC-QTOF-MS identification of porcine-specific peptide in heat treated pork identifies candidate markers for meat species determination.

    Science.gov (United States)

    Sarah, S A; Faradalila, W N; Salwani, M S; Amin, I; Karsani, S A; Sazili, A Q

    2016-05-15

    The purpose of this study was to identify porcine-specific peptide markers from thermally processed meat that could differentiate pork from beef, chevon and chicken meat. In the initial stage, markers from tryptic digested protein of chilled, boiled and autoclaved pork were identified using LC-QTOF-MS. An MRM method was then established for verification. A thorough investigation of LC-QTOF-MS data showed that only seven porcine-specific peptides were consistently detected. Among these peptides, two were derived from lactate dehydrogenase, one from creatine kinase, and four from serum albumin protein. However, MRM could only detect four peptides (EVTEFAK, LVVITAGAR, FVIER and TVLGNFAAFVQK) that were consistently present in pork samples. In conclusion, meat species determination through a tandem mass spectrometry platform shows high potential in providing scientifically valid and reliable results even at peptide level. Besides, the specificity and selectivity offered by the proteomics approach also provide a robust platform for Halal authentication. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. High performance liquid chromatography (HPLC fingerprints and primary structure identification of corn peptides by HPLC-diode array detection and HPLC-electrospray ionization tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Chi Wang

    2016-01-01

    Full Text Available Corn peptides (CPs are reported to have many biological functions, such as facilitating alcohol metabolism, antioxidation, antitumor, antihypertension, and hepatoprotection. To develop a method for quality control, the high-performance liquid chromatography (HPLC system was applied. Twenty-eight common peaks were found in all the CPs of corn samples from Enshi, China, based on which, a fingerprinting chromatogram was established for use in quality control in future research. Subsequently, the major chemical constituents of these common peaks were identified respectively using the HPLC-diode-array detection electrospray ionization tandem mass spectrometry (DAD-ESI-MS/MS system, and 48 peptide fractions were determined ultimately. This was the first time for the majority of these peptides to be reported, and many of them contained amino acids of glutamine (Q, L and A, which might play an important role in the exhibition of the bioactivities of CPs. Many peptides had a similar primary structure to the peptides which had been proven to be bioactive such as facilitating alcohol metabolism, scavenging free radicals, and inhibiting lipid peroxidation. This systematical analysis of the primary structure of CPs facilitated subsequent studies on the relationship between the structures and functions, and could accelerate holistic research on CPs.

  19. Extraction and identification of α-amylase inhibitor peptides from Nephelium lappacheum and Nephelium mutabile seed protein using gastro-digestive enzymes.

    Science.gov (United States)

    Evaristus, Natashya Anak; Wan Abdullah, Wan Nadiah; Gan, Chee-Yuen

    2018-04-01

    The potential of N. lappacheum and N. mutabile seed as a source of α-amylase inhibitor peptides was explored based on the local traditional practice of using the seed. Different gastro-digestive enzymes (i.e. pepsin or chymotrypsin) or a sequential digestion were used to extract the peptides. The effects of digestion time and enzyme to substrate (E:S) ratio on the α-amylase inhibitory activity were investigated. Results showed that chymotrypsin was effective in producing the inhibitor peptides from rambutan seed protein at E:S ratio 1:20 for 1 h, whereas pepsin was more effective for pulasan seed protein under the same condition. A total of 20 and 31 novel inhibitor peptides were identified, respectively. These peptides could bind with the subsites of α-amylase (i.e. Trp58, Trp59, Tyr62, Asp96, Arg195, Asp197, Glu233, His299, Asp300, and His305) and formed a sliding barrier that preventing the formation of enzyme/substrate intermediate leading to lower α-amylase activity. Copyright © 2018 Elsevier Inc. All rights reserved.

  20. Computational identification, characterization and validation of potential antigenic peptide vaccines from hrHPVs E6 proteins using immunoinformatics and computational systems biology approaches.

    Directory of Open Access Journals (Sweden)

    Abbas Khan

    Full Text Available High-risk human papillomaviruses (hrHPVs are the most prevalent viruses in human diseases including cervical cancers. Expression of E6 protein has already been reported in cervical cancer cases, excluding normal tissues. Continuous expression of E6 protein is making it ideal to develop therapeutic vaccines against hrHPVs infection and cervical cancer. Therefore, we carried out a meta-analysis of multiple hrHPVs to predict the most potential prophylactic peptide vaccines. In this study, immunoinformatics approach was employed to predict antigenic epitopes of hrHPVs E6 proteins restricted to 12 Human HLAs to aid the development of peptide vaccines against hrHPVs. Conformational B-cell and CTL epitopes were predicted for hrHPVs E6 proteins using ElliPro and NetCTL. The potential of the predicted peptides were tested and validated by using systems biology approach considering experimental concentration. We also investigated the binding interactions of the antigenic CTL epitopes by using docking. The stability of the resulting peptide-MHC I complexes was further studied by molecular dynamics simulations. The simulation results highlighted the regions from 46-62 and 65-76 that could be the first choice for the development of prophylactic peptide vaccines against hrHPVs. To overcome the worldwide distribution, the predicted epitopes restricted to different HLAs could cover most of the vaccination and would help to explore the possibility of these epitopes for adaptive immunotherapy against HPVs infections.

  1. ESI-MS/MS Identification of a Bradykinin-Potentiating Peptide from Amazon Bothrops atrox Snake Venom Using a Hybrid Qq-oaTOF Mass Spectrometer

    Science.gov (United States)

    Coutinho-Neto, Antonio; Caldeira, Cleópatra A. S.; Souza, Gustavo H. M. F.; Zaqueo, Kayena D.; Kayano, Anderson M.; Silva, Rodrigo S.; Zuliani, Juliana P.; Soares, Andreimar M.; Stábeli, Rodrigo G.; Calderon, Leonardo A.

    2013-01-01

    A bradykinin-potentiating peptide (BPP) from Amazon Bothrops atrox venom with m/z 1384.7386 was identified and characterized by collision induced dissociation (CID) using an ESI-MS/MS spectra obtained in positive ion mode on a hybrid Qq-oaTOF mass spectrometer, Xevo G2 QTof MS (Waters, Manchester, UK). De novo peptide sequence analysis of the CID fragmentation spectra showed the amino acid sequence ZKWPRPGPEIPP, with a pyroglutamic acid and theoretical monoisotopic m/z 1384.7378, which is similar to experimental data, showing a mass accuracy of 0.6 ppm. The peptide is homologous to other BPP from Bothrops moojeni and was named as BPP-BAX12. PMID:23430539

  2. 500 year concrete for a radioactive waste repository

    International Nuclear Information System (INIS)

    Philipose, K.E.

    1988-03-01

    The IRUS (Intrusion resistant underground structure) repository planned at Chalk River for the belowground disposal of low level radioactive waste relies on the durability of concrete for the required 500 year service life. A research program for the IRUS repository to design a durable concrete and also to predict its longevity under the repository environment is in progress. The methodology involves the identification of major degradation agents, and the assessment of the rate of diffusion of corrosive ions and/or the rate of advancement of the reaction front into the concrete. Accelerated test methods are being used on laboratory specimens in conjunction with extrapolation procedures to predict long-term durability from short-term data. The inherent limitations are also examined

  3. Identification of snake bradykinin-potentiating peptides (BPPs)-simile sequences in rat brain--Potential BPP-like precursor protein?

    Science.gov (United States)

    Campeiro, Joana D'Arc; Neshich, Izabella P; Sant'Anna, Osvaldo A; Lopes, Robson; Ianzer, Danielle; Assakura, Marina T; Neshich, Goran; Hayashi, Mirian A F

    2015-08-01

    Bradykinin-potentiating peptides (BPPs) from the South American pit viper snake venom were the first natural inhibitors of the human angiotensin I-converting enzyme (ACE) described. The pioneer characterization of the BPPs precursor from the snake venom glands by our group showed for the first time the presence of the C-type natriuretic peptide (CNP) in this same viper precursor protein. The confirmation of the BPP/CNP expression in snake brain regions correlated with neuroendocrine functions stimulated us to pursue the physiological correlates of these vasoactive peptides in mammals. Notably, several snake toxins were shown to have endogenous physiological correlates in mammals. In the present work, we expressed in bacteria the BPPs domain of the snake venom gland precursor protein, and this purified recombinant protein was used to raise specific polyclonal anti-BPPs antibodies. The correspondent single protein band immune-recognized in adult rat brain cytosol was isolated by 2D-SDS/PAGE and/or HPLC, before characterization by MS fingerprint analysis, which identified this protein as superoxide dismutase (SOD, EC 1.15.1.1), a classically known enzyme with antioxidant activity and important roles in the blood pressure modulation. In silico analysis showed the exposition of the BPP-like peptide sequences on the surface of the 3D structure of rat SOD. These peptides were chemically synthesized to show the BPP-like biological activities in ex vivo and in vivo pharmacological bioassays. Taken together, our data suggest that SOD protein have the potential to be a source for putative BPP-like bioactive peptides, which once released may contribute to the blood pressure control in mammals. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. An update on sORFs.org: a repository of small ORFs identified by ribosome profiling.

    Science.gov (United States)

    Olexiouk, Volodimir; Van Criekinge, Wim; Menschaert, Gerben

    2018-01-04

    sORFs.org (http://www.sorfs.org) is a public repository of small open reading frames (sORFs) identified by ribosome profiling (RIBO-seq). This update elaborates on the major improvements implemented since its initial release. sORFs.org now additionally supports three more species (zebrafish, rat and Caenorhabditis elegans) and currently includes 78 RIBO-seq datasets, a vast increase compared to the three that were processed in the initial release. Therefore, a novel pipeline was constructed that also enables sORF detection in RIBO-seq datasets comprising solely elongating RIBO-seq data while previously, matching initiating RIBO-seq data was necessary to delineate the sORFs. Furthermore, a novel noise filtering algorithm was designed, able to distinguish sORFs with true ribosomal activity from simulated noise, consequently reducing the false positive identification rate. The inclusion of other species also led to the development of an inner BLAST pipeline, assessing sequence similarity between sORFs in the repository. Building on the proof of concept model in the initial release of sORFs.org, a full PRIDE-ReSpin pipeline was now released, reprocessing publicly available MS-based proteomics PRIDE datasets, reporting on true translation events. Next to reporting those identified peptides, sORFs.org allows visual inspection of the annotated spectra within the Lorikeet MS/MS viewer, thus enabling detailed manual inspection and interpretation. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. Repository for fissile materials

    International Nuclear Information System (INIS)

    Gablin, K.A.

    1976-01-01

    A repository for holding and storing fissile or other hazardous materials either under or above the ground is provided by enclosing one or more inner containers, such as standard steel drums, in a larger, corrosion-resistant outer shell, with a layer of foamed polyurethane occupying the space therebetween. The polyurethane foam is free of voids at its interfaces with the inner container and outer shell, and adheres to and reinforces same to provide a stress skin structure. Protection is afforded by the chemical and physical characteristics of the polyurethane foam against destructive influences such as water vapor intrusion, package leakage and damaging effects of the environment, such as freezing, electrolysis, chemical and bacterial action. The outer shell is shaped to conform generally to the shape of the inner container and is made of a tube of bituminized fiber material with endcaps of exterior grade plywood treated with wood preservative. A quantity of fluorescein dye is positioned within the inner container for monitoring each package for leakage

  6. Biospecimen repositories and cytopathology.

    Science.gov (United States)

    Krishnamurthy, Savitri

    2015-03-01

    Biospecimen repositories are important for the advancement of biomedical research. Literature on the potential for biobanking of fine-needle aspiration, gynecologic, and nongynecologic cytology specimens is very limited. The potential for biobanking of these specimens as valuable additional resources to surgically excised tissues appears to be excellent. The cervicovaginal specimens that can be used for biobanking include Papanicolaou-stained monolayer preparations and residual material from liquid-based cytology preparations. Different types of specimen preparations of fine-needle aspiration and nongynecologic specimens, including Papanicolaou-stained and Diff-Quik-stained smears, cell blocks. and dedicated passes/residual material from fine-needle aspiration stored frozen in a variety of solutions, can be used for biobanking. Because of several gaps in knowledge regarding the standard of operative procedures for the procurement, storage, and quality assessment of cytology specimens, further studies as well as national conferences and workshops are needed not only to create awareness but also to facilitate the use of cytopathology specimens for biobanking. © 2014 American Cancer Society.

  7. VHA Data Sharing Agreement Repository

    Data.gov (United States)

    Department of Veterans Affairs — The VHA Data Sharing Agreement Repository serves as a centralized location to collect and report on agreements that share VHA data with entities outside of VA. It...

  8. NIH Common Data Elements Repository

    Data.gov (United States)

    U.S. Department of Health & Human Services — The NIH Common Data Elements (CDE) Repository has been designed to provide access to structured human and machine-readable definitions of data elements that have...

  9. Privacy Impact Assessment (PIA) Repository

    Data.gov (United States)

    Department of Veterans Affairs — This repository contains Privacy Impact Assessments (PIA) that have been vetted/approved. Section 208 of the Electronic Government Act of 2002 (E-Gov Act) requires...

  10. Conceptual design of repository facilities

    International Nuclear Information System (INIS)

    Beale, H.; Engelmann, H.J.; Souquet, G.; Mayence, M.; Hamstra, J.

    1980-01-01

    As part of the European Economic Communities programme of research into underground disposal of radioactive wastes repository design studies have been carried out for application in salt deposits, argillaceous formations and crystalline rocks. In this paper the design aspects of repositories are reviewed and conceptual designs are presented in relation to the geological formations under consideration. Emphasis has been placed on the disposal of vitrified high level radioactive wastes although consideration has been given to other categories of radioactive waste

  11. Tools for Managing Repository Objects

    OpenAIRE

    Banker, Rajiv D.; Isakowitz, Tomas; Kauffman, Robert J.; Kumar, Rachna; Zweig, Dani

    1993-01-01

    working Paper Series: STERN IS-93-46 The past few years have seen the introduction of repository-based computer aided software engineering (CASE) tools which may finally enable us to develop software which is reliable and affordable. With the new tools come new challenges for management: Repository-based CASE changes software development to such an extent that traditional approaches to estimation, performance, and productivity assessment may no longer suffice - if they ever...

  12. Business models for digital repositories

    CERN Document Server

    CERN. Geneva; Bjørnshauge, Lars

    2007-01-01

    Those setting up, or planning to set up, a digital repository may be interested to know more about what has gone before them. What is involved, what is the cost, how many people are needed, how have others made the case to their institution, and how do you get anything into it once it is built? I have recently undertaken a study of European repository business models for the DRIVER project and will present an overview of the findings.

  13. Identification of bradykinin: related peptides from Phyllomedusa nordestina skin secretion using electrospray ionization tandem mass spectrometry after a single-step liquid chromatography

    Directory of Open Access Journals (Sweden)

    K Conceição

    2009-01-01

    Full Text Available Amphibian skin secretions are a source of potential new drugs with medical and biotechnological applications. Rich in peptides produced by holocrine-type serous glands in the integument, these secretions play different roles, either in the regulation of physiological skin functions or in the defense against predators or microorganisms. The aim of the present work was to identify novel peptides with bradykinin-like structure and/or activity present in the skin of Phyllomedusa nordestina. In order to achieve this goal, the crude skin secretion of this frog was pre-fractionated by solid phase extraction and separated by reversed-phase chromatography. The fractions were screened for low-molecular-mass peptides and sequenced by mass spectrometry. It was possible to identify three novel bradykinin-related peptides, namely: KPLWRL-NH2 (Pnor 3, RPLSWLPK (Pnor 5 and VPPKGVSM (Pnor 7 presenting vascular activities as assessed by intravital microscopy. Pnor 3 and Pnor 7 were able to induce vasodilation. On the other hand, Pnor 5 was a potent vasoconstrictor. These effects were reproduced by their synthetic analogues.

  14. Identification of a moronecidin-like antimicrobial peptide in the venomous fish Pterois volitans: Functional and structural study of pteroicidin-α.

    Science.gov (United States)

    Houyvet, Baptiste; Bouchon-Navaro, Yolande; Bouchon, Claude; Goux, Didier; Bernay, Benoît; Corre, Erwan; Zatylny-Gaudin, Céline

    2018-01-01

    The present study characterizes for the first time an antimicrobial peptide in lionfish (Pterois volitans), a venomous fish. Using a peptidomic approach, we identified a mature piscidin in lionfish and called it pteroicidin-α. We detected an amidated form (pteroicidin-α- CONH 2 ) and a non-amidated form (pteroicidin-α-COOH), and then performed their functional and structural study. Interestingly, the two peptides displayed different antibacterial and hemolytic activity levels. Pteroicidin-α-CONH 2 was bactericidal on human pathogens like Staphylococcus aureus or Escherichia coli, as well as on the fish pathogen Aeromonas salmonicida, while pteroicidin-α-COOH only inhibited their growth. Furthermore, the two peptides induced hemolysis of red blood cells from different vertebrates, namely humans, sea bass and lesser-spotted dogfish. Hemolysis occurred with low concentrations of pteroicidin-α-CONH 2 , indicating greater toxicity of the amidated form. Circular dichroism analysis showed that both peptides adopted a helical conformation, yet with a greater α-helix content in pteroicidin-α-CONH 2 . Overall, these results suggest that amidation strongly influences pteroicidin-α by modifying its structure and its physico-chemical characteristics and by increasing its hemolytic activity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Identification of a cDNA encoding a parathyroid hormone-like peptide from a human tumor associated with humoral hypercalcemia of malignancy

    International Nuclear Information System (INIS)

    Mangin, M.; Webb, A.C.; Dreyer, B.E.

    1988-01-01

    Humoral hypercalcemia of malignancy is a common paraneoplastic syndrome that appears to be mediated in many instances by a parathyroid hormone-like peptide. Poly(A) + RNA from a human renal carcinoma associated with this syndrome was enriched by preparative electrophoresis and used to construct an enriched cDNA library in phage λgt10. The library was screened with a codon-preference oligonucleotide synthesized on the basis of a partial N-terminal amino acid sequence from a human tumor-derived peptide, and a 2.0 kilo-base cDNA was identified. The cDNA encodes a 177 amino acid protein consisting of a 36 amino acid leader sequence and a 141 amino acid mature peptide. The first 13 amino acids of the deduced sequence of the mature peptide display strong homology to human PTH, with complete divergence thereafter. RNA blot-hybridization analysis revealed multiple transcripts in mRNA from tumors associated with the humor syndrome and also in mRNA from normal human keratinocytes. Southern blot analysis of genomic DNA from humans and rodents revealed a simple pattern compatible with a single-copy gene. The gene has been mapped to chromosome 12

  16. IAEA safeguards for geological repositories

    International Nuclear Information System (INIS)

    Moran, B.W.

    2005-01-01

    In September. 1988, the IAEA held its first formal meeting on the safeguards requirements for the final disposal of spent fuel and nuclear material-bearing waste. The consensus recommendation of the 43 participants from 18 countries at this Advisory Group Meeting was that safeguards should not terminate of spent fuel even after emplacement in, and closure of, a geologic repository.' As a result of this recommendation, the IAEA initiated a series of consultants' meetings and the SAGOR Programme (Programme for the Development of Safeguards for the Final Disposal of Spent Fuel in Geologic Repositories) to develop an approach that would permit IAEA safeguards to verify the non-diversion of spent fuel from a geologic repository. At the end of this process, in December 1997, a second Advisory Group Meeting, endorsed the generic safeguards approach developed by the SAGOR Programme. Using the SAGOR Programme results and consultants' meeting recommendations, the IAEA Department of Safeguards issued a safeguards policy paper stating the requirements for IAEA safeguards at geologic repositories. Following approval of the safeguards policy and the generic safeguards approach, the Geologic Repository Safeguards Experts Group was established to make recommendations on implementing the safeguards approach. This experts' group is currently making recommendations to the IAEA regarding the safeguards activities to be conducted with respect to Finland's repository programme. (author)

  17. Identification of a novel aFGF-binding peptide with anti-tumor effect on breast cancer from phage display library

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Xiaoyong; Cai, Cuizan [College of Pharmacy, Jinan University, Guangzhou 510632, Guangdong (China); Xiao, Fei [Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, Guangdong (China); Xiong, Yaoling [College of Pharmacy, Jinan University, Guangzhou 510632, Guangdong (China); Huang, Yadong; Zhang, Qihao [Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, Guangdong (China); Xiang, Qi [College of Pharmacy, Jinan University, Guangzhou 510632, Guangdong (China); Lou, Guofeng [Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, Guangdong (China); Lian, Mengyang [College of Pharmacy, Jinan University, Guangzhou 510632, Guangdong (China); Su, Zhijian, E-mail: tjnuszj@jnu.edu.cn [Department of Biopharmaceutical Research and Development Centre, Institute of Biomedicine, Jinan University, Guangzhou 510632, Guangdong (China); Zheng, Qing, E-mail: tzhengq@jnu.edu.cn [College of Pharmacy, Jinan University, Guangzhou 510632, Guangdong (China)

    2014-03-21

    Highlights: • A specific aFGF-binding peptide AP8 was identified from a phage display library. • AP8 could inhibit aFGF-stimulated cell proliferation in a dose-dependent manner. • AP8 arrested the cell cycle at the G0/G1 phase by suppressing Cyclin D1. • AP8 could block the activation of Erk1/2 and Akt kinase. • AP8 counteracted proliferation and cell cycle via influencing PA2G4 and PCNA. - Abstract: It has been reported that acidic fibroblast growth factor (aFGF) is expressed in breast cancer and via interactions with fibroblast growth factor receptors (FGFRs) to promote the stage and grade of the disease. Thus, aFGF/FGFRs have been considered essential targets in breast cancer therapy. We identified a specific aFGF-binding peptide (AGNWTPI, named AP8) from a phage display heptapeptide library with aFGF after four rounds of biopanning. The peptide AP8 contained two (TP) amino acids identical and showed high homology to the peptides of the 182–188 (GTPNPTL) site of high-affinity aFGF receptor FGFR1. Functional analyses indicated that AP8 specifically competed with the corresponding phage clone A8 for binding to aFGF. In addition, AP8 could inhibit aFGF-stimulated cell proliferation, arrested the cell cycle at the G0/G1 phase by increasing PA2G4 and suppressing Cyclin D1 and PCNA, and blocked the aFGF-induced activation of Erk1/2 and Akt kinase in both breast cancer cells and vascular endothelial cells. Therefore, these results indicate that peptide AP8, acting as an aFGF antagonist, is a promising therapeutic agent for the treatment of breast cancer.

  18. Radioactive waste repository of high ecological safety

    International Nuclear Information System (INIS)

    Sobolev, I.; Barinov, A.; Prozorov, L.

    2000-01-01

    With the purpose to construct a radioactive waste repository of high ecological safety and reliable containment, MosNPO 'Radon' specialists have developed an advanced type repository - large diameter well (LBD) one. A project is started for the development of a technology for LDW repository construction and pilot operation of the new repository for 25-30 years. The 2 LDW repositories constructed at the 'Radon' site and the developed monitoring system are described

  19. Demystifying the institutional repository for success

    CERN Document Server

    Buehler, Marianne

    2013-01-01

    Institutional repositories remain key to data storage on campus, fulfilling the academic needs of various stakeholders. Demystifying the Institutional Repository for Success is a practical guide to creating and sustaining an institutional repository through marketing, partnering, and understanding the academic needs of all stakeholders on campus. This title is divided into seven chapters, covering: traditional scholarly communication and open access publishing; the academic shift towards open access; what the successful institutional repository looks like; institutional repository collaboratio

  20. Virtual patient repositories--a comparative analysis.

    Science.gov (United States)

    Küfner, Julia; Kononowicz, Andrzej A; Hege, Inga

    2014-01-01

    Virtual Patients (VPs) are an important component of medical education. One way to reduce the costs for creating VPs is sharing through repositories. We conducted a literature review to identify existing repositories and analyzed the 17 included repositories in regards to the search functions and metadata they provide. Most repositories provided some metadata such as title or description, whereas other data, such as educational objectives, were less frequent. Future research could, in cooperation with the repository provider, investigate user expectations and usage patterns.

  1. Cardioprotective peptides from marine sources.

    Science.gov (United States)

    Harnedy, Padraigín A; FitzGerald, Richard J

    2013-05-01

    Elevated blood pressure or hypertension is one of the fastest growing health problems worldwide. Although the etiology of essential hypertension has a genetic component, dietary factors play an important role. With the high costs and adverse side-effects associated with synthetic antihypertensive drugs and the awareness of the link between diet and health there has been increased focus on identification of food components that may contribute to cardiovascular health. In recent years special interest has been paid to the cardioprotective activity of peptides derived from food proteins including marine proteins. These peptides are latent within the sequence of the parent protein and only become active when released by proteolytic digestion during gastrointestinal digestion or through food processing. Current data on antihypertensive activity of marine-derived protein hydrolysates/peptides in animal and human studies is reviewed herein. Furthermore, products containing protein hydrolysates/peptides from marine origin with antihypertensive effects are discussed.

  2. Watery diarrhea syndrome in an adult with ganglioneuroma-pheochromocytoma: identification of vasoactive intestinal peptide, calcitonin, and catecholamines and assessment of their biologic activity.

    Science.gov (United States)

    Trump, D L; Livingston, J N; Baylin, S B

    1977-10-01

    A case of adult ganglioneuroma-pheochromocytoma with an associated watery diarrhea syndrome is reported. High levels of vasoactive intestinal peptide (VIP) were found in preoperative serum and in tumor tissue. The serum VIP levels fell to normal, and the watery diarrhae syndrome completely ceased following removal of the tumor. In addition to containing VIP, the tumor was rich in catecholamines, and calcitonin. Peptide hormone-containing extracts and catecholamine extracts from the tumor both activated the adenyl cyclase system and increased lipolytic activity in a preparation of isolated rat fat cells. The findings in this patient further link VIP with neural crest tissues, and suggest the importance of determining catecholamine levels in patients with the watery diarrhea syndrome.

  3. Identification of neurotensin-related peptides in human thymic epithelial cell membranes and relationship with major histocompatibility complex class I molecules.

    Science.gov (United States)

    Vanneste, Y; Thome, A N; Vandersmissen, E; Charlet, C; Franchimont, D; Martens, H; Lhiaubet, A M; Schimpff, R M; Rostène, W; Geenen, V

    1997-06-01

    This study shows the expression at the cell surface of human thymic epithelial cells (TEC) of a neurotensin (NT)-like immunoreactivity. NT radio-immunoassay (RIA) revealed that cultured human TEC contain +/-5 ng immunoreactive (ir) NT/10(6) cells, of which 5% is associated with plasma cell membranes. HPLC analysis of NT-ir present in human TEC showed a major peak of NT-ir corresponding to NT1-13. NT-ir was not detected in the supernatant of human TEC cultures. Using an affinity column prepared with a anti-MHC class I monoclonal antibody, NT-ir-related peptides were retained on the column and eluted together with MHC class I-related proteins. According to the elution time on HPLC of these peptides, they correspond to intact NT1-13, as well as to smaller fragments of NT1-13.

  4. Medicago PhosphoProtein Database: a repository for Medicago truncatula phosphoprotein data

    Directory of Open Access Journals (Sweden)

    Christopher M. Rose

    2012-06-01

    Full Text Available The ability of legume crops to fix atmospheric nitrogen via a symbiotic association with soil rhizobia makes them an essential component of many agricultural systems. Initiation of this symbiosis requires protein phosphorylation-mediated signaling in response to rhizobial signals named Nod factors. Medicago truncatula (Medicago is the model system for studying legume biology, making the study of its phosphoproteome essential. Here, we describe the Medicago Phosphoprotein Database (http://phospho.medicago.wisc.edu, a repository built to house phosphoprotein, phosphopeptide, and phosphosite data specific to Medicago. Currently, the Medicago Phosphoprotein Database holds 3,457 unique phosphopeptides that contain 3,404 non-redundant sites of phosphorylation on 829 proteins. Through the web-based interface, users are allowed to browse identified proteins or search for proteins of interest. Furthermore, we allow users to conduct BLAST searches of the database using both peptide sequences and phosphorylation motifs as queries. The data contained within the database are available for download to be investigated at the user’s discretion. The Medicago Phosphoprotein Database will be updated continually with novel phosphoprotein and phosphopeptide identifications, with the intent of constructing an unparalleled compendium of large-scale Medicago phosphorylation data.

  5. Multicenter Evaluation of a New Shortened Peptide Nucleic Acid Fluorescence In Situ Hybridization Procedure for Species Identification of Select Gram-Negative Bacilli from Blood Cultures▿

    Science.gov (United States)

    Morgan, Margie; Marlowe, Elizabeth; Della-Latta, Phyllis; Salimnia, Hossein; Novak-Weekley, Susan; Wu, Fann; Crystal, Benjamin S.

    2010-01-01

    A shortened protocol for two peptide nucleic acid fluorescence in situ hybridization (PNA FISH) assays for the detection of Gram-negative bacilli from positive blood cultures was evaluated in a multicenter trial. There was 100% concordance between the two protocols for each assay (368 of 368 and 370 of 370 results) and 99.7% (367 of 368 and 369 of 370 results) agreement with routine laboratory techniques. PMID:20357212

  6. Purification, identification and molecular mechanism of two dipeptidyl peptidase IV (DPP-IV) inhibitory peptides from Antarctic krill (Euphausia superba) protein hydrolysate.

    Science.gov (United States)

    Ji, Wei; Zhang, Chaohua; Ji, Hongwu

    2017-10-01

    Dipeptidyl peptidase IV (DPP-IV) played an important role in blood glucose regulation. Inhibition of DPP-IV may improve glycemic control in diabetics by preventing the rapid breakdown of incretin hormones and prolonging their physiological action. In this study, Antarctic krill (Euphausia superba) protein was hydrolyzed using animal proteolytic enzymes. The hydrolysate was purified sequentially by ultrafiltration, gel filtration chromatography and reversed phase high-performance liquid chromatography (RP-HPLC). DPP-IV inhibitory activity of the fractions achieved from Antarctic krill protein was determined by DPP-IV screening reagent kit. Two purified peptides were identified by Xevo G2-XS QTof mass spectrometer (QTOF-MS). One peptide purified was Ala-Pro (AP) with IC 50 values of 0.0530mg/mL, the other Ile-Pro-Ala (IPA) with IC 50 values of 0.0370mg/mL. They both exhibited strong DPP-IV inhibitory activity. The molecular docking analysis revealed that DPP-IV inhibition by AP and IPA was mainly due to formation of a strong interaction surface force with the 91-96 and 101-105 amino acids of the DPP-IV. Our results suggested that the protein hydrolysate from Antarctic krill can be considered as a promising natural source of DPP-IV inhibitory peptides in the management of diabetes. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Identification of immunity-related genes in Plutella xylostella in response to fungal peptide destruxin A: RNA-Seq and DGE analysis.

    Science.gov (United States)

    Shakeel, Muhammad; Xu, Xiaoxia; Xu, Jin; Zhu, Xun; Li, Shuzhong; Zhou, Xianqiang; Yu, Jialin; Xu, Xiaojing; Hu, Qiongbo; Yu, Xiaoqiang; Jin, Fengliang

    2017-09-08

    Plutella xylostella has become the major lepidopteran pest of Brassica owing to its strong ability of resistance development to a wide range of insecticides. Destruxin A, a mycotoxin of entomopathogenic fungus, Metarhizium anisopliae, has broad-spectrum insecticidal effects. However, the interaction mechanism of destruxin A with the immune system of P. xylostella at genomic level is still not well understood. Here, we identified 129 immunity-related genes, including pattern recognition receptors, signal modulators, few members of main immune pathways (Toll, Imd, and JAK/STAT), and immune effectors in P. xylostella in response to destruxin A at three different time courses (2 h, 4 h, and 6 h). It is worthy to mention that the immunity-related differentially expressed genes (DEGs) analysis exhibited 30, 78, and 72 up-regulated and 17, 13, and 6 down-regulated genes in P. xylostella after destruxin A injection at 2 h, 4 h, and 6 h, respectively, compared to control. Interestingly, our results revealed that the expression of antimicrobial peptides that play a vital role in insect immune system was up-regulated after the injection of destruxin A. Our findings provide a detailed information on immunity-related DEGs and reveal the potential of P. xylostella to limit the infection of fungal peptide destruxin A by increasing the activity of antimicrobial peptides.

  8. Biological Web Service Repositories Review.

    Science.gov (United States)

    Urdidiales-Nieto, David; Navas-Delgado, Ismael; Aldana-Montes, José F

    2017-05-01

    Web services play a key role in bioinformatics enabling the integration of database access and analysis of algorithms. However, Web service repositories do not usually publish information on the changes made to their registered Web services. Dynamism is directly related to the changes in the repositories (services registered or unregistered) and at service level (annotation changes). Thus, users, software clients or workflow based approaches lack enough relevant information to decide when they should review or re-execute a Web service or workflow to get updated or improved results. The dynamism of the repository could be a measure for workflow developers to re-check service availability and annotation changes in the services of interest to them. This paper presents a review on the most well-known Web service repositories in the life sciences including an analysis of their dynamism. Freshness is introduced in this paper, and has been used as the measure for the dynamism of these repositories. © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  9. Development of site suitability criteria for the high level waste repository for Lawrence Livermore Laboratories

    International Nuclear Information System (INIS)

    1977-06-01

    Results of our mining, geological and geotechnical studies provided in support of the development of site suitability criteria for the high level waste repository are presented. The primary purpose of the work was the identification and development of appropriate geotechnical descriptors and coefficients required for the Site Suitability Repository Model. This model was developed by The Analytic Sciences Corporation (TASC) of Reading, Massachusetts and is not described in this report

  10. Development of site suitability criteria for the high level waste repository for Lawrence Livermore Laboratories

    Energy Technology Data Exchange (ETDEWEB)

    1977-06-01

    Results of our mining, geological and geotechnical studies provided in support of the development of site suitability criteria for the high level waste repository are presented. The primary purpose of the work was the identification and development of appropriate geotechnical descriptors and coefficients required for the Site Suitability Repository Model. This model was developed by The Analytic Sciences Corporation (TASC) of Reading, Massachusetts and is not described in this report.

  11. Harvesting NASA's Common Metadata Repository

    Science.gov (United States)

    Shum, D.; Mitchell, A. E.; Durbin, C.; Norton, J.

    2017-12-01

    As part of NASA's Earth Observing System Data and Information System (EOSDIS), the Common Metadata Repository (CMR) stores metadata for over 30,000 datasets from both NASA and international providers along with over 300M granules. This metadata enables sub-second discovery and facilitates data access. While the CMR offers a robust temporal, spatial and keyword search functionality to the general public and international community, it is sometimes more desirable for international partners to harvest the CMR metadata and merge the CMR metadata into a partner's existing metadata repository. This poster will focus on best practices to follow when harvesting CMR metadata to ensure that any changes made to the CMR can also be updated in a partner's own repository. Additionally, since each partner has distinct metadata formats they are able to consume, the best practices will also include guidance on retrieving the metadata in the desired metadata format using CMR's Unified Metadata Model translation software.

  12. Material control and accountability procedures for a waste isolation repository

    International Nuclear Information System (INIS)

    Jenkins, J.D.; Allen, E.J.; Blakeman, E.D.

    1978-05-01

    The material control and accountability needs of a waste isolation repository are examined. Three levels of control are discussed: (1) item identification and control, (2) tamper indication, and (3) quantitative material assay. A summary of waste characteristics is presented and, based on these, plus a consideration of the accessibility of the various types of waste, material control by item identification and accountability (where the individual waste container is the basic unit) is recommended. Tamper indicating procedures are also recommended for the intermediate and low level waste categories

  13. Recommendations for certification or measurement of reliability for reliable digital archival repositories with emphasis on access

    Directory of Open Access Journals (Sweden)

    Paula Regina Ventura Amorim Gonçalez

    2017-04-01

    Full Text Available Introduction: Considering the guidelines of ISO 16363: 2012 (Space data and information transfer systems -- Audit and certification of trustworthy digital repositories and the text of CONARQ Resolution 39 for certification of Reliable Digital Archival Repository (RDC-Arq, verify the technical recommendations should be used as the basis for a digital archival repository to be considered reliable. Objective: Identify requirements for the creation of Reliable Digital Archival Repositories with emphasis on access to information from the ISO 16363: 2012 and CONARQ Resolution 39. Methodology: For the development of the study, the methodology consisted of an exploratory, descriptive and documentary theoretical investigation, since it is based on ISO 16363: 2012 and CONARQ Resolution 39. From the perspective of the problem approach, the study is qualitative and quantitative, since the data were collected, tabulated, and analyzed from the interpretation of their contents. Results: We presented a set of Checklist Recommendations for reliability measurement and/or certification for RDC-Arq with a clipping focused on the identification of requirements with emphasis on access to information is presented. Conclusions: The right to information as well as access to reliable information is a premise for Digital Archival Repositories, so the set of recommendations is directed to archivists who work in Digital Repositories and wish to verify the requirements necessary to evaluate the reliability of the Digital Repository or still guide the information professional in collecting requirements for repository reliability certification.

  14. Overly Honest Data Repository Development

    Directory of Open Access Journals (Sweden)

    Colleen Fallaw

    2016-10-01

    Full Text Available After a year of development, the library at the University of Illinois at Urbana-Champaign has launched a repository, called the Illinois Data Bank (https://databank.illinois.edu/, to provide Illinois researchers with a free, self-serve publishing platform that centralizes, preserves, and provides persistent and reliable access to Illinois research data. This article presents a holistic view of development by discussing our overarching technical, policy, and interface strategies. By openly presenting our design decisions, the rationales behind those decisions, and associated challenges this paper aims to contribute to the library community's work to develop repository services that meet growing data preservation and sharing needs.

  15. Peptide-Based Vaccinology: Experimental and Computational Approaches to Target Hypervariable Viruses through the Fine Characterization of Protective Epitopes Recognized by Monoclonal Antibodies and the Identification of T-Cell-Activating Peptides

    Directory of Open Access Journals (Sweden)

    Matteo Castelli

    2013-01-01

    Full Text Available Defining immunogenic domains of viral proteins capable of eliciting a protective immune response is crucial in the development of novel epitope-based prophylactic strategies. This is particularly important for the selective targeting of conserved regions shared among hypervariable viruses. Studying postinfection and postimmunization sera, as well as cloning and characterization of monoclonal antibodies (mAbs, still represents the best approach to identify protective epitopes. In particular, a protective mAb directed against conserved regions can play a key role in immunogen design and in human therapy as well. Experimental approaches aiming to characterize protective mAb epitopes or to identify T-cell-activating peptides are often burdened by technical limitations and can require long time to be correctly addressed. Thus, in the last decade many epitope predictive algorithms have been developed. These algorithms are continually evolving, and their use to address the empirical research is widely increasing. Here, we review several strategies based on experimental techniques alone or addressed by in silico analysis that are frequently used to predict immunogens to be included in novel epitope-based vaccine approaches. We will list the main strategies aiming to design a new vaccine preparation conferring the protection of a neutralizing mAb combined with an effective cell-mediated response.

  16. Identification of PrP sequences essential for the interaction between the PrP polymers and Aβ peptide in a yeast-based assay

    OpenAIRE

    Rubel, Aleksandr A; Ryzhova, Tatyana A; Antonets, Kirill S; Chernoff, Yury O; Galkin, Alexey P

    2013-01-01

    Alzheimer disease is associated with the accumulation of oligomeric amyloid β peptide (Aβ), accompanied by synaptic dysfunction and neuronal death. Polymeric form of prion protein (PrP), PrPSc, is implicated in transmissible spongiform encephalopathies (TSEs). Recently, it was shown that the monomeric cellular form of PrP (PrPC), located on the neuron surface, binds Aβ oligomers (and possibly other β-rich conformers) via the PrP23–27 and PrP90–110 segments, acting as Aβ receptor. On the other...

  17. A combined prediction strategy increases identification of peptides bound with high affinity and stability to porcine MHC class I molecules SLA-1*04:01, SLA-2*04:01, and SLA-3*04:01

    DEFF Research Database (Denmark)

    Pedersen, Lasse Eggers; Rasmussen, Michael; Harndahl, Mikkel

    2016-01-01

    constitute an attractive protocol to select target peptides from the vast pool of viral proteome peptides. We have earlier reported the peptide binding motif of the porcine MHC-I molecules SLA-1*04:01 and SLA-2*04:01, identified by an ELISA affinity-based positional scanning combinatorial peptide library...

  18. Hormone action. Part I. Peptide hormones

    International Nuclear Information System (INIS)

    Birnbaumer, L.; O'Malley, B.W.

    1985-01-01

    The major sections of this book on the hormonal action of peptide hormones cover receptor assays, identification of receptor proteins, methods for identification of internalized hormones and hormone receptors, preparation of hormonally responsive cells and cell hybrids, purification of membrane receptors and related techniques, assays of hormonal effects and related functions, and antibodies in hormone action

  19. Identification of a Short Cell-Penetrating Peptide from Bovine Lactoferricin for Intracellular Delivery of DNA in Human A549 Cells.

    Directory of Open Access Journals (Sweden)

    Betty R Liu

    Full Text Available Cell-penetrating peptides (CPPs have been shown to deliver cargos, including protein, DNA, RNA, and nanomaterials, in fully active forms into live cells. Most of the CPP sequences in use today are based on non-native proteins that may be immunogenic. Here we demonstrate that the L5a CPP (RRWQW from bovine lactoferricin (LFcin, stably and noncovalently complexed with plasmid DNA and prepared at an optimal nitrogen/phosphate ratio of 12, is able to efficiently enter into human lung cancer A549 cells. The L5a CPP delivered a plasmid containing the enhanced green fluorescent protein (EGFP coding sequence that was subsequently expressed in cells, as revealed by real-time PCR and fluorescent microscopy at the mRNA and protein levels, respectively. Treatment with calcium chloride increased the level of gene expression, without affecting CPP-mediated transfection efficiency. Zeta-potential analysis revealed that positively electrostatic interactions of CPP/DNA complexes correlated with CPP-mediated transport. The L5a and L5a/DNA complexes were not cytotoxic. This biomimetic LFcin L5a represents one of the shortest effective CPPs and could be a promising lead peptide with less immunogenic for DNA delivery in gene therapy.

  20. Identification of a Short Cell-Penetrating Peptide from Bovine Lactoferricin for Intracellular Delivery of DNA in Human A549 Cells.

    Science.gov (United States)

    Liu, Betty R; Huang, Yue-Wern; Aronstam, Robert S; Lee, Han-Jung

    2016-01-01

    Cell-penetrating peptides (CPPs) have been shown to deliver cargos, including protein, DNA, RNA, and nanomaterials, in fully active forms into live cells. Most of the CPP sequences in use today are based on non-native proteins that may be immunogenic. Here we demonstrate that the L5a CPP (RRWQW) from bovine lactoferricin (LFcin), stably and noncovalently complexed with plasmid DNA and prepared at an optimal nitrogen/phosphate ratio of 12, is able to efficiently enter into human lung cancer A549 cells. The L5a CPP delivered a plasmid containing the enhanced green fluorescent protein (EGFP) coding sequence that was subsequently expressed in cells, as revealed by real-time PCR and fluorescent microscopy at the mRNA and protein levels, respectively. Treatment with calcium chloride increased the level of gene expression, without affecting CPP-mediated transfection efficiency. Zeta-potential analysis revealed that positively electrostatic interactions of CPP/DNA complexes correlated with CPP-mediated transport. The L5a and L5a/DNA complexes were not cytotoxic. This biomimetic LFcin L5a represents one of the shortest effective CPPs and could be a promising lead peptide with less immunogenic for DNA delivery in gene therapy.

  1. Effect of Different Heat Treatments on In Vitro Digestion of Egg White Proteins and Identification of Bioactive Peptides in Digested Products.

    Science.gov (United States)

    Wang, Xuefen; Qiu, Ning; Liu, Yaping

    2018-04-01

    Chicken eggs are ingested by people after a series of processes, but to date, only a few studies have explored the nutrient variations caused by different heat treatments. In this work, the impacts of different heat treatments (4, 56, 65, and 100 °C on the in vitro digestibility of egg white proteins were investigated by hydrolysis with pepsin or pepsin + pancreatin to simulate human gastrointestinal digestion, and the digested products were identified using Nano-LC-ESI-MS/MS. Egg white proteins treated at 65 °C had the highest in vitro pepsin digestibility value whereas the pepsin + pancreatin digestibility increased significantly (P cooking temperature was raised. The molecular weight distribution of the digested products indicated that, when compared to pepsin-treated samples, pepsin + pancreatin-treated samples contained more low-molecular-weight peptides (m/z egg white digested products, especially in samples treated at 4 and 100 °C. These findings may facilitate a better understanding of nutritive values of egg white proteins and their digested products under different cooking temperatures, such as antibacterial and antioxidant peptides identified in the digestion samples treated, respectively at 4 and 100 °C. This study also provided information for improving the applications of eggs in the food industry as well as a theoretical basis for egg consumption. © 2018 Institute of Food Technologists®.

  2. Cancer Epidemiology Data Repository (CEDR)

    Science.gov (United States)

    In an effort to broaden access and facilitate efficient data sharing, the Epidemiology and Genomics Research Program (EGRP) has created the Cancer Epidemiology Data Repository (CEDR), a centralized, controlled-access database, where Investigators can deposit individual-level de-identified observational cancer datasets.

  3. Repository operational criteria comparative analysis

    International Nuclear Information System (INIS)

    Hageman, J.P.; Chowdhury, A.H.

    1994-06-01

    The objective of the ''Repository Operational Criteria (ROC) Feasibility Studies'' (or ROC task) was to conduct comprehensive and integrated analyses of repository design, construction, and operations criteria in 10 CFR Part 60 regulations considering the interfaces among the components of the regulations and impacts of any potential changes to those regulations. The ROC task addresses regulatory criteria and uncertainties related to the preclosure aspects of the geologic repository. Those parts of 10 CFR Part 60 that require routine guidance or minor changes to the rule were addressed in Hageman and Chowdhury, 1992. The ROC task shows a possible need for further regulatory clarity, by major changes to the rule, related to the design bases and siting of a geologic repository operations area and radiological emergency planning in order to assure defense-in-depth. The analyses, presented in this report, resulted in the development and refinement of regulatory concepts and their supporting rationale for recommendations for potential major changes to 10 CFR Pan 0 regulations

  4. NPP Krsko: LILW Repository or Long Term Storage

    International Nuclear Information System (INIS)

    Lokner, V.; Subasic, D.; Levanat, I.

    2008-01-01

    Construction of the facilities for LILW and SF management, as planned in Decommissioning and LILW and SF management program for NPP Krsko, would be a rather expensive and challenging project for such a small nuclear program. In order to accommodate waste arising from a single nuclear power plant, one LILW repository should be constructed before the end of the NPP operation, then one SF dry storage, and finally one geological repository. This requires relatively urgent identification within Slovenian/Croatian territory of three locations that meet the criteria for establishment of such facilities and are acceptable to the local communities. There are very few such potential locations. The siting process for the first of the three facilities is well under way in Slovenia, because the country wants to have its LILW repository in operation by the year 2013. In order to facilitate public acceptance, Slovenian government has introduced financial incentives to local communities for the repository construction and operation. These 'compensations for limited land use' may significantly increase the overall costs of disposal if the repository is in operation for a long period. In the recent years, however, a possibility of long term storage (LTS) is gaining an increased attention in the waste management community, and has already been introduced e.g. in the Netherlands. It is a particularly viable option for limited waste quantities. Disposal remains the final solution, but present technologies have made possible a relatively inexpensive storage up to about hundred years, which can accommodate LILW, HLW and SF from nuclear programs as well as research reactor waste and NORM. Such storage would be a safe and simple temporary solution, encompassing all immediate and near future waste management needs. In addition, it would increase flexibility and reduce financing requirements for the final waste disposal: providing additional time for reduction of radiation emission and heat

  5. Consistency of Network Traffic Repositories: An Overview

    NARCIS (Netherlands)

    Lastdrager, E.; Lastdrager, E.E.H.; Pras, Aiko

    2009-01-01

    Traffc repositories with TCP/IP header information are very important for network analysis. Researchers often assume that such repositories reliably represent all traffc that has been flowing over the network; little thoughts are made regarding the consistency of these repositories. Still, for

  6. Consistency analysis of network traffic repositories

    NARCIS (Netherlands)

    Lastdrager, Elmer; Lastdrager, E.E.H.; Pras, Aiko

    Traffic repositories with TCP/IP header information are very important for network analysis. Researchers often assume that such repositories reliably represent all traffic that has been flowing over the network; little thoughts are made regarding the consistency of these repositories. Still, for

  7. Identification of a cell-penetrating peptide domain from human beta-defensin 3 and characterization of its anti-inflammatory activity

    Directory of Open Access Journals (Sweden)

    Lee JY

    2015-08-01

    Full Text Available Jue Yeon Lee,1,* Jin Sook Suh,2,* Jung Min Kim,1 Jeong Hwa Kim,1 Hyun Jung Park,1 Yoon Jeong Park,1,2 Chong Pyoung Chung1 1Central Research Institute, Nano Intelligent Biomedical Engineering Corporation (NIBEC, Chungcheongbuk-do, Republic of Korea; 2Dental Regenerative Biotechnology, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Republic of Korea *These authors contributed equally to this work Abstract: Human beta-defensins (hBDs are crucial factors of intrinsic immunity that function in the immunologic response to a variety of invading enveloped viruses, bacteria, and fungi. hBDs can cause membrane depolarization and cell lysis due to their highly cationic nature. These molecules participate in antimicrobial defenses and the control of adaptive and innate immunity in every mammalian species and are produced by various cell types. The C-terminal 15-mer peptide within hBD3, designated as hBD3-3, was selected for study due to its cell- and skin-penetrating activity, which can induce anti-inflammatory activity in lipopolysaccharide-treated RAW 264.7 macrophages. hBD3-3 penetrated both the outer membrane of the cells and mouse skin within a short treatment period. Two other peptide fragments showed poorer penetration activity compared to hBD3-3. hBD3-3 inhibited the lipopolysaccharide-induced production of inducible nitric oxide synthase, nitric oxide, and secretory cytokines, such as interleukin-6 and tumor necrosis factor in a concentration-dependent manner. Moreover, hBD3-3 reduced the interstitial infiltration of polymorphonuclear leukocytes in a lung inflammation model. Further investigation also revealed that hBD3-3 downregulated nuclear factor kappa B-dependent inflammation by directly suppressing the degradation of phosphorylated-IκBα and by downregulating active nuclear factor kappa B p65. Our findings indicate that hBD3-3 may be conjugated with drugs of interest to ensure their proper translocation to

  8. Peptide chemistry toolbox - Transforming natural peptides into peptide therapeutics.

    Science.gov (United States)

    Erak, Miloš; Bellmann-Sickert, Kathrin; Els-Heindl, Sylvia; Beck-Sickinger, Annette G

    2018-06-01

    The development of solid phase peptide synthesis has released tremendous opportunities for using synthetic peptides in medicinal applications. In the last decades, peptide therapeutics became an emerging market in pharmaceutical industry. The need for synthetic strategies in order to improve peptidic properties, such as longer half-life, higher bioavailability, increased potency and efficiency is accordingly rising. In this mini-review, we present a toolbox of modifications in peptide chemistry for overcoming the main drawbacks during the transition from natural peptides to peptide therapeutics. Modifications at the level of the peptide backbone, amino acid side chains and higher orders of structures are described. Furthermore, we are discussing the future of peptide therapeutics development and their impact on the pharmaceutical market. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. Identification of ischemia-regulated phosphorylation sites in connexin43: A possible target for the antiarrhythmic peptide analogue rotigaptide (ZP123)

    DEFF Research Database (Denmark)

    Axelsen, Lene Nygaard; Stahlhut, Martin; Mohammed, Shabaz

    2006-01-01

    Previous studies suggest that dephosphorylation of connexin43 (Cx43) is related to uncoupling of gap junction communication, which plays an important role in the genesis of ischemia-induced ventricular tachycardia. We studied changes in Cx43 phosphorylation during global ischemia in the absence...... and presence of the antiarrhythmic peptide analogue rotigaptide (formerly known as ZP123). Phosphorylation analysis was performed on Cx43 purified from isolated perfused rat hearts using matrix-assisted laser desorption/ionization mass spectrometry and liquid chromatography electrospray ionization tandem mass...... of ischemia, the critical time interval where gap junction uncoupling occurs, Ser297 and Ser368 also became fully dephosphorylated. During the same time period, all untreated hearts developed asystole. Treatment with rotigaptide significantly increased the time to ischemia-induced asystole and suppressed...

  10. Proteome identification of the silkworm middle silk gland

    Directory of Open Access Journals (Sweden)

    Jian-ying Li

    2016-03-01

    Full Text Available To investigate the functional differentiation among the anterior (A, middle (M, and posterior (P regions of silkworm middle silk gland (MSG, their proteomes were characterized by shotgun LC–MS/MS analysis with a LTQ-Orbitrap mass spectrometer. To get better proteome identification and quantification, triplicate replicates of mass spectrometry analysis were performed for each sample. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (Vizcaíno et al., 2014 [1] via the PRIDE partner repository (Vizcaino, 2013 [2] with the dataset identifier http://www.ebi.ac.uk/pride/archive/projects/PXD003371. The peptide identifications that were further processed by PeptideProphet program in Trans-Proteomic Pipeline (TPP after database search with Mascot software were also available in .XML format files. Data presented here are related to a research article published in Journal of Proteomics by Li et al. (2015 [3]. Keywords: Bombyx mori, Middle silk gland, Silk protein synthesis, Shotgun proteomics, Label-free

  11. Rock support for nuclear waste repositories

    International Nuclear Information System (INIS)

    Abramson, L.W.; Schmidt, B.

    1984-01-01

    The design of rock support for underground nuclear waste repositories requires consideration of special construction and operation requirements, and of the adverse environmental conditions in which some of the support is placed. While repository layouts resemble mines, design, construction and operation are subject to quality assurance and public scrutiny similar to what is experienced for nuclear power plants. Exploration, design, construction and operation go through phases of review and licensing by government agencies as repositories evolve. This paper discusses (1) the various stages of repository development; (2) the environment that supports must be designed for; (3) the environmental effects on support materials; and (4) alternative types of repository rock support

  12. Peptide de novo sequencing of mixture tandem mass spectra

    DEFF Research Database (Denmark)

    Gorshkov, Vladimir; Hotta, Stéphanie Yuki Kolbeck; Braga, Thiago Verano

    2016-01-01

    they decrease the identification performance using database search engines. De novo sequencing approaches are expected to be even more sensitive to the reduction in mass spectrum quality resulting from peptide precursor co-isolation and thus prone to false identifications. The deconvolution approach matched...... complementary b-, y-ions to each precursor peptide mass, which allowed the creation of virtual spectra containing sequence specific fragment ions of each co-isolated peptide. Deconvolution processing resulted in equally efficient identification rates but increased the absolute number of correctly sequenced...... peptides. The improvement was in the range of 20–35% additional peptide identifications for a HeLa lysate sample. Some correct sequences were identified only using unprocessed spectra; however, the number of these was lower than those where improvement was obtained by mass spectral deconvolution. Tight...

  13. Delivery systems for antimicrobial peptides

    DEFF Research Database (Denmark)

    Nordström, Randi; Malmsten, Martin

    2017-01-01

    Due to rapidly increasing resistance development against conventional antibiotics, finding novel approaches for the treatment of infections has emerged as a key health issue. Antimicrobial peptides (AMPs) have attracted interest in this context, and there is by now a considerable literature...... on the identification such peptides, as well as on their optimization to reach potent antimicrobial and anti-inflammatory effects at simultaneously low toxicity against human cells. In comparison, delivery systems for antimicrobial peptides have attracted considerably less interest. However, such delivery systems...... are likely to play a key role in the development of potent and safe AMP-based therapeutics, e.g., through reducing chemical or biological degradation of AMPs either in the formulation or after administration, by reducing adverse side-effects, by controlling AMP release rate, by promoting biofilm penetration...

  14. [Tissue repositories for research at Sheba Medical Center(SMC].

    Science.gov (United States)

    Cohen, Yehudit; Barshack, Iris; Onn, Amir

    2013-06-01

    Cancer is the number one cause of death in both genders. Breakthroughs in the understanding of cancer biology, the identification of prognostic factors, and the development of new treatments are increasingly dependent on access to human cancer tissues with linked clinicopathological data. Access to human tumor samples and a large investment in translational research are needed to advance this research. The SMC tissue repositories provide researchers with biological materials, which are essential tools for cancer research. SMC tissue repositories for research aim to collect, document and preserve human biospecimens from patients with cancerous diseases. This is in order to provide the highest quality and well annotated biological biospecimens, used as essential tools to achieve the growing demands of scientific research needs. Such repositories are partners in acceLerating biomedical research and medical product development through clinical resources, in order to apply best options to the patients. Following Institutional Review Board approval and signing an Informed Consent Form, the tumor and tumor-free specimens are coLLected by a designated pathologist at the operating room only when there is a sufficient amount of the tumor, in excess of the routine needs. Blood samples are collected prior to the procedure. Other types of specimens collected include ascites fluid, pleural effusion, tissues for Optimal Cutting Temperature [OCT] and primary culture etc. Demographic, clinical, pathologicaL, and follow-up data are collected in a designated database. SMC has already established several organ or disease-specific tissue repositories within different departments. The foundation of tissue repositories requires the concentrated effort of a multidisciplinary team composed of paramedical, medical and scientific professionals. Research projects using these specimens facilitate the development of 'targeted therapy', accelerate basic research aimed at clarifying molecular

  15. Coupled processes in repository sealing

    International Nuclear Information System (INIS)

    Case, J.B.; Kelsall, P.C.

    1985-01-01

    The significance of coupled processes in repository sealing is evaluated. In most repository designs, shaft seals will be located in areas of relatively low temperature perturbation, in which case the coupling of temperature with stress and permeability may be less significant than the coupling between stress and permeability that occurs during excavation. Constitutive relationships between stress and permeability are reviewed for crystalline rock and rocksalt. These provide a basis for predicting the development of disturbed zones near excavations. Field case histories of the degree of disturbance are presented for two contrasting rock types - Stripa granite and Southeastern New Mexico rocksalt. The results of field investigations in both rock types confirm that hydraulic conductivity or permeability is stress dependent, and that shaft seal performance may be related to the degree that stresses are perturbed and restored near the seal

  16. University digital repositories and authors.

    Directory of Open Access Journals (Sweden)

    Alice Keefer

    2008-02-01

    Full Text Available The Open Access movement offers two strategies for making scientific information available without economic, technical or legal obstacles: the publication of articles in OA journals and the deposit by authors of their Works in stable institutional or discipline-based repositories. This article explores the implementation of the second “route” on the part of authors, because it is the strategy that offers the greatest possibility of attaining OA in the short term. However, it does require repositories to exert great effort in informing the authors of the advantages of self-archiving and of the procedures for depositing their work and, even helping them to do so – through services and promotional activities.

  17. Implementing the EuroFIR Document and Data Repositories as accessible resources of food composition information.

    Science.gov (United States)

    Unwin, Ian; Jansen-van der Vliet, Martine; Westenbrink, Susanne; Presser, Karl; Infanger, Esther; Porubska, Janka; Roe, Mark; Finglas, Paul

    2016-02-15

    The EuroFIR Document and Data Repositories are being developed as accessible collections of source documents, including grey literature, and the food composition data reported in them. These Repositories will contain source information available to food composition database compilers when selecting their nutritional data. The Document Repository was implemented as searchable bibliographic records in the Europe PubMed Central database, which links to the documents online. The Data Repository will contain original data from source documents in the Document Repository. Testing confirmed the FoodCASE food database management system as a suitable tool for the input, documentation and quality assessment of Data Repository information. Data management requirements for the input and documentation of reported analytical results were established, including record identification and method documentation specifications. Document access and data preparation using the Repositories will provide information resources for compilers, eliminating duplicated work and supporting unambiguous referencing of data contributing to their compiled data. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Repository development status in Spain

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez Beceiro, Alvaro; Zuloaga, Pablo [ENRESA (Spain)

    2009-06-15

    The paper describes the status of repository development for the different waste categories. Low and Intermediate Waste Disposal facility of El Cabril was commissioned in 1992 and is in normal operation. The major modifications and activities during the last years are the adaptation to waste streams not initially foreseen such as some decommissioning waste or waste from steel industry, and the improvement of its performance assessment, supported by a R and D and refined models. As part of this facility, a new disposal facility specifically intended for very low activity waste has been constructed and commissioned in July 2008. Its design is based on the European Directive for hazardous waste disposal. National policy for Nuclear Spent Fuel and High-Level waste is focused on the development of a centralized storage facility of the vault type, whose site location would be selected through a volunteering process. Meanwhile, with the aim of solving specific problems, three individual storage facilities are in different status at reactor sites. Research on final solution, including some repository aspects as well as separation and transmutation are being carried out in accordance to ENRESA's R and D program. ENRESA has developed conceptual designs for non site specific repositories, both in granite and clay, and has carried out their corresponding performance assessment exercises. (authors)

  19. Geotechnical instrumentation for repository shafts

    International Nuclear Information System (INIS)

    Lentell, R.L.; Byrne, J.

    1993-01-01

    The US Congress passed the Nuclear Waste Policy Act in 1980, which required that three distinctly different geologic media be investigated as potential candidate sites for the permanent disposal of high-level nuclear waste. The three media that were selected for study were basalt (WA), salt (TX, LA, MS, UT), and tuff (NV). Preliminary Exploratory Shaft Facilities (ESF) designs were prepared for seven candidate salt sites, including bedded and domal salt environments. A bedded-salt site was selected in Deaf Smith County, TX for detailed site characterization studies and ESF Final Design. Although Congress terminated the Salt Repository Program in 1988, Final Design for the Deaf Smith ESF was completed, and much of the design rationale can be applied to subsequent deep repository shafts. This paper presents the rationale for the geotechnical instrumentation that was designed for construction and operational performance monitoring of the deep shafts of the in-situ test facility. The instrumentation design described herein can be used as a general framework in designing subsequent instrumentation programs for future high-level nuclear waste repository shafts

  20. Flanking signal and mature peptide residues influence signal peptide cleavage

    Directory of Open Access Journals (Sweden)

    Ranganathan Shoba

    2008-12-01

    Full Text Available Abstract Background Signal peptides (SPs mediate the targeting of secretory precursor proteins to the correct subcellular compartments in prokaryotes and eukaryotes. Identifying these transient peptides is crucial to the medical, food and beverage and biotechnology industries yet our understanding of these peptides remains limited. This paper examines the most common type of signal peptides cleavable by the endoprotease signal peptidase I (SPase I, and the residues flanking the cleavage sites of three groups of signal peptide sequences, namely (i eukaryotes (Euk (ii Gram-positive (Gram+ bacteria, and (iii Gram-negative (Gram- bacteria. Results In this study, 2352 secretory peptide sequences from a variety of organisms with amino-terminal SPs are extracted from the manually curated SPdb database for analysis based on physicochemical properties such as pI, aliphatic index, GRAVY score, hydrophobicity, net charge and position-specific residue preferences. Our findings show that the three groups share several similarities in general, but they display distinctive features upon examination in terms of their amino acid compositions and frequencies, and various physico-chemical properties. Thus, analysis or prediction of their sequences should be separated and treated as distinct groups. Conclusion We conclude that the peptide segment recognized by SPase I extends to the start of the mature protein to a limited extent, upon our survey of the amino acid residues surrounding the cleavage processing site. These flanking residues possibly influence the cleavage processing and contribute to non-canonical cleavage sites. Our findings are applicable in defining more accurate prediction tools for recognition and identification of cleavage site of SPs.

  1. Identification of Proteins and Peptide Biomarkers for Detecting Banned Processed Animal Proteins (PAPs) in Meat and Bone Meal by Mass Spectrometry.

    Science.gov (United States)

    Marbaix, Hélène; Budinger, Dimitri; Dieu, Marc; Fumière, Olivier; Gillard, Nathalie; Delahaut, Philippe; Mauro, Sergio; Raes, Martine

    2016-03-23

    The outbreak of bovine spongiform encephalopathy (BSE) in the United Kingdom in 1986, with processed animal proteins (PAPs) as the main vector of the disease, has led to their prohibition in feed. The progressive release of the feed ban required the development of new analytical methods to determine the exact origin of PAPs from meat and bone meal. We set up a promising MS-based method to determine the species and the source (legal or not) present in PAPs: a TCA-acetone protein extraction followed by a cleanup step, an in-solution tryptic digestion of 5 h (with a 1:20 protein/trypsin ratio), and mass spectrometry analyses, first without any a priori, with a Q-TOF, followed by a targeted triple-quadrupole analysis. Using this procedure, we were able to overcome some of the major limitations of the official methods to analyze PAPs, detecting and identifying prohibited animal products in feedstuffs by the monitoring of peptides specific for cows, pigs, and sheep in PAPs.

  2. Identification and molecular characterization of 48 kDa calcium binding protein as calreticulin from finger millet (Eleusine coracana) using peptide mass fingerprinting and transcript profiling.

    Science.gov (United States)

    Singh, Manoj; Metwal, Mamta; Kumar, Vandana A; Kumar, Anil

    2016-01-30

    Attempts were made to identify and characterize the calcium binding proteins (CaBPs) in grain filling stages of finger millet using proteomics, bioinformatics and molecular approaches. A distinctly observed blue color band of 48 kDa stained by Stains-all was eluted and analyzed as calreticulin (CRT) using nano liquid chromatography-tandem mass spectrometry (nano LC-MS). Based on the top hits of peptide mass fingerprinting results, conserved primers were designed for isolation of the CRT gene from finger millet using calreticulin sequences of different cereals. The deduced nucleotide sequence analysis of 600 bp amplicon showed up to 91% similarity with CRT gene(s) of rice and other plant species and designated as EcCRT1. Transcript profiling of EcCRT1 showed different levels of relative expression at different stages of developing spikes. The higher expression of EcCRT1 transcripts and protein were observed in later stages of developing spikes which might be due to greater translational synthesis of EcCRT1 protein during seed maturation in finger millet. Preferentially higher synthesis of this CaBP during later stages of grain filling may be responsible for the sequestration of calcium in endoplasmic reticulum of finger millet grains. © 2015 Society of Chemical Industry.

  3. Computational Identification and Characterization of a Promiscuous T-Cell Epitope on the Extracellular Protein 85B of Mycobacterium spp. for Peptide-Based Subunit Vaccine Design

    Directory of Open Access Journals (Sweden)

    Md. Saddam Hossain

    2017-01-01

    Full Text Available Tuberculosis (TB is a reemerging disease that remains as a leading cause of morbidity and mortality in humans. To identify and characterize a T-cell epitope suitable for vaccine design, we have utilized the Vaxign server to assess all antigenic proteins of Mycobacterium spp. recorded to date in the Protegen database. We found that the extracellular protein 85B displayed the most robust antigenicity among the proteins identified. Computational tools for identifying T-cell epitopes predicted an epitope, 181-QQFIYAGSLSALLDP-195, that could bind to at least 13 major histocompatibility complexes, revealing the promiscuous nature of the epitope. Molecular docking simulation demonstrated that the epitope could bind to the binding groove of MHC II and MHC I molecules by several hydrogen bonds. Molecular docking analysis further revealed that the epitope had a distinctive binding pattern to all DRB1 and A and B series of MHC molecules and presented almost no polymorphism in its binding site. Moreover, using “Allele Frequency Database,” we checked the frequency of HLA alleles in the worldwide population and found a higher frequency of both class I and II HLA alleles in individuals living in TB-endemic regions. Our results indicate that the identified peptide might be a universal candidate to produce an efficient epitope-based vaccine for TB.

  4. Specific Molecular Signatures for Type II Crustins in Penaeid Shrimp Uncovered by the Identification of Crustin-Like Antimicrobial Peptides in Litopenaeus vannamei

    Science.gov (United States)

    Barreto, Cairé; Coelho, Jaqueline da Rosa; Yuan, Jianbo; Xiang, Jianhai; Perazzolo, Luciane Maria

    2018-01-01

    Crustins form a large family of antimicrobial peptides (AMPs) in crustaceans composed of four sub-groups (Types I-IV). Type II crustins (Type IIa or “Crustins” and Type IIb or “Crustin-like”) possess a typical hydrophobic N-terminal region and are by far the most representative sub-group found in penaeid shrimp. To gain insight into the molecular diversity of Type II crustins in penaeids, we identified and characterized a Type IIb crustin in Litopenaeus vannamei (Crustin-like Lv) and compared Type II crustins at both molecular and transcriptional levels. Although L. vannamei Type II crustins (Crustin Lv and Crustin-like Lv) are encoded by separate genes, they showed a similar tissue distribution (hemocytes and gills) and transcriptional response to the shrimp pathogens Vibrio harveyi and White spot syndrome virus (WSSV). As Crustin Lv, Crustin-like Lv transcripts were found to be present early in development, suggesting a maternal contribution to shrimp progeny. Altogether, our in silico and transcriptional data allowed to conclude that (1) each sub-type displays a specific amino acid signature at the C-terminal end holding both the cysteine-rich region and the whey acidic protein (WAP) domain, and that (2) shrimp Type II crustins evolved from a common ancestral gene that conserved a similar pattern of transcriptional regulation. PMID:29337853

  5. SAPIERR Paves the way towards European regional repository

    International Nuclear Information System (INIS)

    Stefula, V.; McCombie, C.

    2004-01-01

    development to be carried out in future. High profile will be given to so-called co-operative scenario, which is most appropriate for the European context. The identification of the potential host country was deliberately excluded from the scope of SAPIERR project in this first phase. The working group participants will correspond throughout the two year duration of the study and will meet again towards the end of the project at an international seminar in Brussels in October 2005. The objective of the seminar will be to review and disseminate results of the project to a wide audience and to agree on subsequent steps on the way towards the European Regional Repository.(author)

  6. Post-staining electroblotting for efficient and reliable peptide blotting.

    Science.gov (United States)

    Lee, Der-Yen; Chang, Geen-Dong

    2015-01-01

    Post-staining electroblotting has been previously described to transfer Coomassie blue-stained proteins from polyacrylamide gel onto polyvinylidene difluoride (PVDF) membranes. Actually, stained peptides can also be efficiently and reliably transferred. Because of selective staining procedures for peptides and increased retention of stained peptides on the membrane, even peptides with molecular masses less than 2 kDa such as bacitracin and granuliberin R are transferred with satisfactory results. For comparison, post-staining electroblotting is about 16-fold more sensitive than the conventional electroblotting for visualization of insulin on the membrane. Therefore, the peptide blots become practicable and more accessible to further applications, e.g., blot overlay detection or immunoblotting analysis. In addition, the efficiency of peptide transfer is favorable for N-terminal sequence analysis. With this method, peptide blotting can be normalized for further analysis such as blot overlay assay, immunoblotting, and N-terminal sequencing for identification of peptide in crude or partially purified samples.

  7. Identification of single amino acid substitutions (SAAS) in neuraminidase from influenza a virus (H1N1) via mass spectrometry analysis coupled with de novo peptide sequencing.

    Science.gov (United States)

    Peng, Qisheng; Wang, Zijian; Wu, Donglin; Li, Xiaoou; Liu, Xiaofeng; Sun, Wanchun; Liu, Ning

    2016-08-01

    Amino acid substitutions in the neuraminidase of the influenza virus are the main cause of the emergence of resistance to zanamivir or oseltamivir during seasonal influenza treatment; they are the result of non-synonymous mutations in the viral genome that can be successfully detected by polymer chain reaction (PCR)-based approaches. There is always an urgent need to detect variation in amino acid sequences directly at the protein level. Mass spectrometry coupled with de novo sequencing has been explored as an alternative and straightforward strategy for detecting amino acid substitutions, as well - this approach is the primary focus of the present study. Influenza virus (A/Puerto Rico/8/1934 H1N1) propagated in embryonated chicken eggs was purified by ultracentrifugation, followed by PNGase F treatment. The deglycosylated virion was lysed and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The gel band corresponding to neuraminidase was picked up and subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. LC-MS/MS analyses, coupled with manual de novo sequencing, allowed the determination of three amino acid substitutions: R346K, S349 N, and S370I/L, in the neuraminidase from the influenza virus (A/Puerto Rico/8/1934 H1N1), which were located in three mutated peptides of the neuraminidase: YGNGVWIGK, TKNHSSR, and PNGWTETDI/LK, respectively. We found that the amino acid substitutions in the proteins of RNA viruses (including influenza A virus) resulting from non-synonymous gene mutations can indeed be directly analyzed via mass spectrometry, and that manual interpretation of the MS/MS data may be beneficial. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  8. Identification of a second T-cell antigen receptor in human and mouse by an anti-peptide γ-chain-specific monoclonal antibody

    International Nuclear Information System (INIS)

    Ioannides, C.G.; Itoh, K.; Fox, F.E.; Pahwa, R.; Good, R.A.; Platsoucas, C.D.

    1987-01-01

    The authors developed a monoclonal antibody (mAb) (9D7) against a synthetic peptide (P13K) selected from the deduced amino acid sequence of the constant region of the λ chain of the murine T-cell antigen receptor (TCR) (amino acids 118-130). Using this mAb, they identified a putative second TCR expressed on peripheral blood lymphocytes from a patient with severe combined immunodeficiency (SCID) that were propagated in culture with recombinant interleukin 2 (rIL-2) and Con A. This mAb immunoprecipitated two polypeptide chains of 40 and 58 kDa under nonreducing conditions and of 40 and 56 kDa under reducing conditions from 125 I-labeled denatured lysates of T3 + WT31 - lymphocytes expanded in culture from a SCID patient. Chemical crosslinking of 125 I-labeled cells followed by immunoprecipitation with anti-Leu-4 mAb under nonreducing or reducing conditions revealed that the 40- and 56-kDa polypeptide chains were associated with the T3 differentiation antigen. These experiments were done with polyclonal cell populations. Cloned T3 + WT31 - cell populations are required to determine whether the TCR contains two λ polypeptide chains. Using the same 9D7 anti-P18K mAb and immunoblotting analysis, they identified a 35 kDa γ-chain polypeptide under reducing conditions expressed on purified L3T4 - Lyt2 - BALB/c mouse thymocytes. This γ-chain TCR is disulfide linked and has a molecular mass of 80 kDa under nonreducing conditions

  9. Hydrothermal conditions around a radioactive waste repository

    International Nuclear Information System (INIS)

    Thunvik, R.; Braester, C.

    1981-12-01

    Numerical solutions for the hydrothermal conditions around a hard rock repository for nuclear fuel waste are presented. The objective of the present investigation is to illustrate in principle the effect of heat released from a hypothetical radioactive waste repository with regard to anisotropy in the rock permeability. Permeability and porosity are assumed to be constant or to decrease exponentially with depth. The hypothetical repository is situated below a horizontal ground surface or below the crest of a hill, and it is assumed that the water table follows the topography. Major interest in the analysis is directed towards the influence of anisotropy in the permeability on the flow patterns and travel times for water particles, being traced from the repository to the ground surface. The presented results show that anisotropy in the permeability may have a significant influence on the flow conditions around the repository and subsequently also on the travel times from the repository. (Authors)

  10. People's perception of LILW repository

    International Nuclear Information System (INIS)

    Zeleznik, Nadja; Polic, Marko

    2002-01-01

    Full text: Social acceptability of the radioactive waste repository presents a great problem in every country with such a waste. Even if people agree with the need for its construction, the chosen location should be far from their homes (NIMBY). The reasons for such attitudes were attributed to different causes: uneducated public, differences in understanding of radioactivity and risk by experts and lay public, risk communication problems, lack of credibility and social trust, etc. While in earlier days public was blamed for its irrationality, and need for education and information was emphasized, today it is realized that public trust is extremely important if effective risk communication is to be achieved. It is also recognized that it is not so much the content of the risk message itself, as the lack of trust to those responsible for provision of information that is behind this opposition. Perhaps we could apply here Petty and Caciopo's elaboration likelihood model of persuasion, with credibility as a factor in peripheral route of persuasion. Nevertheless also general lowering of social trust should explain social non-agreement. This lack of trust in experts and political institutions is perhaps caused by outwitting public in earlier years, its bad experiences with responsible officials, dangerous accidents (e.g. TMI, Chernobyl), increased influence that professions have over people's welfare, a greater value placed on equality and better educated public, etc. In 1996 the ARAO re-initiated the search for a LILW repository location with a new, so-called combined approach to the site selection, where the technical, geologically led process is combined with participation of local community. In order to get information on people's perception of the LILW repository construction, their willingness to accept it and factors that influence the acceptability, several surveys have been conducted. Groups of experts and lay persons answered the questionnaires. The results of

  11. Underground repository for radioactive wastes

    International Nuclear Information System (INIS)

    Cassibba, R.O.

    1989-01-01

    In the feasibility study for an underground repository in Argentina, the conceptual basis for the final disposal of high activity nuclear waste was set, as well as the biosphere isolation, according to the multiple barrier concept or to the engineering barrier system. As design limit, the container shall act as an engineering barrier, granting the isolation of the radionuclides for approximately 1000 years. The container for reprocessed and vitrified wastes shall have three metallic layers: a stainless steel inner layer, an external one of a metal to be selected and a thick intermediate lead layer preselected due to its good radiological protection and corrosion resistance. Therefore, the study of the lead corrosion behaviour in simulated media of an underground repository becomes necessary. Relevant parameters of the repository system such as temperature, pressure, water flux, variation in salt concentrations and oxidants supply shall be considered. At the same time, a study is necessary on the galvanic effect of lead coupled with different candidate metals for external layer of the container in the same experimental conditions. Also temporal evaluation about the engineering barrier system efficiency is presented in this thesis. It was considered the extrapolated results of corrosion rates and literature data about the other engineering barriers. Taking into account that corrosion is of a generalized type, the integrity of the lead shall be maintained for more than 1000 years and according to temporal evaluation, the multiple barrier concept shall retard the radionuclide dispersion to the biosphere for a period of time between 10 4 and 10 6 years. (Author) [es

  12. Groundwater movements around a repository

    International Nuclear Information System (INIS)

    Burgess, A.

    1977-10-01

    Based on regional models of groundwater flow, the regional hydraulic gradient at depth is equal to the regional topographic gradient. As a result, the equipotentials are near vertical. The permeability distribution with depth influences the groundwater flow patterns. A zone of sluggish flows, the quiescent zone is developed when the permeability decreases with depth. This feature is accentuated when horizontal anisotropy, with the horizontal permeability higher then the vertical permeability, is included. The presence of an inactive zone will be a prerequesite for a satisfactory repository site. The effect of an inclined discontinuity representing a singular geological feature such as a fault plane or shear zone has been modelled. The quiescent zone does not appear to be unduly disturbed by such a feature. However, meaningful quantitative predictions related to the flows in a typical singular feature cannot be made without more specific data on their hydraulic properties. Two dimensional analysis has been made for a site specific section of a candidate repository site at Forsmark, Sweden. The lateral extent of the model was defined by major tectonic features, assumed vertical. Potential gradients and pore velocities have been computed for a range of boundary conditions and assumed material properties. The potential gradients for the model with anisotropic permeability approach the average potential gradient between the boundaries. The result of this study of the initial groundwater conditions will be used as input data for the analyses of the thermomechanical perturbations of the groundwater regime. In the long term, the groundwater flow will return to the initial conditions. The residual effects of the repository on the flow will be discussed in part 2 of this report. (author)

  13. Knowledge repositories for multiple uses

    Science.gov (United States)

    Williamson, Keith; Riddle, Patricia

    1991-01-01

    In the life cycle of a complex physical device or part, for example, the docking bay door of the Space Station, there are many uses for knowledge about the device or part. The same piece of knowledge might serve several uses. Given the quantity and complexity of the knowledge that must be stored, it is critical to maintain the knowledge in one repository, in one form. At the same time, because of quantity and complexity of knowledge that must be used in life cycle applications such as cost estimation, re-design, and diagnosis, it is critical to automate such knowledge uses. For each specific use, a knowledge base must be available and must be in a from that promotes the efficient performance of that knowledge base. However, without a single source knowledge repository, the cost of maintaining consistent knowledge between multiple knowledge bases increases dramatically; as facts and descriptions change, they must be updated in each individual knowledge base. A use-neutral representation of a hydraulic system for the F-111 aircraft was developed. The ability to derive portions of four different knowledge bases is demonstrated from this use-neutral representation: one knowledge base is for re-design of the device using a model-based reasoning problem solver; two knowledge bases, at different levels of abstraction, are for diagnosis using a model-based reasoning solver; and one knowledge base is for diagnosis using an associational reasoning problem solver. It was shown how updates issued against the single source use-neutral knowledge repository can be propagated to the underlying knowledge bases.

  14. National radioactive wasterRepository Mochovce

    International Nuclear Information System (INIS)

    2000-01-01

    In this leaflet the scheme of the Mochovce National radioactive waste repository for the Slovak Republic is presented. The National radioactive waste repository in Mochovce is a surface type storage facility. It is intended for final disposal of solid and solidified low and intermediate radioactive waste produced during the operation of nuclear power plants and institutions located within the territory of the Slovak Republic. The Repository site is situated about 2 km northwest to the Mochovce NPP

  15. Performance assessment of Mochovce repository

    Energy Technology Data Exchange (ETDEWEB)

    Mrskova, A; Hanusik, V [Dept. of Accident Management and Risk Assessment, Vyskumny Ustav Jadrovych Elektrarni, Trnava (Slovakia)

    2000-07-01

    The near-surface disposal site at Mochovce is designed for low-level and intermediate level radioactive waste. It is a vault-type concrete structure housing the reinforced concrete containers as the final waste packages. This paper shortly presents the long-term safety analysis methods applied for the post-closure phase of the repository. The main aim of paper is description of the philosophy of analysis, development of the scenarios, their modeling and comparing of the results of normal evolution scenario, alternative scenario and intruders scenario for some radionuclides. (author)

  16. Performance assessment of Mochovce repository

    International Nuclear Information System (INIS)

    Mrskova, A.; Hanusik, V.

    2000-01-01

    The near-surface disposal site at Mochovce is designed for low-level and intermediate level radioactive waste. It is a vault-type concrete structure housing the reinforced concrete containers as the final waste packages. This paper shortly presents the long-term safety analysis methods applied for the post-closure phase of the repository. The main aim of paper is description of the philosophy of analysis, development of the scenarios, their modeling and comparing of the results of normal evolution scenario, alternative scenario and intruders scenario for some radionuclides. (author)

  17. Office of Geologic Repositories quality assurance plan for high-level radioactive waste repositories

    International Nuclear Information System (INIS)

    1986-08-01

    This document sets forth geologic repository program-wide quality assurance program requirements and defines management's quality assurance responsibilities for the Office of Geologic Repositories and its projects. (LM)

  18. The European Repository Landscape 2008 Inventory of Digital Repositories for Research Output

    CERN Document Server

    Van der Graaf, Maurits

    2009-01-01

    It is widely acknowledged that a common knowledge base for European research is necessary. Research repositories are an important innovation to the scientific information infrastructure. In 2006, digital repositories in the 27 countries of the European we

  19. Workshop: Creating Your Institutional Research Repository

    KAUST Repository

    Grenz, Daryl M.

    2016-11-08

    In 2002, the Scholarly Publishing and Academic Resources Coalition (SPARC) proposed the concept of an institutional repository to simultaneously disrupt and enhance the state of scholarly communications in the academic world. Thirteen years later, thousands of universities and other institutions have answered this call, but many more have not due to gaps in budgets, awareness and, most of all, practical guidance on creating an institutional repository. This workshop provides you with an essential primer on what it takes to establish a fully-functioning institutional repository. Every aspect of the process will be covered, including policies, procedures, staffing guidelines, workflows and repository technologies.

  20. Center for Leadership Development (CLD) Repository

    Data.gov (United States)

    Office of Personnel Management — The Center for Leadership Development Repository stores various data including policies, procedures, governance, guidance, security, and financial documents of the...

  1. Nuclear waste repository design and construction

    International Nuclear Information System (INIS)

    Bohlke, B.M.; Monsees, J.E.

    1987-01-01

    Extensive underground excavation will be required for construction of a mined geologic repository for nuclear waste. Hundreds of thousands of feet of drift will be required based on the conceptual layout design for each candidate nuclear waste repository. Comparison of boring and blasting excavation methods are discussed, as are special design and construction requirements (e.g., quality assurance procedures and performance assessment) for the nuclear waste repository. Comparisons are made between boring and blasting construction methods for the repository designs proposed for salt, volcanic tuff, and basalt

  2. An Optimal Centralized Carbon Dioxide Repository for Florida, USA

    Directory of Open Access Journals (Sweden)

    Brandon Poiencot

    2011-03-01

    Full Text Available For over a decade, the United States Department of Energy, and engineers, geologists, and scientists from all over the world have investigated the potential for reducing atmospheric carbon emissions through carbon sequestration. Numerous reports exist analyzing the potential for sequestering carbon dioxide at various sites around the globe, but none have identified the potential for a statewide system in Florida, USA. In 2005, 83% of Florida’s electrical energy was produced by natural gas, coal, or oil (e.g., fossil fuels, from power plants spread across the state. In addition, only limited research has been completed on evaluating optimal pipeline transportation networks to centralized carbon dioxide repositories. This paper describes the feasibility and preliminary locations for an optimal centralized Florida-wide carbon sequestration repository. Linear programming optimization modeling is used to plan and route an idealized pipeline network to existing Florida power plants. Further analysis of the subsurface geology in these general locations will provide insight into the suitability of the subsurface conditions and the available capacity for carbon sequestration at selected possible repository sites. The identification of the most favorable site(s is also presented.

  3. Aquatic ecosystem characterisation strategy at a repository site

    Energy Technology Data Exchange (ETDEWEB)

    Kangasniemi, Ville; Ikonen, Ari T.K. [Environmental Research and Assessment EnviroCase, Ltd., Hallituskatu 1 D 4, 28100 Pori (Finland); Lahdenperae, Anne-Maj [Saanio and Riekkola Oy, Laulukuja 4, 00420 Helsinki (Finland); Kirkkala, Teija [Pyhaejaervi Institute, Sepaentie 7, 27500 Kauttua (Finland); Koivunen, Sari [Water and Environment Research of South-West Finland, Telekatu 16, 20360 Turku (Finland)

    2014-07-01

    Olkiluoto Island on the western coast of Finland has been selected as a repository site for spent nuclear fuel disposal. According to regulatory requirements, the safety assessment for the repository should have an assessment timeframe of several millennia. Due to the post-glacial land uplift, the relatively shallow sea areas around Olkiluoto Island will change gradually to lakes, rivers and terrestrial areas. As there are no limnic systems at present Olkiluoto site, the reference area was delineated and reference lakes and rivers were selected as an analogue. For the modelling of the transport and accumulation of possible radionuclide releases in the surface environment, aquatic ecosystems were identified and divided into biotopes. Despite the number of available templates, the division of aquatic environment for the biosphere assessment of the Olkiluoto spent fuel repository was necessary to made separately. In this contribution, the processes behind the identification of aquatic ecosystems (e.g. legislation, physical and chemical properties) together with the biotope selection methodology (e.g. light and bottom conditions) and the challenges related to the amount of variable input parameters for each biotope in the modelling are presented. (authors)

  4. Radioactive Waste Repositories Administration - SURAO

    International Nuclear Information System (INIS)

    Kucerka, M.

    1998-01-01

    The Atomic Act specifies, among other things, responsibilities of the government in the field of safe disposal of radioactive wastes. To satisfy this responsibility, the Ministry of Industry and Trade has established the Radioactive Waste Repositories Administration (SURAO). SURAO's major responsibilities include: (a) the preparation, construction, commissioning, operation, and decommissioning of radioactive waste repositories and the monitoring of their environmental impacts; (b) radioactive waste management; (c) spent or irradiated nuclear fuel processing into a form suitable for storage/disposal or reuse; (d) record-keeping of received radioactive wastes and their producers; (e) administration of fund transfers as stipulated by the Atomic Act, Article 27; (f) development of proposals for specification of fees to be paid to the Nuclear Account; (g) responsibility for and coordination of research and development in the field of radioactive waste handling and management; (h) supervision of licensees' margin earmarked for the decommissioning of their facilities; (i) providing services in radioactive waste handling and management; (j) handling and management of radioactive wastes that have been transferred to the Czech Republic from abroad and cannot be sent back; (k) interim administration of radioactive wastes that have become state property. The Statute of the Administration is reproduced in full. (P.A.)

  5. Characteristics of potential repository wastes

    International Nuclear Information System (INIS)

    Notz, K.J.

    1989-01-01

    The Office of Civilian Radioactive Waste Management (OCRWM) is responsible for the spent fuels and other wastes that will be disposed of in a geologic repository. The two major sources of these materials are commercial light-water reactor (LWR) spent fuel and immobilized high-level waste (HLW). Other wastes that may require long-term isolation include non-LWR spent fuels and miscellaneous sources such as activated metals. Detailed characterizations are required for all of these potential repository wastes. These characterizations include physical, chemical, and radiological properties. The latter must take into account decay as a function of time. This information has been extracted from primary data sources, evaluated, and assembled in a Characteristics Data Base which provides data in four formats: hard copy standard reports, menu-driven personal computer (PC) data bases, program-level PC data bases, and mainframe computer files. The Characteristics Data Base provides a standard set of self-consistent data to the various areas of responsibility including systems integration and waste stream analysis, storage, transportation, and geologic disposal. The data will be used for design studies, evaluation of alternatives, and system optimization by OCRWM and supporting contractors. 7 refs., 5 figs., 7 tabs

  6. INIS: Nuclear Grey Literature Repository

    International Nuclear Information System (INIS)

    Savić, Dobrica

    2016-01-01

    As one of the world's largest collections of published information on the peaceful uses of nuclear science and technology, INIS represents an extraordinary example of world cooperation. Currently, as INIS members, 130 countries and 24 international organizations share and allow access to their valuable nuclear information resources, preserving them for future generations and offering a freely available nuclear knowledge repository. Since its creation in 1970, INIS has collected and provided access to more than 3.8 million bibliographic references to publications, documents, technical reports, non-copyrighted documentation, and other grey literature, as well as over a million full texts. Public interest throughout the years in accessing the INIS Collection has been remarkable. This paper deals with the challenges faced by INIS in its endeavour to increase the use, accessibility, usability and expandability of its on-line repository. It also describes document collection, the features and characteristics of implementing a new search engine, as well as the lessons learned. (author)

  7. Nuclear waste repository simulation experiments

    International Nuclear Information System (INIS)

    Rothfuchs, T.; Wieczorek, K.; Feddersen, H.K.; Staupendahl, G.; Coyle, A.J.; Kalia, H.; Eckert, J.

    1986-12-01

    This document is the third joint annual report on the Cooperative German-American 'Brine Migration Tests' that are in progress at the Asse salt mine in the Federal Republic of Germany (FRG). This Government supported mine serves as an underground test facility for research and development (R and D)-work in the field of nuclear waste repository research and simulation experiments. The tests are designed to simulate a nuclear waste repository to measure the effects of heat and gamma radiation on brine migration, salt decrepitation, disassociation of brine, and gases collected. The thermal mechanical behavior of salt, such as room closure, stresses and changes of the properties of salt are measured and compared with predicted behavior. This document covers the following sections: Issues and test objectives: This section presents issues that are investigated by the Brine Migration Test, and the test objectives derived from these issues; test site: This section describes the test site location and geology in the Asse mine; test description: A description of the test configuration, procedures, equipment, and instrumentation is given in this section; actual test chronology: The actual history of the test, in terms of the dates at which major activities occured, is presented in this section. Test results: This section presents the test results observed to data and the planned future work that is needed to complete the test; conclusions and recommendations: This section summarizes the conclusions derived to date regarding the Brine Migration Test. Additional work that would be useful to resolve the issues is discussed. (orig.)

  8. Identification of a novel biomarker candidate, a 4.8-kDa peptide fragment from a neurosecretory protein VGF precursor, by proteomic analysis of cerebrospinal fluid from children with acute encephalopathy using SELDI-TOF-MS

    Directory of Open Access Journals (Sweden)

    Fujino Osamu

    2011-08-01

    Full Text Available Abstract Background Acute encephalopathy includes rapid deterioration and has a poor prognosis. Early intervention is essential to prevent progression of the disease and subsequent neurologic complications. However, in the acute period, true encephalopathy cannot easily be differentiated from febrile seizures, especially febrile seizures of the complex type. Thus, an early diagnostic marker has been sought in order to enable early intervention. The purpose of this study was to identify a novel marker candidate protein differentially expressed in the cerebrospinal fluid (CSF of children with encephalopathy using proteomic analysis. Methods For detection of biomarkers, CSF samples were obtained from 13 children with acute encephalopathy and 42 children with febrile seizure. Mass spectral data were generated by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS technology, which is currently applied in many fields of biological and medical sciences. Diagnosis was made by at least two pediatric neurologists based on the clinical findings and routine examinations. All specimens were collected for diagnostic tests and the remaining portion of the specimens were used for the SELDI-TOF MS investigations. Results In experiment 1, CSF from patients with febrile seizures (n = 28, patients with encephalopathy (n = 8 (including influenza encephalopathy (n = 3, encephalopathy due to rotavirus (n = 1, human herpes virus 6 (n = 1 were used for the SELDI analysis. In experiment 2, SELDI analysis was performed on CSF from a second set of febrile seizure patients (n = 14 and encephalopathy patients (n = 5. We found that the peak with an m/z of 4810 contributed the most to the separation of the two groups. After purification and identification of the 4.8-kDa protein, a 4.8-kDa proteolytic peptide fragment from the neurosecretory protein VGF precursor (VGF4.8 was identified as a novel biomarker for encephalopathy. Conclusions

  9. Low and medium level radioactive waste repository: risk perception

    International Nuclear Information System (INIS)

    Aquino, Afonso Rodrigues de; Bueno, Lilian de Oliveira; Vieira, Martha Marques Ferreira; Fonseca, Edvaldo Roberto Paiva da; Bellintani, Sandra Aparecida

    2009-01-01

    This paper focuses on the risk perception associated to the installation of low and intermediate level radioactive waste (LLRW and ILRW) disposal facilities. The purpose is to give support for the implementation of a repository in Brazil. Public acceptance results from a long term work and trust is vital for the process as it takes long to be conquered but might be shortly lost. Therefore, it is essential to care about the way each step is conducted. The knowledge about the system and the risks, the comprehension about these risks, the commitment with safety, adequate support systems for the project (legislation, involved institutions) and the excellence as a goal to be reached are extremely important parameters. The involvement of all interested parties in the decision-making process is condition for a successful and publicly acceptable implementation of such project. The steps for public acceptance of a repository are summarized as follow: Risk perception: to verify how the local population understand and feel the installation of a repository in the region. Media observatory: to continuously follow the news reaching the region where the repository will be installed, including different media. Local population social/economical/cultural profile identification: to determine the local population social/economical/cultural profile; to conduct a survey to know their expectations, allowing the proposal of compensation and incentives to fully account for their expectations. Finally, the philosophy governing this Project is: on doubt, the public must be heard and only after this public hearing the policies concerning the project shall be formulated. (author)

  10. Low and medium level radioactive waste repository: risk perception

    Energy Technology Data Exchange (ETDEWEB)

    Aquino, Afonso Rodrigues de; Bueno, Lilian de Oliveira; Vieira, Martha Marques Ferreira; Fonseca, Edvaldo Roberto Paiva da; Bellintani, Sandra Aparecida [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)], e-mail: araquino@ipen.br, e-mail: lbueno@ipen.br, e-mail: mmvieira@ipen.br, e-mail: efonseca@ipen.br, e-mail: sbellint@ipen.br

    2009-07-01

    This paper focuses on the risk perception associated to the installation of low and intermediate level radioactive waste (LLRW and ILRW) disposal facilities. The purpose is to give support for the implementation of a repository in Brazil. Public acceptance results from a long term work and trust is vital for the process as it takes long to be conquered but might be shortly lost. Therefore, it is essential to care about the way each step is conducted. The knowledge about the system and the risks, the comprehension about these risks, the commitment with safety, adequate support systems for the project (legislation, involved institutions) and the excellence as a goal to be reached are extremely important parameters. The involvement of all interested parties in the decision-making process is condition for a successful and publicly acceptable implementation of such project. The steps for public acceptance of a repository are summarized as follow: Risk perception: to verify how the local population understand and feel the installation of a repository in the region. Media observatory: to continuously follow the news reaching the region where the repository will be installed, including different media. Local population social/economical/cultural profile identification: to determine the local population social/economical/cultural profile; to conduct a survey to know their expectations, allowing the proposal of compensation and incentives to fully account for their expectations. Finally, the philosophy governing this Project is: on doubt, the public must be heard and only after this public hearing the policies concerning the project shall be formulated. (author)

  11. Human peptide transporters

    DEFF Research Database (Denmark)

    Nielsen, Carsten Uhd; Brodin, Birger; Jørgensen, Flemming Steen

    2002-01-01

    Peptide transporters are epithelial solute carriers. Their functional role has been characterised in the small intestine and proximal tubules, where they are involved in absorption of dietary peptides and peptide reabsorption, respectively. Currently, two peptide transporters, PepT1 and PepT2, wh...

  12. Interpretation of tandem mass spectra of posttranslationally modified peptides

    DEFF Research Database (Denmark)

    Bunkenborg, J.; Matthiesen, R.

    2013-01-01

    spectra and protein database search engines have been developed to match the experimental data to peptide candidates. In most studies there is a schism between discarding perfectly valid data and including nonsensical peptide identifications-this is currently a major bottleneck in data...

  13. Modelling saline intrusion for repository performance assessment

    International Nuclear Information System (INIS)

    Jackson, C.P.

    1989-04-01

    UK Nirex Ltd are currently considering the possibility of disposal of radioactive waste by burial in deep underground repositories. The natural pathway for radionuclides from such a repository to return to Man's immediate environment (the biosphere) is via groundwater. Thus analyses of the groundwater flow in the neighbourhood of a possible repository, and consequent radionuclide transport form an important part of a performance assessment for a repository. Some of the areas in the UK that might be considered as possible locations for a repository are near the coast. If a repository is located in a coastal region seawater may intrude into the groundwater flow system. As seawater is denser than fresh water buoyancy forces acting on the intruding saline water may have significant effects on the groundwater flow system, and consequently on the time for radionuclides to return to the biosphere. Further, the chemistry of the repository near-field may be strongly influenced by the salinity of the groundwater. It is therefore important for Nirex to have a capability for reliably modelling saline intrusion to an appropriate degree of accuracy in order to make performance assessments for a repository in a coastal region. This report describes work undertaken in the Nirex Research programme to provide such a capability. (author)

  14. Reference repository design concept for bedded salt

    Energy Technology Data Exchange (ETDEWEB)

    Carpenter, D.W.; Martin, R.W.

    1980-10-08

    A reference design concept is presented for the subsurface portions of a nuclear waste repository in bedded salt. General geologic, geotechnical, hydrologic and geochemical data as well as descriptions of the physical systems are provided for use on generic analyses of the pre- and post-sealing performance of repositories in this geologic medium. The geology of bedded salt deposits and the regional and repository horizon stratigraphy are discussed. Structural features of salt beds including discontinuities and dissolution features are presented and their effect on repository performance is discussed. Seismic hazards and the potential effects of earthquakes on underground repositories are presented. The effect on structural stability and worker safety during construction from hydrocarbon and inorganic gases is described. Geohydrologic considerations including regional hydrology, repository scale hydrology and several hydrological failure modes are presented in detail as well as the hydrological considerations that effect repository design. Operational phase performance is discussed with respect to operations, ventilation system, shaft conveyances, waste handling and retrieval systems and receival rates of nuclear waste. Performance analysis of the post sealing period of a nuclear repository is discussed, and parameters to be used in such an analysis are presented along with regulatory constraints. Some judgements are made regarding hydrologic failure scenarios. Finally, the design and licensing process, consistent with the current licensing procedure is described in a format that can be easily understood.

  15. Analisis Konten dan Kebijakan Akses Institutional Repository

    Directory of Open Access Journals (Sweden)

    Amirul Ulum

    2016-07-01

    Abstract; Institutional repository has become a major concern of higher education in Indonesia. The number of institutional respository was increased, one of the reason is the ranking web of repositories has been conducted by the Cybermetrics Lab in 2008. At that time, many institutions started to build institutional repository in order to manage the scientific work and also trying to reach the better ranks. Meanwhile, it is an achievement of institution performance which can be promote and increase visibility for the institution. University of Surabaya has also developed the institutional repository and managed by the library. The aims of this study is to analyze the content availability and access policies defined by the University of Surabaya repository  providing services to the academic community and external users. The method used in this study by using observations of the institutional repository University of Surabaya with a literature review to clarify the analysis of the content and access policies. The results of this study indicate that the library's role is has the authority to manage the scientific work of academic community can be done through the institutional repository. However there is still need for library to be proactive to communicate regulations on mandatory deposit of scientific work and create intensive promotion of the institutional repository.

  16. Electronic Repository of Russian Historical Statistics

    NARCIS (Netherlands)

    Tykhonov, Vyacheslav; Kessler, Gijs; Markevich, Andrei; de Vries, Jerry

    2014-01-01

    The Electronic Repository for Russian Historical Statistics brings together data extracted from various published and unpublished sources in one place. Its principal focus is Russian economic and social history of the last three centuries (18th-21st). The repository caters to the needs of the

  17. Decompression of magma into repository tunnels

    NARCIS (Netherlands)

    Bokhove, Onno; Woods, A.W.

    2002-01-01

    It is nontrivial to find and design safe repository sites for nuclear waste. It appears common sense to drill tunnels as repository sites in a mountain in remote and relatively dry regions. However, erosion of the waste canisters by naturally abundant chemicals in the mountains water cycle remains a

  18. Numerical modeling of magma-repository interactions

    NARCIS (Netherlands)

    Bokhove, Onno

    2001-01-01

    This report explains the numerical programs behind a comprehensive modeling effort of magma-repository interactions. Magma-repository interactions occur when a magma dike with high-volatile content magma ascends through surrounding rock and encounters a tunnel or drift filled with either a magmatic

  19. Asset Reuse of Images from a Repository

    Science.gov (United States)

    Herman, Deirdre

    2014-01-01

    According to Markus's theory of reuse, when digital repositories are deployed to collect and distribute organizational assets, they supposedly help ensure accountability, extend information exchange, and improve productivity. Such repositories require a large investment due to the continuing costs of hardware, software, user licenses, training,…

  20. Reference repository design concept for bedded salt

    International Nuclear Information System (INIS)

    Carpenter, D.W.; Martin, R.W.

    1980-01-01

    A reference design concept is presented for the subsurface portions of a nuclear waste repository in bedded salt. General geologic, geotechnical, hydrologic and geochemical data as well as descriptions of the physical systems are provided for use on generic analyses of the pre- and post-sealing performance of repositories in this geologic medium. The geology of bedded salt deposits and the regional and repository horizon stratigraphy are discussed. Structural features of salt beds including discontinuities and dissolution features are presented and their effect on repository performance is discussed. Seismic hazards and the potential effects of earthquakes on underground repositories are presented. The effect on structural stability and worker safety during construction from hydrocarbon and inorganic gases is described. Geohydrologic considerations including regional hydrology, repository scale hydrology and several hydrological failure modes are presented in detail as well as the hydrological considerations that effect repository design. Operational phase performance is discussed with respect to operations, ventilation system, shaft conveyances, waste handling and retrieval systems and receival rates of nuclear waste. Performance analysis of the post sealing period of a nuclear repository is discussed, and parameters to be used in such an analysis are presented along with regulatory constraints. Some judgements are made regarding hydrologic failure scenarios. Finally, the design and licensing process, consistent with the current licensing procedure is described in a format that can be easily understood

  1. Towards Content Development For Institutional Digital Repository ...

    African Journals Online (AJOL)

    The growth in Information and Communication Technology has lead to the emergence of Institutional Digital Repository, a digital archive for the preservation and dissemination of institutional research outputs. Institutional Digital Repositories make possible global dissemination of research outputs through the use of the ...

  2. THPdb: Database of FDA-approved peptide and protein therapeutics.

    Directory of Open Access Journals (Sweden)

    Salman Sadullah Usmani

    Full Text Available THPdb (http://crdd.osdd.net/raghava/thpdb/ is a manually curated repository of Food and Drug Administration (FDA approved therapeutic peptides and proteins. The information in THPdb has been compiled from 985 research publications, 70 patents and other resources like DrugBank. The current version of the database holds a total of 852 entries, providing comprehensive information on 239 US-FDA approved therapeutic peptides and proteins and their 380 drug variants. The information on each peptide and protein includes their sequences, chemical properties, composition, disease area, mode of activity, physical appearance, category or pharmacological class, pharmacodynamics, route of administration, toxicity, target of activity, etc. In addition, we have annotated the structure of most of the protein and peptides. A number of user-friendly tools have been integrated to facilitate easy browsing and data analysis. To assist scientific community, a web interface and mobile App have also been developed.

  3. EEI/UWASTE oversight of the DOE Repository Program by the Repository Information Exchange Team

    International Nuclear Information System (INIS)

    Henkel, C.J.; Supko, E.M.; Schwartz, M.H.

    1993-01-01

    The Utility Nuclear Waste and Transportation Program of the Edison Electric Institute (EEI/UWASTE) has conducted reviews of the US DOE's repository program through its Repository Information Exchange Team (RIET or Team). Eight such reviews have been conducted since 1985 covering topics that include repository program management and control; repository schedule; repository budget; quality assurance; site characterization; repository licensing; environmental issues; and institutional and public information activities. The utility industry has used these repository program reviews as a forum for providing DOE's Office of Civilian Radioactive Waste Management (OCRWM) with comments on the direction of the repository program, advice for future actions regarding quality assurance activities and repository licensing, and suggestions for management and control of the Repository Program. The most significant recommendations made by the utility industry through the RIET are discussed along with any subsequent action by OCRWM in response to or subsequent to utility industry recommendations. The process used by the RIET to develop its recommendations to OCRWM regarding the repository program is also discussed

  4. Overlapping CD8+ and CD4+ T-cell epitopes identification for the progression of epitope-based peptide vaccine from nucleocapsid and glycoprotein of emerging Rift Valley fever virus using immunoinformatics approach.

    Science.gov (United States)

    Adhikari, Utpal Kumar; Rahman, M Mizanur

    2017-12-01

    Rift Valley fever virus (RVFV) is an emergent arthropod-borne zoonotic infectious viral pathogen which causes fatal diseases in the humans and ruminants. Currently, no effective and licensed vaccine is available for the prevention of RVFV infection in endemic as well as in non-endemic regions. So, an immunoinformatics-driven genome-wide screening approach was performed for the identification of overlapping CD8+ and CD4+ T-cell epitopes and also linear B-cell epitopes from the conserved sequences of the nucleocapsid (N) and glycoprotein (G) of RVFV. We identified overlapping 99.39% conserved 1 CD8+ T-cell epitope (MMHPSFAGM) from N protein and 100% conserved 7 epitopes (AVFALAPVV, LAVFALAPV, FALAPVVFA, VFALAPVVF, IAMTVLPAL, FFDWFSGLM, and FLLIYLGRT) from G protein and also identified IL-4 and IFN-γ induced (99.39% conserved) 1 N protein CD4+ T-cell epitope (HMMHPSFAGMVDPSL) and 100% conserved 5 G protein CD4+ T-cell epitopes (LPALAVFALAPVVFA, PALAVFALAPVVFAE, GIAMTVLPALAVFAL, GSWNFFDWFSGLMSW, and FFLLIYLGRTGLSKM). The overlapping CD8+ and CD4+ T-cell epitopes were bound with most conserved HLA-C*12:03 and HLA-DRB1*01:01, respectively with the high binding affinity (kcal/mol). The combined population coverage analysis revealed that the allele frequencies of these epitopes are high in endemic and non-endemic regions. Besides, we found 100% conserved and non-allergenic 2 decamer B-cell epitopes, GVCEVGVQAL and RVFNCIDWVH of G protein had the sequence similarity with the nonamer CD8+ T-cell epitopes, VCEVGVQAL and RVFNCIDWV, respectively. Consequently, these epitopes may be used for the development of epitope-based peptide vaccine against emerging RVFV. However, in vivo and in vitro experiments are required for their efficient use as a vaccine. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Characteristics of potential repository wastes

    International Nuclear Information System (INIS)

    Cowart, C.G.; Notz, K.J.

    1992-10-01

    This report presents the results of a fully documented peer review of DOE/RW-0184, Rev. 1, ''Characteristics of Potential Repository Wastes''. The peer review was chaired and administered by oak Ridge National Laboratory (ORNL) for the Department of Energy (DOE), Office of Civilian Radioactive Waste Management (OCRWM) and was conducted in accordance with OCRWM QA procedure QAAP 3.3 ''Peer Review'' for the purpose of quailing the document for use in OCRWM quality-affecting work. The peer reviewers selected represent a wide range of experience and knowledge particularly suitable for evaluating the subject matter. A total of 596 formal comments were documented by the seven peer review panels, and all were successfully resolved. The peers reached the conclusion that DOE/RW-0184, Rev. 1, is quality determined and suitable for use in quality-affecting work

  6. A global nuclear waste repository

    Science.gov (United States)

    Lin, Wunan

    As a concerned scientist, I think that having a global nuclear waste repository is a reachable goal for human beings. Maybe through this common goal, mankind can begin to treat each other as brothers and sisters. So far, most human activities are framed by national boundaries, which are purely arbitrary. Breaking through these national boundaries will be very beneficial to human beings.Formation of the International Geosphere-Biosphere Program in 1986 indicates a growing awareness on the part of scientists regarding Earth as a system. The Apollo missions gave us a chance to look back at Earth from space. That perspective emphasized that our Earth is just one system: our only home. It is in deed a lonely boat in the high sea of dark space. We must take good care of our “boat.”

  7. Hydrologic issues in repository siting

    International Nuclear Information System (INIS)

    Remson, I.; Gorelick, S.M.

    1982-01-01

    Extrapolation of Darcy's law to the transport of water an solutes in unfractured poorly permeable rocks being studied for nuclear waste disposal is questioned. The hydrologic literature includes numerous references to both non-Darcian flow in dense materials devoid of macrofractures and microfractures and to threshold gradients below which no flow occurs. For such situations to occur, the pore-size range must be small enough so that all pore water is sufficiently close to mineral surfaces to be affected by the surficial forces. Then the flow will be non-Newtonian and non-Darcian, and solute transport will be by molecular diffusion. If fluid transport in very dense unfractured rocks is non-Darcian, useful methods of testing candidate host rocks become apparent. In situ nondestructive pressure testing of canister waste emplacement boreholes in a mined repository can verify the absence of both fracture flow and Darcian flow. 18 references

  8. IDENTIFICATION OF THE CORE RESIDUES OF THE EPITOPE OF A MONOCLONAL-ANTIBODY RAISED AGAINST GLYCOPROTEIN-D OF HERPES-SIMPLEX VIRUS TYPE-1 BY SCREENING OF A RANDOM PEPTIDE LIBRARY

    NARCIS (Netherlands)

    SCHELLEKENS, GA; LASONDER, E; FEIJLBRIEF, M; KOEDIJK, DGAM; DRIJFHOUT, JW; SCHEFFER, AJ; WELLINGWESTER, S; WELLING, GW

    1994-01-01

    Random peptide libraries (RPL) displayed on the surface of a filamentous bacteriophage can be used to identify peptide ligands that interact with target molecules. We have used a 15-amino acid residue RPL displayed on bacteriophage M13 to identify the core residues within the epitope of a monoclonal

  9. How many geologic repositories will be needed

    International Nuclear Information System (INIS)

    Evans, T.J.; Halstead, R.J.

    1987-01-01

    DOE's postponement of site-specific work on the second repository program had rekindled debate over the number of geologic repositories needed for disposal of high level radioactive waste. The multiple repository approach grew out of the March, 1979 IRG report, which recommended co-disposal of civilian and defense HLW in a system of regional repositories. The multiple repository approach was adopted by DOE, and incorporated in the Nuclear Waste Policy Act passed by Congress in December, 1982. Since the late 1970's, the slower than anticipated growth of the nuclear power industry has substantially reduced earlier estimates of the amount of civilian spent fuel which will require geologic disposal. Reactors currently in operation (78.5 GWe) and reactors in the construction pipeline (28 GWe) are expected to discharge about 103,200 MTU of spent fuel by the year 2036, assuming no increase in fuel burnup rate. By the year 2020, defense high level radioactive wastes equivalent to as much as 27,000 MTU could require geologic disposal. Small amounts of high level waste from other sources will also require geologic disposal. Total disposal requirements appear to be less than 140,000 MTU. The five sites nominated for the first repository, as well as hypothetical sites in granite, the host rock under primary consideration for the second repository, all appear capable of accommodating up to 140,000 MTU

  10. Learning frameworks as an alternative to repositories

    DEFF Research Database (Denmark)

    Dalsgaard, Christian

    2005-01-01

    This paper presents the concept of ‘learning frameworks’. The purpose of the paper is to discuss and question collections of digital learning objects in large repositories and to argue for large learning frameworks which organise a number of thematically related digital learning materials. Whereas...... a learning object repository contains all kinds of materials, a learning framework consists of an organisation of materials related to a common theme. Further, a repository consists of single, self-contained objects, whereas a learning framework is an open-ended environment which presents a number...

  11. Geotechnical materials considerations for conceptual repository design in the Palo Duro Basin, Texas

    International Nuclear Information System (INIS)

    Versluis, W.S.; Balderman, M.A.

    1984-01-01

    The Palo Duro Basin is only one of numerous potential repository locations for placement of a nuclear waste repository. Conceptual designs in the Palo Duro Basin involve considerations of the character and properties of the geologic materials found on several sites throughout the Basin. The first consideration presented includes current basin exploration results and interpretations of engineering properties for the basin geologic sequences. The next consideration presented includes identification of the characteristics of rock taken from the geologic sequence of interest through laboratory and field testing. Values for materials properties of representative samples are obtained for input into modeling of the material response to repository placement. Conceptual designs which respond to these geotechnical considerations are discussed. 4 references, 4 figures, 4 tables

  12. A cocoa peptide protects Caenorhabditis elegans from oxidative stress and β-amyloid peptide toxicity.

    Directory of Open Access Journals (Sweden)

    Patricia Martorell

    Full Text Available BACKGROUND: Cocoa and cocoa-based products contain different compounds with beneficial properties for human health. Polyphenols are the most frequently studied, and display antioxidant properties. Moreover, protein content is a very interesting source of antioxidant bioactive peptides, which can be used therapeutically for the prevention of age-related diseases. METHODOLOGY/PRINCIPAL FINDINGS: A bioactive peptide, 13L (DNYDNSAGKWWVT, was obtained from a hydrolyzed cocoa by-product by chromatography. The in vitro inhibition of prolyl endopeptidase (PEP was used as screening method to select the suitable fraction for peptide identification. Functional analysis of 13L peptide was achieved using the transgenic Caenorhabditis elegans strain CL4176 expressing the human Aβ₁₋₄₂ peptide as a pre-clinical in vivo model for Alzheimer's disease. Among the peptides isolated, peptide 13L (1 µg/mL showed the highest antioxidant activity (P≤0.001 in the wild-type strain (N2. Furthermore, 13L produced a significant delay in body paralysis in strain CL4176, especially in the 24-47 h period after Aβ₁₋₄₂ peptide induction (P≤0.0001. This observation is in accordance with the reduction of Aβ deposits in CL4176 by western blot. Finally, transcriptomic analysis in wild-type nematodes treated with 13L revealed modulation of the proteosomal and synaptic functions as the main metabolic targets of the peptide. CONCLUSIONS/SIGNIFICANCE: These findings suggest that the cocoa 13L peptide has antioxidant activity and may reduce Aβ deposition in a C. elegans model of Alzheimer's disease; and therefore has a putative therapeutic potential for prevention of age-related diseases. Further studies in murine models and humans will be essential to analyze the effectiveness of the 13L peptide in higher animals.

  13. Nuclear waste repository characterization: a spatial estimation/identification approach

    International Nuclear Information System (INIS)

    Candy, J.V.; Mao, N.

    1981-03-01

    This paper considers the application of spatial estimation techniques to a groundwater aquifer and geological borehole data. It investigates the adequacy of these techniques to reliably develop contour maps from various data sets. The practice of spatial estimation is discussed and the estimator is then applied to a groundwater aquifer system and a deep geological formation. It is shown that the various statistical models must first be identified from the data and evaluated before reasonable results can be expected

  14. Identification of the IGF-1 processing product human Ec/rodent Eb peptide in various tissues: Evidence for its differential regulation after exercise-induced muscle damage in humans.

    Science.gov (United States)

    Vassilakos, George; Philippou, Anastassios; Koutsilieris, Michael

    2017-02-01

    Insulin-like growth factor-1 (IGF-1) is a pleiotropic factor expressed in various tissues and plays a critical role in skeletal muscle physiology. Alternative splicing of the IGF-1 gene gives rise to different precursor polypeptides (isoforms) which could undergo post-translational cleavage, generating the common mature IGF-1 peptide and different carboxyl terminal extension (E-) peptides, with the fate of the latter being, so far, unknown. The objective if this study was to identify the IGF-1Ec forms or processing product(s), other than mature IGF-1, generated in different human and rodent tissues and particularly in human skeletal muscle after exercise-induced damage. Protein lysates from a wide range of human and rodent tissues were immunoblotted with a rabbit anti-human Ec polyclonal antibody raised against the last 24 amino acids of the C-terminal of the Ec peptide. This antibody can recognize the Ec peptide, both as part of IGF-1Ec and alone, and also the corresponding rodent forms, due to the high homology that the human Ec shares with the rodent Eb. We were able to confirm, for the first time, that the human Ec peptide and its rodent homologous Eb peptide are produced simultaneously with their precursor protein (pro-IGF-1Ec/Eb) in vivo, in a wide range of tissues (e.g. muscle, liver, heart). Proprotein convertase furin digestion of human muscle and liver protein lysates confirmed that the higher molecular form, pro-IGF-1Ec, can be cleaved to produce the free Ec peptide. Furthermore, initial evidence is provided that Ec peptide is differentially regulated during the process of muscle regeneration after exercise-induced damage in humans. The findings of this study possibly imply that the post-translational modification of the IGF-1Ec pro-peptide may regulate the bioavailability and activity of the processing product(s). Copyright © 2016. Published by Elsevier Ltd.

  15. PeptideAtlas

    Data.gov (United States)

    U.S. Department of Health & Human Services — PeptideAtlas is a multi-organism, publicly accessible compendium of peptides identified in a large set of tandem mass spectrometry proteomics experiments. Mass...

  16. Sorption on inactive repository components

    International Nuclear Information System (INIS)

    Gardiner, M.P.; Smith, A.J.; Williams, S.J.

    1990-11-01

    The near-field of an intermediate level/low level radioactive waste repository will contain significant quantities of iron and steel, Magnox and Zircaloy. Their corrosion products may possess significant sorption capacity for radioelements. The sorption of americium and plutonium onto magnesium hydroxide, zirconium hydroxide, colloidal magnetite and colloidal haematite has been studied under conditions typical of the porewater of a cementitious near-field. R D values ≥ 10 5 m g -1 were measured for both actinides on the oxides and hydroxides. These values are at least as great as those measured on crushed 3:1 Blast Furnace Slag/Ordinary Portland cement. Competitive sorption experiments have shown that sorption onto the corrosion products does not take place in preference to that on the cement or the converse. Magnetite and haematite colloids are positively charged in cement-equilibrated water whilst zirconium hydroxide is negatively charged. Crushed cement was found to be positively charged. Simple experiments show that only a small proportion of haematite colloids is potentially mobile through a column of crushed cement. (author)

  17. Sorption on inactive repository components

    International Nuclear Information System (INIS)

    Gardiner, M.P.; Smith, A.J.; Williams, S.J.

    1990-07-01

    The near-field of an intermediate level/low level radioactive waste repository will contain significant quantities of iron and steel, Magnox and Zircaloy. Their corrosion products may possess significant sorption capacity for radioelements. The sorption of americium and plutonium onto magnesium hydroxide, zirconium hydroxide, colloidal magnetite and colloidal haematite has been studied under conditions typical of the porewater of a cementitious near-field. R D values ≥ 10 5 ml g -1 were measured for both actinides on the oxides and hydroxides. These values are at least as great at those measured on crushed 3:1 Blast Furnace Slag/Ordinary Portland Cement. Competitive sorption experiments have shown that sorption onto the corrosion products does not take place in preference to that on the cement or the converse. Magnetite and haematite colloids are positively charged in cement-equilibrated water whilst zirconium hydroxide is negatively charged. Crushed cement was found to be positively charged. Simple experiments show that only a small proportion of haematite colloids is potentially mobile through a column of crushed cement. (author)

  18. Andra's geologic repository monitoring strategy

    International Nuclear Information System (INIS)

    Buschaert, S.; Lesoille, S.; Bertrand, J.; Landais, P.

    2012-01-01

    Document available in extended abstract form only. After having concluded a feasibility study of deep geological disposal for high-level and long-lived radioactive waste in 2005, Andra was charged by the Planning Act no. 2006-739 to design and create an industrial site for geological disposal called Cigeo which must be reversible for at least a century-long period. The French Safety Guide recommends that Andra develop a monitoring program to be implemented at repository construction and conducted until closure, and possibly after closure, with the aim to confirming prior expectations and enhancing knowledge of relevant processes. This abstract focuses on underground structure monitoring. The monitoring system is based on a combination of in-situ instrumentation and nondestructive methods to obtain the required level of reliable performance. To optimize the device distribution, we take into account both the repetitive design of disposal cells and the homogeneity of the rock properties. This resulted in distinguishing pilot disposal cells that are highly instrumented and standard disposal cells where the instrumentation density could be reduced; monitoring will rely mostly on robotic nondestructive evaluations. If monitoring technologies do not comply with all monitoring objectives, real withdrawal tests of high level wastes in some pilot disposal cells are also planned to provide the possibility of carrying out visual inspection, destructive analyses and samplings on construction materials. Such cells are planned to be dismantled because of the potential disturbance of their component performances from the testing process. Based on this overall strategy, Andra has analyzed the technical requirements that must be met by its monitoring equipment. First, these must be able to provide information on key THMCR (Thermal- Hydraulic-Mechanical-Chemical and Radiological) processes, to provide a three-dimensional image of a disposal component's behavior and thus to understand

  19. REPOSITORY RADIATION SHIELDING DESIGN GUIDE

    International Nuclear Information System (INIS)

    M. Haas; E.M. Fortsch

    1997-01-01

    The scope of this document includes radiation safety considerations used in the design of facilities for the Yucca Mountain Site Characterization Project (YMP). The purpose of the Repository Radiation Shielding Design Guide is to document the approach used in the radiological design of the Mined Geologic Disposal System (MGDS) surface and subsurface facilities for the protection of workers, the public, and the environment. This document is intended to ensure that a common methodology is used by all groups that may be involved with Radiological Design. This document will also assist in ensuring the long term survivability of the information basis used for radiological safety design and will assist in satisfying the documentation requirements of the licensing body, the Nuclear Regulatory Commission (NRC). This design guide provides referenceable information that is current and maintained under the YMP Quality Assurance (QA) Program. Furthermore, this approach is consistent with maintaining continuity in spite of a changing design environment. This approach also serves to ensure common inter-disciplinary interpretation and application of data

  20. REPOSITORY RADIATION SHIELDING DESIGN GUIDE

    Energy Technology Data Exchange (ETDEWEB)

    M. Haas; E.M. Fortsch

    1997-09-12

    The scope of this document includes radiation safety considerations used in the design of facilities for the Yucca Mountain Site Characterization Project (YMP). The purpose of the Repository Radiation Shielding Design Guide is to document the approach used in the radiological design of the Mined Geologic Disposal System (MGDS) surface and subsurface facilities for the protection of workers, the public, and the environment. This document is intended to ensure that a common methodology is used by all groups that may be involved with Radiological Design. This document will also assist in ensuring the long term survivability of the information basis used for radiological safety design and will assist in satisfying the documentation requirements of the licensing body, the Nuclear Regulatory Commission (NRC). This design guide provides referenceable information that is current and maintained under the YMP Quality Assurance (QA) Program. Furthermore, this approach is consistent with maintaining continuity in spite of a changing design environment. This approach also serves to ensure common inter-disciplinary interpretation and application of data.

  1. Characteristics of potential repository wastes

    International Nuclear Information System (INIS)

    1992-07-01

    The LWR spent fuels discussed in Volume 1 of this report comprise about 99% of all domestic non-reprocessed spent fuel. In this report we discuss other types of spent fuels which, although small in relative quantity, consist of a number of diverse types, sizes, and compositions. Many of these fuels are candidates for repository disposal. Some non-LWR spent fuels are currently reprocessed or are scheduled for reprocessing in DOE facilities at the Savannah River Site, Hanford Site, and the Idaho National Engineering Laboratory. It appears likely that the reprocessing of fuels that have been reprocessed in the past will continue and that the resulting high-level wastes will become part of defense HLW. However, it is not entirely clear in some cases whether a given fuel will be reprocessed, especially in cases where pretreatment may be needed before reprocessing, or where the enrichment is not high enough to make reprocessing attractive. Some fuels may be canistered, while others may require special means of disposal. The major categories covered in this chapter include HTGR spent fuel from the Fort St. Vrain and Peach Bottom-1 reactors, research and test reactor fuels, and miscellaneous fuels, and wastes generated from the decommissioning of facilities

  2. Economics of mined geologic repositories

    International Nuclear Information System (INIS)

    Hofmann, P.L.; Dippold, D.G.

    1983-01-01

    During 1982, Congress considered legislation to provide for the development of repositories for the disposal of high-level radioactive waste and spent nuclear fuel. The result of this legislative effort was the Nuclear Waste Policy Act of 1982 (NWPA), PL 97-425, signed into law January 7, 1983. An important part of the NWPA was the establishment of special funds in the US Treasury for Waste Disposal and Interim Storage to be financed by user fees to pay for all costs of the program. An initial fee of 1.0 mill per kilowatt-hour was specified. The Secretary was asked to annually review the amount of the fees established... to evaluate whether collection of the fee will provide sufficient revenues to offset the costs... In the event of a prospective fee cost mismatch, the Secretary was asked to propose an adjustment to the fee to insure full cost recovery. A series of studies were sponsored by DOE in 1982 to estimate program costs, to calculate the necessary fees to assure cost recovery, and to address uncertainties that could affect future program costs and consequent fee schedules. A brief summary of the 1982 cost estimates is presented. Sources of key cost uncertainties are discussed and the bases for the cost recovery fee calculations are summarized. 17 references, 2 figures, 3 tables

  3. Repository Closure and Sealing Approach

    International Nuclear Information System (INIS)

    A.T. Watkins

    2000-01-01

    The scope of this analysis will be to develop the conceptual design of the closure seals and their locations in the Subsurface Facilities. The design will be based on the recently established program requirements for transitioning to the Site Recommendation (SR) design as outlined by ''Approach to Implementing the Site Recommendation Baseline'' (Stroupe 2000) and the ''Monitored Geologic Repository Project Description Document'' (CRWMS M andO 1999b). The objective of this analysis will be to assist in providing a description for the Subsurface Facilities System Description Document, Section 2 and finally to document any conclusions reached in order to contribute and provide support to the SR. This analysis is at a conceptual level and is considered adequate to support the SR design. The final closure barriers and seals for the ventilation shafts, and the north and south ramps will require these openings to be permanently sealed to limit excessive air and water inflows and prevent human intrusion. The major tasks identified with closure in this analysis are: (1) Developing the overall subsurface seal layout and identifying design and operational interfaces for the Subsurface Facilities. (2) Summarizing the general site conditions and general rock characteristic with respect to seal location and describing the seal selected. (3) Identify seal construction materials, methodology of construction and strategic locations including design of the seal and plugs. (4) Discussing methods to prevent human intrusion

  4. Building the repositories to serve

    International Nuclear Information System (INIS)

    Lersch, D.

    1994-01-01

    The project to design and build the Superconducting Super Collider (SSC) Laboratory also includes the exciting opportunity to implement client/server information systems. Lab technologists were eager to take advantage of the cost savings inherent in the open systems and a distributed, client server environment and, at the same time, conscious of the need to provide secure repositories for sensitive data as well as a schedule sensitive acquisition strategy for mission critical software. During the first year of project activity, micro-based project management and business support systems were acquired and implemented to support a small study project of less than 400 people allocating contracts of less than $1 million. The transition to modern business systems capable of supporting more than 10,000 participants (world wide) who would be researching and developing the new technologies that would support the world's largest scientific instrument, a 42 Tevatron, superconducting, super collider became a mission critical event. This paper will present the SSC Laboratory's strategy to balance its commitment to open systems, structured query language (SQL) standards and its success with acquiring commercial off the shelf software to support immediate goals. Included will be an outline of the vital roles played by other labs (Livermore, CERN, Brookhaven, Fermi and others) and a discussion of future collaboration potentials to leverage the information activities of all Department of Energy funded labs

  5. Building the repositories to serve

    International Nuclear Information System (INIS)

    Lersch, D.

    1993-04-01

    The project to design and build the Superconducting Super Collider (SSC) Laboratory also includes the exciting opportunity to implement client/server information systems. Lab technologists were eager to take advantage of the cost savings inherent in the open systems and a distributed, client server environment and, at the same time, conscious of the need to provide secure repositories for sensitive data as well as a schedule sensitive acquisition strategy for mission critical software. During the first year of project activity, micro-based project management and business support systems were acquired and implemented to support a small study project of less than 400 people allocating contracts of less than $1 million. The transition to modern business systems capable of supporting more than 10,000 participants (world wide) who would be researching and developing the new technologies that would support the world's largest scientific instrument, a 42 Tevatron, superconducting, super collider became a mission critical event. This paper will present the SSC Laboratory's strategy to balance our commitment to open systems, structured query language (SQL) standards and our success with acquiring commercial off the shelf software (COTS) to support our immediate goals. Included will be an outline of the vital roles played by other labs (Livermore, CERN, Brookhaven, Fermi and others) and a discussion of future collaboration potentials to leverage the information activities of all Department of Energy (DOE) funded labs

  6. Identification of peptides from foot‐and‐mouth disease virus structural proteins bound by class I swine leukocyte antigen (SLA) alleles, SLA‐1*0401 and SLA‐2*0401

    DEFF Research Database (Denmark)

    Pedersen, Lasse Eggers; Harndahl, M.; Nielsen, Morten

    2013-01-01

    within the structural proteins of foot‐and‐mouth disease virus (FMDV), strain A24 were analyzed as candidate T‐cell epitopes. Peptides predicted by the NetMHCpan were tested in ELISA for binding to the SLA‐1*0401 and SLA‐2*0401 major histocompatibility complex class I proteins. Four of the 10 predicted...... FMDV peptides bound to SLA‐2*0401, whereas five of the nine predicted FMDV peptides bound to SLA‐1*0401. These methods provide the characterization of T‐cell epitopes in response to pathogens in more detail. The development of such approaches to analyze vaccine performance will contribute to a more...

  7. Shaft placement in a bedded salt repository

    International Nuclear Information System (INIS)

    Klasi, M.L.

    1982-10-01

    Preferred shaft pillar sizes and shaft locations were determined with respect to the induced thermal stresses in a generic bedded salt repository at a depth of 610 m with a gross thermal loading of 14.8 W/m 2 . The model assumes isotropic material properties, plane strain and linear elastic behavior. Various shaft locations were analyzed over a 25 year period. The thermal results show that for this time span, the stratigraphy is unimportant except for the region immediately adjacent to the repository. The thermomechanical results show that for the given repository depth of 610 m, a minimum central shaft pillar radius of 244 m is required to equal the material strength in the barrier pillar. An assumed constant stress and constant temperature distribution creep model of the central shaft region adjacent to the repository conservatively overestimates a creep closure of 310 mm in a 6.1 m diameter centrally-located shaft

  8. Preliminary design of the repository, stage 2

    International Nuclear Information System (INIS)

    Saanio, T.; Kirkkomaeki, T.; Keto, P.; Kukkola, T.; Raiko, H.

    2007-01-01

    Spent nuclear fuel from Finnish nuclear power plants will be disposed of in deep bedrock in Olkiluoto, Eurajoki. The repository is planned to be excavated at a depth of 400 - 500 metres. Access routes to the repository include a 1:10 inclined access tunnel, and vertical shafts. The fuel is encapsulated in the encapsulation plant above ground and transferred to the repository in the canister lift. Deposition tunnels, central tunnels and technical rooms are excavated at the disposal level. The canisters are deposited in deposition holes that are covered with bentonite blocks. The deposition holes are bored in the floors of the deposition tunnels. The central tunnel system consists of two parallel central tunnels that are inter-connected at certain distances. Two parallel central tunnels improve the fire safety of the rooms and also allow flexible backfilling and closing of the deposition tunnels in stages at the operational phase of the repository. An underground rock characterization facility, ONKALO, is excavated at the disposal level to support and confirm investigations carried out from above ground. ONKALO is designed so that it can later serve as part of the repository. ONKALO excavations were started in 2004. The repository will be excavated in the 2010s and operation will start in 2020. The fifth nuclear power unit makes the operational phase of the repository very long. Parts of the repository will be excavated and closed over the long operational period. The repository can be constructed at one or several levels. The one-storey alternative is the so-called reference alternative in this preliminary design report. The two-storey alternative is also taken into account in the ONKALO designs. The preliminary designs of the repository are presented as located in Olkiluoto. The location of the repository will be revised when more information on the bedrock has been gained. More detailed data of the circumstances will be obtained from above ground investigations

  9. Decision theory applied to radioactive repository construction

    International Nuclear Information System (INIS)

    Heilbron Filho, Paulo Fernando Lavalle; Pontedeiro, Elizabeth May

    2001-01-01

    The objective of this article is to present, through the presentation of an example, the applicability of the decision theory on the selection and construction of a repository for low and intermediate radioactive waste. (author)

  10. Preliminary design of the repository. Stage 2

    International Nuclear Information System (INIS)

    Saanio, T.; Kirkkomaeki, T.; Keto, P.; Kukkola, T.; Raiko, H.

    2007-04-01

    Spent nuclear fuel from Finnish nuclear power plants will be disposed of in deep bedrock in Olkiluoto, Eurajoki. The repository is planned to be excavated at a depth of 400 - 500 metres. Access routes to the repository include a 1:10 inclined access tunnel, and vertical shafts. The fuel is encapsulated in the encapsulation plant above ground and transferred to the repository in the canister lift. Deposition tunnels, central tunnels and technical rooms are excavated at the disposal level. The canisters are deposited in deposition holes that are covered with bentonite blocks. The deposition holes are bored in the floors of the deposition tunnels. The central tunnel system consists of two parallel central tunnels that are inter-connected at certain distances. Two parallel central tunnels improve the fire safety of the rooms and also allow flexible backfilling and closing of the deposition tunnels in stages at the operational phase of the repository. An underground rock characterization facility, ONKALO, is excavated at the disposal level to support and confirm investigations carried out from above ground. ONKALO is designed so that it can later serve as part of the repository. ONKALO excavations were started in 2004. The repository will be excavated in the 2010s and operation will start in 2020. The fifth nuclear power unit makes the operational phase of the repository very long. Parts of the repository will be excavated and closed over the long operational period. The repository can be constructed at one or several levels. The one-storey alternative is the so-called reference alternative in this preliminary design report. The two-storey alternative is also taken into account in the ONKALO designs. The preliminary designs of the repository are presented as located in Olkiluoto. The location of the repository will be revised when more information on the bedrock has been gained. More detailed data of the circumstances will be obtained from above ground investigations

  11. Biospecimen Repository Access and Data Sharing (BRADS)

    Data.gov (United States)

    U.S. Department of Health & Human Services — BRADS is a repository for data and biospecimens from population health research initiatives and clinical or interventional trials designed and implemented by NICHD’s...

  12. NIMH Repository and Genomics Resources (RGR)

    Data.gov (United States)

    U.S. Department of Health & Human Services — The NIMH Repository and Genomics Resource (RGR) stores biosamples, genetic, pedigree and clinical data collected in designated NIMH-funded human subject studies. The...

  13. Evaluasi Website Repositori Institusi Universitas Surabaya

    Directory of Open Access Journals (Sweden)

    Amirul Ulum

    2015-11-01

    Full Text Available The  development  of  institutional  repositories  (IRs  in Indonesia within the broader framework of open access has significant impact on preserving intellectual capital and scholarly communication. Institutional  repositories  play  a  fundamental  role  in  centralizing, preserving,  and  making  accessible  institution’s  intellectual  capital. Evaluation of the system is to determine the functionality the system to meet the users need. Using a descriptive analysis this study wants to evaluate institutional repositories of University of Surabaya. The result is usefull for institution to develop the repository systems.

  14. Workshop: Creating Your Institutional Research Repository

    KAUST Repository

    Grenz, Daryl M.; Baessa, Mohamed A.

    2016-01-01

    In 2002, the Scholarly Publishing and Academic Resources Coalition (SPARC) proposed the concept of an institutional repository to simultaneously disrupt and enhance the state of scholarly communications in the academic world. Thirteen years later

  15. 48 CFR 227.7108 - Contractor data repositories.

    Science.gov (United States)

    2010-10-01

    ... repositories. 227.7108 Section 227.7108 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS... Technical Data 227.7108 Contractor data repositories. (a) Contractor data repositories may be established when permitted by agency procedures. The contractual instrument establishing the data repository must...

  16. Peptide-Carrier Conjugation

    DEFF Research Database (Denmark)

    Hansen, Paul Robert

    2015-01-01

    To produce antibodies against synthetic peptides it is necessary to couple them to a protein carrier. This chapter provides a nonspecialist overview of peptide-carrier conjugation. Furthermore, a protocol for coupling cysteine-containing peptides to bovine serum albumin is outlined....

  17. PH dependent adhesive peptides

    Science.gov (United States)

    Tomich, John; Iwamoto, Takeo; Shen, Xinchun; Sun, Xiuzhi Susan

    2010-06-29

    A novel peptide adhesive motif is described that requires no receptor or cross-links to achieve maximal adhesive strength. Several peptides with different degrees of adhesive strength have been designed and synthesized using solid phase chemistries. All peptides contain a common hydrophobic core sequence flanked by positively or negatively charged amino acids sequences.

  18. Peptide Nucleic Acids

    DEFF Research Database (Denmark)

    2003-01-01

    A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker....

  19. Peptide Nucleic Acids

    DEFF Research Database (Denmark)

    1998-01-01

    A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker....

  20. Peptide Nucleic Acids (PNA)

    DEFF Research Database (Denmark)

    2002-01-01

    A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker....

  1. Peptide Nucleic Acid Synthons

    DEFF Research Database (Denmark)

    2004-01-01

    A novel class of compounds, known as peptide nucleic acids, bind complementary ssDNA and RNA strands more strongly than a corresponding DNA. The peptide nucleic acids generally comprise ligands such as naturally occurring DNA bases attached to a peptide backbone through a suitable linker....

  2. Antimicrobial Peptides in 2014

    Directory of Open Access Journals (Sweden)

    Guangshun Wang

    2015-03-01

    Full Text Available This article highlights new members, novel mechanisms of action, new functions, and interesting applications of antimicrobial peptides reported in 2014. As of December 2014, over 100 new peptides were registered into the Antimicrobial Peptide Database, increasing the total number of entries to 2493. Unique antimicrobial peptides have been identified from marine bacteria, fungi, and plants. Environmental conditions clearly influence peptide activity or function. Human α-defensin HD-6 is only antimicrobial under reduced conditions. The pH-dependent oligomerization of human cathelicidin LL-37 is linked to double-stranded RNA delivery to endosomes, where the acidic pH triggers the dissociation of the peptide aggregate to release its cargo. Proline-rich peptides, previously known to bind to heat shock proteins, are shown to inhibit protein synthesis. A model antimicrobial peptide is demonstrated to have multiple hits on bacteria, including surface protein delocalization. While cell surface modification to decrease cationic peptide binding is a recognized resistance mechanism for pathogenic bacteria, it is also used as a survival strategy for commensal bacteria. The year 2014 also witnessed continued efforts in exploiting potential applications of antimicrobial peptides. We highlight 3D structure-based design of peptide antimicrobials and vaccines, surface coating, delivery systems, and microbial detection devices involving antimicrobial peptides. The 2014 results also support that combination therapy is preferred over monotherapy in treating biofilms.

  3. Chemical risks from nuclear waste repositories

    International Nuclear Information System (INIS)

    Persson, L.

    1988-01-01

    Studies concerning the chemical risks of nuclear waste are reviewed. The radiological toxicity of the material is of primary concern but the potential nonradiological toxicity should not be overlooked as the chemotoxic substances may reach the biosphere from a nuclear waste repository. In the report is concluded that the possible chemotoxic effects of a repository for nuclear waste should be studied as a part of the formal risk assessment of the disposal concept. (author)

  4. Memory provisions for the Manche Surface Repository

    International Nuclear Information System (INIS)

    Dumont, Jean-Noel; Espiet-Subert, Florence

    2015-01-01

    The French La Manche repository site received its last radioactive waste package in 1994. In 2003, the official surveillance phase of the closed repository started under the supervision of Andra (the national industrial operator), the French Nuclear Safety Authority (ASN) and society (e.g. the local municipalities). Florence Espiet explained that information on the existence of the repository, its content, how it was operated and how it works needs to be preserved. It also is planned to review the information periodically for a minimum of 300 years. She described the creation of two documents on memory (a detailed and a summary one), both on permanent paper, and the preservation of the land registration. The latter constitutes 'passive' provisions for preserving memory. In addition, a number of 'active' provisions are and will be put in place: guided visits, exhibitions, partnerships with organisations dealing with memory preservation, and the creation of a think tank. The latter consists of local citizens and politicians, retired employees from Andra and artists that meet several times a year and reflect on memory preservation from the perspective of, for instance, local history, education, arts and rituals. Finally, two types of markers will be used to preserve the repository's memory: i) three herbaria cataloguing the plants growing on the site of the repository, including a very short description of the repository, will be stored at different sites in France; ii) a stele indicating the main characteristics of the repository, potentially linked to an art work, will be erected at the repository

  5. Impact of retrievability of repository design

    International Nuclear Information System (INIS)

    Heijdra, J.J.; Gaag, J. v.d.; Prij, J.

    1995-01-01

    In this paper the impact of the retrievability on the design of the repository will be handled. Retrievability of radioactive waste from a repository in geological formations has received increasing attention during recent years. It is obvious that this retrievability will have consequences in terms of mining engineering, safety and cost. The purpose of the present study is to evaluate cost consequences by comparing two extreme options for retrievable storage. (author). 6 refs., 3 figs

  6. Transfer systems in an underground repository

    International Nuclear Information System (INIS)

    Berg, H.P.; Ehrlich, D.

    1991-01-01

    In addition to logistic problem definitions taking into account the waste types of the wastes to be disposed of and the mining conditions, transport and handling of radioactive wastes in a repository, particularly require the keeping of safety technological marginal conditions mainly resulting from the accident analyses carried out. The realization of these safety technological aspects is described taking the planned Konrad repository as an example. (author)

  7. Implementation of the Brazilian national repository

    International Nuclear Information System (INIS)

    Tello, Cledola Cassia Oliveira de

    2013-01-01

    Ionizing radiation in Brazil is used in electricity generation, medicine, industry, agriculture and for research and development purposes. All these activities can generate radioactive waste. At this point, in Brazil, the use of nuclear energy and radioisotopes justifies the construction of a national repository for radioactive wastes of low and intermediate-level. According to Federal Law No. 10308, Brazilian National Commission for Nuclear Energy (CNEN) is responsible for designing and constructing the intermediate and final storages for radioactive wastes. Additionally, a restriction on the construction of Angra 3 is that the repository is under construction until its operation start, attaining some requirements of the Brazilian Environmental Regulator (IBAMA). The RBMN Project (Repository for Low and Intermediate-Level Radioactive Wastes) aims at the implantation of a National Repository for disposal of low and intermediate-level of radiation wastes. This Project has some aspects that are unique in the Brazilian context, especially referring to the time between its construction and the end of its institutional period. This time is about 360 years, when the area will be released for unrestricted uses. It means that the Repository must be safe and secure for more than three hundred years, which is longer than half of the whole of Brazilian history. This aspect is very new for the Brazilian people, bringing a new dimension to public acceptance. Another point is this will be the first repository in South America, bringing a real challenge for the continent. The current status of the Project is summarized. (author)

  8. The development of safeguards for geological repositories

    International Nuclear Information System (INIS)

    Van der Meer, K.

    2009-01-01

    Traditionally, research and development on geological repositories for High Level Waste (HLW) focuses on the short- and long-term safety aspects of the repository. If the repository will also be used for the disposal of spent fuel, safeguards aspects have to be taken into account. Safety and safeguards requirements may be contradictory; the safety of a geological repository is based on the non-intrusion of the geological containment, while safeguards require regular inspections of position and amount of the spent fuel. Examples to reconcile these contradictory requirements are the use of information required for the safety assessment of the geological repository for safeguards purposes and the adaptation of the safeguards approach to use non-intrusive inspection techniques. The principles of an inspection approach for a geological repository are now generally accepted within the IAEA. The practical applicability of the envisaged inspection techniques is still subject to investigation. It is specifically important for the Belgian situation that an inspection technique can be used in clay, the geological medium in which Belgium intends to dispose its HLW and spent fuel. The work reported in this chapter is the result of an international cooperation in the framework of the IAEA, in which SCK-CEN participates

  9. Monitoring during the stepwise implementation of the Swedish deep repository for spent fuel

    International Nuclear Information System (INIS)

    Baeckblom, Goeran; Almen, Karl-Erik

    2004-03-01

    to: obtain knowledge of undisturbed conditions in nature and their seasonal variations (baseline) in order to identify and evaluate the impact of activities related to the deep repository during different phases, obtain a better understanding of the function of the deep repository system to support the safety account and to test models and assumptions, monitor the environmental impact of the deep repository, provide evidence that the working environment is safe with regard to radiological and non-radiological effects, show that requirements on radioactive waste verification (safeguards) are fulfilled. The monitoring is concerned with bedrock conditions before and during construction and operation of the facilities, active design to engineer the underground facilities based on monitoring results during construction, the safe and environmental construction and operation of facilities. Experience exists from monitoring of the barrier function at SFR, the final repository for low - and intermediate level waste and at the Aespoe Hard Rock Laboratory and monitoring for safeguard at CLAB, the interim storage for spent nuclear fuel. The know-how on monitoring relates to all aspects from design, installation, operation and maintenance to decommissioning of systems. This report provides an overview of the features, processes and parameters that are collected during the site investigations to establish the Primary Baseline conditions for monitoring prior to the construction of the repository. It is also proposed that the monitoring programme should at least entail the following elements: objectives for the monitoring programme, scope for the monitoring (criteria for selection of issues to be monitored, identification of the properties, processes, phenomena and observable quantities to be monitored), identification on what methods to be used, operation of the monitoring system (identification of the duration and frequency of monitoring, including criteria for when monitoring may

  10. Proteomics data exchange and storage: the need for common standards and public repositories.

    Science.gov (United States)

    Jiménez, Rafael C; Vizcaíno, Juan Antonio

    2013-01-01

    Both the existence of data standards and public databases or repositories have been key factors behind the development of the existing "omics" approaches. In this book chapter we first review the main existing mass spectrometry (MS)-based proteomics resources: PRIDE, PeptideAtlas, GPMDB, and Tranche. Second, we report on the current status of the different proteomics data standards developed by the Proteomics Standards Initiative (PSI): the formats mzML, mzIdentML, mzQuantML, TraML, and PSI-MI XML are then reviewed. Finally, we present an easy way to query and access MS proteomics data in the PRIDE database, as a representative of the existing repositories, using the workflow management system (WMS) tool Taverna. Two different publicly available workflows are explained and described.

  11. Muon Tomography for Geological Repositories.

    Science.gov (United States)

    Woodward, D.; Kudryavtsev, V.; Gluyas, J.; Clark, S. J.; Thompson, L. F.; Klinger, J.; Spooner, N. J.; Blackwell, T. B.; Pal, S.; Lincoln, D. L.; Paling, S. M.; Mitchell, C. N.; Benton, C.; Coleman, M. L.; Telfer, S.; Cole, A.; Nolan, S.; Chadwick, P.

    2015-12-01

    Cosmic-ray muons are subatomic particles produced in the upper atmosphere in collisions of primary cosmic rays with atoms in air. Due to their high penetrating power these muons can be used to image the content (primarily density) of matter they pass through. They have already been used to image the structure of pyramids, volcanoes and other objects. Their applications can be extended to investigating the structure of, and monitoring changes in geological formations and repositories, in particular deep subsurface sites with stored CO2. Current methods of monitoring subsurface CO2, such as repeat seismic surveys, are episodic and require highly skilled personnel to operate. Our simulations based on simplified models have previously shown that muon tomography could be used to continuously monitor CO2 injection and migration and complement existing technologies. Here we present a simulation of the monitoring of CO2 plume evolution in a geological reservoir using muon tomography. The stratigraphy in the vicinity of the reservoir is modelled using geological data, and a numerical fluid flow model is used to describe the time evolution of the CO2 plume. A planar detection region with a surface area of 1000 m2 is considered, at a vertical depth of 776 m below the seabed. We find that one year of constant CO2 injection leads to changes in the column density of about 1%, and that the CO2 plume is already resolvable with an exposure time of less than 50 days. The attached figure show a map of CO2 plume in angular coordinates as reconstructed from observed muons. In parallel with simulation efforts, a small prototype muon detector has been designed, built and tested in a deep subsurface laboratory. Initial calibrations of the detector have shown that it can reach the required angular resolution for muon detection. Stable operation in a small borehole within a few months has been demonstrated.

  12. Biopharmaceuticals: From peptide to drug

    Science.gov (United States)

    Hannappel, Margarete

    2017-08-01

    Biologics are therapeutic proteins or peptides that are produced by means of biological processes within living organisms and cells. They are highly specific molecules and play a crucial role as therapeutics for the treatment of severe and chronic diseases (e.g. cancer, rheumatoid arthritis, diabetes, autoimmune disorders). The development of new biologics and biologics-based drugs gains more and more importance in the fight against various diseases. A short overview on biotherapeutical drug development is given. Cone snails are a large group of poisonous, predatory sea snails with more than 700 species. They use a very powerful venom which rapidly inactivates and paralyzes their prey. Most bioactive venom components are small peptides (conotoxins, conopeptides) which are precisely directed towards a specific target (e.g. ion channel, receptors). Due to their small size, their precision and speed of action, naturally occurring cone snail venom peptides represent an attractive source for the identification and design of novel biological drug entities. The Jagna cone snail project is an encouraging initiative to map the ecological variety of cone snails around the island of Bohol (Philippines) and to conserve the biological information for potential future application.

  13. Safety assessment methodology for waste repositories in deep geological formations

    International Nuclear Information System (INIS)

    Chapuis, A.M.; Lewi, J.; Pradel, J.; Queniart, D.; Raimbault, P.; Assouline, M.

    1986-06-01

    The long term safety of a nuclear waste repository relies on the evaluation of the doses which could be transferred to man in the future. This implies a detailed knowledge of the medium where the waste will be confined, the identification of the basic phenomena which govern the migration of the radionuclides and the investigation of all possible scenarios that may affect the integrity of the barriers between the waste and the biosphere. Inside the Institute of protection and nuclear safety of the French Atomic Energy Commission (CEA/IPSN), the Department of the Safety Analysis (DAS) is currently developing a methodology for assessing the safety of future geological waste repositories, and is in charge of the modelling development, while the Department of Technical Protection (DPT) is in charge of the geological experimental studies. Both aspects of this program are presented. The methodology for risk assessment stresses the needs for coordination between data acquisition and model development which should result in the obtention of an efficient tool for safety evaluation. Progress needs to be made in source and geosphere modelling. Much more sophisticated models could be used than the ones which is described; however sensitivity analysis will determine the level of sophistication which is necessary to implement. Participation to international validation programs are also very important for gaining confidence in the approaches which have been chosen

  14. A novel multidimensional protein identification technology approach combining protein size exclusion prefractionation, peptide zwitterion-ion hydrophilic interaction chromatography, and nano-ultraperformance RP chromatography/nESI-MS2 for the in-depth analysis of the serum proteome and phosphoproteome: application to clinical sera derived from humans with benign prostate hyperplasia.

    Science.gov (United States)

    Garbis, Spiros D; Roumeliotis, Theodoros I; Tyritzis, Stavros I; Zorpas, Kostas M; Pavlakis, Kitty; Constantinides, Constantinos A

    2011-02-01

    The current proof-of-principle study was aimed toward development of a novel multidimensional protein identification technology (MudPIT) approach for the in-depth proteome analysis of human serum derived from patients with benign prostate hyperplasia (BPH) using rational chromatographic design principles. This study constituted an extension of our published work relating to the identification and relative quantification of potential clinical biomarkers in BPH and prostate cancer (PCa) tissue specimens. The proposed MudPIT approach encompassed the use of three distinct yet complementary liquid chromatographic chemistries. High-pressure size-exclusion chromatography (SEC) was used for the prefractionation of serum proteins followed by their dialysis exchange and solution phase trypsin proteolysis. The tryptic peptides were then subjected to offline zwitterion-ion hydrophilic interaction chromatography (ZIC-HILIC) fractionation followed by their online analysis with reversed-phase nano-ultraperformance chromatography (RP-nUPLC) hyphenated to nanoelectrospray ionization-tandem mass spectrometry using an ion trap mass analyzer. For the spectral processing, the sequential use of the SpectrumMill, Scaffold, and InsPecT software tools was applied for the tryptic peptide product ion MS(2) spectral processing, false discovery rate (FDR) assessment, validation, and protein identification. This milestone serum analysis study allowed the confident identification of over 1955 proteins (p ≤ 0.05; FDR ≤ 5%) with a broad spectrum of biological and physicochemical properties including secreted, tissue-specific proteins spanning approximately 12 orders of magnitude as they occur in their native abundance levels in the serum matrix. Also encompassed in this proteome was the confident identification of 375 phosphoproteins (p ≤ 0.05; FDR ≤ 5%) with potential importance to cancer biology. To demonstrate the performance characteristics of this novel MudPIT approach, a comparison

  15. Repository environmental parameters and models/methodologies relevant to assessing the performance of high-level waste packages in basalt, tuff, and salt

    Energy Technology Data Exchange (ETDEWEB)

    Claiborne, H.C.; Croff, A.G.; Griess, J.C.; Smith, F.J.

    1987-09-01

    This document provides specifications for models/methodologies that could be employed in determining postclosure repository environmental parameters relevant to the performance of high-level waste packages for the Basalt Waste Isolation Project (BWIP) at Richland, Washington, the tuff at Yucca Mountain by the Nevada Test Site, and the bedded salt in Deaf Smith County, Texas. Guidance is provided on the identify of the relevant repository environmental parameters; the models/methodologies employed to determine the parameters, and the input data base for the models/methodologies. Supporting studies included are an analysis of potential waste package failure modes leading to identification of the relevant repository environmental parameters, an evaluation of the credible range of the repository environmental parameters, and a summary of the review of existing models/methodologies currently employed in determining repository environmental parameters relevant to waste package performance. 327 refs., 26 figs., 19 tabs.

  16. Repository environmental parameters and models/methodologies relevant to assessing the performance of high-level waste packages in basalt, tuff, and salt

    International Nuclear Information System (INIS)

    Claiborne, H.C.; Croff, A.G.; Griess, J.C.; Smith, F.J.

    1987-09-01

    This document provides specifications for models/methodologies that could be employed in determining postclosure repository environmental parameters relevant to the performance of high-level waste packages for the Basalt Waste Isolation Project (BWIP) at Richland, Washington, the tuff at Yucca Mountain by the Nevada Test Site, and the bedded salt in Deaf Smith County, Texas. Guidance is provided on the identify of the relevant repository environmental parameters; the models/methodologies employed to determine the parameters, and the input data base for the models/methodologies. Supporting studies included are an analysis of potential waste package failure modes leading to identification of the relevant repository environmental parameters, an evaluation of the credible range of the repository environmental parameters, and a summary of the review of existing models/methodologies currently employed in determining repository environmental parameters relevant to waste package performance. 327 refs., 26 figs., 19 tabs

  17. Expanding the detectable HLA peptide repertoire using electron-transfer/higher-energy collision dissociation (EThcD)

    NARCIS (Netherlands)

    Mommen, Geert P M; Frese, Christian K; Meiring, Hugo D; van Gaans-van den Brink, Jacqueline; de Jong, Ad P J M; van Els, Cécile A C M; Heck, Albert J R

    2014-01-01

    The identification of peptides presented by human leukocyte antigen (HLA) class I is tremendously important for the understanding of antigen presentation mechanisms under healthy or diseased conditions. Currently, mass spectrometry-based methods represent the best methodology for the identification

  18. [Plant signaling peptides. Cysteine-rich peptides].

    Science.gov (United States)

    Ostrowski, Maciej; Kowalczyk, Stanisław

    2015-01-01

    Recent bioinformatic and genetic analyses of several model plant genomes have revealed the existence of a highly abundant group of signaling peptides that are defined as cysteine-rich peptides (CRPs). CRPs are usually in size between 50 and 90 amino acid residues, they are positively charged, and they contain 4-16 cysteine residues that are important for the correct conformational folding. Despite the structural differences among CRP classes, members from each class have striking similarities in their molecular properties and function. The present review presents the recent progress in research on signaling peptides from several families including: EPF/EPFL, SP11/SCR, PrsS, RALF, LURE, and some other peptides belonging to CRP group. There is convincing evidence indicating multiple roles for these CRPs as signaling molecules during the plant life cycle, ranging from stomata development and patterning, self-incompatibility, pollen tube growth and guidance, reproductive processes, and nodule formation.

  19. A feedback framework for protein inference with peptides identified from tandem mass spectra

    Directory of Open Access Journals (Sweden)

    Shi Jinhong

    2012-11-01

    Full Text Available Abstract Background Protein inference is an important computational step in proteomics. There exists a natural nest relationship between protein inference and peptide identification, but these two steps are usually performed separately in existing methods. We believe that both peptide identification and protein inference can be improved by exploring such nest relationship. Results In this study, a feedback framework is proposed to process peptide identification reports from search engines, and an iterative method is implemented to exemplify the processing of Sequest peptide identification reports according to the framework. The iterative method is verified on two datasets with known validity of proteins and peptides, and compared with ProteinProphet and PeptideProphet. The results have shown that not only can the iterative method infer more true positive and less false positive proteins than ProteinProphet, but also identify more true positive and less false positive peptides than PeptideProphet. Conclusions The proposed iterative method implemented according to the feedback framework can unify and improve the results of peptide identification and protein inference.

  20. A distributed computational search strategy for the identification of diagnostics targets: application to finding aptamer targets for methicillin-resistant staphylococci.

    Science.gov (United States)

    Flanagan, Keith; Cockell, Simon; Harwood, Colin; Hallinan, Jennifer; Nakjang, Sirintra; Lawry, Beth; Wipat, Anil

    2014-06-30

    The rapid and cost-effective identification of bacterial species is crucial, especially for clinical diagnosis and treatment. Peptide aptamers have been shown to be valuable for use as a component of novel, direct detection methods. These small peptides have a number of advantages over antibodies, including greater specificity and longer shelf life. These properties facilitate their use as the detector components of biosensor devices. However, the identification of suitable aptamer targets for particular groups of organisms is challenging. We present a semi-automated processing pipeline for the identification of candidate aptamer targets from whole bacterial genome sequences. The pipeline can be configured to search for protein sequence fragments that uniquely identify a set of strains of interest. The system is also capable of identifying additional organisms that may be of interest due to their possession of protein fragments in common with the initial set. Through the use of Cloud computing technology and distributed databases, our system is capable of scaling with the rapidly growing genome repositories, and consequently of keeping the resulting data sets up-to-date. The system described is also more generically applicable to the discovery of specific targets for other diagnostic approaches such as DNA probes, PCR primers and antibodies.

  1. A distributed computational search strategy for the identification of diagnostics targets: Application to finding aptamer targets for methicillin-resistant staphylococci

    Directory of Open Access Journals (Sweden)

    Flanagan Keith

    2014-06-01

    Full Text Available The rapid and cost-effective identification of bacterial species is crucial, especially for clinical diagnosis and treatment. Peptide aptamers have been shown to be valuable for use as a component of novel, direct detection methods. These small peptides have a number of advantages over antibodies, including greater specificity and longer shelf life. These properties facilitate their use as the detector components of biosensor devices. However, the identification of suitable aptamer targets for particular groups of organisms is challenging. We present a semi-automated processing pipeline for the identification of candidate aptamer targets from whole bacterial genome sequences. The pipeline can be configured to search for protein sequence fragments that uniquely identify a set of strains of interest. The system is also capable of identifying additional organisms that may be of interest due to their possession of protein fragments in common with the initial set. Through the use of Cloud computing technology and distributed databases, our system is capable of scaling with the rapidly growing genome repositories, and consequently of keeping the resulting data sets up-to-date. The system described is also more generically applicable to the discovery of specific targets for other diagnostic approaches such as DNA probes, PCR primers and antibodies.

  2. SUBSURFACE REPOSITORY INTEGRATED CONTROL SYSTEM DESIGN

    International Nuclear Information System (INIS)

    Randle, D.C.

    2000-01-01

    The primary purpose of this document is to develop a preliminary high-level functional and physical control system architecture for the potential repository at Yucca Mountain. This document outlines an overall control system concept that encompasses and integrates the many diverse process and communication systems being developed for the subsurface repository design. This document presents integrated design concepts for monitoring and controlling the diverse set of subsurface operations. The Subsurface Repository Integrated Control System design will be composed of a series of diverse process systems and communication networks. The subsurface repository design contains many systems related to instrumentation and control (I andC) for both repository development and waste emplacement operations. These systems include waste emplacement, waste retrieval, ventilation, radiological and air monitoring, rail transportation, construction development, utility systems (electrical, lighting, water, compressed air, etc.), fire protection, backfill emplacement, and performance confirmation. Each of these systems involves some level of I andC and will typically be integrated over a data communications network throughout the subsurface facility. The subsurface I andC systems will also interface with multiple surface-based systems such as site operations, rail transportation, security and safeguards, and electrical/piped utilities. In addition to the I andC systems, the subsurface repository design also contains systems related to voice and video communications. The components for each of these systems will be distributed and linked over voice and video communication networks throughout the subsurface facility. The scope and primary objectives of this design analysis are to: (1) Identify preliminary system-level functions and interfaces (Section 6.2). (2) Examine the overall system complexity and determine how and on what levels the engineered process systems will be monitored

  3. Fons antic i repositoris universitaris a Espanya

    Directory of Open Access Journals (Sweden)

    Herrera Morillas, José Luis

    2015-12-01

    Full Text Available Es mostra la presència de col·leccions de fons antic en els repositoris de les biblioteques universitàries espanyoles després d'analitzar tots els repositoris. Per a aquesta anàlisi, com a part de la metodologia emprada, s'ha elaborat un model o llista que consta d'onze elements. Del conjunt de les universitats espanyoles, seixanta tenen repositoris, vint-i-vuit dels quals (16,8 % disposen de col·leccions de fons antic. Com que del concepte de repositori institucional no sembla desprendre's que tingui com a finalitat incloure aquest tipus de col·leccions, es reflexiona sobre la peculiaritat que una part dels repositoris universitaris espanyols inclogui col·leccions d'aquestes característiques.Se muestra la presencia de colecciones de fondo antiguo en los repositorios de las bibliotecas universitarias españolas después de analizar todos los repositorios. Para este análisis, como parte de la metodología empleada, se ha elaborado un modelo o lista que consta de once elementos. Del conjunto de las universidades españolas, sesenta cuentan con repositorios y, de estos, veintiocho (16,8 % disponen de colecciones de fondo antiguo. Debido a que del concepto de repositorio institucional no parece desprenderse que tenga como finalidad albergar este tipo de colecciones, se hace una reflexión sobre la peculiaridad de que parte de los repositorios universitarios españoles incluya colecciones de estas características.This paper uses an analysis of the repositories of Spanish universities to identify which institutions contain rare book and manuscript collections. The method used in this analysis involved examining each university on the basis of a list comprising eleven elements. A total of 60 universities were found to have repositories but only 28 (16.8 % of these contained rare book and manuscript collections. In the light of these figures, which suggest that Spanish university repositories do not generally consider the preservation of rare

  4. Coupling fuel cycles with repositories: how repository institutional choices may impact fuel cycle design

    International Nuclear Information System (INIS)

    Forsberg, C.; Miller, W.F.

    2013-01-01

    The historical repository siting strategy in the United States has been a top-down approach driven by federal government decision making but it has been a failure. This policy has led to dispatching fuel cycle facilities in different states. The U.S. government is now considering an alternative repository siting strategy based on voluntary agreements with state governments. If that occurs, state governments become key decision makers. They have different priorities. Those priorities may change the characteristics of the repository and the fuel cycle. State government priorities, when considering hosting a repository, are safety, financial incentives and jobs. It follows that states will demand that a repository be the center of the back end of the fuel cycle as a condition of hosting it. For example, states will push for collocation of transportation services, safeguards training, and navy/private SNF (Spent Nuclear Fuel) inspection at the repository site. Such activities would more than double local employment relative to what was planned for the Yucca Mountain-type repository. States may demand (1) the right to take future title of the SNF so if recycle became economic the reprocessing plant would be built at the repository site and (2) the right of a certain fraction of the repository capacity for foreign SNF. That would open the future option of leasing of fuel to foreign utilities with disposal of the SNF in the repository but with the state-government condition that the front-end fuel-cycle enrichment and fuel fabrication facilities be located in that state

  5. Identification of an immunodominant epitope in glycoproteins B and G of herpes simplex viruses (HSVs) using synthetic peptides as antigens in assay of antibodies to HSV in herpes simplex encephalitis patients.

    Science.gov (United States)

    Bhullar, S S; Chandak, N H; Baheti, N N; Purohit, H J; Taori, G M; Daginawala, H F; Kashyap, R S

    2014-01-01

    Herpes simplex encephalitis (HSE) is a severe viral infection of the central nervous system (CNS). Assay of antibody response is widely used in diagnostics of HSE. The aim of this study was to identify an immunodominant epitope determining the antibody response to herpes simplex viruses (HSVs) in cerebrospinal fluid (CSF) of HSE patients. The synthetic peptides that resembled type-common as well as type-specific domains of glycoproteins B (gB) and G (gG) of these viruses were evaluated for binding with IgM and IgG antibodies in CSF samples from HSE and non-HSE patients in ELISA. The QLHDLRF peptide, derived from gB of HSV was found to be an immunodominant epitope in the IgM and IgG antibody response. The patients with confirmed and suspected HSE showed in ELISA against this peptide 26% and 23% positivities for IgM, 43% and 37% positivities for IgG and 17% and 15% for both IgM and IgG antibodies, respectively. The total positivities of 86% and 75% for both IgM and IgG antibodies were obtained in the patients with confirmed and suspected HSE, respectively. These results demonstrate that a synthetic peptide-based diagnostics of HSE can be an efficient and easily accessible alternative. This is the first report describing the use of synthetic peptides derived from HSVs in diagnostics of HSE using patientsʹ CSF samples.

  6. Analysis of computational vulnerabilities in digital repositories

    Directory of Open Access Journals (Sweden)

    Valdete Fernandes Belarmino

    2015-04-01

    Full Text Available Objective. Demonstrates the results of research that aimed to analyze the computational vulnerabilities of digital directories in public Universities. Argues the relevance of information in contemporary societies like an invaluable resource, emphasizing scientific information as an essential element to constitute scientific progress. Characterizes the emergence of Digital Repositories and highlights its use in academic environment to preserve, promote, disseminate and encourage the scientific production. Describes the main software for the construction of digital repositories. Method. The investigation identified and analyzed the vulnerabilities that are exposed the digital repositories using Penetration Testing running. Discriminating the levels of risk and the types of vulnerabilities. Results. From a sample of 30 repositories, we could examine 20, identified that: 5% of the repositories have critical vulnerabilities, 85% high, 25% medium and 100% lowers. Conclusions. Which demonstrates the necessity to adapt actions for these environments that promote informational security to minimizing the incidence of external and / or internal systems attacks.Abstract Grey Text – use bold for subheadings when needed.

  7. Researching radioactive waste disposal. [Underground repository

    Energy Technology Data Exchange (ETDEWEB)

    Feates, F; Keen, N [UKAEA Research Group, Harwell. Atomic Energy Research Establishment

    1976-02-16

    At present it is planned to use the vitrification process to convert highly radioactive liquid wastes, arising from nuclear power programme, into glass which will be contained in steel cylinders for storage. The UKAEA in collaboration with other European countries is currently assessing the relative suitability of various natural geological structures as final repositories for the vitrified material. The Institute of Geological Sciences has been commissioned to specify the geological criteria that should be met by a rock structure if it is to be used for the construction of a repository though at this stage disposal sites are not being sought. The current research programme aims to obtain basic geological data about the structure of the rocks well below the surface and is expected to continue for at least three years. The results in all the European countries will then be considered so that the United Kingdom can choose a preferred method for isolating their wastes. It is only at that stage that a firm commitment may be made to select a site for a potential repository, when a far more detailed scientific research study will be instituted. Heat transfer problems and chemical effects which may occur within and around repositories are being investigated and a conceptual design study for an underground repository is being prepared.

  8. Towards a spectroscopic standard database for Pu in repository environments

    International Nuclear Information System (INIS)

    Amme, M.

    2005-01-01

    Full text of publication follows: The alteration behaviour of Pu in geological and technical environments is, although of crucial importance for example in final repository assessment procedures for high-level nuclear waste, not sufficiently investigated. Since Pu chemistry differs significantly from U behaviour (mostly due to the different stabilities of the +IV oxidation states of both elements), conclusions based on the uranium analogy cannot be extended to Pu chemistry in many cases. In order to examine precisely the alteration behaviour of Pu under repository storage conditions, customized tools for the spectroscopic identification of the element need to be developed. We are currently constructing systematically a database of Pu compounds and collect their spectra for this purpose. Pu compounds (with the element in the oxidation states +III, +IV, +V, and +VI) are synthesised, mostly by using hydrothermal synthesis techniques [1]. Compounds of high importance for repository studies are: Carbonates, (oxy)hydroxides, silicates, peroxides, and phosphates. The products are characterised by Scanning Electron Microscopy (SEM), X-Ray Photoelectron spectroscopy (XPS), and X-Ray diffraction (XRD). Furthermore, vibrational spectra (Raman and IR) of the substances will be recorded. These will allow the unambiguous identification of chemically similar compounds, once the database is compiled: Although reported for a few cases with nuclear material only, the combined investigation of solids with Raman [2,3] and SEM-EDX [3] in a combined mode, especially when spatially resolving, holds great potential An investigation with accelerator-based XAS techniques is planned within the framework of the Excellence network Actinet 6. [1] Grigorev, M., Bessonov, A., Makarenkov, V., Fedoseev, A., Model of the (PuO 2 ) 2 SiO 4 * 2H 2 O crystal structure, based on powder X-ray diffraction data, Radiochemistry, Vol. 45, No 3 (2003) 257-260. [2] M. Amme, B. Renker, B. Schmid, M. Feth, H

  9. Planning the rad waste repository - Croatian case

    International Nuclear Information System (INIS)

    Kucar Dragicevic, S.; Subasic, D.; Lokner, V.

    1996-01-01

    Radioactive waste is generated in Croatia from various nuclear applications as well as from the Krsko NPP (Slovenian and Croatian joint venture facility). The national programme on radioactive waste management is aimed at straightening existing infrastructure, establishing new (more transparent) system of responsibilities and development of new legislation. The siting of LL/ILW repository is important segments of the whole radioactive waste management cycle. The status and efficiency of the rad waste management infrastructure in the country have the significant influence on all the activities related to the project of repository construction - from the very first phases of preliminary planning and background preparations to advanced phases of the project development. The present status of the Croatian national radioactive waste infrastructure and its influence on the repository project are presented. The role of national legislation and institutional framework are specially discussed. (author)

  10. Augmenting interoperability across repositories architectural ideas

    CERN Multimedia

    CERN. Geneva

    2005-01-01

    The aDORe digital repository architecture designed and implemented by the Los Alamos Research Library is fully standards-based and highly modular, with the various components of the architecture interacting in a protocol-driven manner. Although aDORe was designed for use in the context of the Los Alamos Library, its modular and standards-based design has led to interesting insights regarding possible new levels of interoperability in a federation of heterogeneous repositories. The presentation will discuss these insights, and will illustrate that attractive federations of repositories can be built by introducing rather basic interoperability requirements. The presentation will also show that, once these requirements are met, a powerful service framework that overlays the federation can emerge.

  11. Reducing Psychological Resistance to Digital Repositories

    Directory of Open Access Journals (Sweden)

    Brian Quinn

    2010-06-01

    Full Text Available The potential value of digital repositories is dependent on the cooperation of scholars to deposit their work. Although many researchers have been resistant to submitting their work, the literature on digital repositories contains very little research on the psychology of resistance. This article looks at the psychological literature on resistance and explores what its implications might be for reducing the resistance of scholars to submitting their work to digital repositories. Psychologists have devised many potentially useful strategies for reducing resistance that might be used to address the problem; this article examines these strategies and how they might be applied.

  12. A Repository of Semantic Open EHR Archetypes

    Directory of Open Access Journals (Sweden)

    Fernando Sánchez

    2015-03-01

    Full Text Available This paper describes a repository of openEHR archetypes that have been translated to OWL. In the work presented here, five different CKMs (Clinical Knowledge Managers have been downloaded and the archetypes have been translated to OWL. This translation is based on an existing translator that has been improved to solve programming problems with certain structures. As part of the repository a tool has been developed to keep it always up-to-date. So, any change in one of the CKMs (addition, elimination or even change of an archetype will involve translating the changed archetypes once more. The repository is accessible through a Web interface (http://www.openehr.es/.

  13. Initial design process of the repository

    International Nuclear Information System (INIS)

    Osmanlioglu, A.E.

    2001-01-01

    The concept of the final disposal of high level wastes is to isolate the waste from the biosphere for extremely long periods of time by emplacement of wastes into deep stable geological formations. Several geological formations have been considered as candidate host environments for high level waste disposal and several techniques have been developed for repository design. In this study, interrelationships of main parameters of a general repository design have been defined and effective parameters are shown at each step. Initial design process is based on the long term stability of underground openings as disposal galleries. For this reason, this design process includes two main analyses: mechanical analysis and thermal analysis. Each of the analysis systems is directly related to each other by technical precautions. As a result of this design process, general information about the acceptable depth of the repository, layout and emplacement pattern can be taken. Final design study can be established on the result of initial design process. (author)

  14. Modelling gas generation in radioactive waste repositories

    International Nuclear Information System (INIS)

    Agg, P.J.

    1993-02-01

    In a repository containing low- and intermediate-level waste, gas generation will occur principally by the coupled processes of metal corrosion and microbial degradation of cellulosic waste. This Paper describes a mathematical model design to address gas generation by these mechanisms. The metal corrosion model incorporates a three-stage process encompassing both aerobic and anaerobic corrosion regimes; the microbial degradation model simulates the activities of eight different microbial populations, which are maintained as functions both of pH and of the concentrations of particular chemical species. Gas concentrations have been measured over a period of three years in large-scale drum experiments designed to simulate repository conditions. Model predictions are confirmed against the experimental measurements, and a prediction is then made of gas concentrations and generation rates over an assessment period of one million years in a radioactive waste repository. (author)

  15. Modelling gas generation in radioactive waste repositories

    International Nuclear Information System (INIS)

    Agg, P.J.

    1992-07-01

    In a repository containing low- and intermediate-level waste, gas generation will occur principally by the coupled processes of metal corrosion and microbial degradation of cellulosic waste. This paper describes a mathematical model designed to address gas generation by these mechanisms. The metal corrosion model incorporates a three-stage process encompassing both aerobic and anaerobic corrosion regimes; the microbial degradation model simulates the activities of eight different microbial populations, which are maintained as functions both of pH and of the concentrations of particular chemical species. Gas concentrations have been measured over a period of three years in large-scale drum experiments designed to simulate repository conditions. Model predictions are confirmed against the experimental measurements, and a prediction is then made of gas concentrations and generation rates over an assessment period of one million years in a radioactive waste repository. (Author)

  16. Developing criteria to establish Trusted Digital Repositories

    Science.gov (United States)

    Faundeen, John L.

    2017-01-01

    This paper details the drivers, methods, and outcomes of the U.S. Geological Survey’s quest to establish criteria by which to judge its own digital preservation resources as Trusted Digital Repositories. Drivers included recent U.S. legislation focused on data and asset management conducted by federal agencies spending $100M USD or more annually on research activities. The methods entailed seeking existing evaluation criteria from national and international organizations such as International Standards Organization (ISO), U.S. Library of Congress, and Data Seal of Approval upon which to model USGS repository evaluations. Certification, complexity, cost, and usability of existing evaluation models were key considerations. The selected evaluation method was derived to allow the repository evaluation process to be transparent, understandable, and defensible; factors that are critical for judging competing, internal units. Implementing the chosen evaluation criteria involved establishing a cross-agency, multi-disciplinary team that interfaced across the organization. 

  17. Alternative measure for performance of HLW geologic repository

    International Nuclear Information System (INIS)

    Joonhang, Ahn; Chambre, P.L.

    2001-01-01

    A repository performance model that can show effects of canister multiplicity and repository configuration has been developed. Masses of a radionuclide in the repository and in the far field are proposed as performance measures. Canister multiplicity has significant effects on the release of long-lived radionuclides from the repository. As more canisters are included in the same water stream, the radionuclide concentration in the stream increases, but becomes independent of the number of canisters for sufficiently many canisters. Effects of reduction of radionuclide mass in the repository on the repository performance are clearly observed if the canister multiplicity is taken into account and the mass-based measures are applied. (author)

  18. Investigative study of standards for digital repositories and related services

    CERN Document Server

    Foulonneau, Muriel; Badolato, Anne-Marie

    2008-01-01

    This study is meant for institutional repository managers, service providers, repository software developers and generally, all players taking an active part in the creation of the digital repository infrastructure for e-research and e-learning. It reviews the current standards, protocols and applications in the domain of digital repositories. Special attention is being paid to the interoperability of repositories to enhance the exchange of data in repositories. It aims to stimulate discussion about these topics and supports initiatives for the integration of and, where needed, development of

  19. Analytic framework for peptidomics applied to large-scale neuropeptide identification

    DEFF Research Database (Denmark)

    Secher, Anna; Kelstrup, Christian D; Conde-Frieboes, Kilian W

    2016-01-01

    was integrated with publically available databases. We developed and applied an algorithm that reduces the peptide complexity for identification of biologically relevant peptides. The developed pipeline was applied to rat hypothalamus and identifies thousands of neuropeptides and their post...

  20. Anti-dengue virus serotype 2 activity and mode of action of a novel peptide.

    Science.gov (United States)

    Chew, M-F; Tham, H-W; Rajik, M; Sharifah, S H

    2015-10-01

    To identify a novel antiviral peptide against dengue virus serotype 2 (DENV-2) by screening a phage display peptide library and to evaluate its in vitro antiviral activity and mode of action. A phage display peptide library was biopanned against purified DENV-2 and resulted in the identification and selection of a peptide (peptide gg-ww) for further investigation. ELISA was performed, and peptide gg-ww was shown to possess the highest binding affinity against DENV-2. Thus, peptide gg-ww was synthesized for cytotoxicity and antiviral assays. Virus plaque reduction assay, real-time PCR and immunofluorescence assay were used to investigate the inhibitory effect of peptide gg-ww on DENV-2 infection in Vero cells. Three different assays (pre-, simultaneous and post-treatments assays) were performed to investigate the peptide's mode of action. Results indicated that peptide gg-ww possessed strong antiviral activity with a ~96% inhibition rate, which was achieved at 250 μmol l(-1) . Viral replication was inhibited during a simultaneous treatment assay, indicating that the entry of the virus was impeded by this peptide. Peptide gg-ww displayed antiviral action against DENV-2 by targeting an early stage of viral replication (i.e. during viral entry). Peptide gg-ww may represent a new therapeutic candidate for the treatment of DENV infections and is a potential candidate to be developed as a peptide drug. © 2015 The Society for Applied Microbiology.

  1. The Morsleben radwaste repository. Preparing for decommissioning

    International Nuclear Information System (INIS)

    Mehnert, M.; Schmitt, R.

    2001-06-01

    The publication is intended to illustrate with a brief chronology the history and the present situation of the Morsleben radwaste repository, including specific aspects such as the geology of the site and construction and engineering activities, the particulars of waste form emplacement and log-term storage conditions, topical issues relating to radiological safety during operation and after decommissioning. The brochure is designed for the general audience interested in background information on all aspects of the uses, operation and decommissioning of a radwaste repository in Germany. (orig./CB) [de

  2. Overview of the current CRWMS repository design

    International Nuclear Information System (INIS)

    Daniel, R.B.; Teraoka, G.M.

    1998-01-01

    This paper summarizes the current design for a potential geologic repository for spent fuels and high-level wastes at Yucca Mountain, Nevada. The objective of the paper is to present the key design features of the Mined Geologic Disposal System (MGDS) surface facilities and MGDS subsurface facilities. The paper describes the following: surface layout; waste handling operations design; subsurface design; and the underground transport and emplacement design. A more detailed presentation of key features is provided in the ''Reference design description for a geologic repository'' which is located on the YMP Homepage at www.ymp.gov

  3. GreyGuide Forum and Repository

    OpenAIRE

    Biagioni, Stefania; Farace, Dominic

    2014-01-01

    We present the GreyGuide: an online forum and repository of good practice in the field of grey literature. The launch of the GreyGuide Repository took place in December 2013 at the Fifteenth International Conference on Grey Literature. Since then, the acquisition of both proposed and published good practices are underway. The GreyGuide as an online forum is currently in a developmental stage and is influenced by the changes that have taken place in GreyNet's new infrastructure commencing in J...

  4. Citizen participation in nuclear waste repository siting

    International Nuclear Information System (INIS)

    Howell, R.E.; Olsen, D.

    1982-12-01

    The following study presents a proposed strategy for citizen participation during the planning stages of nuclear waste repository siting. It discusses the issue from the general perspective of citizen participation in controversial issues and in community development. Second, rural institutions and attitudes toward energy development as the context for developing a citizen participation program are examined. Third, major citizen participation techniques and the advantages and disadvantages of each approach for resolving public policy issues are evaluated. Fourth, principles of successful citizen participation are presented. Finally, a proposal for stimulating and sustaining effective responsible citizen participation in nuclear waste repository siting and management is developed

  5. Peptides in melanoma therapy.

    Science.gov (United States)

    Mocellin, Simone

    2012-01-01

    Peptides derived from tumor associated antigens can be utilized to elicit a therapeutically effective immune response against melanoma in experimental models. However, patient vaccination with peptides - although it is often followed by the induction of melanoma- specific T lymphocytes - is rarely associated with tumor response of clinical relevance. In this review I summarize the principles of peptide design as well as the results so far obtained in the clinical setting while treating cutaneous melanoma by means of this active immunotherapy strategy. I also discuss some immunological and methodological issues that might be helpful for the successful development of peptide-based vaccines.

  6. Antimicrobial Peptides in Reptiles

    Science.gov (United States)

    van Hoek, Monique L.

    2014-01-01

    Reptiles are among the oldest known amniotes and are highly diverse in their morphology and ecological niches. These animals have an evolutionarily ancient innate-immune system that is of great interest to scientists trying to identify new and useful antimicrobial peptides. Significant work in the last decade in the fields of biochemistry, proteomics and genomics has begun to reveal the complexity of reptilian antimicrobial peptides. Here, the current knowledge about antimicrobial peptides in reptiles is reviewed, with specific examples in each of the four orders: Testudines (turtles and tortosises), Sphenodontia (tuataras), Squamata (snakes and lizards), and Crocodilia (crocodilans). Examples are presented of the major classes of antimicrobial peptides expressed by reptiles including defensins, cathelicidins, liver-expressed peptides (hepcidin and LEAP-2), lysozyme, crotamine, and others. Some of these peptides have been identified and tested for their antibacterial or antiviral activity; others are only predicted as possible genes from genomic sequencing. Bioinformatic analysis of the reptile genomes is presented, revealing many predicted candidate antimicrobial peptides genes across this diverse class. The study of how these ancient creatures use antimicrobial peptides within their innate immune systems may reveal new understandings of our mammalian innate immune system and may also provide new and powerful antimicrobial peptides as scaffolds for potential therapeutic development. PMID:24918867

  7. Photoaffinity labeling of human serum vitamin D binding protein and chemical cleavage of the labeled protein: Identification of an 11.5-kDa peptide containing the putative 25-hydroxyvitamin D3 binding site

    International Nuclear Information System (INIS)

    Ray, R.; Holick, M.F.; Bouillon, R.; Baelen, H.V.

    1991-01-01

    In this paper, the authors describe photoaffinity labeling and related studies of human serum vitamin D binding protein (hDBP) with 25-hydroxyvitamin D 3 3β-3'-[N-(4-azido-2-nitrophenyl)amino]propyl ether (25-ANE) and its radiolabeled counterpart, i.e., 25-hydroxyvitamin D 3 3β-3'-[N-(4-azido-2-nitro-[3,5- 3 H]phenyl)amino]propyl ether ( 3 H-25-ANE). They have carried out studies to demonstrate that (1) 25-ANE competes with 25-OH-D 3 for the binding site of the latter in hDBP and (2) 3 H-25-ANE is capable of covalently labeling the hDBP molecule when exposed ot UV light. Treatment of a sample of purified hDBP, labeled with 3 H-25-ANE, with BNPS-skatole produced two Coomassie Blue stained peptide fragments, and the majority of the radioactivity was assoicated with the smaller of the two peptide fragments (16.5 kDa). On the other hand, cleavage of the labeled protein with cyanogen bromide produced a peptide (11.5 kDa) containing most of the covalently attached radioactivity. Considering the primary amino acid structure of hDBP, this peptide fragment (11.5 kDa) represents the N-terminus through residue 108 of the intact protein. Thus, the results tentatively identify this segment of the protein containing the binding pocket for 25-OH-D 3

  8. Human Antimicrobial Peptides and Proteins

    Directory of Open Access Journals (Sweden)

    Guangshun Wang

    2014-05-01

    Full Text Available As the key components of innate immunity, human host defense antimicrobial peptides and proteins (AMPs play a critical role in warding off invading microbial pathogens. In addition, AMPs can possess other biological functions such as apoptosis, wound healing, and immune modulation. This article provides an overview on the identification, activity, 3D structure, and mechanism of action of human AMPs selected from the antimicrobial peptide database. Over 100 such peptides have been identified from a variety of tissues and epithelial surfaces, including skin, eyes, ears, mouths, gut, immune, nervous and urinary systems. These peptides vary from 10 to 150 amino acids with a net charge between −3 and +20 and a hydrophobic content below 60%. The sequence diversity enables human AMPs to adopt various 3D structures and to attack pathogens by different mechanisms. While α-defensin HD-6 can self-assemble on the bacterial surface into nanonets to entangle bacteria, both HNP-1 and β-defensin hBD-3 are able to block cell wall biosynthesis by binding to lipid II. Lysozyme is well-characterized to cleave bacterial cell wall polysaccharides but can also kill bacteria by a non-catalytic mechanism. The two hydrophobic domains in the long amphipathic α-helix of human cathelicidin LL-37 lays the basis for binding and disrupting the curved anionic bacterial membrane surfaces by forming pores or via the carpet model. Furthermore, dermcidin may serve as ion channel by forming a long helix-bundle structure. In addition, the C-type lectin RegIIIα can initially recognize bacterial peptidoglycans followed by pore formation in the membrane. Finally, histatin 5 and GAPDH(2-32 can enter microbial cells to exert their effects. It appears that granulysin enters cells and kills intracellular pathogens with the aid of pore-forming perforin. This arsenal of human defense proteins not only keeps us healthy but also inspires the development of a new generation of personalized

  9. Insulin C-peptide test

    Science.gov (United States)

    C-peptide ... the test depends on the reason for the C-peptide measurement. Ask your health care provider if ... C-peptide is measured to tell the difference between insulin the body produces and insulin someone injects ...

  10. Radioactive waste disposal: Recommendations for a repository site selection

    International Nuclear Information System (INIS)

    Cadelli, N.; Orlowski, S.

    1992-01-01

    This report is a guidebook on recommendations for site selection of radioactive waste repository, based on a consensus in european community. This report describes particularly selection criteria and recommendations for radioactive waste disposal in underground or ground repositories. 14 refs

  11. Open DOAR the Directory of Open Access Repositories

    CERN Multimedia

    CERN. Geneva

    2005-01-01

    The last year has seen wide-spread growth in the idea of using open access repositories as a part of a research institution's accepted infrastructure. Policy development from institutions and funding bodies has also supported the growth of the repository network. The next stage of expansion will be in the provision of services and cross-repository facilities and resources. Of course, it is hoped that these will then establish a feed-back loop to encourage repository population and further repository establishment, as the potential of open access to research materials is realised. The growth of repositories has been organic, with a variety of different repositories based in departments, institutions, funding agencies or subject communities, with a range of content, both in type and subject. Existing repositories are expanding their holdings, from eprints to associated research data-sets, or with learning objects and multimedia material. This presentation will look at the development of the Directory of Open Ac...

  12. The Vital Role of Free Access in Supporting Digital Repositories

    African Journals Online (AJOL)

    pc

    2018-03-05

    Mar 5, 2018 ... the scientific research forward, stressing that the free access to the scientific .... links in the digital repositories for the scientific journals publishers not to mention .... psychology, physics and encryption. Moreover, repositories.

  13. Visual querying and analysis of large software repositories

    NARCIS (Netherlands)

    Voinea, Lucian; Telea, Alexandru

    We present a software framework for mining software repositories. Our extensible framework enables the integration of data extraction from repositories with data analysis and interactive visualization. We demonstrate the applicability of the framework by presenting several case studies performed on

  14. Geohydrological simulation of a deep coastal repository

    International Nuclear Information System (INIS)

    Follin, S.

    1995-12-01

    This conceptual-numerical study treats the dewatering and resaturation phases associated with the construction, use and closure of a coastal nuclear waste repository located at depth in sparsely fractured Baltic Shield rocks. The main objective is to simulate the extent and duration of saline intrusion for a reasonable set of geohydrological assumptions. Long-term changes in the chemical environment associated with saline intrusion may affect the properties of the buffer zone material (bentonite). The first part of the study deals with history matching of a simple model geometry and the second part treats the dewatering and resaturation phases of the simulated repository. The history matching supports the standpoint that the occurrence of saline ground water reflects an ongoing but incomplete Holocene flushing of the Baltic Shield. The drawdown after fifty years of dewatering is highly dependent on the permeability of the excavated damaged zone. If the permeability close the repository is unaltered the entire region between the top side of the model and the repository is more or less partially saturated at the end of the simulation period. The simulations of a fifty year long recovery period suggest that the distribution between fresh and saline ground waters may be quite close to the conditions prior to the dewatering phase already after fifty years of closure despite an incomplete pressure recovery, which is an interesting result considering the objective of the study. 12 refs

  15. Unifying Learning Object Repositories in MACE

    NARCIS (Netherlands)

    Prause, Christian; Ternier, Stefaan; De Jong, Tim; Apelt, Stefan; Scholten, Marius; Wolpers, Martin; Eisenhauer, Markus; Vandeputte, Bram; Specht, Marcus; Duval, Erik

    2007-01-01

    Prause, C., Ternier, S., De Jong, T., Apelt, S., Scholten, M., Wolpers, M., et al. (2007). Unifying Learning Object Repositories in MACE. In D. Massart, J.-N. Colin & F. V. Assche (Eds.). Proceedings of the First International Workshop on Learning Object Discovery & Exchange (LODE'07). September,

  16. Repository Services for Outcome-based Learning

    NARCIS (Netherlands)

    Totschnig, Michael; Derntl, Michael; Gutiérrez, Israel; Najjar, Jad; Klemke, Roland; Klerkx, Joris; Duval, Erik; Müller, Franz

    2010-01-01

    Totschnig, M., Derntl, M., Gutiérrez, I., Najjar, J., Klemke, R., Klerkx, J., Duval, E., & Müller, F. (2010). Repository Services for Outcome-based Learning. Fourth International Workshop on Search and Exchange of e-le@rning Materials (SE@M’10). September, 27-28, 2010, Barcelona, Spain.

  17. Design of repository sealing systems - 1981

    International Nuclear Information System (INIS)

    Ellison, R.D.; Shukla, D.K.; Kelsall, P.C.; D'Appolonia Consulting Engineers, Albuquerque, NM)

    1982-01-01

    Isolating nuclear waste in geologic repositories will require the sealing of penetrations such as access shafts and tunnels, disposal rooms, and exploration boreholes. This paper discusses seal designs developed for a repository in bedded salt referenced to the stratigraphy of southeastern New Mexico. Designs are based on a multiple component concept whereby individual components are designed for a specific function and location. For a repository in salt the major function of the seals is to exclude groundwater inflow. Two main types of component are included for this purpose: (1) bulk-heads are dense concrete structures keyed into the walls of the penetration and are intended to reduce flow at the interface between the seal and the salt; (2) backfills are granular materials compacted in place in the penetration. In the repository the major backfill material is crushed salt, which is expected to consolidate and recrystallize as the rooms close in response to salt creep. Densely compacted clays will be used as backfill in the shafts closer to potential sources of water inflow. 22 references, 10 figures, 1 table

  18. Grey Guide Repository: presentation and demo

    OpenAIRE

    Biagioni, Stefania; Carlesi, Carlo; Schopfel, Joachim; Farace, Dominic; Frantzen, Jerry

    2013-01-01

    The goal of this project is to develop an open source repository of good practices in the field of grey literature. That which originated in monographic form will now open and expand to include content from the global grey literature community. Such practices will range from the production and processing of grey literature through to its distribution, uses, and preservation.

  19. The Sellafield repository project information programme

    Energy Technology Data Exchange (ETDEWEB)

    Curd, P J [United Kingdom Nirex Limited (United Kingdom)

    1993-07-01

    The Sellafield Repository Project Information programme has been guided by formal research and by feedback through members of the team. Progress has been made and a significant majority of local people support the project and feel it will benefit the area. (author)

  20. The Sellafield repository project information programme

    International Nuclear Information System (INIS)

    Curd, P.J.

    1993-01-01

    The Sellafield Repository Project Information programme has been guided by formal research and by feedback through members of the team. Progress has been made and a significant majority of local people support the project and feel it will benefit the area. (author)

  1. Local groundwater depression around a repository

    International Nuclear Information System (INIS)

    Thunvik, R.

    1978-01-01

    Local Groundwater Depression around a Repository. A two-dimensional flow analysis was made to study the effect on the groundwater table due to drainage of the storage tunnels during the construction resp. operation period. The net accretion to the phreatic surface was assumed evenly distributed in space and time. Numerical examples with equipotentials and consecutive positions of the phreatic surface are presented

  2. Data deposit into the ASEP repository

    Czech Academy of Sciences Publication Activity Database

    Chmelařová, Zdeňka; Doleželová, Jana

    2018-01-01

    Roč. 14, Special winter issue (2018), s. 44-49 ISSN 1574-1796 Institutional support: RVO:67985971 Keywords : data repositories * ASEP * Library of the Czech Academy of Sciences * Czech Academy of Sciences Subject RIV: AF - Documentation, Librarianship, Information Studies

  3. Business process model repositories : framework and survey

    NARCIS (Netherlands)

    Yan, Z.; Dijkman, R.M.; Grefen, P.W.P.J.

    2009-01-01

    Large organizations often run hundreds or even thousands of business processes. Managing such large collections of business processes is a challenging task. Intelligent software can assist in that task by providing common repository functions such as storage, search and version management. They can

  4. A framework for business process model repositories

    NARCIS (Netherlands)

    Yan, Z.; Grefen, P.W.P.J.; Muehlen, zur M.; Su, J.

    2010-01-01

    Large organizations often run hundreds or even thousands of business processes. Managing such large collections of business processes is a challenging task. Intelligent software can assist in that task by providing common repository functions such as storage, search and version management. They can

  5. Protein interaction networks by proteome peptide scanning.

    Directory of Open Access Journals (Sweden)

    Christiane Landgraf

    2004-01-01

    Full Text Available A substantial proportion of protein interactions relies on small domains binding to short peptides in the partner proteins. Many of these interactions are relatively low affinity and transient, and they impact on signal transduction. However, neither the number of potential interactions mediated by each domain nor the degree of promiscuity at a whole proteome level has been investigated. We have used a combination of phage display and SPOT synthesis to discover all the peptides in the yeast proteome that have the potential to bind to eight SH3 domains. We first identified the peptides that match a relaxed consensus, as deduced from peptides selected by phage display experiments. Next, we synthesized all the matching peptides at high density on a cellulose membrane, and we probed them directly with the SH3 domains. The domains that we have studied were grouped by this approach into five classes with partially overlapping specificity. Within the classes, however, the domains display a high promiscuity and bind to a large number of common targets with comparable affinity. We estimate that the yeast proteome contains as few as six peptides that bind to the Abp1 SH3 domain with a dissociation constant lower than 100 microM, while it contains as many as 50-80 peptides with corresponding affinity for the SH3 domain of Yfr024c. All the targets of the Abp1 SH3 domain, identified by this approach, bind to the native protein in vivo, as shown by coimmunoprecipitation experiments. Finally, we demonstrate that this strategy can be extended to the analysis of the entire human proteome. We have developed an approach, named WISE (whole interactome scanning experiment, that permits rapid and reliable identification of the partners of any peptide recognition module by peptide scanning of a proteome. Since the SPOT synthesis approach is semiquantitative and provides an approximation of the dissociation constants of the several thousands of interactions that are

  6. Peptide Vaccines for Leishmaniasis

    Directory of Open Access Journals (Sweden)

    Rory C. F. De Brito

    2018-05-01

    Full Text Available Due to an increase in the incidence of leishmaniases worldwide, the development of new strategies such as prophylactic vaccines to prevent infection and decrease the disease have become a high priority. Classic vaccines against leishmaniases were based on live or attenuated parasites or their subunits. Nevertheless, the use of whole parasite or their subunits for vaccine production has numerous disadvantages. Therefore, the use of Leishmania peptides to design more specific vaccines against leishmaniases seems promising. Moreover, peptides have several benefits in comparison with other kinds of antigens, for instance, good stability, absence of potentially damaging materials, antigen low complexity, and low-cost to scale up. By contrast, peptides are poor immunogenic alone, and they need to be delivered correctly. In this context, several approaches described in this review are useful to solve these drawbacks. Approaches, such as, peptides in combination with potent adjuvants, cellular vaccinations, adenovirus, polyepitopes, or DNA vaccines have been used to develop peptide-based vaccines. Recent advancements in peptide vaccine design, chimeric, or polypeptide vaccines and nanovaccines based on particles attached or formulated with antigenic components or peptides have been increasingly employed to drive a specific immune response. In this review, we briefly summarize the old, current, and future stands on peptide-based vaccines, describing the disadvantages and benefits associated with them. We also propose possible approaches to overcome the related weaknesses of synthetic vaccines and suggest future guidelines for their development.

  7. Peptide Vaccines for Leishmaniasis.

    Science.gov (United States)

    De Brito, Rory C F; Cardoso, Jamille M De O; Reis, Levi E S; Vieira, Joao F; Mathias, Fernando A S; Roatt, Bruno M; Aguiar-Soares, Rodrigo Dian D O; Ruiz, Jeronimo C; Resende, Daniela de M; Reis, Alexandre B

    2018-01-01

    Due to an increase in the incidence of leishmaniases worldwide, the development of new strategies such as prophylactic vaccines to prevent infection and decrease the disease have become a high priority. Classic vaccines against leishmaniases were based on live or attenuated parasites or their subunits. Nevertheless, the use of whole parasite or their subunits for vaccine production has numerous disadvantages. Therefore, the use of Leishmania peptides to design more specific vaccines against leishmaniases seems promising. Moreover, peptides have several benefits in comparison with other kinds of antigens, for instance, good stability, absence of potentially damaging materials, antigen low complexity, and low-cost to scale up. By contrast, peptides are poor immunogenic alone, and they need to be delivered correctly. In this context, several approaches described in this review are useful to solve these drawbacks. Approaches, such as, peptides in combination with potent adjuvants, cellular vaccinations, adenovirus, polyepitopes, or DNA vaccines have been used to develop peptide-based vaccines. Recent advancements in peptide vaccine design, chimeric, or polypeptide vaccines and nanovaccines based on particles attached or formulated with antigenic components or peptides have been increasingly employed to drive a specific immune response. In this review, we briefly summarize the old, current, and future stands on peptide-based vaccines, describing the disadvantages and benefits associated with them. We also propose possible approaches to overcome the related weaknesses of synthetic vaccines and suggest future guidelines for their development.

  8. Peptide Nucleic Acids

    DEFF Research Database (Denmark)

    2004-01-01

    A novel class of compounds known as peptide nucleic acids, bind complementary DNA and RNA strands, and generally do so more strongly than the corresponding DNA or RNA strands while exhibiting increased sequence specificity and solubility. The peptide nucleic acids comprise ligands selected from...

  9. The repository ecology an approach to understanding repository and service interactions

    CERN Document Server

    CERN. Geneva; Hagemann, Melissa

    2007-01-01

    An increasing number of university institutions and other organisations are deciding to deploy repositories and a growing number of formal and informal distributed services are supporting or capitalising on the information these repositories provide. Despite reasonably well understood technical architectures, early majority adopters may struggle to articulate their place within the actualities of a wider information environment. The idea of a repository ecology provides developers and administrators with a useful way of articulating and analysing their place in the information environment, and the technical and organisational interactions they have, or are developing, with other parts of such an environment. This presentation will provide an overview of the concept of a repository ecology and examine some examples from the domains of scholarly communications and elearning.

  10. The repository ecology: an approach to understanding repository and service interactions

    CERN Multimedia

    CERN. Geneva

    2007-01-01

    An increasing number of university institutions and other organisations are deciding to deploy repositories and a growing number of formal and informal distributed services are supporting or capitalising on the information these repositories provide. Despite reasonably well understood technical architectures, early majority adopters may struggle to articulate their place within the actualities of a wider information environment. The idea of a repository ecology provides developers and administrators with a useful way of articulating and analysing their place in the information environment, and the technical and organisational interactions they have, or are developing, with other parts of such an environment. This presentation will provide an overview of the concept of a repository ecology and examine some examples from the domains of scholarly communications and elearning. View John Robertson's biography

  11. Desiderata for healthcare integrated data repositories based on architectural comparison of three public repositories.

    Science.gov (United States)

    Huser, Vojtech; Cimino, James J

    2013-01-01

    Integrated data repositories (IDRs) are indispensable tools for numerous biomedical research studies. We compare three large IDRs (Informatics for Integrating Biology and the Bedside (i2b2), HMO Research Network's Virtual Data Warehouse (VDW) and Observational Medical Outcomes Partnership (OMOP) repository) in order to identify common architectural features that enable efficient storage and organization of large amounts of clinical data. We define three high-level classes of underlying data storage models and we analyze each repository using this classification. We look at how a set of sample facts is represented in each repository and conclude with a list of desiderata for IDRs that deal with the information storage model, terminology model, data integration and value-sets management.

  12. Towards a Swedish repository for spent fuel

    International Nuclear Information System (INIS)

    Ahlstroem, P.-E.

    1997-01-01

    Nuclear power is producing electricity for the benefit of society but is also leaving radioactive residues behind. It is our responsibility to handle these residues in a safe and proper manner. The development of a system for handling spent fuel from nuclear power plants has proceeded in steps. The same is true for the actual construction of facilities and will continue to be the case for the final repository for spent fuel and other types of long-lived wastes. The primary objective in constructing the repository will be to isolate and contain the radioactive waste. In case the isolation fails for some reason the multibarrier system should retain and retard the radionuclides that might come into contact with the groundwater. A repository is now planned to be built in two steps where the first step will include deposition of about 400 canisters with spent fuel. This first step should be finished in about 20 years from now and be followed by an extensive evaluation of the results from not only this particular step but also from the development of alternative routes before deciding on how to proceed. A special facility to encapsulate the spent fuel is also required. Such an encapsulation plant is proposed to be constructed as an extension of the existing interim storage CLAB. Finding a site for the repository is a critical issue in the implementation of any repository. The siting process started a few years ago and made some progress but is by no means yet completed. It will go on at least into the early part of the next decade. When the present nuclear power plants begin to be due for retirement there should also be some facilities in place to take permanent care of the long-lived radioactive residues. Progress in siting will be a prerequisite for success in our responsibility to make progress towards a safe permanent solution of the waste issue. (orig.)

  13. Making proteomics data accessible and reusable: current state of proteomics databases and repositories.

    Science.gov (United States)

    Perez-Riverol, Yasset; Alpi, Emanuele; Wang, Rui; Hermjakob, Henning; Vizcaíno, Juan Antonio

    2015-03-01

    Compared to other data-intensive disciplines such as genomics, public deposition and storage of MS-based proteomics, data are still less developed due to, among other reasons, the inherent complexity of the data and the variety of data types and experimental workflows. In order to address this need, several public repositories for MS proteomics experiments have been developed, each with different purposes in mind. The most established resources are the Global Proteome Machine Database (GPMDB), PeptideAtlas, and the PRIDE database. Additionally, there are other useful (in many cases recently developed) resources such as ProteomicsDB, Mass Spectrometry Interactive Virtual Environment (MassIVE), Chorus, MaxQB, PeptideAtlas SRM Experiment Library (PASSEL), Model Organism Protein Expression Database (MOPED), and the Human Proteinpedia. In addition, the ProteomeXchange consortium has been recently developed to enable better integration of public repositories and the coordinated sharing of proteomics information, maximizing its benefit to the scientific community. Here, we will review each of the major proteomics resources independently and some tools that enable the integration, mining and reuse of the data. We will also discuss some of the major challenges and current pitfalls in the integration and sharing of the data. © 2014 The Authors. PROTEOMICS published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Design approaches for access plugs in a basalt repository

    International Nuclear Information System (INIS)

    O'Rourke, J.; Allirot, D.; O'Connor, K.

    1982-01-01

    This paper describes research, laboratory testing, and analytical approaches taken toward the development of designs for sealing boreholes, shafts, and tunnels penetrating from ground surface to a deep, mined nuclear waste repository in basalt. A material selection process leading to identification of preferred sealing materials is discussed, and the laboratory testing program to define the geochemical and geotechnical performance of these materials is described. Analysis of the environmental conditions in the Columbia Plateau basalt flows leads to identification of tentative design criteria for plug systems. These design criteria include performance of the plug as a hydraulic barrier and as a radionuclide barrier. An important problem for effective performance of a plug system as a hydraulic barrier is shown to be a potentially disturbed zone surrounding the excavation in the stressed and jointed host rock. An idealized one-dimensional numerical model is described for analyzing the performance of the plug as a barrier to radionuclide transport. The preliminary analyses led to the conclusion that the composition and dimensions of practical candidate plugs can satisfy both hydraulic and radionuclide barrier criteria. Examples of candidate designs are shown for boreholes, shafts, and tunnels. 9 references, 6 figures, 6 tables

  15. Investigative study of standards for Digital Repositories and related services

    NARCIS (Netherlands)

    Foulonneau, Muriel; André, Francis

    2007-01-01

    This study is meant for institutional repository managers, service providers, repository software developers and generally, all players taking an active part in the creation of the digital repository infrastructure for e-research and e-learning. It reviews the current standards, protocols and

  16. Prestudy Oskarshamn. Tourism in Oskarshamn with or without a repository

    International Nuclear Information System (INIS)

    Nordblom, C.J.; Foghagen, C.

    1998-11-01

    Consequences for the tourism at Oskarshamn from siting a spent fuel repository in the community are studied. Four questionnaire/interview enquires were performed, and the analysis of the results show that no noticeable effects are expected. Still, an uncertainty about the impact of a repository is felt by the public, and more thorough information about the repository and its components is needed

  17. 10 CFR 51.67 - Environmental information concerning geologic repositories.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 2 2010-01-01 2010-01-01 false Environmental information concerning geologic repositories... information concerning geologic repositories. (a) In lieu of an environmental report, the Department of Energy... connection with any geologic repository developed under Subtitle A of Title I, or under Title IV, of the...

  18. Semantic Linking of Learning Object Repositories to DBpedia

    Science.gov (United States)

    Lama, Manuel; Vidal, Juan C.; Otero-Garcia, Estefania; Bugarin, Alberto; Barro, Senen

    2012-01-01

    Large-sized repositories of learning objects (LOs) are difficult to create and also to maintain. In this paper we propose a way to reduce this drawback by improving the classification mechanisms of the LO repositories. Specifically, we present a solution to automate the LO classification of the Universia repository, a collection of more than 15…

  19. Institutional Repositories in Indian Universities and Research Institutes: A Study

    Science.gov (United States)

    Krishnamurthy, M.; Kemparaju, T. D.

    2011-01-01

    Purpose: The purpose of this paper is to report on a study of the institutional repositories (IRs) in use in Indian universities and research institutes. Design/methodology/approach: Repositories in various institutions in India were accessed and described in a standardised way. Findings: The 20 repositories studied covered collections of diverse…

  20. The Use of Digital Repositories for Enhancing Teacher Pedagogical Performance

    Science.gov (United States)

    Cohen, Anat; Kalimi, Sharon; Nachmias, Rafi

    2013-01-01

    This research examines the usage of local learning material repositories at school, as well as related teachers' attitudes and training. The study investigates the use of these repositories for enhancing teacher performance and assesses whether the assimilation of the local repositories increases their usage of and contribution to by teachers. One…

  1. 48 CFR 227.7207 - Contractor data repositories.

    Science.gov (United States)

    2010-10-01

    ... repositories. 227.7207 Section 227.7207 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS... Computer Software and Computer Software Documentation 227.7207 Contractor data repositories. Follow 227.7108 when it is in the Government's interests to have a data repository include computer software or to...

  2. Institutional Repositories as Infrastructures for Long-Term Preservation

    Science.gov (United States)

    Francke, Helena; Gamalielsson, Jonas; Lundell, Björn

    2017-01-01

    Introduction: The study describes the conditions for long-term preservation of the content of the institutional repositories of Swedish higher education institutions based on an investigation of how deposited files are managed with regards to file format and how representatives of the repositories describe the functions of the repositories.…

  3. The Lincoln Repository presentation: ten reasons why you should put a copy of your work in the Repository

    OpenAIRE

    Stainthorp, Paul

    2010-01-01

    Slideshow presentation created to promote the Lincoln Repository to staff at the University of Lincoln. Consists of ten reasons why academic authors should consider depositing copies of their work in the Repository.

  4. The Drosophila melanogaster PeptideAtlas facilitates the use of peptide data for improved fly proteomics and genome annotation

    Directory of Open Access Journals (Sweden)

    King Nichole L

    2009-02-01

    Full Text Available Abstract Background Crucial foundations of any quantitative systems biology experiment are correct genome and proteome annotations. Protein databases compiled from high quality empirical protein identifications that are in turn based on correct gene models increase the correctness, sensitivity, and quantitative accuracy of systems biology genome-scale experiments. Results In this manuscript, we present the Drosophila melanogaster PeptideAtlas, a fly proteomics and genomics resource of unsurpassed depth. Based on peptide mass spectrometry data collected in our laboratory the portal http://www.drosophila-peptideatlas.org allows querying fly protein data observed with respect to gene model confirmation and splice site verification as well as for the identification of proteotypic peptides suited for targeted proteomics studies. Additionally, the database provides consensus mass spectra for observed peptides along with qualitative and quantitative information about the number of observations of a particular peptide and the sample(s in which it was observed. Conclusion PeptideAtlas is an open access database for the Drosophila community that has several features and applications that support (1 reduction of the complexity inherently associated with performing targeted proteomic studies, (2 designing and accelerating shotgun proteomics experiments, (3 confirming or questioning gene models, and (4 adjusting gene models such that they are in line with observed Drosophila peptides. While the database consists of proteomic data it is not required that the user is a proteomics expert.

  5. Current Status of Deep Geological Repository Development

    International Nuclear Information System (INIS)

    Budnitz, R J

    2005-01-01

    This talk provided an overview of the current status of deep-geological-repository development worldwide. Its principal observation is that a broad consensus exists internationally that deep-geological disposal is the only long-term solution for disposition of highly radioactive nuclear waste. Also, it is now clear that the institutional and political aspects are as important as the technical aspects in achieving overall progress. Different nations have taken different approaches to overall management of their highly radioactive wastes. Some have begun active programs to develop a deep repository for permanent disposal: the most active such programs are in the United States, Sweden, and Finland. Other countries (including France and Russia) are still deciding on whether to proceed quickly to develop such a repository, while still others (including the UK, China, Japan) have affirmatively decided to delay repository development for a long time, typically for a generation of two. In recent years, a major conclusion has been reached around the world that there is very high confidence that deep repositories can be built, operated, and closed safely and can meet whatever safety requirements are imposed by the regulatory agencies. This confidence, which has emerged in the last few years, is based on extensive work around the world in understanding how repositories behave, including both the engineering aspects and the natural-setting aspects, and how they interact together. The construction of repositories is now understood to be technically feasible, and no major barriers have been identified that would stand in the way of a successful project. Another major conclusion around the world is that the overall cost of a deep repository is not as high as some had predicted or feared. While the actual cost will not be known in detail until the costs are incurred, the general consensus is that the total life-cycle cost will not exceed a few percent of the value of the

  6. Design aspects of the alpha repository. II. Conceptual layouts of underground storage facilities

    International Nuclear Information System (INIS)

    Grams, W.H.

    1975-01-01

    Five conceptual repository layouts are presented: linear repository, 1 panel; bow tie repository, 2 panels; maltese cross repository, 4 panels; snowflake repository; 5 panels, and sash window repository, 8 panels. The layouts are compared with respect to excavation requirements, haulage distances, ventilation flow path designs, and safety features

  7. Thermal analyses of spent nuclear fuel repository

    International Nuclear Information System (INIS)

    Ikonen, K.

    2003-06-01

    This report contains the temperature dimensioning of the KBS-3V type 1- or 2-panel repository based on the rock properties measured from the Olkiluoto investigations. The report describes first the development of a calculation methodology for the thermal analysis of a repository for nuclear fuel. The disposed canisters produce residual heat due to decay (or disintegration) of radioactive products. The decay heat is conducted to surrounding rock mass. The methods were applied to determine the effect of different parameters on the highest canister temperature and to support the planning, dimensioning and operation of the repository. The thermal diffusivity of the rock is low and the heat released from the canisters is spread into the surrounding rock volume quite slowly causing thermal gradient in the rock close to canisters and the canister temperature is increased remarkably. The maximum temperature on the canister surface is limited to the design temperature of +100 deg C. However, due to uncertainties in thermal analysis parameters (like scattering in rock conductivity) the allowable calculated maximum canister temperature is set to 90 deg C causing a safety margin of 10 deg C. The allowable temperature is controlled by the spacing between adjacent canisters, adjacent tunnels and the distance between separate panels of the repository and the pre-cooling time affecting power of the canisters. Because of the fact that the disposal operation takes several decades, the moment of disposal of an individual canister in addition to the location has an influence on the maximum temperature in the canister. Also, a second disposal panel in the repository has a thermal interaction with the other panel. This interaction is expressed after a few decades at the strongest. It became apparent that the temperature of canister surfaces can be determined by analytic line heat source model much more efficiently than by numerical analysis, if the analytic model is first verified and

  8. Safety analysis of the VLJ repository

    International Nuclear Information System (INIS)

    Vieno, T.; Nordman, H.

    1991-05-01

    The VLJ repository is an underground disposal facility for the low and medium level waste generated at the Olkiluoto nuclear power plant. The repository is located within 1 km from TVO I and TVO II (2 x 710 MWe) BWR's on the Olkiluoto island at the west coast of Finland. It contains two rock silos excavated at the depth of 60...100 meters in the bedrock. Low level waste will be disposed of in a shotcreted rock silo. For bituminized medium level waste, a separate silo of reinforced concrete has been built inside the shotcreted rock silo. The post-closure safety analysis has been done for the Final Safety Analysis Report (FSAR) of the VLJ repository. In addition to the normal evolution scenario, several disturbed evolution and accident scenarios have been analysed. In the reference scenario, radio-nuclides are assumed to be released from the bituminized waste within 500 years, the concrete silo is assumed to gradually disintegrate and finally to collapse at 5 000 years, all concrete in the silo is assumed to be also chemically depleted within 6 000 years, and all the seals of the repository are assumed to deteriorate within 12 000 years. The ability of alone natural barriers to restrict the release of radionuclides into the biosphere has been evaluated by means of scenarios where the degradation of engineered barriers has been assumed to take place at a still faster rate. In one of the disturbed evolution scenarios it has been assumed that the concrete silo for medium level waste is severely impaired immediately after sealing of the repository. Effects of gas generation and consequences of human intrusion have been evaluated, too. The results of the safety analysis show that radiation doses of any significance are caused only if a well is bored in the vicinity of the repository or if the groundwater discharge spot is inhabited and used for cultivation. In the reference scenario the maximum expectation value of the individual dose rate is 0.3 mSv/a

  9. What do we mean by a cold repository?

    International Nuclear Information System (INIS)

    Halsey, W.G.

    1994-01-01

    The topic of thermal loading of a potential repository at Yucca Mountain in Nevada has been the subject of intense discussion within the project technical community. While terms such as ''Hot Repository'' and ''Cold Repository'' are frequently used, they have not been clearly defined. In particular, the definition of a cold repository has remained the opinion of each individual. This has led to confusion and misunderstanding. In this paper, a number of observed definitions for a cold repository are discussed along with the technical implications, assumptions and inconsistencies. Finally, a common language is suggested

  10. Reformulating Component Identification as Document Analysis Problem

    NARCIS (Netherlands)

    Gross, H.G.; Lormans, M.; Zhou, J.

    2007-01-01

    One of the first steps of component procurement is the identification of required component features in large repositories of existing components. On the highest level of abstraction, component requirements as well as component descriptions are usually written in natural language. Therefore, we can

  11. Tandem MS Analysis of Selenamide-Derivatized Peptide Ions

    Science.gov (United States)

    Zhang, Yun; Zhang, Hao; Cui, Weidong; Chen, Hao

    2011-09-01

    Our previous study showed that selenamide reagents such as ebselen and N-(phenylseleno)phthalimide (NPSP) can be used for selective and rapid derivatization of protein/peptide thiols in high conversion yield. This paper reports the systematic investigation of MS/MS dissociation behaviors of selenamide-derivatized peptide ions upon collision induced dissociation (CID) and electron transfer dissociation (ETD). In the positive ion mode, derivatized peptide ions exhibit tag-dependent CID dissociation pathways. For instance, ebselen-derivatized peptide ions preferentially undergo Se-S bond cleavage upon CID to produce a characteristic fragment ion, the protonated ebselen ( m/z 276), which allows selective identification of thiol peptides from protein digest as well as selective detection of thiol proteins from protein mixture using precursor ion scan (PIS). In contrast, NPSP-derivatized peptide ions retain their phenylselenenyl tags during CID, which is useful in sequencing peptides and locating cysteine residues. In the negative ion CID mode, both types of tags are preferentially lost via the Se-S cleavage, analogous to the S-S bond cleavage during CID of disulfide-containing peptide anions. In consideration of the convenience in preparing selenamide-derivatized peptides and the similarity of Se-S of the tag to the S-S bond, we also examined ETD of the derivatized peptide ions to probe the mechanism for electron-based ion dissociation. Interestingly, facile cleavage of Se-S bond occurs to the peptide ions carrying either protons or alkali metal ions, while backbone cleavage to form c/z ions is severely inhibited. These results are in agreement with the Utah-Washington mechanism proposed for depicting electron-based ion dissociation processes.

  12. Ventilation System Strategy for a Prospective Korean Radioactive Waste Repository

    International Nuclear Information System (INIS)

    Kim, Jin; Kwon, Sang Ki

    2005-01-01

    In the stage of conceptual design for the construction and operation of the geologic repository for radioactive wastes, it is important to consider a repository ventilation system which serves the repository working environment, hygiene and safety of the public at large, and will allow safe maintenance like moisture content elimination in repository for the duration of the repositories life, construction/operation/closure, also allowing safe waste transportation and emplacement. This paper describes the possible ventilation system design criteria and requirements for the prospective Korean radioactive waste repositories with emphasis on the underground rock cavity disposal method in the both cases of low and medium-level and high-level wastes. It was found that the most important concept is separate ventilation systems for the construction (development) and waste emplacement (storage) activities. In addition, ventilation network system modeling, natural ventilation, ventilation monitoring systems and real time ventilation simulation, and fire simulation and emergency system in the repository are briefly discussed.

  13. Di peptides of the Zeyhera digitalis roots; Dipeptideo das raizes de Zeyhera digitalis

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Dalva Trevisan; Silva, Rosely Barbosa da [Universidade Estadual de Londrina, PR (Brazil). Dept. de Quimica; Oliveira, Alaide Braga de [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Dept. de Quimica; Isobe, Minoru [Nagoya Univ. (Japan)

    1991-12-31

    Systematic investigation using {sup 1} H NMR have been done on the Zeyhera digitalis roots for isolation, purification, and identification of substances with biological or medical potential activities , such as di peptides compounds 5 figs.

  14. Di peptides of the Zeyhera digitalis roots; Dipeptideo das raizes de Zeyhera digitalis

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Dalva Trevisan; Silva, Rosely Barbosa da [Universidade Estadual de Londrina, PR (Brazil). Dept. de Quimica; Oliveira, Alaide Braga de [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Dept. de Quimica; Isobe, Minoru [Nagoya Univ. (Japan)

    1992-12-31

    Systematic investigation using {sup 1} H NMR have been done on the Zeyhera digitalis roots for isolation, purification, and identification of substances with biological or medical potential activities , such as di peptides compounds 5 figs.

  15. Digital Repository of Research Institutes – RCIN

    Directory of Open Access Journals (Sweden)

    Kamila Kaczyńska

    2014-03-01

    Full Text Available The paper describes the project of Digital Repository of Scientific Institutes RCIN and presents opportunities for promoting science by digitization and sharing them on the Internet. The Repository has been created by the 16 Scientific Institutes in Warsaw, Krakow and Bialowieza to modernize the science-research and IT infrastructure, to increase digital resources of mathematical, technical, natural and medical sciences, and to popularize and promote of Polish science. That dissemination and popularization of science affects its development and competitiveness in the international arena and it allows transfer of research results to the economy. In addition, Institutes of RCIN providing contemporary and archival materials of science, support the intellectual capital of Polish science and raise awareness of professional literature of search on the Internet. Project RCIN is implemented in the years 2010–2014 and financing is provided by the funds of the European Fund of Regional Development.

  16. Geological study of radioactive waste repositories

    International Nuclear Information System (INIS)

    Oyama, Takahiro; Kitano, Koichi

    1987-01-01

    The investigation of the stability and the barrier efficiency of the deep underground radioactive waste repositories become a subject of great concern. The purpose of this paper is to gather informations on the geology, engineering geology and hydrogeology in deep galleries in Japan. Conclusion can be summarised as follows: (1) The geological structure of deep underground is complicated. (2) Stress in deep underground is greatly affected by crustal movement. (3) Rock-burst phenomena occur in the deep underground excavations. (4) In spite of deep underground, water occasionally gush out from the fractured zone of rock mass. These conclusion will be useful for feasibility study of underground waste disposal and repositories in Japan. (author)

  17. The German quality system for waste repositories

    International Nuclear Information System (INIS)

    Beckmerhagen, I.; Berg, H.P.; Brennecke, P.

    1993-01-01

    The Bundesamt fuer Strahlenschutz (BfS)--Federal Office for Radiation protection--has to guarantee that the requirements resulting from different regulations concerning planning, design, construction, operation and decommissioning of a waste repository are fulfilled. In addition, the results of the safety assessments lead to nuclear-specific requirements on the design of the plant as well as to requirements on the radioactive waste packages intended to be disposed of. Therefore, the implementation of a quality assurance (QA) and quality control (QC) system is an essential task in order to ensure that the designed quality is achieved so that the necessary precaution against damage is taken. In this paper, a detailed description of QA and QC to be applied to the planned Konrad repository as well as the basic principles and the present status of the waste package QC are indicated and discussed

  18. Summary of repository siting models. Final report

    International Nuclear Information System (INIS)

    Thomas, S.D.; Ross, B.; Mercer, J.W.

    1982-07-01

    This report is the first in a series of reports that will provide critical reviews and summaries of computer programs that can be used to analyze the potential performance of a high-level radioactive waste repository. The computer programs identified address the following phenomena: saturated and unsaturated subsurface flow, heat transport, solute transport, surface water runoff, geomechanical interactions, and geochemical interactions. The report identifies 183 computer programs that can be used to analyze a repository site and provides a summary description of 31 computer programs. The summary descriptions can be used: to assist in code evaluation, to facilitate code comparison, to determine applicability of codes to specific problems, to identify code deficiencies, and to provide a screening mechanism for code selection

  19. Numerical modeling capabilities to predict repository performance

    International Nuclear Information System (INIS)

    1979-09-01

    This report presents a summary of current numerical modeling capabilities that are applicable to the design and performance evaluation of underground repositories for the storage of nuclear waste. The report includes codes that are available in-house, within Golder Associates and Lawrence Livermore Laboratories; as well as those that are generally available within the industry and universities. The first listing of programs are in-house codes in the subject areas of hydrology, solute transport, thermal and mechanical stress analysis, and structural geology. The second listing of programs are divided by subject into the following categories: site selection, structural geology, mine structural design, mine ventilation, hydrology, and mine design/construction/operation. These programs are not specifically designed for use in the design and evaluation of an underground repository for nuclear waste; but several or most of them may be so used

  20. Use of modeling in repository licensing

    International Nuclear Information System (INIS)

    McGarry, J.M. III; Echols, F.S.

    1995-01-01

    A review of the regulatory history of the Nuclear Regulatory Commission (NRC) regulations applicable to the licensing of a geologic repository, as well as a review of NRC administrative (licensing) decisions and federal case law, support the NRC's use of simplified models, in appropriate circumstances, which provide well-documented and reasonably conservative bounding assumptions, together with the use of expert judgement, natural analogues, and other aids to supplement available information, in reaching its reasonable assurance determination whether the public health and safety will be adequately protected if the Yucca Mountain, Nevada site should be licensed for development as a geologic repository. Specific examples are provided to assist the reader to better understand how such qualitative concepts as open-quote reasonable assurance close-quote, open-quote reasonably conservative close-quote, and open-quote adequate close-quote protection are used in an administrative context to resolve technical issues