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Sample records for sav3 based replicon

  1. A highly pathogenic porcine reproductive and respiratory syndrome virus candidate vaccine based on Japanese encephalitis virus replicon system.

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    Hu, Pingsheng; Chen, Xiaoming; Huang, Lihong; Liu, Shukai; Zang, Fuyu; Xing, Jinchao; Zhang, Youyue; Liang, Jiaqi; Zhang, Guihong; Liao, Ming; Qi, Wenbao

    2017-01-01

    In the swine industry, porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease which causes heavy economic losses worldwide. Effective prevention and disease control is an important issue. In this study, we described the construction of a Japanese encephalitis virus (JEV) DNA-based replicon with a cytomegalovirus (CMV) promoter based on the genome of Japanese encephalitis live vaccine virus SA14-14-2, which is capable of offering a potentially novel way to develop and produce vaccines against a major pathogen of global health. This JEV DNA-based replicon contains a large deletion in the structural genes (C-prM-E). A PRRSV GP5/M was inserted into the deletion position of JEV DNA-based replicons to develop a chimeric replicon vaccine candidate for PRRSV. The results showed that BALB/c mice models with the replicon vaccines pJEV-REP-G-2A-M-IRES and pJEV-REP-G-2A-M stimulated antibody responses and induced a cellular immune response. Analysis of ELSA data showed that vaccination with the replicon vaccine expressing GP5/M induced a better antibodies response than traditional DNA vaccines. Therefore, the results suggested that this ectopic expression system based on JEV DNA-based replicons may represent a useful molecular platform for various biological applications, and the JEV DNA-based replicons expressing GP5/M can be further developed into a novel, safe vaccine candidate for PRRS.

  2. In vitro adaptation of SAV3 in cell culture correlates with reduced in vivo replication capacity and virulence to Atlantic salmon (Salmo salar L.) parr.

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    Petterson, Elin; Guo, Tz-Chun; Evensen, Øystein; Haugland, Øyvind; Mikalsen, Aase B

    2015-10-01

    Salmonid alphavirus (SAV) is the causative agent of pancreas disease affecting Atlantic salmon and rainbow trout and causes a major burden to the aquaculture industry. This study describes a Norwegian subtype SAV3 virus isolate (SAV3-H10) subjected to serial passages in Chinook salmon embryo cells (CHSE-214) followed by Asian Grouper skin cells (AGK). Two passages from CHSE and one after transfer to AGK cells were chosen for further investigation, based on variation in degree and development of cytopathic effect (CPE). After plaque purification, several in vitro studies were performed. Cell viability after infection, viral replication and ability to cause morphological changes in CHSE and AGK cells was studied for the three isolates. The AGK-transferred isolate was identified with the strongest abilities to reduce cell viability, replicate more and cause more CPE in cell culture when compared with the early and late CHSE-grown isolates. Subsequently, the isolates were tested in an experimental fish challenge, showing higher viral load and higher pathological score for the least cell-cultured isolate. Full-length sequencing of the viral genome of the three isolates revealed divergence in four amino acid positions and the AGK-grown isolate also had a 3 nt deletion in the 3'UTR. In conclusion, we show that cell culture of SAV3-H10 selects for strains inducing earlier CPE in vitro with increased viral replication. In vivo, the effect is reversed, with lower replication levels and lower pathology scores in target organs. This study outlines a path to identify potential virulence motifs of SAV3.

  3. Immunogenicity of a DNA-launched replicon-based canine parvovirus DNA vaccine expressing VP2 antigen in dogs.

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    Dahiya, Shyam S; Saini, Mohini; Kumar, Pankaj; Gupta, Praveen K

    2012-10-01

    A replicon-based DNA vaccine encoding VP2 gene of canine parvovirus (CPV) was developed by cloning CPV-VP2 gene into a replicon-based DNA vaccine vector (pAlpha). The characteristics of a replicon-based DNA vaccine like, self-amplification of transcripts and induction of apoptosis were analyzed in transfected mammalian cells. When the pAlpha-CPV-VP2 was injected intradermal as DNA-launched replicon-based DNA vaccine in dogs, it induced CPV-specific humoral and cell mediated immune responses. The virus neutralization antibody and lymphocyte proliferative responses were higher than conventional CPV DNA vaccine and commercial CPV vaccine. These results indicated that DNA-launched replicon-based CPV DNA vaccine was effective in inducing both CPV-specific humoral and cellular immune responses and can be considered as effective alternative to conventional CPV DNA vaccine and commercial CPV vaccine. Crown Copyright © 2012. Published by Elsevier India Pvt Ltd. All rights reserved.

  4. Alphavirus vector-based replicon particles expressing multivalent cross-protective Lassa virus glycoproteins.

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    Wang, Min; Jokinen, Jenny; Tretyakova, Irina; Pushko, Peter; Lukashevich, Igor S

    2018-01-29

    Lassa virus (LASV) is the most prevalent rodent-borne arenavirus circulated in West Africa. With population at risk from Senegal to Nigeria, LASV causes Lassa fever and is responsible for thousands of deaths annually. High genetic diversity of LASV is one of the challenges for vaccine R&D. We developed multivalent virus-like particle vectors (VLPVs) derived from the human Venezuelan equine encephalitis TC-83 IND vaccine (VEEV) as the next generation of alphavirus-based bicistronic RNA replicon particles. The genes encoding VEEV structural proteins were replaced with LASV glycoproteins (GPC) from distantly related clades I and IV with individual 26S promoters. Bicistronic RNA replicons encoding wild-type LASV GPC (GPCwt) and C-terminally deleted, non-cleavable modified glycoprotein (ΔGPfib), were encapsidated into VLPV particles using VEEV capsid and glycoproteins provided in trans. In transduced cells, VLPVs induced simultaneous expression of LASV GPCwt and ΔGPfib from 26S alphavirus promoters. LASV ΔGPfib was predominantly expressed as trimers, accumulated in the endoplasmic reticulum, induced ER stress and apoptosis promoting antigen cross-priming. VLPV vaccines were immunogenic and protective in mice and upregulated CD11c + /CD8 + dendritic cells playing the major role in cross-presentation. Notably, VLPV vaccination resulted in induction of cross-reactive multifunctional T cell responses after stimulation of immune splenocytes with peptide cocktails derived from LASV from clades I-IV. Multivalent RNA replicon-based LASV vaccines can be applicable for first responders, international travelers visiting endemic areas, military and lab personnel. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Development of expression vectors for Escherichia coli based on the pCR2 replicon

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    Deb J K

    2007-05-01

    Full Text Available Abstract Background Recent developments in metabolic engineering and the need for expanded compatibility required for co-expression studies, underscore the importance of developing new plasmid vectors with properties such as stability and compatibility. Results We utilized the pCR2 replicon of Corynebacterium renale, which harbours multiple plasmids, for constructing a range of expression vectors. Different antibiotic-resistance markers were introduced and the vectors were found to be 100% stable over a large number of generations in the absence of selection pressure. Compatibility of this plasmid was studied with different Escherichia coli plasmid replicons viz. pMB1 and p15A. It was observed that pCR2 was able to coexist with these E.coli plasmids for 60 generations in the absence of selection pressure. Soluble intracellular production was checked by expressing GFP under the lac promoter in an expression plasmid pCR2GFP. Also high level production of human IFNγ was obtained by cloning the h-IFNγ under a T7 promoter in the expression plasmid pCR2-IFNγ and using a dual plasmid heat shock system for expression. Repeated sub-culturing in the absence of selection pressure for six days did not lead to any fall in the production levels post induction, for both GFP and h-IFNγ, demonstrating that pCR2 is a useful plasmid in terms of stability and compatibility. Conclusion We have constructed a series of expression vectors based on the pCR2 replicon and demonstrated its high stability and sustained expression capacity, in the absence of selection pressure which will make it an efficient tool for metabolic engineering and co-expression studies, as well as for scale up of expression.

  6. Role of innate signalling pathways in the immunogenicity of alphaviral replicon-based vaccines

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    Chen Margaret

    2011-01-01

    Full Text Available Abstract Background Alphaviral replicon-based vectors induce potent immune responses both when given as viral particles (VREP or as DNA (DREP. It has been suggested that the strong immune stimulatory effect induced by these types of vectors is mediated by induction of danger signals and activation of innate signalling pathways due to the replicase activity. To investigate the innate signalling pathways involved, mice deficient in either toll-like receptors or downstream innate signalling molecules were immunized with DREP or VREP. Results We show that the induction of a CD8+ T cell response did not require functional TLR3 or MyD88 signalling. However, IRF3, converging several innate signalling pathways and important for generation of pro-inflammatory cytokines and type I IFNs, was needed for obtaining a robust primary immune response. Interestingly, type I interferon (IFN, induced by most innate signalling pathways, had a suppressing effect on both the primary and memory T cell responses after DREP and VREP immunization. Conclusions We show that alphaviral replicon-based vectors activate multiple innate signalling pathways, which both activate and restrict the induced immune response. These results further show that there is a delicate balance in the strength of innate signalling and induction of adaptive immune responses that should be taken into consideration when innate signalling molecules, such as type I IFNs, are used as vaccine adjuvant.

  7. Inhibitors of alphavirus entry and replication identified with a stable Chikungunya replicon cell line and virus-based assays.

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    Leena Pohjala

    Full Text Available Chikungunya virus (CHIKV, an alphavirus, has recently caused epidemic outbreaks and is therefore considered a re-emerging pathogen for which no effective treatment is available. In this study, a CHIKV replicon containing the virus replicase proteins together with puromycin acetyltransferase, EGFP and Renilla luciferase marker genes was constructed. The replicon was transfected into BHK cells to yield a stable cell line. A non-cytopathic phenotype was achieved by a Pro718 to Gly substitution and a five amino acid insertion within non-structural protein 2 (nsP2, obtained through selection for stable growth. Characterization of the replicon cell line by Northern blotting analysis revealed reduced levels of viral RNA synthesis. The CHIKV replicon cell line was validated for antiviral screening in 96-well format and used for a focused screen of 356 compounds (natural compounds and clinically approved drugs. The 5,7-dihydroxyflavones apigenin, chrysin, naringenin and silybin were found to suppress activities of EGFP and Rluc marker genes expressed by the CHIKV replicon. In a concomitant screen against Semliki Forest virus (SFV, their anti-alphaviral activity was confirmed and several additional inhibitors of SFV with IC₅₀ values between 0.4 and 24 µM were identified. Chlorpromazine and five other compounds with a 10H-phenothiazinyl structure were shown to inhibit SFV entry using a novel entry assay based on a temperature-sensitive SFV mutant. These compounds also reduced SFV and Sindbis virus-induced cytopathic effect and inhibited SFV virion production in virus yield experiments. Finally, antiviral effects of selected compounds were confirmed using infectious CHIKV. In summary, the presented approach for discovering alphaviral inhibitors enabled us to identify potential lead structures for the development of alphavirus entry and replication phase inhibitors as well as demonstrated the usefulness of CHIKV replicon and SFV as biosafe surrogate

  8. A 96-well based analysis of replicon elimination with the HCV NS5A replication complex inhibitor daclatasvir.

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    O'Boyle, Donald R; Nower, Peter T; Sun, Jin-Hua; Fridell, Robert; Wang, Chunfu; Valera, Lourdes; Gao, Min

    2013-10-01

    A 96-well based replicon elimination and colony formation assay is presented for comparing the resistance barrier of the hepatitis C virus (HCV) NS5A replication complex inhibitor daclatasvir (DCV, BMS-790052) on three HCV genotypes (gts) in a proof of concept experimental protocol. The 96-well assay format provides both individual colony as well as population characterization and is readily applicable to other HCV direct-acting antiviral agents (DAAs). The assay provides an assessment of HCV replication levels over a 5log10 range by measuring a luciferase reporter resident in the HCV replicons. Individual colony status can be measured with a separate and compatible resazurin assay to assess relative host cell fitness following inhibitor treatments. The methods employed are non-toxic and leave intact isolatable colonies that can be used for phenotyping and genotyping. The utility of the assay is demonstrated by the identification and isolation of resistant variants as well as in the ranking of the relative resistance barrier for the replication complex inhibitor DCV for gts 1a, 1b and 2a. The format provides a quantitative ranking based upon luciferase activity and has the ability to monitor DAA resistance development over time for large numbers of compounds. Copyright © 2013. Published by Elsevier B.V.

  9. Alphavirus replicon-based adjuvants enhance the immunogenicity and effectiveness of Fluzone ® in rhesus macaques.

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    Carroll, Timothy D; Matzinger, Shannon R; Barro, Mario; Fritts, Linda; McChesney, Michael B; Miller, Christopher J; Johnston, Robert E

    2011-01-29

    Venezuelan equine encephalitis virus replicon particles (VRP) without a transgene (null VRP) have been used to adjuvant effective humoral [1], cellular [2], and mucosal [3] immune responses in mice. To assess the adjuvant activity of null VRP in the context of a licensed inactivated influenza virus vaccine, rhesus monkeys were immunized with Fluzone(®) alone or Fluzone(®) mixed with null VRP and then challenged with a human seasonal influenza isolate, A/Memphis/7/2001 (H1N1). Compared to Fluzone(®) alone, Fluzone(®)+null VRP immunized animals had stronger influenza-specific CD4(+) T cell responses (4.4 fold) with significantly higher levels of virus-specific IFN-γ (7.6 fold) and IL-2 (5.3 fold) producing CD4+ T cells. Fluzone(®)+null VRP immunized animals also had significantly higher plasma anti-influenza IgG (pVRP immunization was 1.2 log greater (pVRP immunized monkeys had a significantly lower level of viral replication (pVRP immunized monkeys immediately after challenge. There were significant inverse correlations between influenza RNA levels in tracheal lavages and plasma anti-influenza HI and IgG anti-influenza antibody titers prior to challenge. These results demonstrate that null VRP dramatically improve both the immunogenicity and protection elicited by a licensed inactivated influenza vaccine. Copyright © 2010 Elsevier Ltd. All rights reserved.

  10. RNA-seq based transcriptome analysis of hepatitis E virus (HEV and hepatitis B virus (HBV replicon transfected Huh-7 cells.

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    Neetu Jagya

    Full Text Available Pathogenesis of hepatitis B virus (HBV and hepatitis E virus (HEV infection is as varied as they appear similar; while HBV causes an acute and/or chronic liver disease and hepatocellular carcinoma, HEV mostly causes an acute self-limiting disease. In both infections, host responses are crucial in disease establishment and/or virus clearance. In the wake of worsening prognosis described during HEV super-infection over chronic HBV hepatitis, we investigated the host responses by studying alterations in gene expression in liver cells (Huh-7 cell line by transfection with HEV replicon only (HEV-only, HBV replicon only (HBV-only and both HBV and HEV replicons (HBV+HEV. Virus replication was validated by strand-specific real-time RT-PCR for HEV and HBsAg ELISA of the culture supernatants for HBV. Indirect immunofluorescence for the respective viral proteins confirmed infection. Transcription profiling was carried out by RNA Sequencing (RNA-Seq analysis of the poly-A enriched RNA from the transfected cells. Averages of 600 million bases within 5.6 million reads were sequenced in each sample and ∼15,800 genes were mapped with at least one or more reads. A total of 461 genes in HBV+HEV, 408 in HBV-only and 306 in HEV-only groups were differentially expressed as compared to mock transfection control by two folds (p<0.05 or more. Majority of the significant genes with altered expression clustered into immune-associated, signal transduction, and metabolic process categories. Differential gene expression of functionally important genes in these categories was also validated by real-time RT-PCR based relative gene-expression analysis. To our knowledge, this is the first report of in vitro replicon transfected RNA-Seq based transcriptome analysis to understand the host responses against HEV and HBV.

  11. Model-driven engineering of gene expression from RNA replicons.

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    Beal, Jacob; Wagner, Tyler E; Kitada, Tasuku; Azizgolshani, Odisse; Parker, Jordan Moberg; Densmore, Douglas; Weiss, Ron

    2015-01-16

    RNA replicons are an emerging platform for engineering synthetic biological systems. Replicons self-amplify, can provide persistent high-level expression of proteins even from a small initial dose, and, unlike DNA vectors, pose minimal risk of chromosomal integration. However, no quantitative model sufficient for engineering levels of protein expression from such replicon systems currently exists. Here, we aim to enable the engineering of multigene expression from more than one species of replicon by creating a computational model based on our experimental observations of the expression dynamics in single- and multireplicon systems. To this end, we studied fluorescent protein expression in baby hamster kidney (BHK-21) cells using a replicon derived from Sindbis virus (SINV). We characterized expression dynamics for this platform based on the dose-response of a single species of replicon over 50 h and on a titration of two cotransfected replicons expressing different fluorescent proteins. From this data, we derive a quantitative model of multireplicon expression and validate it by designing a variety of three-replicon systems, with profiles that match desired expression levels. We achieved a mean error of 1.7-fold on a 1000-fold range, thus demonstrating how our model can be applied to precisely control expression levels of each Sindbis replicon species in a system.

  12. Molecular characterization and PCR-based replicon typing of multidrug resistant Shigella sonnei isolates from an outbreak in Thimphu, Bhutan.

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    Ruekit, Sirigade; Wangchuk, Sonam; Dorji, Tshering; Tshering, Kinzang Pem; Pootong, Piyarat; Nobthai, Panida; Serichantalergs, Oralak; Poramathikul, Kamonporn; Bodhidatta, Ladaporn; Mason, Carl Jeffries

    2014-02-20

    Shigella species are an important cause of diarrhea in developing countries. These bacteria normally acquire their antibiotic resistance via several different mobile genetic elements including plasmids, transposons, and integrons involving gene cassettes. During a diarrhea surveillance study in Thimphu, Bhutan in June and July, 2011, Shigella sonnei were isolated more frequently than expected. This study describes the antibiotic resistance of these S. sonnei isolates. A total of 29 S. sonnei isolates from Thimphu, Bhutan was characterized for antimicrobial susceptibility by disc diffusion assay and minimum inhibitory concentration (MIC) assay. All isolates were tested by pulsed-field gel electrophoresis (PFGE) with restriction enzyme XbaI and were tested for plasmid. The plasmid patterns and the PFGE patterns were analyzed by Bionumerics software. DNA sequencing was performed on amplified products for gyraseA gene and class 1 and class 2 integrons. S. sonnei isolates were classified for incompatibility of plasmids by PCR-based replicon typing (PBRT). These S. sonnei were resistant to multiple drugs like ciprofloxacin, nalidixic acid, trimethoprim-sulfamethoxazole, streptomycin, and tetracycline but susceptible to azithromycin. All isolates had class 2 integrons dfrA1, sat1 and aadA1 genes. Two point mutations in Gyrase A subunit at position Ser83Leu and Asp87Gly were detected in these quinolone resistant isolates. The plasmid and PFGE patterns of S. sonnei isolates suggested a clonal relationship of the isolates. All isolates carried common ColE plasmid. ColE plasmid co-resided with B/O plasmid (nine isolates) or I1 plasmid (one isolate). The characteristics of 29 S. sonnei isolates from Thimphu, Bhutan in June and July, 2011 are identical in PFGE, plasmid and resistance pattern. This study suggests that these recent S. sonnei isolates are clonally related and multidrug-resistant.

  13. An oral Sindbis virus replicon-based DNA vaccine containing VP2 gene of canine parvovirus delivered by Escherichia coli elicits immune responses in dogs.

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    Dahiya, S S; Saini, M; Kumar, P; Gupta, P K

    2011-01-01

    A Sindbis virus replicon-based DNA vaccine containing VP2 gene of canine parvovirus (CPV) was delivered by Escherichia coli to elicit immune responses. The orally immunized dogs developed CPV-specific serum IgG and virus neutralizing antibody responses. The cellular immune responses analyzed using lymphocyte proliferation test and flow cytometry indicated CPV-specific sensitization of both CD3+CD4+ and CD3+CD8+ lymphocytes. This study demonstrated that the oral CPV DNA vaccine delivered by E. coli can be considered as a promising approach for vaccination of dogs against CPV.

  14. Cytomegalovirus replicon-based regulation of gene expression in vitro and in vivo.

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    Hermine Mohr

    Full Text Available There is increasing evidence for a connection between DNA replication and the expression of adjacent genes. Therefore, this study addressed the question of whether a herpesvirus origin of replication can be used to activate or increase the expression of adjacent genes. Cell lines carrying an episomal vector, in which reporter genes are linked to the murine cytomegalovirus (MCMV origin of lytic replication (oriLyt, were constructed. Reporter gene expression was silenced by a histone-deacetylase-dependent mechanism, but was resolved upon lytic infection with MCMV. Replication of the episome was observed subsequent to infection, leading to the induction of gene expression by more than 1000-fold. oriLyt-based regulation thus provided a unique opportunity for virus-induced conditional gene expression without the need for an additional induction mechanism. This principle was exploited to show effective late trans-complementation of the toxic viral protein M50 and the glycoprotein gO of MCMV. Moreover, the application of this principle for intracellular immunization against herpesvirus infection was demonstrated. The results of the present study show that viral infection specifically activated the expression of a dominant-negative transgene, which inhibited viral growth. This conditional system was operative in explant cultures of transgenic mice, but not in vivo. Several applications are discussed.

  15. RNA Replicons - A New Approach for Influenza Virus Immunoprophylaxis

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    Gert Zimmer

    2010-01-01

    Full Text Available RNA replicons are derived from either positive- or negative-strand RNA viruses. They represent disabled virus vectors that are not only avirulent, but also unable to revert to virulence. Due to autonomous RNA replication, RNA replicons are able to drive high level, cytosolic expression of recombinant antigens stimulating both the humoral and the cellular branch of the immune system. This review provides an update on the available literature covering influenza virus vaccines based on RNA replicons. The pros and cons of these vaccine strategies will be discussed and future perspectives disclosed.

  16. A vesicular stomatitis virus replicon-based bioassay for the rapid and sensitive determination of multi-species type I interferon.

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    Marianne Berger Rentsch

    Full Text Available Type I interferons (IFN comprise a family of cytokines that signal through a common cellular receptor to induce a plethora of genes with antiviral and other activities. Recombinant IFNs are used for the treatment of hepatitis C virus infection, multiple sclerosis, and certain malignancies. The capability of type I IFN to suppress virus replication and resultant cytopathic effects is frequently used to measure their bioactivity. However, these assays are time-consuming and require appropriate biosafety containment. In this study, an improved IFN assay is presented which is based on a recombinant vesicular stomatitis virus (VSV replicon encoding two reporter proteins, firefly luciferase and green fluorescent protein. The vector lacks the essential envelope glycoprotein (G gene of VSV and is propagated on a G protein-expressing transgenic cell line. Several mammalian and avian cells turned out to be susceptible to infection with the complemented replicon particles. Infected cells readily expressed the reporter proteins at high levels five hours post infection. When human fibroblasts were treated with serial dilutions of human IFN-β prior to infection, reporter expression was accordingly suppressed. This method was more sensitive and faster than a classical IFN bioassay based on VSV cytopathic effects. In addition, the antiviral activity of human IFN-λ (interleukin-29, a type III IFN, was determined on Calu-3 cells. Both IFN-β and IFN-λ were acid-stable, but only IFN-β was resistant to alkaline treatment. The antiviral activities of canine, porcine, and avian type I IFN were analysed with cell lines derived from the corresponding species. This safe bioassay will be useful for the rapid and sensitive quantification of multi-species type I IFN and potentially other antiviral cytokines.

  17. Chromosomal replicons of higher plants

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    Van' t Hof, J.

    1987-03-16

    This brief discussion of replicons of higher plants offers a glimpse into the properties of chromosomal DNA replication. It gives evidence that the S phase of unrelated plant species is comprised of temporally ordered replicon families that increase in number with genome size. This orderly process, which assures a normal inheritance of genetic material to recipient daughter cells, is maintained at the level of replicon clusters by two mutually exclusive mechanisms, one involving the rate at which single replicons replicate their allotment of DNA, and another by means of the tempo-pause. The same two mechanisms are used by cells to alter the pattern of chromosomal DNA replication just prior to and during normal development. Both mechanisms are genetically determined and produce genetic effects when disturbed of disrupted by additional non-conforming DNAs. Further insight into how these two mechanisms operate requires more molecular information about the nature of replicons and the factors that govern when a replicon family replicates. Plant material is a rich and ideal source for this information just awaiting exploitation. 63 refs.

  18. Alphavirus replicon-based enhancement of mucosal and systemic immunity is linked to the innate response generated by primary immunization.

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    Tonkin, Daniel R; Jorquera, Patricia; Todd, Tracie; Beard, Clayton W; Johnston, Robert E; Barro, Mario

    2010-04-19

    Venezuelan equine encephalitis virus replicon particles (VRP) function as an effective systemic, cellular and mucosal adjuvant when codelivered with antigen, and show promise for use as a component in new and existing human vaccine formulations. We show here that VRP are effective at low dose and by intramuscular delivery, two useful features for implementation of VRP as a vaccine adjuvant. In mice receiving a prime and boost with antigen, we found that VRP are required in prime only to produce a full adjuvant effect. This outcome indicates that the events triggered during prime with VRP are sufficient to establish the nature and magnitude of the immune response to a second exposure to antigen. Events induced by VRP in the draining lymph node after prime include robust secretion of many inflammatory cytokines, upregulation of CD69 on leukocytes, and increased cellularity, with a disproportionate increase of a cell population expressing CD11c, CD11b, and F4/80. We show that antigen delivered 24h after administration of VRP does not benefit from an adjuvant effect, indicating that the events which are critical to VRP-mediated adjuvant activity occur within the first 24h. Further studies of the events induced by VRP will help elucidate the mechanism of VRP adjuvant activity and will advance the safe implementation of this adjuvant in human vaccines. Copyright 2010 Elsevier Ltd. All rights reserved.

  19. Transient Expression of Lumbrokinase (PI239 in Tobacco (Nicotiana tabacum Using a Geminivirus-Based Single Replicon System Dissolves Fibrin and Blood Clots

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    Alexia Dickey

    2017-01-01

    Full Text Available Lumbrokinases, a group of fibrinolytic enzymes extracted from earthworm, have been widely used to prevent and treat various cardiovascular diseases. They specifically target fibrin to effectively degrade thrombi without major side effects. Plant expression systems are becoming potential alternative expression platforms for producing pharmaceutical proteins. In this work, a lumbrokinase (PI239 was produced from a plant system. Both wild-type (WT and plant codon-optimized (OP PI239 gene sequences were synthesized and cloned into a geminivirus-based single-vector DNA replicon system. Both vectors were independently expressed in tobacco (Nicotiana tabacum leaves transiently by agroinfiltration. Overexpressed PI239 resulted in sudden tissue necrosis 3 days after infiltration. Remaining proteins were purified through His-tag affinity chromatography and analyzed with SDS-PAGE and Western blot methods. Purified PI239 successfully degraded artificial fibrin with relative activity of 13,400 U/mg when compared with commercial lumbrokinase product. In vitro tests demonstrated that plant-derived PI239 dissolved human blood clots and that the plant expression system is capable of producing functional PI239.

  20. Classical Swine Fever Virus-Rluc Replicons

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    Risager, Peter Christian; Belsham, Graham J.; Rasmussen, Thomas Bruun

    Classical swine fever virus (CSFV) is the etiologic agent of the severe porcine disease, classical swine fever. Unraveling the molecular determinants of efficient replication is crucial for gaining proper knowledge of the pathogenic traits of this virus. Monitoring the replication competence with...... type CSFV-Paderborn strain, as judged by qRT-PCR, was picked as the candidate and could potentially be useful as a tool for further downstream applications including investigation of CSFV non-structural proteins involvement in viral replication....... cells can be achieved using autonomously replicating genome constructs (replicons) containing a reporter gene that expresses a readily quantifiable enzyme. Here, a newly implemented cloning technique was applied to genome modification of the full-length CSFV cDNA previously inserted into a single......-copy bacterial artificial chromosome (BAC). This technique, the Red/ET counter-selection method, is based upon homologous recombination, thus obviating the need for internal restriction sites. Several CSFV replicons with deletions in regions encoding structural viral proteins considered non-essential for RNA...

  1. SCH 503034, a mechanism-based inhibitor of hepatitis C virus NS3 protease, suppresses polyprotein maturation and enhances the antiviral activity of alpha interferon in replicon cells.

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    Malcolm, B A; Liu, R; Lahser, F; Agrawal, S; Belanger, B; Butkiewicz, N; Chase, R; Gheyas, F; Hart, A; Hesk, D; Ingravallo, P; Jiang, C; Kong, R; Lu, J; Pichardo, J; Prongay, A; Skelton, A; Tong, X; Venkatraman, S; Xia, E; Girijavallabhan, V; Njoroge, F G

    2006-03-01

    Cleavage of the hepatitis C virus (HCV) polyprotein by the viral NS3 protease releases functional viral proteins essential for viral replication. Recent studies by Foy and coworkers strongly suggest that NS3-mediated cleavage of host factors may abrogate cellular response to alpha interferon (IFN-alpha) (E. Foy, K. Li, R. Sumpter, Jr., Y.-M. Loo, C. L. Johnson, C. Wang, P. M. Fish, M. Yoneyama, T. Fujita, S. M. Lemon, and M. Gale, Jr., Proc. Natl. Acad. Sci. USA 102:2986-2991, 2005, and E. Foy, K. Li, C. Wang, R. Sumpter, Jr., M. Ikeda, S. M. Lemon, and M. Gale, Jr., Science 300:1145-1148, 2003). Blockage of NS3 protease activity therefore is expected to inhibit HCV replication by both direct suppression of viral protein production as well as by restoring host responsiveness to IFN. Using structure-assisted design, a ketoamide inhibitor, SCH 503034, was generated which demonstrated potent (overall inhibition constant, 14 nM) time-dependent inhibition of the NS3 protease in cell-free enzyme assays as well as robust in vitro activity in the HCV replicon system, as monitored by immunofluorescence and real-time PCR analysis. Continuous exposure of replicon-bearing cell lines to six times the 90% effective concentration of SCH 503034 for 15 days resulted in a greater than 4-log reduction in replicon RNA. The combination of SCH 503034 with IFN was more effective in suppressing replicon synthesis than either compound alone, supporting the suggestion of Foy and coworkers that combinations of IFN with protease inhibitors would lead to enhanced therapeutic efficacy.

  2. Veterinary Replicon Vaccines

    NARCIS (Netherlands)

    Hikke, Mia C.; Pijlman, Gorben P.

    2017-01-01

    Vaccination is essential in livestock farming and in companion animal ownership. Nucleic acid vaccines based on DNA or RNA provide an elegant alternative to those classical veterinary vaccines that have performed suboptimally. Recent advances in terms of rational design, safety, and efficacy have

  3. DNA Replicons for Plant Genome Engineering[W][OPEN

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    Baltes, Nicholas J.; Gil-Humanes, Javier; Cermak, Tomas; Atkins, Paul A.; Voytas, Daniel F.

    2014-01-01

    Sequence-specific nucleases enable facile editing of higher eukaryotic genomic DNA; however, targeted modification of plant genomes remains challenging due to ineffective methods for delivering reagents for genome engineering to plant cells. Here, we use geminivirus-based replicons for transient expression of sequence-specific nucleases (zinc-finger nucleases, transcription activator–like effector nucleases, and the clustered, regularly interspaced, short palindromic repeat/Cas system) and delivery of DNA repair templates. In tobacco (Nicotiana tabacum), replicons based on the bean yellow dwarf virus enhanced gene targeting frequencies one to two orders of magnitude over conventional Agrobacterium tumefaciens T-DNA. In addition to the nuclease-mediated DNA double-strand breaks, gene targeting was promoted by replication of the repair template and pleiotropic activity of the geminivirus replication initiator proteins. We demonstrate the feasibility of using geminivirus replicons to generate plants with a desired DNA sequence modification. By adopting a general plant transformation method, plantlets with a desired DNA change were regenerated in genome engineering. PMID:24443519

  4. High-efficiency gene targeting in hexaploid wheat using DNA replicons and CRISPR/Cas9.

    Science.gov (United States)

    Gil-Humanes, Javier; Wang, Yanpeng; Liang, Zhen; Shan, Qiwei; Ozuna, Carmen V; Sánchez-León, Susana; Baltes, Nicholas J; Starker, Colby; Barro, Francisco; Gao, Caixia; Voytas, Daniel F

    2017-03-01

    The ability to edit plant genomes through gene targeting (GT) requires efficient methods to deliver both sequence-specific nucleases (SSNs) and repair templates to plant cells. This is typically achieved using Agrobacterium T-DNA, biolistics or by stably integrating nuclease-encoding cassettes and repair templates into the plant genome. In dicotyledonous plants, such as Nicotinana tabacum (tobacco) and Solanum lycopersicum (tomato), greater than 10-fold enhancements in GT frequencies have been achieved using DNA virus-based replicons. These replicons transiently amplify to high copy numbers in plant cells to deliver abundant SSNs and repair templates to achieve targeted gene modification. In the present work, we developed a replicon-based system for genome engineering of cereal crops using a deconstructed version of the wheat dwarf virus (WDV). In wheat cells, the replicons achieve a 110-fold increase in expression of a reporter gene relative to non-replicating controls. Furthermore, replicons carrying CRISPR/Cas9 nucleases and repair templates achieved GT at an endogenous ubiquitin locus at frequencies 12-fold greater than non-viral delivery methods. The use of a strong promoter to express Cas9 was critical to attain these high GT frequencies. We also demonstrate gene-targeted integration by homologous recombination (HR) in all three of the homoeoalleles (A, B and D) of the hexaploid wheat genome, and we show that with the WDV replicons, multiplexed GT within the same wheat cell can be achieved at frequencies of ~1%. In conclusion, high frequencies of GT using WDV-based DNA replicons will make it possible to edit complex cereal genomes without the need to integrate GT reagents into the genome. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  5. Characterization of cell lines stably transfected with rubella virus replicons

    Energy Technology Data Exchange (ETDEWEB)

    Tzeng, Wen-Pin; Xu, Jie [Department of Biology, Georgia State University, P.O. Box 4010, Atlanta GA 30302-4010 (United States); Frey, Teryl K., E-mail: tfrey@gsu.edu [Department of Biology, Georgia State University, P.O. Box 4010, Atlanta GA 30302-4010 (United States)

    2012-07-20

    Rubella virus (RUBV) replicons expressing a drug resistance gene and a gene of interest were used to select cell lines uniformly harboring the replicon. Replicons expressing GFP and a virus capsid protein GFP fusion (C-GFP) were compared. Vero or BHK cells transfected with either replicon survived drug selection and grew into a monolayer. However, survival was {approx}9-fold greater following transfection with the C-GFP-replicon than with the GFP-expressing replicon and while the C-GFP-replicon cells grew similarly to non-transfected cells, the GFP-replicon cells grew slower. Neither was due to the ability of the CP to enhance RNA synthesis but survival during drug selection was correlated with the ability of CP to inhibit apoptosis. Additionally, C-GFP-replicon cells were not cured of the replicon in the absence of drug selection. Interferon-alpha suppressed replicon RNA and protein synthesis, but did not cure the cells, explaining in part the ability of RUBV to establish persistent infections.

  6. In vitro and in vivo characterization of microRNA-targeted alphavirus replicon and helper RNAs.

    Science.gov (United States)

    Kamrud, Kurt I; Coffield, V McNeil; Owens, Gary; Goodman, Christin; Alterson, Kim; Custer, Max; Murphy, Michael A; Lewis, Whitney; Timberlake, Sarah; Wansley, Elizabeth K; Berglund, Peter; Smith, Jonathan

    2010-08-01

    Alphavirus-based replicon vector systems (family Togaviridae) have been developed as expression vectors with demonstrated potential in vaccine development against both infectious diseases and cancer. The single-cycle nature of virus-like replicon particles (VRP), generated by supplying the structural proteins from separate replicable helper RNAs, is an attractive safety component of these systems. MicroRNAs (miRNAs) have emerged as important cellular RNA regulation elements. Recently, miRNAs have been employed as a mechanism to attenuate or restrict cellular tropism of replication-competent viruses, such as oncolytic adenoviruses, vesicular stomatitis virus, and picornaviruses as well as nonreplicating lentiviral and adenoviral vectors. Here, we describe the incorporation of miRNA-specific target sequences into replicable alphavirus helper RNAs that are used in trans to provide the structural proteins required for VRP production. VRP were found to be efficiently produced using miRNA-targeted helper RNAs if miRNA-specific inhibitors were introduced into cells during VRP production. In the absence of such inhibitors, cellular miRNAs were capable of downregulating helper RNA replication in vitro. When miRNA targets were incorporated into a replicon RNA, cellular miRNAs were capable of downregulating replicon RNA replication upon delivery of VRP into animals, demonstrating activity in vivo. These data provide the first example of miRNA-specific repression of alphavirus replicon and helper RNA replication and demonstrate the feasibility of miRNA targeting of expression vector helper functions that are provided in trans.

  7. PCR-based plasmid typing in Enterococcus faecium strains reveals widely distributed pRE25-, pRUM-, pIP501-and pHT beta-related replicons associated with glycopeptide resistance and stabilizing toxin-antitoxin systems

    DEFF Research Database (Denmark)

    Rosvoll, T.C.S.; Pedersen, T.; Sletvold, H.

    2010-01-01

    A PCR-based typing scheme was applied to identify plasmids in an epidemiologically and geographically diverse strain collection of Enterococcus faecium (n=93). Replicon types of pRE25 (n=56), pRUM (n=41), pIP501 (n=17) and pHT beta (n=14) were observed in 83% of the strains, while pS86, pCF10, p...

  8. Recombination-ready Sindbis replicon expression vectors for transgene expression

    Directory of Open Access Journals (Sweden)

    Olson Ken E

    2007-10-01

    Full Text Available Abstract Background Sindbis viruses have been widely used as tools to study gene function in cells. Despite the utility of these systems, the construction and production of alphavirus replicons is time consuming and inefficient due to potential additional restriction sites within the insert region and lack of directionality for insert ligation. In this report, we present a system useful for producing recombinant Sindbis replicons that uses lambda phage recombination technology to rapidly and specifically construct replicon expression plasmids that contain insert regions in the desired orientation. Results Recombination of the gene of interest with the replicon plasmid resulted in nearly 100% recombinants, each of which contained a correctly orientated insert. Replicons were easily produced in cell culture and packaged into pseudo-infectious viral particles. Insect and mammalian cells infected with pseudo-infectious viral particles expressed various transgenes at high levels. Finally, inserts from persistently replicating replicon RNA were easily isolated and recombined back into entry plasmids for sequencing and subsequent analysis. Conclusion Replication-ready replicon expression plasmids make the use of alphavirus replicons fast and easy as compared to traditional replicon production methods. This system represents a significant step forward in the utility and ease of use of alphavirus replicons in the study of gene function.

  9. Monitoring the determinants of efficient viral replication using Classical Swine Fever Virus-reporter replicons

    DEFF Research Database (Denmark)

    Risager, Peter Christian; Everett, Helen; Crooke, Helen

    2012-01-01

    of the CSFV genome within cells can be achieved using autonomously replicating constructs (replicons) containing a reporter gene that expresses a readily quantifiable enzyme. Here, a newly implemented cloning technique was applied to genome modification of the fulllength CSFV cDNA previously inserted...... into a single-copy bacterial artificial chromosome (BAC). This technique, the Red/ET counter-selection method, is based upon homologous recombination, thus obviating the need for internal restriction sites or complex cloning strategies. Several CSFV replicons with deletions in regions encoding virus structural...... proteins considered non-essential for RNA replication were constructed and these deletions were replaced with an in-frame insertion of the Renilla luciferase (Rluc) sequence. RNA transcripts from these replicons should be translated as a single functional open reading frame. Full-genome cDNAs (~10-12,3 kb...

  10. Replicon RNA Viral Vectors as Vaccines

    Directory of Open Access Journals (Sweden)

    Kenneth Lundstrom

    2016-11-01

    Full Text Available Single-stranded RNA viruses of both positive and negative polarity have been used as vectors for vaccine development. In this context, alphaviruses, flaviviruses, measles virus and rhabdoviruses have been engineered for expression of surface protein genes and antigens. Administration of replicon RNA vectors has resulted in strong immune responses and generation of neutralizing antibodies in various animal models. Immunization of mice, chicken, pigs and primates with virus-like particles, naked RNA or layered DNA/RNA plasmids has provided protection against challenges with lethal doses of infectious agents and administered tumor cells. Both prophylactic and therapeutic efficacy has been achieved in cancer immunotherapy. Moreover, recombinant particles and replicon RNAs have been encapsulated by liposomes to improve delivery and targeting. Replicon RNA vectors have also been subjected to clinical trials. Overall, immunization with self-replicating RNA viruses provides high transient expression levels of antigens resulting in generation of neutralizing antibody responses and protection against lethal challenges under safe conditions.

  11. Replicon typing of plasmids encoding resistance to newer beta-lactams.

    Science.gov (United States)

    Carattoli, Alessandra; Miriagou, Vivi; Bertini, Alessia; Loli, Alexandra; Colinon, Celine; Villa, Laura; Whichard, Jean M; Rossolini, Gian Maria

    2006-07-01

    Polymerase chain reaction-based replicon typing represents a novel method to describe the dissemination and follow the evolution of resistance plasmids. We used this approach to study 26 epidemiologically unrelated Enterobacteriaceae and demonstrate the dominance of incompatibility (Inc) A/C or Inc N-related plasmids carrying some emerging resistance determinants to extended-spectrum cephalosporins and carbapenems.

  12. Replicon Typing of Plasmids Encoding Resistance to Newer β-Lactams

    Science.gov (United States)

    Miriagou, Vivi; Bertini, Alessia; Loli, Alexandra; Colinon, Celine; Villa, Laura; Whichard, Jean M.; Rossolini, Gian Maria

    2006-01-01

    Polymerase chain reaction–based replicon typing represents a novel method to describe the dissemination and follow the evolution of resistance plasmids. We used this approach to study 26 epidemiologically unrelated Enterobacteriaceae and demonstrate the dominance of incompatibility (Inc) A/C or Inc N-related plasmids carrying some emerging resistance determinants to extended-spectrum cephalosporins and carbapenems. PMID:16836838

  13. Transfer of tetracycline resistance gene (tetr) between replicons in ...

    African Journals Online (AJOL)

    Transfer of tetracycline resistance gene (tetr) between replicons in some enteric bacteria of diarrhoeal origin from some hospitals in South-South, Nigeria. ... Attempt was made to transfer the tetr gene from one replicon to the other within the same species and from one genus to the other. The rate of intra-species transfer of ...

  14. Identification, Characterization, and Application of the Replicon Region of the Halophilic Temperate Sphaerolipovirus SNJ1.

    Science.gov (United States)

    Wang, Yuchen; Sima, Linshan; Lv, Jie; Huang, Suiyuan; Liu, Ying; Wang, Jiao; Krupovic, Mart; Chen, Xiangdong

    2016-07-15

    The temperate haloarchaeal virus SNJ1 displays lytic and lysogenic life cycles. During the lysogenic cycle, the virus resides in its host, Natrinema sp. strain J7-1, in the form of an extrachromosomal circular plasmid, pHH205. In this study, a 3.9-kb region containing seven predicted genes organized in two operons was identified as the minimal replicon of SNJ1. Only RepA, encoded by open reading frame 11-12 (ORF11-12), was found to be essential for replication, and its expression increased during the lytic cycle. Sequence analysis suggested that RepA is a distant homolog of HUH endonucleases, a superfamily that includes rolling-circle replication initiation proteins from various viruses and plasmids. In addition to RepA, two genetic elements located within both termini of the 3.9-kb replicon were also required for SNJ1 replication. SNJ1 genome and SNJ1 replicon-based shuttle vectors were present at 1 to 3 copies per chromosome. However, the deletion of ORF4 significantly increased the SNJ1 copy number, suggesting that the product of ORF4 is a negative regulator of SNJ1 abundance. Shuttle vectors based on the SNJ1 replicon were constructed and validated for stable expression of heterologous proteins, both in J7 derivatives and in Natrinema pallidum JCM 8980(T), suggesting their broad applicability as genetic tools for Natrinema species. Archaeal viruses exhibit striking morphological diversity and unique gene content. In this study, the minimal replicon of the temperate haloarchaeal virus SNJ1 was identified. A number of ORFs and genetic elements controlling virus genome replication, maintenance, and copy number were characterized. In addition, based on the replicon, a novel expression shuttle vector has been constructed and validated for protein expression and purification in Natrinema sp. CJ7 and Natrinema pallidum JCM 8980(T) This study not only provided mechanistic and functional insights into SNJ1 replication but also led to the development of useful genetic

  15. Prolonged transgene expression in glomeruli using an EBV replicon vector system combined with HVJ liposomes.

    Science.gov (United States)

    Tsujie, M; Isaka, Y; Nakamura, H; Kaneda, Y; Imai, E; Hori, M

    2001-04-01

    Various gene transfer vectors as well as delivery systems have been developed; however, many problems remain to be solved. We already achieved a technique to introduce genes into glomerular mesangial cells by hemagglutinating virus of Japan (HVJ) liposome-mediated gene transfer via renal artery. The main limitation of this method is the transient transgene expression. For long-term gene expression in glomeruli, Epstein-Barr virus (EBV) replicon-based plasmid was employed, containing the latent viral DNA replication origin (oriP) and EBV nuclear antigen-1 (EBNA-1), which are the minimum EBV component of transgene-nuclear retention. To examine the effect of EBV replicon apparatus on the duration of transgene expression in glomeruli in vivo, the EBV replicon vector pEBActLuc, and the control plasmid vector pActLuc were adopted. These plasmid vectors were transferred into the kidney via renal artery by using artificial viral envelope (AVE)-type HVJ liposome method, and glomerular luciferase activities were analyzed at various time points after transfection. On day 4, pEBActLuc and pActLuc transfer resulted in equal glomerular luciferase activity, and the luciferase gene expression was sustained for at least 56 days in glomeruli transfected with pEBActLuc, whereas it was reduced on seven days in glomeruli transfected with pActLuc. The combination of EBV replicon apparatus and HVJ liposomes appears to be a powerful tool for long-term gene expression in vivo, and furthermore, it may be a promising new therapeutic method for the progression of renal disease.

  16. A Polyprotein-Expressing Salmonid Alphavirus Replicon Induces Modest Protection in Atlantic Salmon (Salmo Salar Against Infectious Pancreatic Necrosis

    Directory of Open Access Journals (Sweden)

    Azila Abdullah

    2015-01-01

    Full Text Available Vaccination is an important strategy for the control and prevention of infectious pancreatic necrosis (IPN in farmed Atlantic salmon (Salmo salar in the post-smolt stage in sea-water. In this study, a heterologous gene expression system, based on a replicon construct of salmonid alphavirus (SAV, was used for in vitro and in vivo expression of IPN virus proteins. The large open reading frame of segment A, encoding the polyprotein NH2-pVP2-VP4-VP3-COOH, as well as pVP2, were cloned and expressed by the SAV replicon in Chinook salmon embryo cells (CHSE-214 and epithelioma papulosum cyprini (EPC cells. The replicon constructs pSAV/polyprotein (pSAV/PP and pSAV/pVP2 were used to immunize Atlantic salmon (Salmo salar by a single intramuscular injection and tested in a subsequent IPN virus (IPNV challenge trial. A low to moderate protection against IPN was observed in fish immunized with the replicon vaccine that encoded the pSAV/PP, while the pSAV/pVP2 construct was not found to induce protection.

  17. Development and evaluation of a replicon particle vaccine expressing the E2 glycoprotein of bovine viral diarrhea virus (BVDV in cattle

    Directory of Open Access Journals (Sweden)

    Loy John Dustin

    2013-01-01

    Full Text Available Abstract Background Bovine viral diarrhea virus is one of the most significant and costly viral pathogens of cattle worldwide. Alphavirus-derived replicon particles have been shown to be safe and highly effective vaccine vectors against a variety of human and veterinary pathogens. Replicon particles are non-propagating, DIVA compatible, and can induce both humoral and cell mediated immune responses. This is the first experiment to demonstrate that Alphavirus-based replicon particles can be utilized in a standard prime/boost vaccination strategy in calves against a commercially significant bovine pathogen. Findings Replicon particles that express bovine viral diarrhea virus sub-genotype 1b E2 glycoprotein were generated and expression was confirmed in vitro using polyclonal and monoclonal antibodies specific to E2. Vaccine made from particles was generated in Vero cells and administered to BVDV free calves in a prime/boost regimen at two dosage levels. Vaccination resulted in neutralizing antibody titers that cross-neutralized both type 1 and type 2 BVD genotypes following booster vaccination. Additionally, high dose vaccine administration demonstrated some protection from clinical disease and significantly reduced the degree of leukopenia caused by viral infection. Conclusions Replicon particle vaccines administered in a prime/boost regimen expressing BVDV E2 glycoprotein can induce cross-neutralizing titers, reduce leukopenia post challenge, and mitigate clinical disease in calves. This strategy holds promise for a safe and effective vaccine to BVDV.

  18. Analysis of Venezuelan equine encephalitis replicon particles packaged in different coats.

    Science.gov (United States)

    Kamrud, Kurt I; Alterson, Kim D; Andrews, Chasity; Copp, Laura O; Lewis, Whitney C; Hubby, Bolyn; Patel, Deepa; Rayner, Jonathan O; Talarico, Todd; Smith, Jonathan F

    2008-07-16

    The Venezuelan equine encephalitis (VEE) virus replicon system was used to produce virus-like replicon particles (VRP) packaged with a number of different VEE-derived glycoprotein (GP) coats. The GP coat is believed to be responsible for the cellular tropism noted for VRP and it is possible that different VEE GP coats may have different affinities for cells. We examined VRP packaged in four different VEE GP coats for their ability to infect cells in vitro and to induce both humoral and cellular immune responses in vivo. The VRP preparations were characterized to determine both infectious units (IU) and genome equivalents (GE) prior to in vivo analysis. VRP packaged with different VEE GP coats demonstrated widely varying GE/IU ratios based on Vero cell infectivity. BALB/c mice were immunized with the different VRP based on equal GE titers and the humoral and cellular responses to the expressed HIV gag gene measured. The magnitude of the immune responses measured in mice revealed small but significant differences between different GP coats when immunization was based on GE titers. We suggest that care should be taken when alternative coat proteins are used to package vector-based systems as the titers determined by cell culture infection may not represent accurate particle numbers and in turn may not accurately represent actual in vivo dose.

  19. Analysis of Venezuelan equine encephalitis replicon particles packaged in different coats.

    Directory of Open Access Journals (Sweden)

    Kurt I Kamrud

    2008-07-01

    Full Text Available The Venezuelan equine encephalitis (VEE virus replicon system was used to produce virus-like replicon particles (VRP packaged with a number of different VEE-derived glycoprotein (GP coats. The GP coat is believed to be responsible for the cellular tropism noted for VRP and it is possible that different VEE GP coats may have different affinities for cells. We examined VRP packaged in four different VEE GP coats for their ability to infect cells in vitro and to induce both humoral and cellular immune responses in vivo.The VRP preparations were characterized to determine both infectious units (IU and genome equivalents (GE prior to in vivo analysis. VRP packaged with different VEE GP coats demonstrated widely varying GE/IU ratios based on Vero cell infectivity. BALB/c mice were immunized with the different VRP based on equal GE titers and the humoral and cellular responses to the expressed HIV gag gene measured. The magnitude of the immune responses measured in mice revealed small but significant differences between different GP coats when immunization was based on GE titers.We suggest that care should be taken when alternative coat proteins are used to package vector-based systems as the titers determined by cell culture infection may not represent accurate particle numbers and in turn may not accurately represent actual in vivo dose.

  20. Plasmid Replicons from Pseudomonas Are Natural Chimeras of Functional, Exchangeable Modules

    Science.gov (United States)

    Bardaji, Leire; Añorga, Maite; Ruiz-Masó, José A.; del Solar, Gloria; Murillo, Jesús

    2017-01-01

    Plasmids are a main factor for the evolution of bacteria through horizontal gene exchange, including the dissemination of pathogenicity genes, resistance to antibiotics and degradation of pollutants. Their capacity to duplicate is dependent on their replication determinants (replicon), which also define their bacterial host range and the inability to coexist with related replicons. We characterize a second replicon from the virulence plasmid pPsv48C, from Pseudomonas syringae pv. savastanoi, which appears to be a natural chimera between the gene encoding a newly described replication protein and a putative replication control region present in the widespread family of PFP virulence plasmids. We present extensive evidence of this type of chimerism in structurally similar replicons from species of Pseudomonas, including environmental bacteria as well as plant, animal and human pathogens. We establish that these replicons consist of two functional modules corresponding to putative control (REx-C module) and replication (REx-R module) regions. These modules are functionally separable, do not show specificity for each other, and are dynamically exchanged among replicons of four distinct plasmid families. Only the REx-C module displays strong incompatibility, which is overcome by a few nucleotide changes clustered in a stem-and-loop structure of a putative antisense RNA. Additionally, a REx-C module from pPsv48C conferred replication ability to a non-replicative chromosomal DNA region containing features associated to replicons. Thus, the organization of plasmid replicons as independent and exchangeable functional modules is likely facilitating rapid replicon evolution, fostering their diversification and survival, besides allowing the potential co-option of appropriate genes into novel replicons and the artificial construction of new replicon specificities. PMID:28243228

  1. The hepatitis C virus replicon presents a higher barrier to resistance to nucleoside analogs than to nonnucleoside polymerase or protease inhibitors.

    Science.gov (United States)

    McCown, Matthew F; Rajyaguru, Sonal; Le Pogam, Sophie; Ali, Samir; Jiang, Wen-Rong; Kang, Hyunsoon; Symons, Julian; Cammack, Nick; Najera, Isabel

    2008-05-01

    Specific inhibitors of hepatitis C virus (HCV) replication that target the NS3/4A protease (e.g., VX-950) or the NS5B polymerase (e.g., R1479/R1626, PSI-6130/R7128, NM107/NM283, and HCV-796) have advanced into clinical development. Treatment of patients with VX-950 or HCV-796 rapidly selected for drug-resistant variants after a 14-day monotherapy treatment period. However, no viral resistance was identified after monotherapy with R1626 (prodrug of R1479) or NM283 (prodrug of NM107) after 14 days of monotherapy. Based upon the rapid selection of resistance to the protease and nonnucleoside inhibitors during clinical trials and the lack of selection of resistance to the nucleoside inhibitors, we used the replicon system to determine whether nucleoside inhibitors demonstrate a higher genetic barrier to resistance than protease and nonnucleoside inhibitors. Treatment of replicon cells with nucleoside inhibitors at 10 and 15 times the 50% effective concentration resulted in clearance of the replicon, while treatment with a nonnucleoside or protease inhibitor selected resistant colonies. In combination, the presence of a nucleoside inhibitor reduced the frequency of colonies resistant to the other classes of inhibitors. These results indicate that the HCV replicon presents a higher barrier to the selection of resistance to nucleoside inhibitors than to nonnucleoside or protease inhibitors. Furthermore, the combination of a nonnucleoside or protease inhibitor with a nucleoside polymerase inhibitor could have a clear clinical benefit through the delay of resistance emergence.

  2. Characterization of pMC11, a plasmid with dual origins of replication isolated from Lactobacillus casei MCJ and construction of shuttle vectors with each replicon

    DEFF Research Database (Denmark)

    Chen, Zhengjun; Lin, Jinzhong; Ma, Chengjie

    2014-01-01

    %. These vectors were employed to express a green fluorescent protein (GFP) using the promoter of S-layer protein SlpA from Lactobacillus acidophilus. And a growth-phase regulated expression of GFP was observed in different strains. In conclusion, these shuttle vectors provide efficient genetic tools for DNA......Many lactic acid bacteria carry different plasmids, particularly those that replicate via a theta mechanism. Here we describe Lactobacillus casei MCJ(CCTCC AB20130356), a new isolate that contains pMC11, carrying two distinct theta-type replicons. Each replicon contained an iteron in the origin...... of replication (oriV1 or oriV2) and a gene coding for the replicase (RepA_1 or RepB_1), both of which are essential for plasmid replication. Escherichia coli/Lactobacillus shuttle vectors were constructed with each replicon, yielding pEL5.7 and pEL5.6 that are based on oriV2 and oriV1 replicons, respectively...

  3. Potentiation of anthrax vaccines using protective antigen-expressing viral replicon vectors.

    Science.gov (United States)

    Wang, Hai-Chao; An, Huai-Jie; Yu, Yun-Zhou; Xu, Qing

    2015-02-01

    DNA vaccines require improvement for human use because they are generally weak stimulators of the immune system in humans. The efficacy of DNA vaccines can be improved using a viral replicon as vector to administer antigen of pathogen. In this study, we comprehensively evaluated the conventional non-viral DNA, viral replicon DNA or viral replicon particles (VRP) vaccines encoding different forms of anthrax protective antigen (PA) for specific immunity and protective potency against anthrax. Our current results clearly suggested that these viral replicon DNA or VRP vaccines derived from Semliki Forest virus (SFV) induced stronger PA-specific immune responses than the conventional non-viral DNA vaccines when encoding the same antigen forms, which resulted in potent protection against challenge with the Bacillus anthracis strain A16R. Additionally, the naked PA-expressing SFV replicon DNA or VRP vaccines without the need for high doses or demanding particular delivery regimens elicited robust immune responses and afforded completely protective potencies, which indicated the potential of the SFV replicon as vector of anthrax vaccines for use in clinical application. Therefore, our results suggest that these PA-expressing SFV replicon DNA or VRP vaccines may be suitable as candidate vaccines against anthrax. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Engineering a CTL-Tailored Replicon RNA Vaccine against PRRSV

    DEFF Research Database (Denmark)

    Welner, Simon; Werder, Simea; Nielsen, Morten

    The development of vaccines against porcine reproductive and respiratory syndrome virus (PRRSV) has been hampered by the high mutation rate and the multiple immunoevasive strategies of the virus. With the overall aim of designing a broad coverage vaccine that induces an effective CTL response...... detection in the presence of a proteasome inhibitor. Finally, a vaccination-challenge experiment using 18 SLA-matched pigs is currently being conducted until July 2016 in which a test group and a control group are being vaccinated twice with VRPs expressing PRRSV epitopes and non-sense control epitopes...... will be available for IVIS. This study exemplifies how bioinformatics epitope prediction, recombinant SLA molecules and RNA virus replicon design can be used to engineer a replicating non-propagating vaccine tailored to deliver conserved and immunogenic CTL epitopes....

  5. Co-delivery of antigen and IL-12 by Venezuelan equine encephalitis virus replicon particles enhances antigen-specific immune responses and anti-tumor effects

    OpenAIRE

    Osada, Takuya; Berglund, Peter; Morse, Michael A.; Hubby, Bolyn; Lewis, Whitney; Niedzwiecki, Donna; Hobeika, Amy; Burnett, Bruce; Devi, Gayathri R.; Clay, Timothy M.; Smith, Jonathan; Lyerly, H. Kim

    2012-01-01

    We recently demonstrated that Venezuelan equine encephalitis (VEE) virus-based replicon particles (VRP) encoding tumor antigens could break tolerance in the immunomodulatory environment of advanced cancer. We hypothesized that local injection of VRP expressing Interleukin-12 (IL-12) at the site of injections of VRP-based cancer vaccines would enhance the tumor-antigen-specific T cell and antibody responses and anti-tumor efficacy. Mice were immunized with VRP encoding the human tumor-associat...

  6. Replicon typing of plasmids carrying blaCTX-M-1 in Enterobacteriaceae of animal, environmental and human origin

    Directory of Open Access Journals (Sweden)

    Katrin eZurfluh

    2014-10-01

    Full Text Available Objectives: The aim of this work was to determine the plasmid replicon profiles of a collection of blaCTX-M-1-positive enterobacterial strains. The isolates originated from chicken in the production pyramid, healthy food-producing animals at slaughter (chicken, calves and pigs, chicken retail meat, environmental isolates originating from water bodies, and isolates from humans. A selection of IncI and IncN plasmids were characterized by multilocus sequence typing in order to determine their epidemiological relatedness. Methods: Transconjugants of 74 blaCTX-M-1-positive isolates were analysed by PCR-based replicon typing and by PCR-based plasmid multilocus sequence typing.Results: The incompatibility groups detected among the blaCTX-M-1-harboring plasmids included IncI1, IncN, IncHI1B, IncF, IncFIIS, IncFIB and IncB/O, with plasmid lineage IncI1/ST3 predominating in isolates from chicken and from humans. Lineage IncN/ST1 was detected mainly in isolates from pigs. For the first time, blaCTX-M-1 genes encoded on IncHI1 plasmids were detected in isolates from cattle and from water bodies.Conclusions: This study identifies plasmid lineages that are contributing to the dissemination of blaCTX-M-1 genes in the food chain, the environment, and humans.

  7. A personal reflection on the replicon theory: from R1 plasmid to replication timing regulation in human cells.

    Science.gov (United States)

    Masai, Hisao

    2013-11-29

    Fifty years after the Replicon Theory was originally presented, detailed mechanistic insight into prokaryotic replicons has been obtained and rapid progress is being made to elucidate the more complex regulatory mechanisms of replicon regulation in eukaryotic cells. Here, I present my personal perspectives on how studies of model replicons have contributed to our understanding of the basic mechanisms of DNA replication as well as the evolution of replication regulation in human cells. I will also discuss how replication regulation contributes to the stable maintenance of the genome and how disruption of replication regulation leads to human diseases. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Venezuelan equine encephalitis virus replicon particle vaccine protects nonhuman primates from intramuscular and aerosol challenge with ebolavirus.

    Science.gov (United States)

    Herbert, Andrew S; Kuehne, Ana I; Barth, James F; Ortiz, Ramon A; Nichols, Donald K; Zak, Samantha E; Stonier, Spencer W; Muhammad, Majidat A; Bakken, Russell R; Prugar, Laura I; Olinger, Gene G; Groebner, Jennifer L; Lee, John S; Pratt, William D; Custer, Max; Kamrud, Kurt I; Smith, Jonathan F; Hart, Mary Kate; Dye, John M

    2013-05-01

    There are no vaccines or therapeutics currently approved for the prevention or treatment of ebolavirus infection. Previously, a replicon vaccine based on Venezuelan equine encephalitis virus (VEEV) demonstrated protective efficacy against Marburg virus in nonhuman primates. Here, we report the protective efficacy of Sudan virus (SUDV)- and Ebola virus (EBOV)-specific VEEV replicon particle (VRP) vaccines in nonhuman primates. VRP vaccines were developed to express the glycoprotein (GP) of either SUDV or EBOV. A single intramuscular vaccination of cynomolgus macaques with VRP expressing SUDV GP provided complete protection against intramuscular challenge with SUDV. Vaccination against SUDV and subsequent survival of SUDV challenge did not fully protect cynomolgus macaques against intramuscular EBOV back-challenge. However, a single simultaneous intramuscular vaccination with VRP expressing SUDV GP combined with VRP expressing EBOV GP did provide complete protection against intramuscular challenge with either SUDV or EBOV in cynomolgus macaques. Finally, intramuscular vaccination with VRP expressing SUDV GP completely protected cynomolgus macaques when challenged with aerosolized SUDV, although complete protection against aerosol challenge required two vaccinations with this vaccine.

  9. Alphavirus replicon particles acting as adjuvants promote CD8+ T cell responses to co-delivered antigen.

    Science.gov (United States)

    Thompson, Joseph M; Whitmore, Alan C; Staats, Herman F; Johnston, Robert E

    2008-08-05

    Alphavirus replicon particles induce strong antibody and CD8+ T cell responses to expressed antigens in numerous experimental systems. We have recently demonstrated that Venezuelan equine encephalitis virus replicon particles (VRP) possess adjuvant activity for systemic and mucosal antibody responses. In this report, we demonstrate that VRP induced an increased and balanced serum IgG subtype response to co-delivered antigen, with simultaneous induction of antigen-specific IgG1 and IgG2a antibodies, and increased both systemic and mucosal antigen-specific CD8+ T cell responses, as measured by an IFN-gamma ELISPOT assay. Additionally, VRP further increased antigen-specific T cell immunity in an additive fashion following co-delivery with the TLR ligand, CpG DNA. VRP infection led to recruitment of CD8+ T cells into the mucosal compartment, possibly utilizing the mucosal homing receptor, as this integrin was upregulated on CD8+ T cells in the draining lymph node of VRP-infected animals, where VRP-infected dendritic cells reside. This newly recognized ability of VRP to mediate increased T cell response towards co-delivered antigen provides the potential to both define the molecular basis of alphavirus-induced immunity, and improve alphavirus-based vaccines.

  10. Characterization of pMC11, a plasmid with dual origins of replication isolated from Lactobacillus casei MCJ and construction of shuttle vectors with each replicon.

    Science.gov (United States)

    Chen, Zhengjun; Lin, Jinzhong; Ma, Chengjie; Zhao, Shumiao; She, Qunxin; Liang, Yunxiang

    2014-07-01

    Many lactic acid bacteria carry different plasmids, particularly those that replicate via a theta mechanism. Here we describe Lactobacillus casei MCJ(CCTCC AB20130356), a new isolate that contains pMC11, carrying two distinct theta-type replicons. Each replicon contained an iteron in the origin of replication (oriV1 or oriV2) and a gene coding for the replicase (RepA_1 or RepB_1), both of which are essential for plasmid replication. Escherichia coli/Lactobacillus shuttle vectors were constructed with each replicon, yielding pEL5.7 and pEL5.6 that are based on oriV2 and oriV1 replicons, respectively. These plasmids showed distinct properties: pEL5.7 was capable of replicating in L. casei MCJΔ1 and Lactobacillus delbrueckii subsp. lactic LBCH-1 but failed to do so in two other tested lactobacilli strains whereas pEL5.6 replicated in three different strains, including L. casei MCJΔ1, L. casei NJ, Lactobacillus paracasei LPC-37 and L. delbrueckii subsp. lactic LBCH-1. Plasmid stability was studied: pEL5.6 and pEL5.7 were very stably maintained in L. casei, as the loss rate was lower than 1 % per generation. pEL5.7 was also stable in L. delbrueckii subsp. lactic LBCH-1 with the loss rate estimated to be 3 %. These vectors were employed to express a green fluorescent protein (GFP) using the promoter of S-layer protein SlpA from Lactobacillus acidophilus. And a growth-phase regulated expression of GFP was observed in different strains. In conclusion, these shuttle vectors provide efficient genetic tools for DNA cloning and heterologous gene expression in lactobacilli.

  11. Robust production of virus-like particles and monoclonal antibodies with geminiviral replicon vectors in lettuce.

    Science.gov (United States)

    Lai, Huafang; He, Junyun; Engle, Michael; Diamond, Michael S; Chen, Qiang

    2012-01-01

    Pharmaceutical protein production in plants has been greatly promoted by the development of viral-based vectors and transient expression systems. Tobacco and related Nicotiana species are currently the most common host plants for the generation of plant-made pharmaceutical proteins (PMPs). Downstream processing of target PMPs from these plants, however, is hindered by potential technical and regulatory difficulties owing to the presence of high levels of phenolics and toxic alkaloids. Here, we explored the use of lettuce, which grows quickly yet produces low levels of secondary metabolites and viral vector-based transient expression systems to develop a robust PMP production platform. Our results showed that a geminiviral replicon system based on the bean yellow dwarf virus permits high-level expression in lettuce of virus-like particles (VLP) derived from the Norwalk virus capsid protein and therapeutic monoclonal antibodies (mAbs) against Ebola and West Nile viruses. These vaccine and therapeutic candidates can be readily purified from lettuce leaves with scalable processing methods while fully retaining functional activity. Furthermore, this study also demonstrated the feasibility of using commercially produced lettuce for high-level PMP production. This allows our production system to have access to unlimited quantities of inexpensive plant material for large-scale production. These results establish a new production platform for biological pharmaceutical agents that are effective, safe, low cost, and amenable to large-scale manufacturing. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

  12. Vesicular stomatitis virus replicon expressing the VP2 outer capsid protein of bluetongue virus serotype 8 induces complete protection of sheep against challenge infection.

    Science.gov (United States)

    Kochinger, Stefanie; Renevey, Nathalie; Hofmann, Martin A; Zimmer, Gert

    2014-06-13

    Bluetongue virus (BTV) is an arthropod-borne pathogen that causes an often fatal, hemorrhagic disease in ruminants. Different BTV serotypes occur throughout many temperate and tropical regions of the world. In 2006, BTV serotype 8 (BTV-8) emerged in Central and Northern Europe for the first time. Although this outbreak was eventually controlled using inactivated virus vaccines, the epidemic caused significant economic losses not only from the disease in livestock but also from trade restrictions. To date, BTV vaccines that allow simple serological discrimination of infected and vaccinated animals (DIVA) have not been approved for use in livestock. In this study, we generated recombinant RNA replicon particles based on single-cycle vesicular stomatitis virus (VSV) vectors. Immunization of sheep with infectious VSV replicon particles expressing the outer capsid VP2 protein of BTV-8 resulted in induction of BTV-8 serotype-specific neutralizing antibodies. After challenge with a virulent BTV-8 strain, the vaccinated animals neither developed signs of disease nor showed viremia. In contrast, immunization of sheep with recombinant VP5 - the second outer capsid protein of BTV - did not confer protection. Discrimination of infected from vaccinated animals was readily achieved using an ELISA for detection of antibodies against the VP7 antigen. These data indicate that VSV replicon particles potentially represent a safe and efficacious vaccine platform with which to control future outbreaks by BTV-8 or other serotypes, especially in previously non-endemic regions where discrimination between vaccinated and infected animals is crucial.

  13. Molecular characterization of a DNA fragment harboring the replicon of pBMB165 from Bacillus thuringiensis subsp. tenebrionis

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    Yu Ziniu

    2006-10-01

    Full Text Available Abstract Background Bacillus thuringiensis belongs to the Bacillus cereus sensu lato group of Gram-positive and spore-forming bacteria. Most isolates of B. thuringiensis can bear many endogenous plasmids, and the number and size of these plasmids can vary widely among strains or subspecies. As far as we know, the replicon of the plasmid pBMB165 is the first instance of a plasmid replicon being isolated from subsp. tenebrionis and characterized. Results A 20 kb DNA fragment containing a plasmid replicon was isolated from B. thuringiensis subsp. tenebrionis YBT-1765 and characterized. By Southern blot analysis, this replicon region was determined to be located on pBMB165, the largest detected plasmid (about 82 kb of strain YBT-1765. Deletion analysis revealed that a replication initiation protein (Rep165, an origin of replication (ori165 and an iteron region were required for replication. In addition, two overlapping ORFs (orf6 and orf10 were found to be involved in stability control of plasmid. Sequence comparison showed that the replicon of pBMB165 was homologous to the pAMβ1 family replicons, indicating that the pBMB165 replicon belongs to this family. The presence of five transposable elements or remnants thereof in close proximity to and within the replicon control region led us to speculate that genetic exchange and recombination are potentially responsible for the divergence among the replicons of this plasmid family. Conclusion The replication and stability features of the pBMB165 from B. thuringiensis subsp. tenebrionis YBT-1765 were identified. Of particular interest is the homology and divergence shared between the pBMB165 replicon and other pAMβ1 family replicons.

  14. Molecular characterization of a DNA fragment harboring the replicon of pBMB165 from Bacillus thuringiensis subsp. tenebrionis.

    Science.gov (United States)

    Huang, Junyan; Guo, Suxia; Mahillon, Jacques; Van der Auwera, Géraldine A; Wang, Li; Han, Dongmei; Yu, Ziniu; Sun, Ming

    2006-10-23

    Bacillus thuringiensis belongs to the Bacillus cereus sensu lato group of Gram-positive and spore-forming bacteria. Most isolates of B. thuringiensis can bear many endogenous plasmids, and the number and size of these plasmids can vary widely among strains or subspecies. As far as we know, the replicon of the plasmid pBMB165 is the first instance of a plasmid replicon being isolated from subsp. tenebrionis and characterized. A 20 kb DNA fragment containing a plasmid replicon was isolated from B. thuringiensis subsp. tenebrionis YBT-1765 and characterized. By Southern blot analysis, this replicon region was determined to be located on pBMB165, the largest detected plasmid (about 82 kb) of strain YBT-1765. Deletion analysis revealed that a replication initiation protein (Rep165), an origin of replication (ori165) and an iteron region were required for replication. In addition, two overlapping ORFs (orf6 and orf10) were found to be involved in stability control of plasmid. Sequence comparison showed that the replicon of pBMB165 was homologous to the pAMbeta1 family replicons, indicating that the pBMB165 replicon belongs to this family. The presence of five transposable elements or remnants thereof in close proximity to and within the replicon control region led us to speculate that genetic exchange and recombination are potentially responsible for the divergence among the replicons of this plasmid family. The replication and stability features of the pBMB165 from B. thuringiensis subsp. tenebrionis YBT-1765 were identified. Of particular interest is the homology and divergence shared between the pBMB165 replicon and other pAMbeta1 family replicons.

  15. Analysis of classical swine fever virus RNA replication determinants using replicons

    DEFF Research Database (Denmark)

    Risager, Peter Christian; Fahnøe, Ulrik; Gullberg, Maria

    2013-01-01

    Self-replicating RNAs (replicons), with or without reporter gene sequences, derived from the genome of the Paderborn strain of classical swine fever virus (CSFV) have been produced. The full-length viral cDNA, propagated within a bacterial artificial chromosome (BAC), was modified by targeted...... recombination within E. coli. RNA transcripts were produced in vitro and introduced into cells by electroporation. The translation and replication of the replicon RNAs could be followed by the accumulation of luciferase (from Renilla reniformis or Gaussia princeps) protein expression (where appropriate...

  16. Persistent replication of the GBV-C subgenomic replicons in Huh7 cells.

    Science.gov (United States)

    Cao, Ming-Mei; Ren, Hao; Zhao, Ping; Pan, Wei; Chen, Qiu-Li; Qi, Zhong-Tian

    2009-05-01

    Studies of GB virus type C (GBV-C) replication in vitro have been limited because of poor growth of GBV-C in cell culture. In order to address the infection of GBV-C, two GBV-C subgenomic replicons (GBCrepEGFP and GBCrepTNF) were developed from a GBV-C full-length genomic cDNA. The viral replication, protein expression and the production of virus-like particles were evaluated in human hepatoma cell line Huh7. The results showed that the established GBCrepEGFP and GBCrepTNF replicons could be replicated autonomously and expressed in cell culture for at least 2 months and 1 month respectively. The replicon RNA could assemble RNA-containing structures in the HuhEH cells expressing GBV-C structural proteins. It suggests that a cell line supporting the replication of GBV-C was established. This replicon system might be used to understand better the biology of GBV-C.

  17. Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice.

    Science.gov (United States)

    Boylan, Brendan T; Moreira, Fernando R; Carlson, Tim W; Bernard, Kristen A

    2017-02-01

    Half of the human population is at risk of infection by an arthropod-borne virus. Many of these arboviruses, such as West Nile, dengue, and Zika viruses, infect humans by way of a bite from an infected mosquito. This infectious inoculum is insect cell-derived giving the virus particles distinct qualities not present in secondary infectious virus particles produced by infected vertebrate host cells. The insect cell-derived particles differ in the glycosylation of virus structural proteins and the lipid content of the envelope, as well as their induction of cytokines. Thus, in order to accurately mimic the inoculum delivered by arthropods, arboviruses should be derived from arthropod cells. Previous studies have packaged replicon genome in mammalian cells to produce replicon particles, which undergo only one round of infection, but no studies exist packaging replicon particles in mosquito cells. Here we optimized the packaging of West Nile virus replicon genome in mosquito cells and produced replicon particles at high concentration, allowing us to mimic mosquito cell-derived viral inoculum. These particles were mature with similar genome equivalents-to-infectious units as full-length West Nile virus. We then compared the mosquito cell-derived particles to mammalian cell-derived particles in mice. Both replicon particles infected skin at the inoculation site and the draining lymph node by 3 hours post-inoculation. The mammalian cell-derived replicon particles spread from the site of inoculation to the spleen and contralateral lymph nodes significantly more than the particles derived from mosquito cells. This in vivo difference in spread of West Nile replicons in the inoculum demonstrates the importance of using arthropod cell-derived particles to model early events in arboviral infection and highlights the value of these novel arthropod cell-derived replicon particles for studying the earliest virus-host interactions for arboviruses.

  18. 5’ and 3’ untranslated regions strongly enhance performance of geminiviral replicons in Nicotiana benthamiana leaves

    Directory of Open Access Journals (Sweden)

    Andrew G. Diamos

    2016-02-01

    Full Text Available We previously reported a recombinant protein production system based on a geminivirus replicon that yields high levels of vaccine antigens and monoclonal antibodies in plants. The bean yellow dwarf virus (BeYDV replicon generates massive amounts of DNA copies, which engage the plant transcription machinery. However, we noticed a disparity between transcript level and protein production, suggesting that mRNAs could be more efficiently utilized. In this study, we systematically evaluated genetic elements from human, viral, and plant sources for their potential to improve the BeYDV system. The tobacco extensin terminator enhanced transcript accumulation and protein production compared to other commonly used terminators, indicating that efficient transcript processing plays an important role in recombinant protein production. Evaluation of human-derived 5’ untranslated regions (UTRs indicated that many provided high levels of protein production, supporting their cross-kingdom function. Among the viral 5’ UTRs tested, we found the greatest enhancement with the tobacco mosaic virus omega leader. An analysis of the 5’ UTRs from the Arabidopsis thaliana and Nicotinana benthamiana photosystem I K genes found that they were highly active when truncated to include only the near upstream region, providing a dramatic enhancement of transgene production that exceeded that of the tobacco mosaic virus omega leader. The tobacco Rb7 matrix attachment region (MAR inserted downstream from the gene of interest provided significant enhancement, which was correlated with a reduction in plant cell death. Evaluation of agrobacterium strains found that EHA105 enhanced protein production and reduced cell death compared to LBA4301 and GV3101. We used these improvements to produce Norwalk virus capsid protein at >20% total soluble protein, corresponding to 1.8mg/g leaf fresh weight, more than twice the highest level ever reported in a plant system. We also produced

  19. Efficient replication of genotype 3a and 4a hepatitis C virus replicons in human hepatoma cells

    DEFF Research Database (Denmark)

    Saeed, Mohsan; Scheel, Troels K H; Gottwein, Judith M

    2012-01-01

    to express a chimeric fusion protein of firefly luciferase and neomycin phosphotransferase to yield stable replicon-expressing cells. Using these constructs, the inhibitory effects of beta interferon (IFN-β), an NS3 protease inhibitor, and an NS5B nucleoside polymerase inhibitor were readily detected...... culture adaptive mutations originally reported for genotype 1b replicons. RNA replication was confirmed by quantitative reverse transcription-PCR and detection of viral protein. Sequencing of multiple independent replicon clones revealed the presence of additional nonsynonymous mutations. Interestingly......, all potentially adaptive mutations mapped to the NS3 protein. These mutations, when introduced back into original constructs, substantially increased colony formation efficiency. To make these replicons useful for high-throughput screening and evaluation of antiviral compounds, they were modified...

  20. Infected dendritic cells are sufficient to mediate the adjuvant activity generated by Venezuelan equine encephalitis virus replicon particles

    OpenAIRE

    Tonkin, Daniel R; Whitmore, Alan; Johnston, Robert E; Barro, Mario

    2012-01-01

    Replicon particles derived from Venezuelan equine encephalitis virus (VEE) are infectious non-propagating particles which act as a safe and potent systemic, mucosal, and cellular adjuvant when delivered with antigen. VEE and VEE replicon particles (VRP) can target multiple cell types including dendritic cells (DCs). The role of these cell types in VRP adjuvant activity has not been previously evaluated, and for these studies we focused on the contribution of DCs to the response to VRP. By ana...

  1. Chromatin structural changes precede replication in initiated replicons during inhibition of DNA elongation

    Energy Technology Data Exchange (ETDEWEB)

    D' Anna, J.A.; Grady, D.L.; Tobey, R.A.

    1988-01-01

    Partial inhibition of replicative DNA synthesis by hydroxyurea or other agents produces changes in the composition and structure of bulk chromatin. We have begun to investigate the structural changes in specific regions of the genome using synchronized cells and cloned genomic probes. Current results indicate changes in chromatin structure occur preferentially in initiated replicons and can precede the replication fork during inhibition of DNA elongation. 4 refs., 2 figs.

  2. Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon

    Directory of Open Access Journals (Sweden)

    Luftig Ronald

    2007-09-01

    Full Text Available Abstract Background Alpha interferon in combination with ribavirin is the standard therapy for hepatitis C virus infection. Unfortunately, a significant number of patients fail to eradicate their infection with this regimen. The mechanisms of IFN-resistance are unclear. The aim of this study was to determine the contribution of host cell factors to the mechanisms of interferon resistance using replicon cell lines. Results HCV replicons with high and low activation of the IFN-promoter were cultured for a prolonged period of time in the presence of interferon-alpha (IFN-alpha2b. Stable replicon cell lines with resistant phenotype were isolated and characterized by their ability to continue viral replication in the presence of IFN-alpha. Interferon resistant cell colonies developed only in replicons having lower activation of the IFN promoter and no resistant colonies arose from replicons that exhibit higher activation of the IFN promoter. Individual cell clones were isolated and nine IFN resistant cell lines were established. HCV RNA and protein levels in these cells were not altered by IFN- alpha2b. Reduced signaling and IFN-resistant phenotype was found in all Huh-7 cell lines even after eliminating HCV, suggesting that cellular factors are involved. Resistant phenotype in the replicons is not due to lack of interferon receptor expression. All the cell lines show defect in the JAK-STAT signaling and phosphorylation of STAT 1 and STAT 2 proteins were strongly inhibited due to reduced expression of Tyk2 and Jak-1 protein. Conclusion This in vitro study provides evidence that altered expression of the Jak-Stat signaling proteins can cause IFN resistance using HCV replicon cell clones.

  3. Self-replicating Replicon-RNA Delivery to Dendritic Cells by Chitosan-nanoparticles for Translation In Vitro and In Vivo

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    Kenneth C McCullough

    2014-01-01

    Full Text Available Self-amplifying replicon RNA (RepRNA possesses high potential for increasing antigen load within dendritic cells (DCs. The major aim of the present work was to define how RepRNA delivered by biodegradable, chitosan-based nanoparticulate delivery vehicles (nanogel-alginate (NGA interacts with DCs, and whether this could lead to translation of the RepRNA in the DCs. Although studies employed virus replicon particles (VRPs, there are no reports on biodegradable, nanoparticulate vehicle delivery of RepRNA. VRP studies employed cytopathogenic agents, contrary to DC requirements—slow processing and antigen retention. We employed noncytopathogenic RepRNA with NGA, demonstrating for the first time the efficiency of RepRNA association with nanoparticles, NGA delivery to DCs, and RepRNA internalization by DCs. RepRNA accumulated in vesicular structures, with patterns typifying cytosolic release. This promoted RepRNA translation, in vitro and in vivo. Delivery and translation were RepRNA concentration-dependent, occurring in a kinetic manner. Including cationic lipids with chitosan during nanoparticle formation enhanced delivery and translation kinetics, but was not required for translation of immunogenic levels in vivo. This work describes for the first time the characteristics associated with chitosan-nanoparticle delivery of self-amplifying RepRNA to DCs, leading to translation of encoded foreign genes, namely influenza virus hemagglutinin and nucleoprotein.

  4. Recombinant Kunjin virus replicon vaccines induce protective T-cell immunity against human papillomavirus 16 E7-expressing tumour.

    Science.gov (United States)

    Herd, Karen A; Harvey, Tracey; Khromykh, Alexander A; Tindle, Robert W

    2004-02-20

    The persistence of the E7 oncoprotein in transformed cells in human papillomavirus (HPV)-associated cervical cancer provides a tumour-specific antigen to which immunotherapeutic strategies may be directed. Self-replicating RNA (replicon) vaccine vectors derived from the flavivirus Kunjin (KUN) have recently been reported to induce T-cell immunity. Here, we report that inclusion of a CTL epitope of HPV16 E7 protein into a polyepitope encoded by a KUN vector induced E7-directed T-cell responses and protected mice against challenge with an E7-expressing epithelial tumour. We found replicon RNA packaged into virus-like particles to be more effective than naked replicon RNA or plasmid DNA constructed to allow replicon RNA transcription in vivo. Protective immunity was induced although the E7 CTL epitope was subdominant in the context of other CTL epitopes in the polyepitope. The results demonstrate the efficacy of the KUN replicon vector system for inducing protective immunity directed towards a virally encoded human tumour-specific antigen, and for inducing multi-epitopic CTL responses.

  5. Replication of a hepatitis C virus replicon clone in mouse cells

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    Chisari Francis V

    2006-10-01

    Full Text Available Abstract Background Hepatitis C Virus (HCV is a significant public health burden and small animal models are needed to study the pathology and immunobiology of the virus. In effort to develop experimental HCV mouse models, we screened a panel of HCV replicons to identify clones capable of replicating in mouse hepatocytes. Results We report the establishment of stable HCV replication in mouse hepatocyte and fibroblast cell lines using replicons derived from the JFH-1 genotype 2a consensus sequence. Viral RNA replication efficiency in mouse cells was comparable to that observed in human Huh-7 replicon cells, with negative-strand HCV RNA and the viral NS5A protein being readily detected by Northern and Western Blot analysis, respectively. Although HCV replication was established in the absence of adaptive mutations that might otherwise compromise the in vitro infectivity of the JFH-1 clone, no infectious virus was detected when the culture medium from full length HCV RNA replicating mouse cells was titrated on Huh-7 cells, suggesting that the mouse cells were unable to support production of infectious progeny viral particles. Consistent with an additional block in viral entry, infectious JFH-1 particles produced in Huh-7 cells were not able to establish detectable HCV RNA replication in naïve mouse cells. Conclusion Thus, this report expands the repertoire of HCV replication systems and possibly represents a step toward developing mouse models of HCV replication, but it also highlights that other species restrictions might continue to make the development of a purely murine HCV infectious model challenging.

  6. Characterization of Rhizobium grahamii extrachromosomal replicons and their transfer among rhizobia.

    Science.gov (United States)

    Althabegoiti, María Julia; Ormeño-Orrillo, Ernesto; Lozano, Luis; Torres Tejerizo, Gonzalo; Rogel, Marco Antonio; Mora, Jaime; Martínez-Romero, Esperanza

    2014-01-08

    Rhizobium grahamii belongs to a new phylogenetic group of rhizobia together with Rhizobium mesoamericanum and other species. R. grahamii has a broad-host-range that includes Leucaena leucocephala and Phaseolus vulgaris, although it is a poor competitor for P. vulgaris nodulation in the presence of Rhizobium etli or Rhizobium phaseoli strains. This work analyzed the genome sequence and transfer properties of R. grahamii plasmids. Genome sequence was obtained from R. grahamii CCGE502 type strain isolated from Dalea leporina in Mexico. The CCGE502 genome comprises one chromosome and two extrachromosomal replicons (ERs), pRgrCCGE502a and pRgrCCGE502b. Additionally, a plasmid integrated in the CCGE502 chromosome was found. The genomic comparison of ERs from this group showed that gene content is more variable than average nucleotide identity (ANI). Well conserved nod and nif genes were found in R. grahamii and R. mesoamericanum with some differences. R. phaseoli Ch24-10 genes expressed in bacterial cells in roots were found to be conserved in pRgrCCGE502b. Regarding conjugative transfer we were unable to transfer the R. grahamii CCGE502 symbiotic plasmid and its megaplasmid to other rhizobial hosts but we could transfer the symbiotic plasmid to Agrobacterium tumefaciens with transfer dependent on homoserine lactones. Variable degrees of nucleotide identity and gene content conservation were found among the different R. grahamii CCGE502 replicons in comparison to R. mesoamericanum genomes. The extrachromosomal replicons from R. grahamii were more similar to those found in phylogenetically related Rhizobium species. However, limited similarities of R. grahamii CCGE502 symbiotic plasmid and megaplasmid were observed in other more distant Rhizobium species. The set of conserved genes in R. grahamii comprises some of those that are highly expressed in R. phaseoli on plant roots, suggesting that they play an important role in root colonization.

  7. Identification, Characterization, and Application of the Replicon Region of the Halophilic Temperate Sphaerolipovirus SNJ1

    OpenAIRE

    Wang, Yuchen; Sima, Linshan; Lv, Jie; Huang, Suiyuan; Liu, Ying; Wang, Jiao; Krupovic, Mart; Chen, Xiangdong

    2016-01-01

    The temperate haloarchaeal virus SNJ1 displays lytic and lysogenic life cycles. During the lysogenic cycle, the virus resides in its host, Natrinema sp. strain J7-1, in the form of an extrachromosomal circular plasmid, pHH205. In this study, a 3.9-kb region containing seven predicted genes organized in two operons was identified as the minimal replicon of SNJ1. Only RepA, encoded by open reading frame 11-12 (ORF11-12), was found to be essential for replication, and its expression increased du...

  8. Microarray analysis identifies a common set of cellular genes modulated by different HCV replicon clones

    Directory of Open Access Journals (Sweden)

    Gerosolimo Germano

    2008-06-01

    Full Text Available Abstract Background Hepatitis C virus (HCV RNA synthesis and protein expression affect cell homeostasis by modulation of gene expression. The impact of HCV replication on global cell transcription has not been fully evaluated. Thus, we analysed the expression profiles of different clones of human hepatoma-derived Huh-7 cells carrying a self-replicating HCV RNA which express all viral proteins (HCV replicon system. Results First, we compared the expression profile of HCV replicon clone 21-5 with both the Huh-7 parental cells and the 21-5 cured (21-5c cells. In these latter, the HCV RNA has been eliminated by IFN-α treatment. To confirm data, we also analyzed microarray results from both the 21-5 and two other HCV replicon clones, 22-6 and 21-7, compared to the Huh-7 cells. The study was carried out by using the Applied Biosystems (AB Human Genome Survey Microarray v1.0 which provides 31,700 probes that correspond to 27,868 human genes. Microarray analysis revealed a specific transcriptional program induced by HCV in replicon cells respect to both IFN-α-cured and Huh-7 cells. From the original datasets of differentially expressed genes, we selected by Venn diagrams a final list of 38 genes modulated by HCV in all clones. Most of the 38 genes have never been described before and showed high fold-change associated with significant p-value, strongly supporting data reliability. Classification of the 38 genes by Panther System identified functional categories that were significantly enriched in this gene set, such as histones and ribosomal proteins as well as extracellular matrix and intracellular protein traffic. The dataset also included new genes involved in lipid metabolism, extracellular matrix and cytoskeletal network, which may be critical for HCV replication and pathogenesis. Conclusion Our data provide a comprehensive analysis of alterations in gene expression induced by HCV replication and reveal modulation of new genes potentially useful

  9. Molecular characterization of a DNA fragment harboring the replicon of pBMB165 from Bacillus thuringiensis subsp tenebrionis

    OpenAIRE

    Yu Ziniu; Han Dongmei; Wang Li; Van der Auwera Géraldine A; Mahillon Jacques; Guo Suxia; Huang Junyan; Sun Ming

    2006-01-01

    Abstract Background Bacillus thuringiensis belongs to the Bacillus cereus sensu lato group of Gram-positive and spore-forming bacteria. Most isolates of B. thuringiensis can bear many endogenous plasmids, and the number and size of these plasmids can vary widely among strains or subspecies. As far as we know, the replicon of the plasmid pBMB165 is the first instance of a plasmid replicon being isolated from subsp. tenebrionis and characterized. Results A 20 kb DNA fragment containing a plasmi...

  10. Alphavirus replicon particles expressing TRP-2 provide potent therapeutic effect on melanoma through activation of humoral and cellular immunity.

    Directory of Open Access Journals (Sweden)

    Francesca Avogadri

    2010-09-01

    Full Text Available Malignant melanoma is the deadliest form of skin cancer and is refractory to conventional chemotherapy and radiotherapy. Therefore alternative approaches to treat this disease, such as immunotherapy, are needed. Melanoma vaccine design has mainly focused on targeting CD8+ T cells. Activation of effector CD8+ T cells has been achieved in patients, but provided limited clinical benefit, due to immune-escape mechanisms established by advanced tumors. We have previously shown that alphavirus-based virus-like replicon particles (VRP simultaneously activate strong cellular and humoral immunity against the weakly immunogenic melanoma differentiation antigen (MDA tyrosinase. Here we further investigate the antitumor effect and the immune mechanisms of VRP encoding different MDAs.VRP encoding different MDAs were screened for their ability to prevent the growth of the B16 mouse transplantable melanoma. The immunologic mechanisms of efficacy were investigated for the most effective vaccine identified, focusing on CD8+ T cells and humoral responses. To this end, ex vivo immune assays and transgenic mice lacking specific immune effector functions were used. The studies identified a potent therapeutic VRP vaccine, encoding tyrosinase related protein 2 (TRP-2, which provided a durable anti-tumor effect. The efficacy of VRP-TRP2 relies on a novel immune mechanism of action requiring the activation of both IgG and CD8+ T cell effector responses, and depends on signaling through activating Fcγ receptors.This study identifies a VRP-based vaccine able to elicit humoral immunity against TRP-2, which plays a role in melanoma immunotherapy and synergizes with tumor-specific CD8+ T cell responses. These findings will aid in the rational design of future immunotherapy clinical trials.

  11. Alphavirus replicon particles expressing TRP-2 provide potent therapeutic effect on melanoma through activation of humoral and cellular immunity.

    Science.gov (United States)

    Avogadri, Francesca; Merghoub, Taha; Maughan, Maureen F; Hirschhorn-Cymerman, Daniel; Morris, John; Ritter, Erika; Olmsted, Robert; Houghton, Alan N; Wolchok, Jedd D

    2010-09-10

    Malignant melanoma is the deadliest form of skin cancer and is refractory to conventional chemotherapy and radiotherapy. Therefore alternative approaches to treat this disease, such as immunotherapy, are needed. Melanoma vaccine design has mainly focused on targeting CD8+ T cells. Activation of effector CD8+ T cells has been achieved in patients, but provided limited clinical benefit, due to immune-escape mechanisms established by advanced tumors. We have previously shown that alphavirus-based virus-like replicon particles (VRP) simultaneously activate strong cellular and humoral immunity against the weakly immunogenic melanoma differentiation antigen (MDA) tyrosinase. Here we further investigate the antitumor effect and the immune mechanisms of VRP encoding different MDAs. VRP encoding different MDAs were screened for their ability to prevent the growth of the B16 mouse transplantable melanoma. The immunologic mechanisms of efficacy were investigated for the most effective vaccine identified, focusing on CD8+ T cells and humoral responses. To this end, ex vivo immune assays and transgenic mice lacking specific immune effector functions were used. The studies identified a potent therapeutic VRP vaccine, encoding tyrosinase related protein 2 (TRP-2), which provided a durable anti-tumor effect. The efficacy of VRP-TRP2 relies on a novel immune mechanism of action requiring the activation of both IgG and CD8+ T cell effector responses, and depends on signaling through activating Fcγ receptors. This study identifies a VRP-based vaccine able to elicit humoral immunity against TRP-2, which plays a role in melanoma immunotherapy and synergizes with tumor-specific CD8+ T cell responses. These findings will aid in the rational design of future immunotherapy clinical trials.

  12. Replicon-Helper Systems from Attenuated Venezuelan Equine Encephalitis Virus: Expression of Heterologous Genes in Vitro and Immunization Against Heterologous Pathogens in Vivo

    National Research Council Canada - National Science Library

    Pushko, Peter

    1997-01-01

    ...) or the Lassa virus nucleocapsid (N) gene, and upon transfection into eukaryotic cells by electroportation, these replicon RNAs directed the efficient, high-level synthesis of the HA or N proteins...

  13. Development of dengue virus replicons expressing HIV-1 gp120 and other heterologous genes: a potential future tool for dual vaccination against dengue virus and HIV

    Directory of Open Access Journals (Sweden)

    Dayton Andrew I

    2001-11-01

    Full Text Available Abstract Background Toward the goals of providing an additional vector to add to the armamentarium available to HIV vaccinologists and of creating a bivalent vaccine effective against dengue virus and HIV, we have attempted to create vectors which express dengue virus non-structural proteins and HIV immunogens. Previously we reported the successful construction of dengue virus replicons which lack structural genes necessary for virion release and spreading infection in culture but which can replicate intracellularly and abundantly produce dengue non-structural proteins. Here we attempted to express heterologous genetic material from these replicons. Results We cloned into a Δpre-M/E dengue virus replicon genes for either green fluorescent protein (GFP, HIV gp160 or HIV gp120 and tested the ability of these constructs to express dengue virus proteins as well as the heterologous proteins in tissue culture after transfection of replicon RNA. Conclusions Heterologous proteins were readily expressed from these constructs. GFP and gp120 demonstrated minimal or no toxicity. Gp160 expressing replicons were found to express proteins abundantly at 36 hours post transfection, but after 50 hrs of transfection, few replicon positive cells could be found despite the presence of cellular debris positive for replicon proteins. This suggested that gp160 expressed from dengue virus replicons is considerably more toxic than either GFP or gp120. The successful expression of heterologous proteins, including HIV gp120 for long periods in culture suggests this vector system may be useful as a vaccine vector, given appropriate delivery methods.

  14. Isolation of broad-host-range replicons from marine sediment bacteria.

    Science.gov (United States)

    Sobecky, P A; Mincer, T J; Chang, M C; Toukdarian, A; Helinski, D R

    1998-08-01

    Naturally occurring plasmids isolated from heterotrophic bacterial isolates originating from coastal California marine sediments were characterized by analyzing their incompatibility and replication properties. Previously, we reported on the lack of DNA homology between plasmids from the culturable bacterial population of marine sediments and the replicon probes specific for a number of well-characterized incompatibility and replication groups (P. A. Sobecky, T. J. Mincer, M. C. Chang, and D. R. Helinski, Appl. Environ. Microbiol. 63:888-895, 1997). In the present study we isolated 1.8- to 2.3-kb fragments that contain functional replication origins from one relatively large (30-kb) and three small (marine isolates. 16S rRNA sequence analyses indicated that the four plasmid-bearing marine isolates belonged to the alpha and gamma subclasses of the class Proteobacteria. Three of the marine sediment isolates are related to the gamma-3 subclass organisms Vibrio splendidus and Vibrio fischeri, while the fourth isolate may be related to Roseobacter litoralis. Sequence analysis of the plasmid replication regions revealed the presence of features common to replication origins of well-characterized plasmids from clinical bacterial isolates, suggesting that there may be similar mechanisms for plasmid replication initiation in the indigenous plasmids of gram-negative marine sediment bacteria. In addition to replication in Escherichia coli DH5alpha and C2110, the host ranges of the plasmid replicons, designated repSD41, repSD121, repSD164, and repSD172, extended to marine species belonging to the genera Achromobacter, Pseudomonas, Serratia, and Vibrio. While sequence analysis of repSD41 and repSD121 revealed considerable stretches of homology between the two fragments, these regions do not display incompatibility properties against each other. The replication origin repSD41 was detected in 5% of the culturable plasmid-bearing marine sediment bacterial isolates, whereas the

  15. Deep sequencing uncovers numerous small RNAs on all four replicons of the plant pathogen Agrobacterium tumefaciens

    Science.gov (United States)

    Wilms, Ina; Overlöper, Aaron; Nowrousian, Minou; Sharma, Cynthia M.; Narberhaus, Franz

    2012-01-01

    Agrobacterium species are capable of interkingdom gene transfer between bacteria and plants. The genome of Agrobacterium tumefaciens consists of a circular and a linear chromosome, the At-plasmid and the Ti-plasmid, which harbors bacterial virulence genes required for tumor formation in plants. Little is known about promoter sequences and the small RNA (sRNA) repertoire of this and other α-proteobacteria. We used a differential RNA sequencing (dRNA-seq) approach to map transcriptional start sites of 388 annotated genes and operons. In addition, a total number of 228 sRNAs was revealed from all four Agrobacterium replicons. Twenty-two of these were confirmed by independent RNA gel blot analysis and several sRNAs were differentially expressed in response to growth media, growth phase, temperature or pH. One sRNA from the Ti-plasmid was massively induced under virulence conditions. The presence of 76 cis-antisense sRNAs, two of them on the reverse strand of virulence genes, suggests considerable antisense transcription in Agrobacterium. The information gained from this study provides a valuable reservoir for an in-depth understanding of sRNA-mediated regulation of the complex physiology and infection process of Agrobacterium. PMID:22336765

  16. Antimicrobial resistance and plasmid replicons in Yersinia enterocolitica strains isolated in Brazil in 30 years.

    Science.gov (United States)

    Frazão, Miliane R; Andrade, Leonardo N; Darini, Ana L C; Falcão, Juliana P

    Some studies evaluated the resistance profile of the Y. enterocolitica strains isolated in diverse countries. However, in Brazil the isolation and the study of Y. enterocolitica are not common and therefore information about the antimicrobial resistance profile of this species in this country is scarce. Therefore, the aim of this study was to evaluate the antimicrobial resistance of Y. enterocolitica of biotypes 1A, 2 and 4 isolated from clinical and non-clinical sources between 1979 and 2012, in Brazil. This study showed that some Yersinia enterocolitica of different biotypes remain susceptible to antimicrobials used for gastroenteritis treatment. Moreover, neither acquired resistance genes nor diversity of plasmids replicons were found; however, variation in the in vitro intrinsic resistant pattern was detected, except the non-resistance to cefoxitin in all strains. Notwithstanding, due to epidemiological link between Y. enterocolitica and the pork production chain, monitoring plasmid acquired resistance in Y. enterocolitica could also be considered for antimicrobial resistance control purposes and food safety measures. Copyright © 2017 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  17. Replicon Particles of Venezuelan Equine Encephalitis Virus as a Reductionist Murine Model for Encephalitis▿

    Science.gov (United States)

    Schäfer, Alexandra; Whitmore, Alan C.; Konopka, Jennifer L.; Johnston, Robert E.

    2009-01-01

    Venezuelan equine encephalitis virus (VEE) replicon particles (VRP) were used to model the initial phase of VEE-induced encephalitis in the mouse brain. VRP can target and infect cells as VEE, but VRP do not propagate beyond the first infected cell due to the absence of the structural genes. Direct intracranial inoculation of VRP into mice induced acute encephalitis with signs similar to the neuronal phase of wild-type VEE infection and other models of virus-induced encephalitis. Using the previously established VRP-mRNP tagging system, a new method to distinguish the host responses in infected cells from those in uninfected bystander cell populations, we detected a robust and rapid innate immune response in the central nervous system (CNS) by infected neurons and uninfected bystander cells. Moreover, this innate immune response in the CNS compromised blood-brain barrier integrity, created an inflammatory response, and directed an adaptive immune response characterized by proliferation and activation of microglia cells and infiltration of inflammatory monocytes, in addition to CD4+ and CD8+ T lymphocytes. Taken together, these data suggest that a naïve CNS has an intrinsic potential to induce an innate immune response that could be crucial to the outcome of the infection by determining the composition and dynamics of the adaptive immune response. Furthermore, these results establish a model for neurotropic virus infection to identify host and viral factors that contribute to invasion of the brain, the mechanism(s) whereby the adaptive immune response can clear the infection, and the role of the host innate response in these processes. PMID:19225006

  18. Venezuelan equine encephalitis replicon particles can induce rapid protection against foot-and-mouth disease virus.

    Science.gov (United States)

    Diaz-San Segundo, Fayna; Dias, Camila C A; Moraes, Mauro P; Weiss, Marcelo; Perez-Martin, Eva; Owens, Gary; Custer, Max; Kamrud, Kurt; de los Santos, Teresa; Grubman, Marvin J

    2013-05-01

    We have previously shown that delivery of the porcine type I interferon gene (poIFN-α/β) with a replication-defective human adenovirus vector (adenovirus 5 [Ad5]) can sterilely protect swine challenged with foot-and-mouth disease virus (FMDV) 1 day later. However, the need of relatively high doses of Ad5 limits the applicability of such a control strategy in the livestock industry. Venezuelan equine encephalitis virus (VEE) empty replicon particles (VRPs) can induce rapid protection of mice against either homologous or, in some cases, heterologous virus challenge. As an alternative approach to induce rapid protection against FMDV, we have examined the ability of VRPs containing either the gene for green fluorescent protein (VRP-GFP) or poIFN-α (VRP-poIFN-α) to block FMDV replication in vitro and in vivo. Pretreatment of swine or bovine cell lines with either VRP significantly inhibited subsequent infection with FMDV as early as 6 h after treatment and for at least 120 h posttreatment. Furthermore, mice pretreated with either 10(7) or 10(8) infectious units of VRP-GFP and challenged with a lethal dose of FMDV 24 h later were protected from death. Protection was induced as early as 6 h after treatment and lasted for at least 48 h and correlated with induction of an antiviral response and production of IFN-α. By 6 h after treatment several genes were upregulated, and the number of genes and the level of induction increased at 24 h. Finally, we demonstrated that the chemokine IP-10, which is induced by IFN-α and VRP-GFP, is directly involved in protection against FMDV.

  19. Replicon particles of Venezuelan equine encephalitis virus as a reductionist murine model for encephalitis.

    Science.gov (United States)

    Schäfer, Alexandra; Whitmore, Alan C; Konopka, Jennifer L; Johnston, Robert E

    2009-05-01

    Venezuelan equine encephalitis virus (VEE) replicon particles (VRP) were used to model the initial phase of VEE-induced encephalitis in the mouse brain. VRP can target and infect cells as VEE, but VRP do not propagate beyond the first infected cell due to the absence of the structural genes. Direct intracranial inoculation of VRP into mice induced acute encephalitis with signs similar to the neuronal phase of wild-type VEE infection and other models of virus-induced encephalitis. Using the previously established VRP-mRNP tagging system, a new method to distinguish the host responses in infected cells from those in uninfected bystander cell populations, we detected a robust and rapid innate immune response in the central nervous system (CNS) by infected neurons and uninfected bystander cells. Moreover, this innate immune response in the CNS compromised blood-brain barrier integrity, created an inflammatory response, and directed an adaptive immune response characterized by proliferation and activation of microglia cells and infiltration of inflammatory monocytes, in addition to CD4(+) and CD8(+) T lymphocytes. Taken together, these data suggest that a naïve CNS has an intrinsic potential to induce an innate immune response that could be crucial to the outcome of the infection by determining the composition and dynamics of the adaptive immune response. Furthermore, these results establish a model for neurotropic virus infection to identify host and viral factors that contribute to invasion of the brain, the mechanism(s) whereby the adaptive immune response can clear the infection, and the role of the host innate response in these processes.

  20. The contribution of type I interferon signaling to immunity induced by alphavirus replicon vaccines.

    Science.gov (United States)

    Thompson, Joseph M; Whitmore, Alan C; Staats, Herman F; Johnston, Robert

    2008-09-15

    The type I interferon (IFN) system is critical for protecting the mammalian host from numerous virus infections and plays a key role in shaping the antiviral adaptive immune response. In this report, the importance of type I IFN signaling was assessed in a mouse model of alphavirus-induced humoral immune induction. Venezuelan equine encephalitis virus replicon particles (VRP) expressing the hemagglutinin (HA) gene from influenza virus (HA-VRP) were used to vaccinate both wildtype (wt) and IFN alpha/beta receptor knockout (RKO) mice. HA-VRP vaccination induced equivalent levels of flu-specific systemic IgG, mucosal IgG, and systemic IgA antibodies in both wt and IFN RKO mice. In contrast, HA-VRP vaccination of IFN RKO mice failed to induce significant levels of flu-specific mucosal IgA antibodies at multiple mucosal surfaces. In the VRP adjuvant system, co-delivery of null VRP with ovalbumin (OVA) protein significantly increased the levels of OVA-specific serum IgG, fecal IgG, and fecal IgA antibodies in both wt and RKO mice, suggesting that type I IFN signaling plays a less significant role in the VRP adjuvant effect. Taken together, these results suggest that (1) at least in regard to IFN signaling, the mechanisms which regulate alphavirus-induced immunity differ when VRP are utilized as expression vectors as opposed to adjuvants, and (2) type I IFN signaling is required for the induction of mucosal IgA antibodies directed against VRP-expressed antigen. These results shed new light on the regulatory networks which promote immune induction, and specifically mucosal immune induction, with alphavirus vaccine vectors.

  1. Effects of short RNA structural analogues against hepatitis C virus genotypes 2, 3 and 4 in replicon cells.

    Science.gov (United States)

    Elshaffei, Ismail M; Gupta, Nidhi; Wu, Catherine H; Wu, David C; Hammad, Lamiaa N; Abo-Elmatty, Dina M; Mesbah, Noha M; Wu, George Y

    2015-08-01

    To determine whether computer-predicted short RNA structural analogues could inhibit hepatitis C virus (HCV) genotype 2a, 3a and 4a replication in cultured cells. Short RNA sequences, X12, X12a and X12b, designed to be identical in secondary structure to the X region in the 3'-untranslated region (3'-UTR) of the HCV 1b genome, as well as shorter stem-loop components of X region, were inserted into a plasmid and transfected into separate Huh7.5 human hepatoma cells stably transfected with subgenomic replicons for genotypes 2a, 3a and 4a. All replicons included a firefly luciferase reporter gene. After 48 h of plasmid transfection, the inhibition of HCV replication was determined by HCV RNA isolation and quantification by real-time polymerase chain reaction and luciferase assays. All the secondary structural analogues to genotype 1b X region cross-inhibited genotype 2a, 3a and 4a replicons. The maximum inhibition by genotype 1b X region structural analogues was obtained against genotype 2a cells in which X12, X12a and X12b inhibited replication by 30%, 63% and 72%, respectively (P < 0.05 for all), compared to an unrelated hepatitis B viral analogue. Despite substantial sequence dissimilarity, HCV RNA genotype 1b X region analogues cross-inhibited the replication of HCV genotypes 2a, 3a and 4a. Particular conformations and not the sequence of the stem-loops of the X region are involved in HCV replication. © 2015 Chinese Medical Association Shanghai Branch, Chinese Society of Gastroenterology, Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine and Wiley Publishing Asia Pty Ltd.

  2. In vivo DNA cloning and adjacent gene fusing with a mini-Mu-lac bacteriophage containing a plasmid replicon.

    OpenAIRE

    Groisman, E A; Castilho, B A; Casadaban, M J

    1984-01-01

    A mini-Mu bacteriophage containing a high copy number plasmid replicon was constructed to clone genes in vivo. A chloramphenicol resistance gene for independent selection and the lacZYA operon to form gene fusions were also incorporated into this phage. This mini-Mu element can transpose at a high frequency when derepressed, and it can be complemented by a helper Mu prophage for lytic growth. DNA sequences that are flanked by two copies of this mini-Mu can be packaged along with them. After i...

  3. A vaccinia virus recombinant transcribing an alphavirus replicon and expressing alphavirus structural proteins leads to packaging of alphavirus infectious single cycle particles.

    Directory of Open Access Journals (Sweden)

    Juana M Sánchez-Puig

    Full Text Available Poxviruses and Alphaviruses constitute two promising viral vectors that have been used extensively as expression systems, or as vehicles for vaccine purposes. Poxviruses, like vaccinia virus (VV are well-established vaccine vectors having large insertion capacity, excellent stability, and ease of administration. In turn, replicons derived from Alphaviruses like Semliki Forest virus (SFV are potent protein expression and immunization vectors but stocks are difficult to produce and maintain. In an attempt to demonstrate the use of a Poxvirus as a means for the delivery of small vaccine vectors, we have constructed and characterized VV/SFV hybrid vectors. A SFV replicon cDNA was inserted in the VV genome and placed under the control of a VV early promoter. The replicon, transcribed from the VV genome as an early transcript, was functional, and thus capable of initiating its own replication and transcription. Further, we constructed a VV recombinant additionally expressing the SFV structural proteins under the control of a vaccinia synthetic early/late promoter. Infection with this recombinant produced concurrent transcription of the replicon and expression of SFV structural proteins, and led to the generation of replicon-containing SFV particles that were released to the medium and were able to infect additional cells. This combined VV/SFV system in a single virus allows the use of VV as a SFV delivery vehicle in vivo. The combination of two vectors, and the possibility of generating in vivo single-cycle, replicon containing alphavirus particles, may open new strategies in vaccine development or in the design of oncolytic viruses.

  4. Antiviral Activity and Resistance Analysis of NS3/4A Protease Inhibitor Grazoprevir and NS5A Inhibitor Elbasvir in Hepatitis C Virus GT4 Replicons.

    Science.gov (United States)

    Asante-Appiah, Ernest; Curry, Stephanie; McMonagle, Patricia; Ingravallo, Paul; Chase, Robert; Nickle, David; Qiu, Ping; Howe, Anita; Lahser, Frederick C

    2017-07-01

    Although genotype 4 (GT4)-infected patients represent a minor overall percentage of the global hepatitis C virus (HCV)-infected population, the high prevalence of the genotype in specific geographic regions coupled with substantial sequence diversity makes it an important genotype to study for antiviral drug discovery and development. We evaluated two direct-acting antiviral agents-grazoprevir, an HCV NS3/4A protease inhibitor, and elbasvir, an HCV NS5A inhibitor-in GT4 replicons prior to clinical studies in this genotype. Following a bioinformatics analysis of available GT4 sequences, a set of replicons bearing representative GT4 clinical isolates was generated. For grazoprevir, the 50% effective concentration (EC50) against the replicon bearing the reference GT4a (ED43) NS3 protease and NS4A was 0.7 nM. The median EC50 for grazoprevir against chimeric replicons encoding NS3/4A sequences from GT4 clinical isolates was 0.2 nM (range, 0.11 to 0.33 nM; n = 5). The difficulty in establishing replicons bearing NS3/4A resistance-associated substitutions was substantially overcome with the identification of a G162R adaptive substitution in NS3. Single NS3 substitutions D168A/V identified from de novo resistance selection studies reduced grazoprevir antiviral activity by 137- and 47-fold, respectively, in the background of the G162R replicon. For elbasvir, the EC50 against the replicon bearing the reference full-length GT4a (ED43) NS5A gene was 0.0002 nM. The median EC50 for elbasvir against chimeric replicons bearing clinical isolates from GT4 was 0.0007 nM (range, 0.0002 to 34 nM; n = 14). De novo resistance selection studies in GT4 demonstrated a high propensity to suppress the emergence of amino acid substitutions that confer high-potency reductions to elbasvir. Phenotypic characterization of the NS5A amino acid substitutions identified (L30F, L30S, M31V, and Y93H) indicated that they conferred 15-, 4-, 2.5-, and 7.5-fold potency losses, respectively, to elbasvir. The

  5. Enhanced potency of individual and bivalent DNA replicon vaccines or conventional DNA vaccines by formulation with aluminum phosphate.

    Science.gov (United States)

    Yu, Yun-Zhou; Wang, Wen-Bin; Li, Na; Wang, Shuang; Yu, Wei-Yuan; Sun, Zhi-Wei

    2010-11-01

    DNA vaccines against botulinum neurotoxin (BoNTs) induce protective humoral immune responses in mouse model, but when compared with conventional vaccines such as toxoid and protein vaccines, DNA vaccines often induce lower antibody level and protective efficacy and are still necessary to increase their potency. In this study we evaluated the potency of aluminum phosphate as an adjuvant of DNA vaccines to enhance antibody responses and protective efficacy against botulinum neurotoxin serotypes A and B in Balb/c mice. The administration of these individual and bivalent plasmid DNA replicon vaccines against botulinum neurotoxin serotypes A and B in the presence of aluminum phosphate improved both antibody responses and protective efficacy. Furthermore, formulation of conventional plasmid DNA vaccines encoding the same Hc domains of botulinum neurotoxin serotypes A and B with aluminum phosphate adjuvant increased both antibody responses and protective efficacy. These results indicate aluminum phosphate is an effective adjuvant for these two types of DNA vaccines (i.e., plasmid DNA replicon vaccines and conventional plasmid DNA vaccines), and the vaccine formulation described here may be an excellent candidate for further vaccine development against botulinum neurotoxins. Copyright © 2010 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.

  6. Quantitative Proteomics Analysis of the Hepatitis C Virus Replicon High-Permissive and Low-Permissive Cell Lines.

    Directory of Open Access Journals (Sweden)

    Fei Ye

    Full Text Available Chronic hepatitis C virus (HCV infection is one of the leading causes of severe hepatitis. The molecular mechanisms underlying HCV replication and pathogenesis remain unclear. The development of the subgenome replicon model system significantly enhanced study of HCV. However, the permissiveness of the HCV subgenome replicon greatly differs among different hepatoma cell lines. Proteomic analysis of different permissive cell lines might provide new clues in understanding HCV replication. In this study, to detect potential candidates that might account for the differences in HCV replication. Label-free and iTRAQ labeling were used to analyze the differentially expressed protein profiles between Huh7.5.1 wt and HepG2 cells. A total of 4919 proteins were quantified in which 114 proteins were commonly identified as differentially expressed by both quantitative methods. A total of 37 differential proteins were validated by qRT-PCR. The differential expression of Glutathione S-transferase P (GSTP1, Ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1, carboxylesterase 1 (CES1, vimentin, Proteasome activator complex subunit1 (PSME1, and Cathepsin B (CTSB were verified by western blot. And over-expression of CTSB or knock-down of vimentin induced significant changes to HCV RNA levels. Additionally, we demonstrated that CTSB was able to inhibit HCV replication and viral protein translation. These results highlight the potential role of CTSB and vimentin in virus replication.

  7. High-level rapid production of full-size monoclonal antibodies in plants by a single-vector DNA replicon system.

    Science.gov (United States)

    Huang, Zhong; Phoolcharoen, Waranyoo; Lai, Huafang; Piensook, Khanrat; Cardineau, Guy; Zeitlin, Larry; Whaley, Kevin J; Arntzen, Charles J; Mason, Hugh S; Chen, Qiang

    2010-05-01

    Plant viral vectors have great potential in rapid production of important pharmaceutical proteins. However, high-yield production of hetero-oligomeric proteins that require the expression and assembly of two or more protein subunits often suffers problems due to the "competing" nature of viral vectors derived from the same virus. Previously we reported that a bean yellow dwarf virus (BeYDV)-derived, three-component DNA replicon system allows rapid production of single recombinant proteins in plants (Huang et al., 2009. Biotechnol Bioeng 103: 706-714). In this article, we report further development of this expression system for its application in high-yield production of oligomeric protein complexes including monoclonal antibodies (mAbs) in plants. We showed that the BeYDV replicon system permits simultaneous efficient replication of two DNA replicons and thus, high-level accumulation of two recombinant proteins in the same plant cell. We also demonstrated that a single vector that contains multiple replicon cassettes was as efficient as the three-component system in driving the expression of two distinct proteins. Using either the non-competing, three-vector system or the multi-replicon single vector, we produced both the heavy and light chain subunits of a protective IgG mAb 6D8 against Ebola virus GP1 (Wilson et al., 2000. Science 287: 1664-1666) at 0.5 mg of mAb per gram leaf fresh weight within 4 days post-infiltration of Nicotiana benthamiana leaves. We further demonstrated that full-size tetrameric IgG complex containing two heavy and two light chains was efficiently assembled and readily purified, and retained its functionality in specific binding to inactivated Ebola virus. Thus, our single-vector replicon system provides high-yield production capacity for hetero-oligomeric proteins, yet eliminates the difficult task of identifying non-competing virus and the need for co-infection of multiple expression modules. The multi-replicon vector represents a

  8. Partial Protection against Porcine Influenza A Virus by a Hemagglutinin-Expressing Virus Replicon Particle Vaccine in the Absence of Neutralizing Antibodies.

    Science.gov (United States)

    Ricklin, Meret E; Vielle, Nathalie J; Python, Sylvie; Brechbühl, Daniel; Zumkehr, Beatrice; Posthaus, Horst; Zimmer, Gert; Summerfield, Artur

    2016-01-01

    This work was initiated by previous reports demonstrating that mismatched influenza A virus (IAV) vaccines can induce enhanced disease, probably mediated by antibodies. Our aim was, therefore, to investigate if a vaccine inducing opsonizing but not neutralizing antibodies against the hemagglutinin (HA) of a selected heterologous challenge virus would enhance disease or induce protective immune responses in the pig model. To this end, we immunized pigs with either whole inactivated virus (WIV)-vaccine or HA-expressing virus replicon particles (VRP) vaccine based on recombinant vesicular stomatitis virus (VSV). Both types of vaccines induced virus neutralizing and opsonizing antibodies against homologous virus as shown by a highly sensitive plasmacytoid dendritic cell-based opsonization assay. Opsonizing antibodies showed a broader reactivity against heterologous IAV compared with neutralizing antibodies. Pigs immunized with HA-recombinant VRP vaccine were partially protected from infection with a mismatched IAV, which was not neutralized but opsonized by the immune sera. The VRP vaccine reduced lung lesions, lung inflammatory cytokine responses, serum IFN-α responses, and viral loads in the airways. Only the VRP vaccine was able to prime IAV-specific IFNγ/TNFα dual secreting CD4(+) T cells detectable in the peripheral blood. In summary, this work demonstrates that with the virus pair selected, a WIV vaccine inducing opsonizing antibodies against HA which lack neutralizing activity, is neither protective nor does it induce enhanced disease in pigs. In contrast, VRP-expressing HA is efficacious vaccines in swine as they induced both potent antibodies and T-cell immunity resulting in a broader protective value.

  9. Co-delivery of antigen and IL-12 by Venezuelan equine encephalitis virus replicon particles enhances antigen-specific immune responses and anti-tumor effects

    Science.gov (United States)

    Osada, Takuya; Berglund, Peter; Morse, Michael A.; Hubby, Bolyn; Lewis, Whitney; Niedzwiecki, Donna; Hobeika, Amy; Burnett, Bruce; Devi, Gayathri R.; Clay, Timothy M.; Smith, Jonathan; Lyerly, H. Kim

    2013-01-01

    We recently demonstrated that Venezuelan equine encephalitis (VEE) virus-based replicon particles (VRP) encoding tumor antigens could break tolerance in the immunomodulatory environment of advanced cancer. We hypothesized that local injection of VRP expressing Interleukin-12 (IL-12) at the site of injections of VRP-based cancer vaccines would enhance the tumor-antigen-specific T cell and antibody responses and anti-tumor efficacy. Mice were immunized with VRP encoding the human tumor-associated antigen, carcinoembryonic antigen (CEA) (VRP-CEA(6D)) and VRP-IL-12 was also administered at the same site or at a distant location. CEA-specific T cell and antibody responses were measured. To determine antitumor activity, mice were implanted with MC38-CEA-2 cells and immunized with VRP-CEA with and without VRP-IL-12 and tumor growth and mouse survival were measured. VRP-IL-12 greatly enhanced CEA-specific T cell and antibody responses when combined with VRP-CEA(6D) vaccination. VRP IL-12 was superior to IL-12 protein at enhancing immune responses. Vaccination with VRP-CEA(6D) plus VRP-IL-12 was superior to VRP-CEA(6D) or VRP-IL-12 alone in inducing anti-tumor activity and prolonging survival in tumor-bearing mice. Importantly, local injection of VRP-IL-12 at the VRP-CEA(6D) injection site provided more potent activation of CEA-specific immune responses than VRP-IL-12 injected at a distant site from the VRP-CEA injections. Together, this study shows that VRP-IL-12 enhances vaccination with VRP-CEA(6D) and was more effective at activating CEA-specific T cell responses when locally expressed at the vaccine site. Clinical trials evaluating the adjuvant effect of VRP-IL-12 at enhancing the immunogenicity of cancer vaccines are warranted. PMID:22488274

  10. Co-delivery of antigen and IL-12 by Venezuelan equine encephalitis virus replicon particles enhances antigen-specific immune responses and antitumor effects.

    Science.gov (United States)

    Osada, Takuya; Berglund, Peter; Morse, Michael A; Hubby, Bolyn; Lewis, Whitney; Niedzwiecki, Donna; Yang, Xiao Yi; Hobeika, Amy; Burnett, Bruce; Devi, Gayathri R; Clay, Timothy M; Smith, Jonathan; Kim Lyerly, H

    2012-11-01

    We recently demonstrated that Venezuelan equine encephalitis virus-based replicon particle (VRPs) encoding tumor antigens could break tolerance in the immunomodulatory environment of advanced cancer. We hypothesized that local injection of VRP-expressing interleukin-12 (IL-12) at the site of injections of VRP-based cancer vaccines would enhance the tumor-antigen-specific T cell and antibody responses and antitumor efficacy. Mice were immunized with VRP encoding the human tumor-associated antigen, carcinoembryonic antigen (CEA) (VRP-CEA(6D)), and VRP-IL-12 was also administered at the same site or at a distant location. CEA-specific T cell and antibody responses were measured. To determine antitumor activity, mice were implanted with MC38-CEA-2 cells and immunized with VRP-CEA with and without VRP-IL-12, and tumor growth and mouse survival were measured. VRP-IL-12 greatly enhanced CEA-specific T cell and antibody responses when combined with VRP-CEA(6D) vaccination. VRP-IL-12 was superior to IL-12 protein at enhancing immune responses. Vaccination with VRP-CEA(6D) plus VRP-IL-12 was superior to VRP-CEA(6D) or VRP-IL-12 alone in inducing antitumor activity and prolonging survival in tumor-bearing mice. Importantly, local injection of VRP-IL-12 at the VRP-CEA(6D) injection site provided more potent activation of CEA-specific immune responses than that of VRP-IL-12 injected at a distant site from the VRP-CEA injections. Together, this study shows that VRP-IL-12 enhances vaccination with VRP-CEA(6D) and was more effective at activating CEA-specific T cell responses when locally expressed at the vaccine site. Clinical trials evaluating the adjuvant effect of VRP-IL-12 at enhancing the immunogenicity of cancer vaccines are warranted.

  11. Vaccination with recombinant RNA replicon particles protects chickens from H5N1 highly pathogenic avian influenza virus.

    Science.gov (United States)

    Halbherr, Stefan J; Brostoff, Terza; Tippenhauer, Merve; Locher, Samira; Berger Rentsch, Marianne; Zimmer, Gert

    2013-01-01

    Highly pathogenic avian influenza viruses (HPAIV) of subtype H5N1 not only cause a devastating disease in domestic chickens and turkeys but also pose a continuous threat to public health. In some countries, H5N1 viruses continue to circulate and evolve into new clades and subclades. The rapid evolution of these viruses represents a problem for virus diagnosis and control. In this work, recombinant vesicular stomatitis virus (VSV) vectors expressing HA of subtype H5 were generated. To comply with biosafety issues the G gene was deleted from the VSV genome. The resulting vaccine vector VSV*ΔG(HA) was propagated on helper cells providing the VSV G protein in trans. Vaccination of chickens with a single intramuscular dose of 2×10⁸ infectious replicon particles without adjuvant conferred complete protection from lethal H5N1 infection. Subsequent application of the same vaccine strongly boosted the humoral immune response and completely prevented shedding of challenge virus and transmission to sentinel birds. The vaccine allowed serological differentiation of infected from vaccinated animals (DIVA) by employing a commercially available ELISA. Immunized chickens produced antibodies with neutralizing activity against multiple H5 viruses representing clades 1, 2.2, 2.5, and low-pathogenic avian influenza viruses (classical clade). Studies using chimeric H1/H5 hemagglutinins showed that the neutralizing activity was predominantly directed against the globular head domain. In summary, these results suggest that VSV replicon particles are safe and potent DIVA vaccines that may help to control avian influenza viruses in domestic poultry.

  12. Infected dendritic cells are sufficient to mediate the adjuvant activity generated by Venezuelan equine encephalitis virus replicon particles.

    Science.gov (United States)

    Tonkin, Daniel R; Whitmore, Alan; Johnston, Robert E; Barro, Mario

    2012-06-22

    Replicon particles derived from Venezuelan equine encephalitis virus (VEE) are infectious non-propagating particles which act as a safe and potent systemic, mucosal, and cellular adjuvant when delivered with antigen. VEE and VEE replicon particles (VRP) can target multiple cell types including dendritic cells (DCs). The role of these cell types in VRP adjuvant activity has not been previously evaluated, and for these studies we focused on the contribution of DCs to the response to VRP. By analysis of VRP targeting in the draining lymph node, we found that VRP induced rapid recruitment of TNF-secreting monocyte-derived inflammatory dendritic cells. VRP preferentially infected these inflammatory DCs as well as classical DCs and macrophages, with less efficient infection of other cell types. DC depletion suggested that the interaction of VRP with classical DCs was required for recruitment of inflammatory DCs, induction of high levels of many cytokines, and for stable transport of VRP to the draining lymph node. Additionally, in vitro-infected DCs enhanced antigen-specific responses by CD4 and CD8 T cells. By transfer of VRP-infected DCs into mice we showed that these DCs generated an inflammatory state in the draining lymph node similar to that achieved by VRP injection. Most importantly, VRP-infected DCs were sufficient to establish robust adjuvant activity in mice comparable to that produced by VRP injection. These findings indicate that VRP infect, recruit and activate both classical and inflammatory DCs, and those DCs become mediators of the VRP adjuvant activity. Published by Elsevier Ltd.

  13. Molecular smallpox vaccine delivered by alphavirus replicons elicits protective immunity in mice and non-human primates.

    Science.gov (United States)

    Hooper, Jay W; Ferro, Anthony M; Golden, Joseph W; Silvera, Peter; Dudek, Jeanne; Alterson, Kim; Custer, Max; Rivers, Bryan; Morris, John; Owens, Gary; Smith, Jonathan F; Kamrud, Kurt I

    2009-12-11

    Naturally occurring smallpox was eradicated as a result of successful vaccination campaigns during the 1960s and 1970s. Because of its highly contagious nature and high mortality rate, smallpox has significant potential as a biological weapon. Unfortunately, the current vaccine for orthopoxviruses is contraindicated for large portions of the population. Thus, there is a need for new, safe, and effective orthopoxvirus vaccines. Alphavirus replicon vectors, derived from strains of Venezuelan equine encephalitis virus, are being used to develop alternatives to the current smallpox vaccine. Here, we demonstrated that virus-like replicon particles (VRPs) expressing the vaccinia virus A33R, B5R, A27L, and L1R genes elicited protective immunity in mice comparable to vaccination with live-vaccinia virus. Furthermore, cynomolgus macaques vaccinated with a combination of the four poxvirus VRPs (4pox-VRP) developed antibody responses to each antigen. These antibody responses were able to neutralize and inhibit the spread of both vaccinia virus and monkeypox virus. Macaques vaccinated with 4pox-VRP, flu HA VRP (negative control), or live-vaccinia virus (positive control) were challenged intravenously with 5 x 10(6)pfu of monkeypox virus 1 month after the second VRP vaccination. Four of the six negative control animals succumbed to monkeypox and the remaining two animals demonstrated either severe or grave disease. Importantly, all 10 macaques vaccinated with the 4pox-VRP vaccine survived without developing severe disease. These findings revealed that a single-boost VRP smallpox vaccine shows promise as a safe alternative to the currently licensed live-vaccinia virus smallpox vaccine.

  14. Single-dose immunization with virus replicon particles confers rapid robust protection against Rift Valley fever virus challenge.

    Science.gov (United States)

    Dodd, Kimberly A; Bird, Brian H; Metcalfe, Maureen G; Nichol, Stuart T; Albariño, César G

    2012-04-01

    Rift Valley fever virus (RVFV) causes outbreaks of severe disease in people and livestock throughout Africa and the Arabian Peninsula. The potential for RVFV introduction outside the area of endemicity highlights the need for fast-acting, safe, and efficacious vaccines. Here, we demonstrate a robust system for the reverse genetics generation of a RVF virus replicon particle (VRP(RVF)) vaccine candidate. Using a mouse model, we show that VRP(RVF) immunization provides the optimal balance of safety and single-dose robust efficacy. VRP(RVF) can actively synthesize viral RNA and proteins but lacks structural glycoprotein genes, preventing spread within immunized individuals and reducing the risk of vaccine-induced pathogenicity. VRP(RVF) proved to be completely safe following intracranial inoculation of suckling mice, a stringent test of vaccine safety. Single-dose subcutaneous immunization with VRP(RVF), although it is highly attenuated, completely protected mice against a virulent RVFV challenge dose which was 100,000-fold greater than the 50% lethal dose (LD(50)). Robust protection from lethal challenge was observed by 24 h postvaccination, with 100% protection induced in as little as 96 h. We show that a single subcutaneous VRP(RVF) immunization initiated a systemic antiviral state followed by an enhanced adaptive response. These data contrast sharply with the much-reduced survivability and immune responses observed among animals immunized with nonreplicating viral particles, indicating that replication, even if confined to the initially infected cells, contributes substantially to protective efficacy at early and late time points postimmunization. These data demonstrate that replicon vaccines successfully bridge the gap between safety and efficacy and provide insights into the kinetics of antiviral protection from RVFV infection.

  15. Vaccination with recombinant RNA replicon particles protects chickens from H5N1 highly pathogenic avian influenza virus.

    Directory of Open Access Journals (Sweden)

    Stefan J Halbherr

    Full Text Available Highly pathogenic avian influenza viruses (HPAIV of subtype H5N1 not only cause a devastating disease in domestic chickens and turkeys but also pose a continuous threat to public health. In some countries, H5N1 viruses continue to circulate and evolve into new clades and subclades. The rapid evolution of these viruses represents a problem for virus diagnosis and control. In this work, recombinant vesicular stomatitis virus (VSV vectors expressing HA of subtype H5 were generated. To comply with biosafety issues the G gene was deleted from the VSV genome. The resulting vaccine vector VSV*ΔG(HA was propagated on helper cells providing the VSV G protein in trans. Vaccination of chickens with a single intramuscular dose of 2×10⁸ infectious replicon particles without adjuvant conferred complete protection from lethal H5N1 infection. Subsequent application of the same vaccine strongly boosted the humoral immune response and completely prevented shedding of challenge virus and transmission to sentinel birds. The vaccine allowed serological differentiation of infected from vaccinated animals (DIVA by employing a commercially available ELISA. Immunized chickens produced antibodies with neutralizing activity against multiple H5 viruses representing clades 1, 2.2, 2.5, and low-pathogenic avian influenza viruses (classical clade. Studies using chimeric H1/H5 hemagglutinins showed that the neutralizing activity was predominantly directed against the globular head domain. In summary, these results suggest that VSV replicon particles are safe and potent DIVA vaccines that may help to control avian influenza viruses in domestic poultry.

  16. Burkholderia xernovorans LB400 harbors a multi-replicon, 9.73-Mbp genome shaped for versatility

    Energy Technology Data Exchange (ETDEWEB)

    Chain, Patrick S. G. [Lawrence Livermore National Laboratory (LLNL); Denef, Vincent [University of California, Berkeley; Konstantinidis, Konstantinos T [Michigan State University, East Lansing; Vergez, Lisa [Lawrence Livermore National Laboratory (LLNL); Agullo, Loreine [Universidad Tecnica Federico Santa Maria, Casilla 110-V; Reyes, Valeria Latorre [Universidad Tecnica Federico Santa Maria, Casilla 110-V; Hauser, Loren John [ORNL; Cordova, Macarena [Universidad Tecnica Federico Santa Maria, Casilla 110-V; Gomez, Luis [Universidad Tecnica Federico Santa Maria, Casilla 110-V; Gonzalez, Myriam [Universidad Tecnica Federico Santa Maria, Casilla 110-V; Land, Miriam L [ORNL; Lao, Victoria [Lawrence Livermore National Laboratory (LLNL); Larimer, Frank W [ORNL; LiPuma, John J [University of Michigan; Mahenthiralingam, Eshwar [Cardiff University, Wales; Malfatti, Stephanie [Lawrence Livermore National Laboratory (LLNL); Marx, Christopher J [Harvard University; Parnell, J Jacob [Michigan State University, East Lansing; Ramette, Alban [Michigan State University, East Lansing; Richardson, P M [U.S. Department of Energy, Joint Genome Institute; Seeger, Michael [Universidad Tecnica Federico Santa Maria, Casilla 110-V; Smith, Daryl [University of British Columbia, Vancouver; Spilker, Theodore [University of Michigan; Sul, Woo Jun [Michigan State University, East Lansing; Tsoi, Tamara V [Michigan State University, East Lansing; Zhulin, Igor B [University of Tennessee, Knoxville (UTK) & Oak Ridge National Laboratory (ORNL); Tiedje, James M. [Michigan State University, East Lansing

    2006-01-01

    Burkholderia xenovorans LB400 (LB400), a well studied, effective polychlorinated biphenyl-degrader, has one of the two largest known bacterial genomes and is the first nonpathogenic Burkholderia isolate sequenced. From an evolutionary perspective, we find significant differences in functional specialization between the three replicons of LB400, as well as a more relaxed selective pressure for genes located on the two smaller vs. the largest replicon. High genomic plasticity, diversity, and specialization within the Burkholderia genus are exemplified by the conservation of only 44% of the genes between LB400 and Burkholderia cepacia complex strain 383. Even among four B. xenovorans strains, genome size varies from 7.4 to 9.73 Mbp. The latter is largely explained by our findings that >20% of the LB400 sequence was recently acquired by means of lateral gene transfer. Although a range of genetic factors associated with in vivo survival and intercellular interactions are present, these genetic factors are likely related to niche breadth rather than determinants of pathogenicity. The presence of at least eleven 'central aromatic' and twenty 'peripheral aromatic' pathways in LB400, among the highest in any sequenced bacterial genome, supports this hypothesis. Finally, in addition to the experimentally observed redundancy in benzoate degradation and formaldehyde oxidation pathways, the fact that 17.6% of proteins have a better LB400 paralog than an ortholog in a different genome highlights the importance of gene duplication and repeated acquirement, which, coupled with their divergence, raises questions regarding the role of paralogs and potential functional redundancies in large-genome microbes.

  17. Structure and Immunogenicity of Alternative Forms of the Simian Immunodeficiency Virus Gag Protein Expressed Using Venezuelan Equine Encephalitis Virus Replicon Particles

    OpenAIRE

    Cecil, Chad; West, Ande; Collier, Martha; Jurgens, Christy; Madden, Victoria; Whitmore, Alan; Johnston, Robert; Moore, Dominic T.; Swanstrom, Ronald; Davis, Nancy L.

    2007-01-01

    Venezuelan equine encephalitis virus replicon particles (VRP) were engineered to express different forms of SIV Gag to compare expression in vitro, formation of intra- and extracellular structures and induction of humoral and cellular immunity in mice. The three forms examined were full-length myristylated SIV Gag (Gagmyr+), full-length Gag lacking the myristylation signal (Gagmyr-), or a truncated form of Gagmyr- comprising only the matrix and capsid domains (MA/CA). Comparison of VRP-infect...

  18. Dual Roles of Two Isoforms of Autophagy-related Gene ATG10 in HCV-Subgenomic replicon Mediated Autophagy Flux and Innate Immunity.

    Science.gov (United States)

    Zhao, Qiong; Hu, Zhan-Ying; Zhang, Jing-Pu; Jiang, Jian-Dong; Ma, Yuan-Yuan; Li, Jian-Rui; Peng, Zong-Gen; Chen, Jin-Hua

    2017-09-12

    Autophagy and immune response are two defense systems that human-body uses against viral infection. Previous studies documented that some viral mechanisms circumvented host immunity mechanisms and hijacked autophagy for its replication and survival. Here, we focus on interactions between autophagy mechanism and innate-immune-response in HCV-subgenomic replicon cells to find a mechanism linking the two pathways. We report distinct effects of two autophagy-related protein ATG10s on HCV-subgenomic replication. ATG10, a canonical long isoform in autophagy process, can facilitate HCV-subgenomic replicon amplification by promoting autophagosome formation and by combining with and detaining autophagosomes in cellular periphery, causing impaired autophagy flux. ATG10S, a non-canonical short isoform of ATG10 proteins, can activate expression of IL28A/B and immunity genes related to viral ds-RNA including ddx-58, tlr-3, tlr-7, irf-3 and irf-7, and promote autophagolysosome formation by directly combining and driving autophagosomes to perinuclear region where lysosomes gather, leading to lysosomal degradation of HCV-subgenomic replicon in HepG2 cells. ATG10S also can suppress infectious HCV virion replication in Huh7.5 cells. Another finding is that IL28A protein directly conjugates ATG10S and helps autophagosome docking to lysosomes. ATG10S might be a new host factor against HCV replication, and as a target for screening chemicals with new anti-virus mechanisms.

  19. The tra locus of streptomycete plasmid pIJ101 mediates efficient transfer of a circular but not a linear version of the same replicon.

    Science.gov (United States)

    Wang, Jing; Pettis, Gregg S

    2010-09-01

    Conjugal transfer of circular plasmids in Streptomyces involves a unique mechanism employing few plasmid-encoded loci and the transfer of double-stranded DNA by an as yet uncharacterized intercellular route. Efficient transfer of the circular streptomycete plasmid pIJ101 requires only two plasmid loci: the pIJ101 tra gene, and as a cis-acting function known as clt. Here, we compared the ability of the pIJ101 transfer apparatus to promote conjugal transfer of circular versus linear versions of the same replicon. While the pIJ101 tra locus readily transferred the circular form of the replicon, the linear version was transferred orders of magnitude less efficiently and all plasmids isolated from the transconjugants were circular, regardless of their original configuration in the donor. Additionally, relatively rare circularization of linear plasmids was detectable in the donor cells, which is consistent with the notion that this event was a prerequisite for transfer by TraB(pIJ101). Linear versions of this same replicon did transfer efficiently, in that configuration, from strains containing the conjugative linear plasmid SLP2. Our data indicate that functions necessary and sufficient for transfer of circular DNA were insufficient for transfer of a related linear DNA molecule. The results here suggest that the conjugation mechanisms of linear versus circular DNA in Streptomyces spp. are inherently different and/or that efficient transfer of linear DNA requires additional components.

  20. Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Mingrui; Du, Yinming; Jiang, Wenyan; Chang, Wei-Lun; Yang, Shang-Tian [Ohio State Univ., Columbus, OH (United States). William G. Lowrie Dept. of Chemical and Biomolecular Engineering; Tang, I-Ching [Bioprocessing Innovative Company, Dublin, OH (United States)

    2012-01-15

    Clostridium tyrobutyricum ATCC 25755 can produce butyric acid, acetic acid, and hydrogen as the main products from various carbon sources. In this study, C. tyrobutyricum was used as a host to produce n-butanol by expressing adhE2 gene under the control of a native thiolase promoter using four different conjugative plasmids (pMTL82151, 83151, 84151, and 85151) each with a different replicon (pBP1 from C. botulinum NCTC2916, pCB102 from C. butyricum, pCD6 from Clostridium difficile, and pIM13 from Bacillus subtilis). The effects of different replicons on transformation efficiency, plasmid stability, adhE2 expression and aldehyde/alcohol dehydrogenase activities, and butanol production by different mutants of C. tyrobutyricum were investigated. Among the four plasmids and replicons studied, pMTL82151 with pBP1 gave the highest transformation efficiency, plasmid stability, gene expression, and butanol biosynthesis. Butanol production from various substrates, including glucose, xylose, mannose, and mannitol were then investigated with the best mutant strain harboring adhE2 in pMTL82151. A high butanol titer of 20.5 g/L with 0.33 g/g yield and 0.32 g/L h productivity was obtained with mannitol as the substrate in batch fermentation with pH controlled at {proportional_to}6.0. (orig.)

  1. Acute infection with venezuelan equine encephalitis virus replicon particles catalyzes a systemic antiviral state and protects from lethal virus challenge.

    Science.gov (United States)

    Konopka, Jennifer L; Thompson, Joseph M; Whitmore, Alan C; Webb, Drue L; Johnston, Robert E

    2009-12-01

    The host innate immune response provides a critical first line of defense against invading pathogens, inducing an antiviral state to impede the spread of infection. While numerous studies have documented antiviral responses within actively infected tissues, few have described the earliest innate response induced systemically by infection. Here, utilizing Venezuelan equine encephalitis virus (VEE) replicon particles (VRP) to limit infection to the initially infected cells in vivo, a rapid activation of the antiviral response was demonstrated not only within the murine draining lymph node, where replication was confined, but also within distal tissues. In the liver and brain, expression of interferon-stimulated genes was detected by 1 to 3 h following VRP footpad inoculation, reaching peak expression of >100-fold over that in mock-infected animals. Moreover, mice receiving a VRP footpad inoculation 6, 12, or 24 h prior to an otherwise lethal VEE footpad challenge were completely protected from death, including a drastic reduction in challenge virus titers. VRP pretreatment also provided protection from intranasal VEE challenge and extended the average survival time following intracranial challenge. Signaling through the interferon receptor was necessary for antiviral gene induction and protection from VEE challenge. However, VRP pretreatment failed to protect mice from a heterologous, lethal challenge with vesicular stomatitis virus, yet conferred protection following challenge with influenza virus. Collectively, these results document a rapid modulation of the host innate response within hours of infection, capable of rapidly alerting the entire animal to pathogen invasion and leading to protection from viral disease.

  2. Small high-yielding binary Ti vectors pLSU with co-directional replicons for Agrobacterium tumefaciens-mediated transformation of higher plants.

    Science.gov (United States)

    Lee, Seokhyun; Su, Guiying; Lasserre, Eric; Aghazadeh, Monty Arta; Murai, Norimoto

    2012-05-01

    Small high-yielding binary Ti vectors of Agrobacterium tumefaciens were constructed to increase the cloning efficiency and plasmid yield in Escherichia coli and A. tumefaciens for transformation of higher plants. We reduced the size of the binary vector backbone to 4566bp with ColE1 replicon (715bp) for E. coli and VS1 replicon (2654bp) for A. tumefaciens, a bacterial kanamycin resistance gene (999bp), and the T-DNA region (152bp). The binary Ti vectors with the truncated VS1 replicon were stably maintained with more than 98% efficiency in A. tumefaciens without antibiotic selection for 4 days of successive transfers. The transcriptional direction of VS1 replicon can be the same as that of ColE1 replicon (co-directional transcription), or opposite (head-on transcription) as in the case of widely used vectors (pPZP or pCambia). New binary vectors with co-directional transcription yielded in E. coli up to four-fold higher transformation frequency than those with the head-on transcription. In A. tumefaciens the effect of co-directional transcription is still positive in up to 1.8-fold higher transformation frequency than that of head-on transcription. Transformation frequencies of new vectors are over six-fold higher than those of pCambia vector in A. tumefaciens. DNA yields of new vectors were three to five-fold greater than pCambia in E. coli. The proper functions of the new T-DNA borders and new plant selection marker genes were confirmed after A. tumefaciens-mediated transformation of tobacco leaf discs, resulting in virtually all treated leaf discs transformed and induced calli. Genetic analysis of kanamycin resistance trait among the progeny showed that the kanamycin resistance and sensitivity traits were segregated into the 3:1 ratio, indicating that the kanamycin resistance genes were integrated stably into a locus or closely linked loci of the nuclear chromosomal DNA of the primary transgenic tobacco plants and inherited to the second generation. © 2012

  3. SH2 modified STAT1 induces HLA-I expression and improves IFN-γ signaling in IFN-α resistant HCV replicon cells.

    Directory of Open Access Journals (Sweden)

    Bret Poat

    2010-09-01

    Full Text Available We have developed multiple stable cell lines containing subgenomic HCV RNA that are resistant to treatment with interferon alpha (IFN-α. Characterization of these IFN-α resistant replicon cells showed defects in the phosphorylation and nuclear translocation of STAT1 and STAT2 proteins due to a defective Jak-STAT pathway.In this study, we have developed an alternative strategy to overcome interferon resistance in a cell culture model by improving intracellular STAT1 signaling. An engineered STAT1-CC molecule with double cysteine substitutions in the Src-homology 2 (SH2 domains of STAT1 (at Ala-656 and Asn-658 efficiently phosphorylates and translocates to the nucleus of IFN-resistant cells in an IFN-γ dependent manner. Transfection of a plasmid clone containing STAT1-CC significantly activated the GAS promoter compared to wild type STAT1 and STAT3. The activity of the engineered STAT1-CC is dependent upon the phosphorylation of tyrosine residue 701, since the construct with a substituted phenylalanine residue at position 701 (STAT1-CC-Y701F failed to activate GAS promoter in the replicon cells. Intracellular expression of STAT1-CC protein showed phosphorylation and nuclear translocation in the resistant cell line after IFN-γ treatment. Transient transfection of STAT1-CC plasmid clone into an interferon resistant cell line resulted in inhibition of viral replication and viral clearance in an IFN-γ dependent manner. Furthermore, the resistant replicon cells transfected with STAT1-CC constructs significantly up regulated surface HLA-1 expression when compared to the wild type and Y to F mutant controls.These results suggest that modification of the SH2 domain of the STAT1 molecule allows for improved IFN-γ signaling through increased STAT1 phosphorylation, nuclear translocation, HLA-1 surface expression, and prolonged interferon antiviral gene activation.

  4. 3D replicon distributions arise from stochastic initiation and domino-like DNA replication progression.

    Science.gov (United States)

    Löb, D; Lengert, N; Chagin, V O; Reinhart, M; Casas-Delucchi, C S; Cardoso, M C; Drossel, B

    2016-04-07

    DNA replication dynamics in cells from higher eukaryotes follows very complex but highly efficient mechanisms. However, the principles behind initiation of potential replication origins and emergence of typical patterns of nuclear replication sites remain unclear. Here, we propose a comprehensive model of DNA replication in human cells that is based on stochastic, proximity-induced replication initiation. Critical model features are: spontaneous stochastic firing of individual origins in euchromatin and facultative heterochromatin, inhibition of firing at distances below the size of chromatin loops and a domino-like effect by which replication forks induce firing of nearby origins. The model reproduces the empirical temporal and chromatin-related properties of DNA replication in human cells. We advance the one-dimensional DNA replication model to a spatial model by taking into account chromatin folding in the nucleus, and we are able to reproduce the spatial and temporal characteristics of the replication foci distribution throughout S-phase.

  5. Replicon properties of chromosomal DNA fibers and the duration of DNA synthesis of sunflower root-tip meristem cells at different temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Van' t Hof, J.; Bjerknes, C.A.; Clinton, J.H.

    1978-01-01

    Chromosomal DNA fiber autoradiography was used to examine the replicon properties of root-tip meristem cells of Helianthus annuus intact seedlings grown at temperatures from 10 to 38/sup 0/C and those of root-tip cells grown in vitro at 23/sup 0/. The average replicon size was approximately 22 ..mu..m and it did not change with temperature nor when the roots were grown in culture. The average fork rate was 6 ..mu..m/h at 10/sup 0/ and it rose gradually to 12 ..mu..m/h at 38/sup 0/. The responses of replication fork movement and of the duration of S to temperature were of three types: those in which change in fork rate was primarily (more than 90%) responsible for change in the duration of S, those in which the fork rate remained constant while S increased nearly twofold, and those in which the duration of S increased even though the replication forks were moving faster. The first type of response listed was observed at temperatures from 20 to 35/sup 0/, the second type listed was observed at 10 to 15/sup 0/, and the third, was produced at 38/sup 0/.

  6. Virus replicon particles expressing porcine reproductive and respiratory syndrome virus proteins elicit immune priming but do not confer protection from viremia in pigs.

    Science.gov (United States)

    Eck, Melanie; Durán, Margarita García; Ricklin, Meret E; Locher, Samira; Sarraseca, Javier; Rodríguez, María José; McCullough, Kenneth C; Summerfield, Artur; Zimmer, Gert; Ruggli, Nicolas

    2016-02-19

    Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of one of the most devastating and economically significant viral disease of pigs worldwide. The vaccines currently available on the market elicit only limited protection. Recombinant vesicular stomatitis virus (VSV) replicon particles (VRP) have been used successfully to induce protection against influenza A virus (IAV) in chickens and bluetongue virus in sheep. In this study, VSV VRP expressing the PRRSV envelope proteins GP5, M, GP4, GP3, GP2 and the nucleocapsid protein N, individually or in combination, were generated and evaluated as a potential vector vaccine against PRRSV infection. High level expression of the recombinant PRRSV proteins was demonstrated in cell culture. However, none of the PRRSV antigens expressed from VRP, with the exception of the N protein, did induce any detectable antibody response in pigs before challenge infection with PRRSV. After challenge however, the antibody responses against GP5, GP4 and GP3 appeared in average 2 weeks earlier than in pigs vaccinated with the empty control VRP. No reduction of viremia was observed in the vaccinated group compared with the control group. When pigs were co-vaccinated with VRP expressing IAV antigens and VRP expressing PRRSV glycoproteins, only antibody responses to the IAV antigens were detectable. These data show that the VSV replicon vector can induce immune responses to heterologous proteins in pigs, but that the PRRSV envelope proteins expressed from VSV VRP are poorly immunogenic. Nevertheless, they prime the immune system for significantly earlier B-cell responses following PRRSV challenge infection.

  7. Chimeric classical swine fever (CSF)-Japanese encephalitis (JE) viral replicon as a non-transmissible vaccine candidate against CSF and JE infections.

    Science.gov (United States)

    Yang, Zhenhua; Wu, Rui; Li, Robert W; Li, Ling; Xiong, Zhongliang; Zhao, Haizhong; Guo, Deyin; Pan, Zishu

    2012-04-01

    A trans-complemented chimeric CSF-JE virus replicon was constructed using an infectious cDNA clone of the CSF virus (CSFV) Alfort/187 strain. The CSFV E2 gene was deleted, and a fragment containing the region encoding a truncated envelope protein (tE, amino acid 292-402, domain III) of JE virus (JEV) was inserted into the resultant plasmid, pA187delE2, to generate the recombinant cDNA clone pA187delE2/JEV-tE. Porcine kidney 15 (PK15) cells that constitutively express the CSFV E2p7 proteins were then transfected with in vitro-transcribed RNA from pA187delE2/JEV-tE. As a result, the chimeric CSF-JE virus replicon particle (VRP), rv187delE2/JEV-tE, was rescued. In a mouse model, immunization with the chimeric CSF-JE VRP induced strong production of JEV-specific antibody and conferred protection against a lethal JEV challenge. Pigs immunized with CSF-JE VRP displayed strong anti-CSFV and anti-JEV antibody responses and protection against CSFV and JEV challenge infections. Our evidence suggests that E2-complemented CSF-JE VRP not only has potential as a live-attenuated non-transmissible vaccine candidate against CSF and JE but also serves as a potential DIVA (Differentiating Infected from Vaccinated Animals) vaccine for CSF in pigs. Together, our data suggest that the non-transmissible chimeric VRP expressing foreign antigenic proteins may represent a promising strategy for bivalent DIVA vaccine design. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. The Composite 259-kb Plasmid of Martelella mediterranea DSM 17316(T)-A Natural Replicon with Functional RepABC Modules from Rhodobacteraceae and Rhizobiaceae.

    Science.gov (United States)

    Bartling, Pascal; Brinkmann, Henner; Bunk, Boyke; Overmann, Jörg; Göker, Markus; Petersen, Jörn

    2017-01-01

    A multipartite genome organization with a chromosome and many extrachromosomal replicons (ECRs) is characteristic for Alphaproteobacteria. The best investigated ECRs of terrestrial rhizobia are the symbiotic plasmids for legume root nodulation and the tumor-inducing (Ti) plasmid of Agrobacterium tumefaciens. RepABC plasmids represent the most abundant alphaproteobacterial replicon type. The currently known homologous replication modules of rhizobia and Rhodobacteraceae are phylogenetically distinct. In this study, we surveyed type-strain genomes from the One Thousand Microbial Genomes (KMG-I) project and identified a roseobacter-specific RepABC-type operon in the draft genome of the marine rhizobium Martelella mediterranea DSM 17316(T). PacBio genome sequencing demonstrated the presence of three circular ECRs with sizes of 593, 259, and 170-kb. The rhodobacteral RepABC module is located together with a rhizobial equivalent on the intermediate sized plasmid pMM259, which likely originated in the fusion of a pre-existing rhizobial ECR with a conjugated roseobacter plasmid. Further evidence for horizontal gene transfer (HGT) is given by the presence of a roseobacter-specific type IV secretion system on the 259-kb plasmid and the rhodobacteracean origin of 62% of the genes on this plasmid. Functionality tests documented that the genuine rhizobial RepABC module from the Martelella 259-kb plasmid is only maintained in A. tumefaciens C58 (Rhizobiaceae) but not in Phaeobacter inhibens DSM 17395 (Rhodobacteraceae). Unexpectedly, the roseobacter-like replication system is functional and stably maintained in both host strains, thus providing evidence for a broader host range than previously proposed. In conclusion, pMM259 is the first example of a natural plasmid that likely mediates genetic exchange between roseobacters and rhizobia.

  9. The Composite 259-kb Plasmid of Martelella mediterranea DSM 17316T–A Natural Replicon with Functional RepABC Modules from Rhodobacteraceae and Rhizobiaceae

    Science.gov (United States)

    Bartling, Pascal; Brinkmann, Henner; Bunk, Boyke; Overmann, Jörg; Göker, Markus; Petersen, Jörn

    2017-01-01

    A multipartite genome organization with a chromosome and many extrachromosomal replicons (ECRs) is characteristic for Alphaproteobacteria. The best investigated ECRs of terrestrial rhizobia are the symbiotic plasmids for legume root nodulation and the tumor-inducing (Ti) plasmid of Agrobacterium tumefaciens. RepABC plasmids represent the most abundant alphaproteobacterial replicon type. The currently known homologous replication modules of rhizobia and Rhodobacteraceae are phylogenetically distinct. In this study, we surveyed type-strain genomes from the One Thousand Microbial Genomes (KMG-I) project and identified a roseobacter-specific RepABC-type operon in the draft genome of the marine rhizobium Martelella mediterranea DSM 17316T. PacBio genome sequencing demonstrated the presence of three circular ECRs with sizes of 593, 259, and 170-kb. The rhodobacteral RepABC module is located together with a rhizobial equivalent on the intermediate sized plasmid pMM259, which likely originated in the fusion of a pre-existing rhizobial ECR with a conjugated roseobacter plasmid. Further evidence for horizontal gene transfer (HGT) is given by the presence of a roseobacter-specific type IV secretion system on the 259-kb plasmid and the rhodobacteracean origin of 62% of the genes on this plasmid. Functionality tests documented that the genuine rhizobial RepABC module from the Martelella 259-kb plasmid is only maintained in A. tumefaciens C58 (Rhizobiaceae) but not in Phaeobacter inhibens DSM 17395 (Rhodobacteraceae). Unexpectedly, the roseobacter-like replication system is functional and stably maintained in both host strains, thus providing evidence for a broader host range than previously proposed. In conclusion, pMM259 is the first example of a natural plasmid that likely mediates genetic exchange between roseobacters and rhizobia. PMID:28983283

  10. Three clustered origins of replication in a promiscuous-plasmid replicon and their differential use in a PolA+ strain and a delta PolA strain of Escherichia coli K-12.

    OpenAIRE

    Banerjee, S. K.; Luck, B T; Kim, H Y; Iyer, V. N.

    1992-01-01

    A 1,197-bp region of the broad-host-range plasmid pCU1 is adequate for its replication. Analysis of replicating molecules containing this region reveals three clustered origins of vegetative replication and replication proceeds bidirectionally from each in a theta mode. In an Escherichia coli polymerase I deletion mutant, utilization of one of these three origins was not detected. The potentiality for origin utilization may therefore be a determinant of replicon host range.

  11. Three clustered origins of replication in a promiscuous-plasmid replicon and their differential use in a PolA+ strain and a delta PolA strain of Escherichia coli K-12.

    Science.gov (United States)

    Banerjee, S K; Luck, B T; Kim, H Y; Iyer, V N

    1992-12-01

    A 1,197-bp region of the broad-host-range plasmid pCU1 is adequate for its replication. Analysis of replicating molecules containing this region reveals three clustered origins of vegetative replication and replication proceeds bidirectionally from each in a theta mode. In an Escherichia coli polymerase I deletion mutant, utilization of one of these three origins was not detected. The potentiality for origin utilization may therefore be a determinant of replicon host range.

  12. PacBio SMRT assembly of a complex multi-replicon genome reveals chlorocatechol degradative operon in a region of genome plasticity.

    Science.gov (United States)

    Ricker, N; Shen, S Y; Goordial, J; Jin, S; Fulthorpe, R R

    2016-07-25

    We have sequenced a Burkholderia genome that contains multiple replicons and large repetitive elements that would make it inherently difficult to assemble by short read sequencing technologies. We illustrate how the integrated long read correction algorithms implemented through the PacBio Single Molecule Real-Time (SMRT) sequencing technology successfully provided a de novo assembly that is a reasonable estimate of both the gene content and genome organization without making any further modifications. This assembly is comparable to related organisms assembled by more labour intensive methods. Our assembled genome revealed regions of genome plasticity for further investigation, one of which harbours a chlorocatechol degradative operon highly homologous to those previously identified on globally ubiquitous plasmids. In an ideal world, this assembly would still require experimental validation to confirm gene order and copy number of repeated elements. However, we submit that particularly in instances where a polished genome is not the primary goal of the sequencing project, PacBio SMRT sequencing provides a financially viable option for generating a biologically relevant genome estimate that can be utilized by other researchers for comparative studies. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. A phase I dose escalation trial of vaccine replicon particles (VRP) expressing prostate-specific membrane antigen (PSMA) in subjects with prostate cancer.

    Science.gov (United States)

    Slovin, Susan F; Kehoe, Marissa; Durso, Robert; Fernandez, Celina; Olson, William; Gao, Jian P; Israel, Robert; Scher, Howard I; Morris, Stephen

    2013-01-30

    PSMA-VRP is a propagation defective, viral replicon vector system encoding PSMA under phase I evaluation for patients with castration resistant metastatic prostate cancer (CRPC). The product is derived from an attenuated strain of the alphavirus, Venezuelan Equine Encephalitis (VEE) virus, and incorporates multiple redundant safety features. In this first in human trial, two cohorts of 3 patients with CRPC metastatic to bone were treated with up to five doses of either 0.9×10(7)IU or 0.36×10(8)IU of PSMA-VRP at weeks 1, 4, 7, 10 and 18, followed by an expansion cohort of 6 patients treated with 0.36×10(8)IU of PSMA-VRP at weeks 1, 4, 7, 10 and 18. No toxicities were observed. In the first dose cohort, no PSMA specific cellular immune responses were seen but weak PSMA-specific signals were observed by ELISA. The remaining 9 patients, which included the higher cohort and the extension cohort, had no PSMA specific cellular responses. PSMA-VRP was well-tolerated at both doses. While there did not appear to be clinical benefit nor robust immune signals at the two doses studied, neutralizing antibodies were produced by both cohorts suggesting that dosing was suboptimal. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Immunogenic and replicative properties of classical swine fever virus replicon particles modified to induce IFN-α/β and carry foreign genes.

    Science.gov (United States)

    Suter, Rolf; Summerfield, Artur; Thomann-Harwood, Lisa J; McCullough, Kenneth C; Tratschin, Jon-Duri; Ruggli, Nicolas

    2011-02-04

    Virus replicon particles (VRP) are genetically engineered infectious virions incapable of generating progeny virus due to partial or complete deletion of at least one structural gene. VRP fulfil the criteria of a safe vaccine and gene delivery system. With VRP derived from classical swine fever virus (CSF-VRP), a single intradermal vaccination protects from disease. Spreading of the challenge virus in the host is however not completely abolished. Parameters that are critical for immunogenicity of CSF-VRP are not well characterized. Considering the importance of type I interferon (IFN-α/β) to immune defence development, we generated IFN-α/β-inducing VRP to determine how this would influence vaccine efficacy. We also evaluated the effect of co-expressing granulocyte macrophage colony-stimulating factor (GM-CSF) in the vaccine context. The VRP were capable of long-term replication in cell culture despite the presence of IFN-α/β. In vivo, RNA replication was essential for the induction of an immune response. IFN-α/β-inducing and GM-CSF-expressing CSF-VRP were similar to unmodified VRP in terms of antibody and peripheral T-cell responses, and in reducing the blood levels of challenge virus RNA. Importantly, the IFN-α/β-inducing VRP did show increased efficacy over the unmodified VRP in terms of B-cell and T-cell responses, when tested with secondary immune responses by in vitro restimulation assay. Copyright © 2010 Elsevier Ltd. All rights reserved.

  15. The Combination of Grazoprevir, a Hepatitis C Virus (HCV) NS3/4A Protease Inhibitor, and Elbasvir, an HCV NS5A Inhibitor, Demonstrates a High Genetic Barrier to Resistance in HCV Genotype 1a Replicons.

    Science.gov (United States)

    Lahser, Frederick C; Bystol, Karin; Curry, Stephanie; McMonagle, Patricia; Xia, Ellen; Ingravallo, Paul; Chase, Robert; Liu, Rong; Black, Todd; Hazuda, Daria; Howe, Anita Y M; Asante-Appiah, Ernest

    2016-05-01

    The selection of resistance-associated variants (RAVs) against single agents administered to patients chronically infected with hepatitis C virus (HCV) necessitates that direct-acting antiviral agents (DAAs) targeting multiple viral proteins be developed to overcome failure resulting from emergence of resistance. The combination of grazoprevir (formerly MK-5172), an NS3/4A protease inhibitor, and elbasvir (formerly MK-8742), an NS5A inhibitor, was therefore studied in genotype 1a (GT1a) replicon cells. Both compounds were independently highly potent in GT1a wild-type replicon cells, with 90% effective concentration (EC90) values of 0.9 nM and 0.006 nM for grazoprevir and elbasvir, respectively. No cross-resistance was observed when clinically relevant NS5A and NS3 RAVs were profiled against grazoprevir and elbasvir, respectively. Kinetic analyses of HCV RNA reduction over 14 days showed that grazoprevir and elbasvir inhibited prototypic NS5A Y93H and NS3 R155K RAVs, respectively, with kinetics comparable to those for the wild-type GT1a replicon. In combination, grazoprevir and elbasvir interacted additively in GT1a replicon cells. Colony formation assays with a 10-fold multiple of the EC90 values of the grazoprevir-elbasvir inhibitor combination suppressed emergence of resistant colonies, compared to a 100-fold multiple for the independent agents. The selected resistant colonies with the combination harbored RAVs that required two or more nucleotide changes in the codons. Mutations in the cognate gene caused greater potency losses for elbasvir than for grazoprevir. Replicons bearing RAVs identified from resistant colonies showed reduced fitness for several cell lines and may contribute to the activity of the combination. These studies demonstrate that the combination of grazoprevir and elbasvir exerts a potent effect on HCV RNA replication and presents a high genetic barrier to resistance. The combination of grazoprevir and elbasvir is currently approved for

  16. Potato virus X and Tobacco mosaic virus-based vectors compatible with the Gateway-TM cloning system

    NARCIS (Netherlands)

    Lacorte, C.C.; Ribeiro, S.G.; Lohuis, H.; Goldbach, R.W.; Prins, M.W.

    2010-01-01

    Virus-based expression vectors are important tools for high-level production of foreign proteins and for gene function analysis through virus induced gene silencing. To exploit further their advantages as fast, high yield replicons, a set of vectors was produced by converting and adapting Potato

  17. Alphavirus replicon DNA expressing HIV antigens is an excellent prime for boosting with recombinant modified vaccinia Ankara (MVA or with HIV gp140 protein antigen.

    Directory of Open Access Journals (Sweden)

    Maria L Knudsen

    Full Text Available Vaccination with DNA is an attractive strategy for induction of pathogen-specific T cells and antibodies. Studies in humans have shown that DNA vaccines are safe, but their immunogenicity needs further improvement. As a step towards this goal, we have previously demonstrated that immunogenicity is increased with the use of an alphavirus DNA-launched replicon (DREP vector compared to conventional DNA vaccines. In this study, we investigated the effect of varying the dose and number of administrations of DREP when given as a prime prior to a heterologous boost with poxvirus vector (MVA and/or HIV gp140 protein formulated in glucopyranosyl lipid A (GLA-AF adjuvant. The DREP and MVA vaccine constructs encoded Env and a Gag-Pol-Nef fusion protein from HIV clade C. One to three administrations of 0.2 μg DREP induced lower HIV-specific T cell and IgG responses than the equivalent number of immunizations with 10 μg DREP. However, the two doses were equally efficient as a priming component in a heterologous prime-boost regimen. The magnitude of immune responses depended on the number of priming immunizations rather than the dose. A single low dose of DREP prior to a heterologous boost resulted in greatly increased immune responses compared to MVA or protein antigen alone, demonstrating that a mere 0.2 μg DREP was sufficient for priming immune responses. Following a DREP prime, T cell responses were expanded greatly by an MVA boost, and IgG responses were also expanded when boosted with protein antigen. When MVA and protein were administered simultaneously following multiple DREP primes, responses were slightly compromised compared to administering them sequentially. In conclusion, we have demonstrated efficient priming of HIV-specific T cell and IgG responses with a low dose of DREP, and shown that the priming effect depends on number of primes administered rather than dose.

  18. A CGMMV genome-replicon vector with partial sequences of coat protein gene efficiently expresses GFP in Nicotiana benthamiana.

    Science.gov (United States)

    Jailani, A Abdul Kader; Solanki, Vikas; Roy, Anirban; Sivasudha, T; Mandal, Bikash

    2017-04-02

    A highly infectious clone of Cucumber green mottle mosaic virus (CGMMV), a cucurbit-infecting tobamovirus was utilized for designing of gene expression vectors. Two versions of vector were examined for their efficacy in expressing the green fluorescent protein (GFP) in Nicotiana benthamiana. When the GFP gene was inserted at the stop codon of coat protein (CP) gene of the CGMMV genome without any read-through codon, systemic expression of GFP, as well as virion formation and systemic symptoms expression were obtained in N. benthamiana. The qRT-PCR analysis showed 23 fold increase of GFP over actin at 10days post inoculation (dpi), which increased to 45 fold at 14dpi and thereafter the GFP expression was significantly declined. Further, we show that when the most of the CP sequence is deleted retaining only the first 105 nucleotides, the shortened vector containing GFP in frame of original CP open reading frame (ORF) resulted in 234 fold increase of GFP expression over actin at 5dpi in N. benthamiana without the formation of virions and disease symptoms. Our study demonstrated that a simple manipulation of CP gene in the CGMMV genome while preserving the translational frame of CP resulted in developing a virus-free, rapid and efficient foreign protein expression system in the plant. The CGMMV based vectors developed in this study may be potentially useful for the production of edible vaccines in cucurbits. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. [The interrelation between changes in the structural organization of replicon clusters, a retarded fork displacement rate and the high level of spontaneous SCEs in form II of xeroderma pigmentosum].

    Science.gov (United States)

    Barenfel'd, L S; Nergadze, S G; Pleskach, N M; Mikhel'son, V M

    1992-01-01

    A cytogenetic observation, that the sister chromatid exchanges (SCE) occur 3 times more frequently in a special form of xeroderma pigmentosum--XPII than in the norm, prompted a study of DNA replication in this rare disease. Using DNA fiber autoradiography, the rate of fork movement and the frequency of initiation in the adjacent clusters of replicons were estimated. The rate of fork movement was significantly slower than that in classical XP and in normal cells. Here evidence was provided on another defect in DNA replication in XPII that involves a significantly decreased number of simultaneously operating adjacent clusters of replicons, which results in a decreased rate of DNA chain-growth. According to the Painter replication model for SCE, the exchanges arise due to double-strand DNA breaks occurring on the border between two adjacent clusters, respectively, completely and partially replicated. A retarded fork-displacement rate together with a decreased rate of DNA-chain growth may cause this situation to persist longer than in the norm. Thus, our data provide a further support of the replication model for SCE. A similar combination of cytogenetic and molecular defects has been obtained earlier in the Bloom syndrome cells.

  20. Alphavirus-Based Vaccines.

    Science.gov (United States)

    Lundstrom, Kenneth

    2016-01-01

    Alphavirus vectors based on Semliki Forest virus, Sindbis virus, and Venezuelan equine encephalitis virus have been widely applied for vaccine development. Naked RNA replicons, recombinant viral particles, and layered DNA vectors have been subjected to immunization in preclinical animal models with antigens for viral targets and tumor antigens. Moreover, a limited number of clinical trials have been conducted in humans. Vaccination with alphavirus vectors has demonstrated efficient immune responses and has showed protection against challenges with lethal doses of virus and tumor cells, respectively. Moreover, vaccines have been developed against alphaviruses causing epidemics such as Chikungunya virus.

  1. Kunjin virus replicons: an RNA-based, non-cytopathic viral vector system for protein production, vaccine and gene therapy applications

    NARCIS (Netherlands)

    Pijlman, G.P.; Suhrbier, A.; Khromykh, A.A.

    2006-01-01

    The application of viral vectors for gene expression and delivery is rapidly evolving, with several entering clinical trials. However, a number of issues, including safety, gene expression levels, cell selectivity and antivector immunity, are driving the search for new vector systems. A number of

  2. Characterization of a Minimal pKW2124 Replicon from Weissella cibaria KLC140 and Its Application for the Construction of the Weissella Expression Vector pKUCm1

    Directory of Open Access Journals (Sweden)

    Hye-Jin eKu

    2015-02-01

    Full Text Available A 2.1-kb plasmid was previously isolated from Weissella cibaria KLC140 in kimchi and cloned into pUC19 along with the slpA and gfp genes, resulting in an 8.6-kb pKWCSLGFP construct for use as a novel surface display vector. To reduce the size of the vector, the minimal replicon of pKW2124 was determined. The pKW2124 plasmid contains a putative origin of replication (ori, a potential ribosomal binding site (RBS, and the repA gene encoding a plasmid replication protein. To conduct the minimal replicon experiment, three different PCR products (MR1, ori + RBS + repA; MR2, RBS + repA; MR2’, repA; MR3, fragment of repA were obtained and cloned into pUC19 (pKUCm1, pKUCm2, and pKUCm3, respectively containing the chloramphenicol acetyltransferase (CAT gene. These three constructed vectors were electroporated into W. confusa ATCC 10881 with different transformation efficiencies of 1.5×105 CFU/μg, 1.3×101 CFU/μg, and no transformation, respectively, suggesting that the putative ori, RBS, and repA gene are essential for optimum plasmid replication. Subsequent segregational plasmid stability testing of pKUCm1 and pKUCm2 showed that the vector pKUCm1 is highly stable up to 100 generations but pKUCm2 was completely lost after 60 generations, suggesting that the putative ori may be important for plasmid stability in the host strain. In addition, a host range test of pKUCm1 revealed that it has a broad host range spectrum including Weissella, Lactococcus, Leuconostoc, and even Lactobacillus. To verify the application of pKUCm1, the β-galactosidase gene and its promoter region from W. cibaria KSD1 were cloned in the vector, resulting in pKUGal. Expression of the β-galactosidase gene was confirmed using blue-white screening after IPTG induction. The small and stable pKUGal vector will be useful for gene transfer, expression, and manipulation in the Weissella genome and in other lactic acid bacteria.

  3. Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1

    Science.gov (United States)

    Ku, Hye-Jin; Park, Myeong Soo; Lee, Ju-Hoon

    2015-01-01

    A 2.1-kb plasmid was previously isolated from Weissella cibaria KLC140 in kimchi and cloned into pUC19 along with the slpA and gfp genes, resulting in an 8.6-kb pKWCSLGFP construct for use as a novel surface display vector. To reduce the size of the vector, the minimal replicon of pKW2124 was determined. The pKW2124 plasmid contains a putative origin of replication (ori), a potential ribosomal binding site (RBS), and the repA gene encoding a plasmid replication protein. To conduct the minimal replicon experiment, four different PCR products (MR1, ori+RBS+repA; MR2, RBS+repA; MR2’, repA; MR3, fragment of repA) were obtained and cloned into pUC19 (pKUCm1, pKUCm2, pKUCm2’, and pKUCm3, respectively) containing the chloramphenicol acetyltransferase (CAT) gene. These constructed vectors were electroporated into W. confusa ATCC 10881 with different transformation efficiencies of 1.5 × 105 CFU/μg, 1.3 × 101 CFU/μg, and no transformation, respectively, suggesting that the putative ori, RBS, and repA gene are essential for optimum plasmid replication. Subsequent segregational plasmid stability testing of pKUCm1 and pKUCm2 showed that the vector pKUCm1 is highly stable up to 100 generations but pKUCm2 was completely lost after 60 generations, suggesting that the putative ori may be important for plasmid stability in the host strain. In addition, a host range test of pKUCm1 revealed that it has a broad host range spectrum including Weissella, Lactococcus, Leuconostoc, and even Lactobacillus. To verify the application of pKUCm1, the β-galactosidase gene and its promoter region from W. cibaria KSD1 were cloned in the vector, resulting in pKUGal. Expression of the β-galactosidase gene was confirmed using blue-white screening after IPTG induction. The small and stable pKUGal vector will be useful for gene transfer, expression, and manipulation in the Weissella genome and in other lactic acid bacteria. PMID:25691882

  4. Structure-based design, synthesis, and biological evaluation of 1,1-dioxoisothiazole and benzo[b]thiophene-1,1-dioxide derivatives as novel inhibitors of hepatitis C virus NS5B polymerase.

    Science.gov (United States)

    Kim, Sun Hee; Tran, Martin T; Ruebsam, Frank; Xiang, Alan X; Ayida, Benjamin; McGuire, Helen; Ellis, David; Blazel, Julie; Tran, Chinh V; Murphy, Douglas E; Webber, Stephen E; Zhou, Yuefen; Shah, Amit M; Tsan, Mei; Showalter, Richard E; Patel, Rupal; Gobbi, Alberto; LeBrun, Laurie A; Bartkowski, Darian M; Nolan, Thomas G; Norris, Daniel A; Sergeeva, Maria V; Kirkovsky, Leo; Zhao, Qiang; Han, Qing; Kissinger, Charles R

    2008-07-15

    A novel series of HCV NS5B polymerase inhibitors comprising 1,1-dioxoisothiazoles and benzo[b]thiophene-1,1-dioxides were designed, synthesized, and evaluated. SAR studies guided by structure-based design led to the identification of a number of potent NS5B inhibitors with nanomolar IC(50) values. The most potent compound exhibited IC(50) less than 10nM against the genotype 1b HCV polymerase and EC(50) of 70 nM against a genotype 1b replicon in cell culture. The DMPK properties of selected compounds were also evaluated.

  5. Transfer of tetracycline resistance gene (tet ) between replicons in ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-09-17

    Sep 17, 2008 ... (71.4%), gentamicin (43.5%), nalidixic acid (38.3%) and nitrofurantoin (7.7%; Table 2). The results of antibiotic susceptibility test also indicated that E. coli showed higher rates of resistance to most of the antibiotics. A total of 110 (71.4%) isolates were resistant to tetracy- cline. Tetracycline resistant gene tetr ...

  6. Type I Interferons are essential for the efficacy of replicase-based DNA vaccines.

    Science.gov (United States)

    Leitner, Wolfgang W; Bergmann-Leitner, Elke S; Hwang, Leroy N; Restifo, Nicholas P

    2006-06-12

    The immunogenicity and efficacy of nucleic acid vaccines can be greatly enhanced when antigen production is under the control of an alphaviral replicase enzyme. However, replicase-mediated mRNA overproduction does not necessarily result in enhanced antigen level. Instead, the strong adaptive immune response of alphavirus replicon-based vectors is due to their production of double-stranded RNA (dsRNA) intermediates, which trigger innate immunity. Because viral infections are known to trigger innate immune responses that lead to the rapid production of Type I Interferons (IFNs), namely IFN-alpha and IFN-beta, we investigated the role of Type I IFNs in the enhanced immunogenicity of replicase-based DNA vaccines. In vitro, cells transfected with replicase-based plasmids produce significantly more Type I IFNs than cells transfected with a conventional DNA plasmid. In vivo, replicase-based DNA vaccines yield stronger humoral responses in the absence of Type I IFN signaling but the lack of this signaling pathway in IFN-alphabeta receptor-/- (knockout) mice abolishes T cell mediated efficacy against tumors of both conventional and alphavirus replicase-based DNA vaccines. Moreover, the co-delivery of an IFNalpha-encoding plasmid significantly improved the efficacy of a weakly immunogenic conventional plasmid. These results suggest a central role for Type I IFNs in the mechanism of replicase-based DNA vaccines and indicate that vaccines can be enhanced by enabling their capacity to triggering innate anti-viral defense pathways.

  7. Genome-based genetic tool development for Bacillus methanolicus: theta- and rolling circle-replicating plasmids for inducible gene expression and application to methanol-based cadaverine production

    Directory of Open Access Journals (Sweden)

    Marta Irla

    2016-09-01

    Full Text Available Bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene. For independent complementary gene expression control, the heterologous xylose-inducible system from B. megaterium was employed and a two-plasmid gene expression system was developed. Four different replicons for expression vectors were compared with respect to their copy number and stability. As an application example, methanol-based production of cadaverine was shown to be improved from 11.3 g/L to 17.5 g/L when a heterologous lysine decarboxylase gene cadA was expressed from a theta-replicating rather than a rolling-circle replicating vector. The current work on inducible promoter systems and compatible theta- or rolling circle-replicating vectors is an important extension of the poorly developed B. methanolicus genetic toolbox, valuable for genetic engineering and further exploration of this bacterium.

  8. Viral evasion of a bacterial suicide system by RNA-based molecular mimicry enables infectious altruism.

    Directory of Open Access Journals (Sweden)

    Tim R Blower

    Full Text Available Abortive infection, during which an infected bacterial cell commits altruistic suicide to destroy the replicating bacteriophage and protect the clonal population, can be mediated by toxin-antitoxin systems such as the Type III protein-RNA toxin-antitoxin system, ToxIN. A flagellum-dependent bacteriophage of the Myoviridae, ΦTE, evolved rare mutants that "escaped" ToxIN-mediated abortive infection within Pectobacterium atrosepticum. Wild-type ΦTE encoded a short sequence similar to the repetitive nucleotide sequence of the RNA antitoxin, ToxI, from ToxIN. The ΦTE escape mutants had expanded the number of these "pseudo-ToxI" genetic repeats and, in one case, an escape phage had "hijacked" ToxI from the plasmid-borne toxIN locus, through recombination. Expression of the pseudo-ToxI repeats during ΦTE infection allowed the phage to replicate, unaffected by ToxIN, through RNA-based molecular mimicry. This is the first example of a non-coding RNA encoded by a phage that evolves by selective expansion and recombination to enable viral suppression of a defensive bacterial suicide system. Furthermore, the ΦTE escape phages had evolved enhanced capacity to transduce replicons expressing ToxIN, demonstrating virus-mediated horizontal transfer of genetic altruism.

  9. Tattoo Delivery of a Semliki Forest Virus-Based Vaccine Encoding Human Papillomavirus E6 and E7

    Science.gov (United States)

    van de Wall, Stephanie; Walczak, Mateusz; van Rooij, Nienke; Hoogeboom, Baukje-Nynke; Meijerhof, Tjarko; Nijman, Hans W.; Daemen, Toos

    2015-01-01

    The skin is an attractive organ for immunization because of the presence of antigen-presenting cells. Intradermal delivery via tattooing has demonstrated superior vaccine immunogenicity of DNA vaccines in comparison to conventional delivery methods. In this study, we explored the efficacy of tattoo injection of a tumor vaccine based on recombinant Semliki Forest virus replicon particles (rSFV) targeting human papillomavirus (HPV). Tattoo injection of rSFV particles resulted in antigen expression in both the skin and draining lymph nodes. In comparison with intramuscular injection, the overall antigen expression determined at the site of administration and draining lymph nodes was 10-fold lower upon tattoo injection. Delivery of SFV particles encoding the E6 and E7 antigens of human papillomavirus type 16 (SFVeE6,7) via tattooing resulted in HPV-specific cytotoxic T cells and in vivo therapeutic antitumor response. Strikingly, despite the observed lower overall transgene expression, SFVeE6,7 delivered via tattoo injection resulted in higher or equal levels of immune responses as compared to intramuscular injection. The intrinsic immunogenic potential of tattooing provides a benefit for immunotherapy based on an alphavirus. PMID:26343186

  10. Tattoo Delivery of a Semliki Forest Virus-Based Vaccine Encoding Human Papillomavirus E6 and E7

    Directory of Open Access Journals (Sweden)

    Stephanie van de Wall

    2015-03-01

    Full Text Available The skin is an attractive organ for immunization because of the presence of antigen-presenting cells. Intradermal delivery via tattooing has demonstrated superior vaccine immunogenicity of DNA vaccines in comparison to conventional delivery methods. In this study, we explored the efficacy of tattoo injection of a tumor vaccine based on recombinant Semliki Forest virus replicon particles (rSFV targeting human papillomavirus (HPV. Tattoo injection of rSFV particles resulted in antigen expression in both the skin and draining lymph nodes. In comparison with intramuscular injection, the overall antigen expression determined at the site of administration and draining lymph nodes was 10-fold lower upon tattoo injection. Delivery of SFV particles encoding the E6 and E7 antigens of human papillomavirus type 16 (SFVeE6,7 via tattooing resulted in HPV-specific cytotoxic T cells and in vivo therapeutic antitumor response. Strikingly, despite the observed lower overall transgene expression, SFVeE6,7 delivered via tattoo injection resulted in higher or equal levels of immune responses as compared to intramuscular injection. The intrinsic immunogenic potential of tattooing provides a benefit for immunotherapy based on an alphavirus.

  11. Novel plasmid-based genetic tools for the study of promoters and terminators in Streptococcus pneumoniae and Enterococcus faecalis.

    Science.gov (United States)

    Ruiz-Cruz, Sofía; Solano-Collado, Virtu; Espinosa, Manuel; Bravo, Alicia

    2010-11-01

    Promoter-probe and terminator-probe plasmid vectors make possible to rapidly examine whether particular sequences function as promoter or terminator signals in various genetic backgrounds and under diverse environmental stimuli. At present, such plasmid-based genetic tools are very scarce in the Gram-positive pathogenic bacteria Streptococcus pneumoniae and Enterococcus faecalis. Hence, we developed novel promoter-probe and terminator-probe vectors based on the Streptococcus agalactiae pMV158 plasmid, which replicates autonomously in numerous Gram-positive bacteria. As reporter gene, a gfp allele encoding a variant of the green fluorescent protein was used. These genetic tools were shown to be suitable to assess the activity of promoters and terminators (both homologous and heterologous) in S. pneumoniae and E. faecalis. In addition, the promoter-probe vector was shown to be a valuable tool for the analysis of regulated promoters in vivo, such as the promoter of the pneumococcal fuculose kinase gene. These new plasmid vectors will be very useful for the experimental verification of predicted promoter and terminator sequences, as well as for the construction of new inducible-expression vectors. Given the promiscuity exhibited by the pMV158 replicon, these vectors could be used in a variety of Gram-positive bacteria. Copyright © 2010 Elsevier B.V. All rights reserved.

  12. Individual and bivalent vaccines against botulinum neurotoxin serotypes A and B using DNA-based Semliki Forest virus vectors.

    Science.gov (United States)

    Yu, Yunzhou; Yu, Jiyu; Li, Na; Wang, Shuang; Yu, Weiyuan; Sun, Zhiwei

    2009-10-19

    We evaluated individual and bivalent replicon vaccines against Clostridiumbotulinum neurotoxin serotypes A (BoNT/A) or B (BoNT/B). The DNA replicon vaccine (pSCARSBHc) encoding the Hc domain of BoNT/B (BHc) induced better responses and protection against BoNT/B mouse challenge than conventional DNA vaccine. The dual-expressing DNA vaccine (pSCARSA/BHc) protected similarly to a DNA replicon vaccine mixture (pSCARSAHc+pSCARSBHc). Additionally, recombinant SFV particles, VRP-AHc or VRP-BHc, protected mice from high-dose BoNT/A or BoNT/B challenge, respectively. Mice given either dual-expressing VRP-A/BHc or mixture of VRP-AHc and VRP-BHc were protected from challenge with serotype A/B mixtures. These data justify further testing in other animals or humans.

  13. Prevalence estimation of tick-borne encephalitis virus (TBEV) antibodies in dogs from Finland using novel dog anti-TBEV IgG MAb-capture and IgG immunofluorescence assays based on recombinant TBEV subviral particles.

    Science.gov (United States)

    Levanov, Lev; Vera, Cristina Pérez; Vapalahti, Olli

    2016-07-01

    Tick-borne encephalitis (TBE) is one of the most dangerous human neurological infections occurring in Europe and Northern parts of Asia with thousands of cases and millions vaccinated against it. The risk of TBE might be assessed through analyses of the samples taken from wildlife or from animals which are in close contact with humans. Dogs have been shown to be a good sentinel species for these studies. Serological assays for diagnosis of TBE in dogs are mainly based on purified and inactivated TBEV antigens. Here we describe novel dog anti-TBEV IgG monoclonal antibody (MAb)-capture assay which is based on TBEV prME subviral particles expressed in mammalian cells from Semliki Forest virus (SFV) replicon as well as IgG immunofluorescence assay (IFA) which is based on Vero E6 cells transfected with the same SFV replicon. We further demonstrate their use in a small-scale TBEV seroprevalence study of dogs representing different regions of Finland. Altogether, 148 dog serum samples were tested by novel assays and results were compared to those obtained with a commercial IgG enzyme immunoassay (EIA), hemagglutination inhibition test and IgG IFA with TBEV infected cells. Compared to reference tests, the sensitivities of the developed assays were 90-100% and the specificities of the two assays were 100%. Analysis of the dog serum samples showed a seroprevalence of 40% on Åland Islands and 6% on Southwestern archipelago of Finland. In conclusion, a specific and sensitive EIA and IFA for the detection of IgG antibodies in canine sera were developed. Based on these assays the seroprevalence of IgG antibodies in dogs from different regions of Finland was assessed and was shown to parallel the known human disease burden as the Southwestern archipelago and Åland Islands in particular had considerable dog TBEV antibody prevalence and represent areas with high risk of TBE for humans. Copyright © 2016 Elsevier GmbH. All rights reserved.

  14. Production of Human papillomavirus pseudovirions in plants and their use in pseudovirion-based neutralisation assays in mammalian cells.

    Science.gov (United States)

    Lamprecht, Renate L; Kennedy, Paul; Huddy, Suzanne M; Bethke, Susanne; Hendrikse, Megan; Hitzeroth, Inga I; Rybicki, Edward P

    2016-02-08

    Human papillomaviruses (HPV) cause cervical cancer and have recently also been implicated in mouth, laryngeal and anogenital cancers. There are three commercially available prophylactic vaccines that show good efficacy; however, efforts to develop second-generation vaccines that are more affordable, stable and elicit a wider spectrum of cross-neutralising immunity are still ongoing. Testing antisera elicited by current and candidate HPV vaccines for neutralizing antibodies is done using a HPV pseudovirion (PsV)-based neutralisation assay (PBNA). PsVs are produced by transfection of mammalian cell cultures with plasmids expressing L1 and L2 capsid proteins, and a reporter gene plasmid, a highly expensive process. We investigated making HPV-16 PsVs in plants, in order to develop a cheaper alternative. The secreted embryonic alkaline phosphatase (SEAP) reporter gene and promoter were cloned into a geminivirus-derived plant expression vector, in order to produce circular dsDNA replicons. This was co-introduced into Nicotiana benthamiana plants with vectors expressing L1 and L2 via agroinfiltration, and presumptive PsVs were purified. The PsVs contained DNA, and could be successfully used for PBNA with anti-HPV antibodies. This is the first demonstration of the production of mammalian pseudovirions in plants, and the first demonstration of the potential of plants to make DNA vaccines.

  15. An alphavirus vector-based tetravalent dengue vaccine induces a rapid and protective immune response in macaques that differs qualitatively from immunity induced by live virus infection.

    Science.gov (United States)

    White, Laura J; Sariol, Carlos A; Mattocks, Melissa D; Wahala M P B, Wahala; Yingsiwaphat, Vorraphun; Collier, Martha L; Whitley, Jill; Mikkelsen, Rochelle; Rodriguez, Idia V; Martinez, Melween I; de Silva, Aravinda; Johnston, Robert E

    2013-03-01

    Despite many years of research, a dengue vaccine is not available, and the more advanced live attenuated vaccine candidate in clinical trials requires multiple immunizations with long interdose periods and provides low protective efficacy. Here, we report important contributions to the development of a second-generation dengue vaccine. First, we demonstrate that a nonpropagating vaccine vector based on Venezuelan equine encephalitis virus replicon particles (VRP) expressing two configurations of dengue virus E antigen (subviral particles [prME] and soluble E dimers [E85]) successfully immunized and protected macaques against dengue virus, while antivector antibodies did not interfere with a booster immunization. Second, compared to prME-VRP, E85-VRP induced neutralizing antibodies faster, to higher titers, and with improved protective efficacy. Third, this study is the first to map antigenic domains and specificities targeted by vaccination versus natural infection, revealing that, unlike prME-VRP and live virus, E85-VRP induced only serotype-specific antibodies, which predominantly targeted EDIII, suggesting a protective mechanism different from that induced by live virus and possibly live attenuated vaccines. Fourth, a tetravalent E85-VRP dengue vaccine induced a simultaneous and protective response to all 4 serotypes after 2 doses given 6 weeks apart. Balanced responses and protection in macaques provided further support for exploring the immunogenicity and safety of this vaccine candidate in humans.

  16. Heterologous Prime-Boost Immunizations with a Virosomal and an Alphavirus Replicon Vaccine

    NARCIS (Netherlands)

    Walczak, Mateusz; de Mare, Arjan; Riezebos-Brilman, Annelies; Regts, Joke; Hoogeboom, Baukje-Nynke; Visser, Jeroen T.; Fiedler, Marc; Jansen-Duerr, Pidder; van der Zee, Ate G. J.; Nijman, Hans W.; Wilschut, Jan; Daemen, Toos

    2011-01-01

    Heterologous prime-boost immunization strategies in general establish higher frequencies of antigen-specific T lymphocytes than homologous prime-boost protocols or single immunizations. We developed virosomes and recombinant Semliki Forest virus (rSFV) as antigen delivery systems, each capable of

  17. Toward Viral Vaccine Development: A Modified Venezuelan Equine Encephalitis Replicon as Strategy for Optimizing Immunogenicity

    Science.gov (United States)

    2007-04-13

    Morrissy, J. White, and B. T. Eaton. 2002. A rapid immune plaque assay for the detection of Hendra and Nipah viruses and anti- virus antibodies. J...HIV-1 envelope glycoprotein and to the fusion (F) and attachment (G) glycoproteins of two zoonotic pathogens, Hendra and Nipah viruses , in a murine...VII. Henipaviruses Introduction: Henipavirus is a genus of the family Paramyxoviridae, order Mononegavirales containing two members, Hendra virus

  18. Cellular expression of a functional nodavirus RNA replicon from vaccinia virus vectors.

    Science.gov (United States)

    Ball, L A

    1992-04-01

    RNA replication provides a powerful means for the amplification of RNA, but to date it has been found to occur naturally only among RNA viruses. In an attempt to harness this process for the amplification of heterologous mRNAs, both an RNA replicase and its corresponding RNA templates have been expressed in functional form, using vaccinia virus-bacteriophage T7 RNA polymerase vectors. Plasmids were constructed which contained in 5'-to-3' order (i) a bacteriophage T7 promoter; (ii) a full-length cDNA encoding either the RNA replicase (RNA 1) or the coat protein (RNA 2) of flock house virus (FHV), (iii) a cDNA sequence that encoded the self-cleaving ribozyme of satellite tobacco ringspot virus, and (iv) a T7 transcriptional terminator. Both in vitro and in vivo, circular plasmids of this structure were transcribed by T7 RNA polymerase to produce RNAs with sizes that closely resembled those of the two authentic FHV genomic RNAs, RNA 1 and RNA 2. In baby hamster kidney cells that expressed authentic FHV RNA replicase, the RNA 2 (coat protein) transcripts were accurately replicated. Moreover, the RNA 1 (replicase) transcripts directed the synthesis of an enzyme that could replicate not only authentic virion-derived FHV RNA but also the plasmid-derived transcripts themselves. Under the latter conditions, replicative amplification of the RNA transcripts ensued and resulted in a high rate of synthesis of the encoded proteins. This successful expression from a DNA vector of the complex biological process of RNA replication will greatly facilitate studies of its mechanism and is a major step towards the goal of harnessing RNA replication for mRNA amplification.

  19. Statin (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor-based therapy for hepatitis C virus (HCV infection-related diseases in the era of direct-acting antiviral agents [version 3; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Sara Sobhy Kishta

    2017-01-01

    Full Text Available Recent improvements have been made in the treatment of hepatitis C virus (HCV infection with the introduction of direct-acting antiviral agents (DAAs. However, despite successful viral clearance, many patients continue to have HCV-related disease progression. Therefore, new treatments must be developed to achieve viral clearance and prevent the risk of HCV-related diseases. In particular, the use of pitavastatin together with DAAs may improve the antiviral efficacy as well as decrease the progression of liver fibrosis and the incidence of HCV-related hepatocellular carcinoma. To investigate the management methods for HCV-related diseases using pitavastatin and DAAs, clinical trials should be undertaken. However, concerns have been raised about potential drug interactions between statins and DAAs. Therefore, pre-clinical trials using a replicon system, human hepatocyte-like cells, human neurons and human cardiomyocytes from human-induced pluripotent stem cells should be conducted. Based on these pre-clinical trials, an optimal direct-acting antiviral agent could be selected for combination with pitavastatin and DAAs. Following the pre-clinical trial, the combination of pitavastatin and the optimal direct-acting antiviral agent should be compared to other combinations of DAAs (e.g., sofosbuvir and velpatasvir according to the antiviral effect on HCV infection, HCV-related diseases and cost-effectiveness.

  20. A cell-based assay for RNA synthesis by the HCV polymerase reveals new insights on mechanism of polymerase inhibitors and modulation by NS5A.

    Directory of Open Access Journals (Sweden)

    C T Ranjith-Kumar

    Full Text Available RNA synthesis by the genotype 1b hepatitis C virus (HCV polymerase (NS5B transiently expressed in Human embryonic kidney 293T cells or liver hepatocytes was found to robustly stimulate RIG-I-dependent luciferase production from the interferon β promoter in the absence of exogenously provided ligand. This cell-based assay, henceforth named the 5BR assay, could be used to examine HCV polymerase activity in the absence of other HCV proteins. Mutations that decreased de novo initiated RNA synthesis in biochemical assays decreased activation of RIG-I signaling. In addition, NS5B that lacks the C-terminal transmembrane helix but remains competent for RNA synthesis could activate RIG-I signaling. The addition of cyclosporine A to the cells reduced luciferase levels without affecting agonist-induced RIG-I signaling. Furthermore, non-nucleoside inhibitor benzothiadiazines (BTDs that bind within the template channel of the 1b NS5B were found to inhibit the readout from the 5BR assay. Mutation M414T in NS5B that rendered the HCV replicon resistant to BTD was also resistant to BTDs in the 5BR assay. Co-expression of the HCV NS5A protein along with NS5B and RIG-I was found to inhibit the readout from the 5BR assay. The inhibition by NS5A was decreased with the removal of the transmembrane helix in NS5B. Lastly, NS5B from all six major HCV genotypes showed robust activation of RIG-I in the 5BR assay. In summary, the 5BR assay could be used to validate inhibitors of the HCV polymerase as well as to elucidate requirements for HCV-dependent RNA synthesis.

  1. Efficient single tobamoviral vector-based bioproduction of broadly neutralizing anti-HIV-1 monoclonal antibody VRC01 in Nicotiana benthamiana plants and utility of VRC01 in combination microbicides.

    Science.gov (United States)

    Hamorsky, Krystal Teasley; Grooms-Williams, Tiffany W; Husk, Adam S; Bennett, Lauren J; Palmer, Kenneth E; Matoba, Nobuyuki

    2013-05-01

    Broadly neutralizing monoclonal antibodies (bnMAbs) may offer powerful tools for HIV-1 preexposure prophylaxis, such as topical microbicides. However, this option is hampered due to expensive MAb biomanufacturing based on mammalian cell culture. To address this issue, we developed a new production system for bnMAb VRC01 in Nicotiana benthamiana plants using a tobamovirus replicon vector. Unlike conventional two-vector-based expression, this system was designed to overexpress full-length IgG1 from a single polypeptide by means of kex2p-like enzyme recognition sites introduced between the heavy and light chains. An enzyme-linked immunosorbent assay (ELISA) revealed that gp120-binding VRC01 IgG1 was maximally accumulated on 5 to 7 days following vector inoculation, yielding ~150 mg of the bnMAb per kg of fresh leaf material. The plant-made VRC01 (VRC01p) was efficiently purified by protein A affinity followed by hydrophobic-interaction chromatography. ELISA, surface plasmon resonance, and an HIV-1 neutralization assay demonstrated that VRC01p has gp120-binding affinity and HIV-1-neutralization capacity virtually identical to the human-cell-produced counterpart. To advance VRC01p's use in topical microbicides, we analyzed combinations of the bnMAb with other microbicide candidates holding distinct antiviral mechanisms in an HIV-1 neutralization assay. VRC01p exhibited clear synergy with the antiviral lectin griffithsin, the CCR5 antagonist maraviroc, and the reverse transcriptase inhibitor tenofovir in multiple CCR5-tropic HIV-1 strains from clades A, B, and C. In summary, VRC01p is amenable to robust, rapid, and large-scale production and may be developed as an active component in combination microbicides with other anti-HIV agents such as antiviral lectins, CCR5 antagonists, and reverse transcriptase inhibitors.

  2. Identification of the minimal replicon of plasmid pMEA300 of the methylotrophic actinomycete Amycolatopsis methanolica

    NARCIS (Netherlands)

    Vrijbloed, J.W.; Jelínková, M.; Hessels, G.I.; Dijkhuizen, L.

    1995-01-01

    The actinomycete Amycolatopsis methanolica contains a 13.3 kb plasmid (pMEA300), capable of enhancing the spontaneous mutation frequency of its host. Depending on the growth medium pMEA300 is not only maintained as an integrated element but can additionally be present as a multicopy, autonomously

  3. EXPRESSION AND SELF-ASSEMBLY OF NORWALK VIRUS CAPSID PROTEIN FROM VENEZUELAN EQUINE ENCEPHALITIS VIRUS REPLICONS. (R826139)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  4. Antimicrobial resistance and plasmid replicons in Yersinia enterocolitica strains isolated in Brazil in 30 years.(Brief communication)

    National Research Council Canada - National Science Library

    Falcao, Juliana P; Frazao, Miliane R; Andrade, Leonardo N; Darini, Ana L.C

    2017-01-01

    .... However, in Brazil the isolation and the study of Y. enterocolitica are not common and therefore information about the antimicrobial resistance profile of this species in this country is scarce...

  5. Superior protection conferred by inactivated whole virus vaccine over subunit and DNA vaccines against salmonid alphavirus infection in Atlantic salmon (Salmo salar L.).

    Science.gov (United States)

    Xu, Cheng; Mutoloki, Stephen; Evensen, Øystein

    2012-06-06

    Salmonid alphavirus 3 (SAV-3) is an emerging pathogen in Norwegian salmon farming and causes severe annual losses. We studied the immunogenicity and protective ability of subunit and DNA vaccines based on E1 and E2 spike proteins of salmonid alphavirus subtype 3 (SAV-3), and compared these to an experimental inactivated, whole virus (IWV) vaccine in Atlantic salmon. The antigens were delivered as water-in-oil emulsions for the subunit and inactivated vaccines and non-formulated for the DNA vaccines. The IWV and the E2 subunit prime-boost groups had circulating neutralizing antibodies at challenge, correlating with high protection against lethal challenge and 3-log(10) reduction of virus titer in heart for the IWV group. Prime-boost with E1 subunit vaccine also conferred significant protection against mortality, but did not correlate with neutralizing antibody levels. Protection against pathology in internal organs was only seen for the IWV group. Prime-boost with E1 and E2 DNA vaccines showed marginal protection in terms of reduction of viral replication in target organs and protection against mortality was not different from controls. The IWV group showed significant upregulation of IFNγ and IL2 mRNA expression at 4 weeks post challenge possibly indicating that other mechanisms in addition to antibody responses play a role in mediating protection against infection. This is the first report comparing the immunogenicity and protection against mortality for IWV vaccines and spike protein subunit and DNA vaccines against salmonid alphavirus infection in Atlantic salmon. The IWV vaccine has superior immunogenicity over sub-unit and DNA vaccines. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Directory of Open Access Journals (Sweden)

    Sabrina R.A. Queiroz

    2013-01-01

    Full Text Available RNA replicon derived from Flavivirus genome is a valuable tool for studying viral replication independent of virion assembly and maturation, besides being a great potencial for heterologous gene expression. In this study we described the construction of subgenomic replicons of yellow fever virus by yeast-based homologous recombination technique. The plasmid containing the yellow fever 17D strain replicon (pBSC-repYFV-17D, previously characterized, was handled to heterologous expression of the green fluorescent protein (repYFV-17D-GFP and firefly luciferase (repYFV-17D-Luc reporter genes. Both replicons were constructed by homologous recombination between the linearized vector pBSC-repYFV-17D and the PCR product containing homologous 25 nucleotides ends incorporated into PCR primers. The genomic organization of these constructs is similar to repYFV-17D, but with insertion of the reporter gene between the remaining 63 N-terminal nucleotides of the capsid protein and 72 C-terminal nucleotides of the E protein. The replicons repYFV-17D-GFP and repYFV-17D-Luc showed efficient replication and expression of the reporter genes. The yeast-based homologous recombination technique used in this study proved to be applicable for manipulation of the yellow fever virus genome in order to construct subgenomic replicons.O replicon de RNA derivado do genoma de Flavivirus é uma ferramenta valiosa para o estudo de replicação viral independente da montagem e da maturação do virion, além de possuir um grande potencial para expressão de genes heterólogos. Neste estudo nós descrevemos a construção de replicons subgenômicos do vírus da febre amarela utilizando a técnica de recombinação homóloga em levedura. O plasmídeo contendo o replicon do vírus febre amarela cepa 17D (pBSC-repYFV-17D, caracterizado anteriormente, foi manipulado para a expressão heteróloga dos genes repórteres green fluorescent protein (repYFV-17D-GFP e firelly luciferase (rep

  7. Lotus Base

    DEFF Research Database (Denmark)

    Mun, Terry; Bachmann, Asger; Gupta, Vikas

    2016-01-01

    exploration of Lotus genomic and transcriptomic data. Equally important are user-friendly in-browser tools designed for data visualization and interpretation. Here, we present Lotus Base, which opens to the research community a large, established LORE1 insertion mutant population containing an excess of 120...... such data, allowing users to construct, visualize, and annotate co-expression gene networks. Lotus Base takes advantage of modern advances in browser technology to deliver powerful data interpretation for biologists. Its modular construction and publicly available application programming interface enable...... developers to tap into the wealth of integrated Lotus data. Lotus Base is freely accessible at: https://lotus.au.dk....

  8. Structure-activity relationships of proline modifications around the tetracyclic-indole class of NS5A inhibitors.

    Science.gov (United States)

    Tong, Ling; Yu, Wensheng; Coburn, Craig A; Chen, Lei; Selyutin, Oleg; Zeng, Qingbei; Dwyer, Michael P; Nair, Anilkumar G; Shankar, Bandarpalle B; Kim, Seong Heon; Yang, De-Yi; Rosenblum, Stuart B; Ruck, Rebecca T; Davies, Ian W; Hu, Bin; Zhong, Bin; Hao, Jinglai; Ji, Tao; Zan, Shuai; Liu, Rong; Agrawal, Sony; Carr, Donna; Curry, Stephanie; McMonagle, Patricia; Bystol, Karin; Lahser, Frederick; Ingravallo, Paul; Chen, Shiying; Asante-Appiah, Ernest; Kozlowski, Joseph A

    2016-11-01

    We describe the impact of proline modifications, in our tetracyclic-indole based series of nonstructural protein 5A (NS5A) inhibitors, to their replicon profiles. This work identified NS5A inhibitors with an improved and flattened resistance profiles. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Characterization of blaCMY-2 plasmids in Salmonella and Escherichia coli isolates from food animals in Canada.

    Science.gov (United States)

    Martin, Laura C; Weir, Emily K; Poppe, Cornelis; Reid-Smith, Richard J; Boerlin, Patrick

    2012-02-01

    One hundred thirty-four bla(CMY-2) plasmids from Salmonella and Escherichia coli strains from animals and food in Canada were characterized. Five plasmid groups were identified based on replicon type and restriction profiles. Three groups contained E. coli plasmids only. IncA/C plasmids included most multiresistant plasmids and all those of bovine origin.

  10. Sultam Thiourea Inhibition of West Nile Virus▿

    Science.gov (United States)

    Barklis, Eric; Still, Amelia; Sabri, Mohammad I.; Hirsch, Alec J.; Nikolich-Zugich, Janko; Brien, James; Dhenub, Tenzin Choesang; Scholz, Isabel; Alfadhli, Ayna

    2007-01-01

    We have identified sultam thioureas as novel inhibitors of West Nile virus (WNV) replication. One such compound inhibited WNV, with a 50% effective concentration of 0.7 μM, and reduced reporter expression from cells that harbored a WNV-based replicon. Our results demonstrate that sultam thioureas can block a postentry, preassembly step of WNV replication. PMID:17452483

  11. Touch BASE

    CERN Multimedia

    Antonella Del Rosso

    2015-01-01

    In a recent Nature article (see here), the BASE collaboration reported the most precise comparison of the charge-to-mass ratio of the proton to its antimatter equivalent, the antiproton. This result is just the beginning and many more challenges lie ahead.   CERN's AD Hall, where the BASE experiment is set-up. The Baryon Antibaryon Symmetry Experiment (BASE) was approved in June 2013 and was ready to take data in August 2014. During these 14 months, the BASE collaboration worked hard to set up its four cryogenic Penning traps, which are the heart of the whole experiment. As their name indicates, these magnetic devices are used to trap antiparticles – antiprotons coming from the Antiproton Decelerator – and particles of matter – negative hydrogen ions produced in the system by interaction with a degrader that slows the antiprotons down, allowing scientists to perform their measurements. “We had very little time to set up the wh...

  12. Web based foundry knowledge base

    Directory of Open Access Journals (Sweden)

    A. Stawowy

    2009-01-01

    Full Text Available The main assumptions and functions of proposed Foundry Knowledge Base (FKB are presented in this paper. FKB is a framework forinformation exchange of casting products and manufacturing methods. We use CMS (Content Management System to develope andmaintain our web-based system. The CastML – XML dialect developed by authors for description of casting products and processes – isused as a tool for information interchange between ours and outside systems, while SQL is used to store and edit knowledge rules and alsoto solve the basic selection problems in the rule-based module. Besides the standard functions (companies data, news, events, forums and media kit, our website contains a number of nonstandard functions; the intelligent search module based on expert system is the main advantage of our solution. FKB is to be a social portal which content will be developed by foundry community.

  13. based conservation

    African Journals Online (AJOL)

    http://dx.doi.org/10.4314/mcd.v10i2.1. Increasing women's par- ticipation in community- based conservation: key to success? Ensuring that both men and women benefit equitably from conservation and development programs is likely to increase the long-term success of both conservation and development goals. However ...

  14. Base Language

    Science.gov (United States)

    Chandrasekhar, A.

    1978-01-01

    A brief discussion of the role of contrastive linguistics and transformational generative grammar in studying transfer and interference in language learning. The base language is not necessarily the mother tongue. It is the language known by the learner which most closely resembles the new language being learned. (AMH)

  15. Based Study

    African Journals Online (AJOL)

    A prospective community-based study of pregnant women in rural communi- ties of south central Ethiopia ... pregnancy outcome were stillbirth (19/1000 births), perinatal mortality (45/1000 births) and neonatal mortality (37/1000 live births). Poor birth ..... ated widi maternal under-nutrition and age (teen- agers giving birdi to ...

  16. Bioactive Cembrane Derivatives from the Indian Ocean Soft Coral, Sinularia kavarattiensis

    Directory of Open Access Journals (Sweden)

    Katja-Emilia Lillsunde

    2014-07-01

    Full Text Available Marine organisms and their metabolites represent a unique source of potential pharmaceutical substances. In this study, we examined marine-derived substances for their bioactive properties in a cell-based Chikungunya virus (CHIKV replicon model and for in vitro anti-inflammatory activity. In the screening of a marine sample library, crude extracts from the Indian soft coral, Sinularia kavarattiensis, showed promising activity against the CHIKV replicon. Bioassay-guided chemical fractionation of S. kavarattiensis resulted in the isolation of six known norcembranoids (1–6 and one new compound, named kavaranolide (7. The structures were elucidated on the basis of NMR and MS spectroscopic data. Compounds 1–3 and 5–7 were evaluated for their replicon-inhibiting potential in the CHIKV model by using a luminescence-based detection technique and live cell imaging. Compounds 1 and 2 showed moderate inhibition of the CHIKV replicon, but imaging studies also revealed cytotoxic properties. Moreover, the effects of the isolated compounds on primary microglial cells, an experimental model for neuroinflammation, were evaluated. Compound 2 was shown to modulate the immune response in microglial cells and to possess potential anti-inflammatory properties by dose-dependently reducing the release of pro- and anti-inflammatory cytokines.

  17. Adapted J6/JFH1-Based Hepatitis C Virus Recombinants with Genotype-Specific NS4A Show Similar Efficacies against Lead Protease Inhibitors, Alpha Interferon, and a Putative NS4A Inhibitor

    Science.gov (United States)

    Gottwein, Judith M.; Jensen, Sanne B.; Serre, Stéphanie B. N.; Ghanem, Lubna; Scheel, Troels K. H.; Jensen, Tanja B.; Krarup, Henrik; Uzcategui, Nathalie; Mikkelsen, Lotte S.

    2013-01-01

    To facilitate studies of hepatitis C virus (HCV) NS4A, we aimed at developing J6/JFH1-based recombinants with genotype 1- to 7-specific NS4A proteins. We developed efficient culture systems expressing NS4A proteins of genotypes (isolates) 1a (H77 and TN), 1b (J4), 2a (J6), 4a (ED43), 5a (SA13), 6a (HK6a), and 7a (QC69), with peak infectivity titers of ∼3.5 to 4.5 log10 focus-forming units per ml. Except for genotype 2a (J6), growth depended on adaptive mutations identified in long-term culture. Genotype 1a, 1b, and 4a recombinants were adapted by amino acid substitutions F772S (p7) and V1663A (NS4A), while 5a, 6a, and 7a recombinants required additional substitutions in the NS3 protease and/or NS4A. We demonstrated applicability of the developed recombinants for study of antivirals. Genotype 1 to 7 NS4A recombinants showed similar responses to the protease inhibitors telaprevir (VX-950), boceprevir (Sch503034), simeprevir (TMC435350), danoprevir (ITMN-191), and vaniprevir (MK-7009), to alpha interferon 2b, and to the putative NS4A inhibitor ACH-806. The efficacy of ACH-806 was lower than that of protease inhibitors and was not influenced by changes at amino acids 1042 and 1065 (in the NS3 protease), which have been suggested to mediate resistance to ACH-806 in replicons. Genotype 1a, 1b, and 2a recombinants showed viral spread under long-term treatment with ACH-806, without acquisition of resistance mutations in the NS3-NS4A region. Relatively high concentrations of ACH-806 inhibited viral assembly, but not replication, in a single-cycle production assay. The developed HCV culture systems will facilitate studies benefitting from expression of genotype-specific NS4A in a constant backbone in the context of the complete viral replication cycle, including functional studies and evaluations of the efficacy of antivirals. PMID:24060868

  18. Clonal diversity of extended-spectrum beta-lactamase producing Escherichia coli isolates in fecal samples of wild animals.

    Science.gov (United States)

    Cristóvão, Filipe; Alonso, Carla Andrea; Igrejas, Gilberto; Sousa, Margarida; Silva, Vanessa; Pereira, José Eduardo; Lozano, Carmen; Cortés-Cortés, Gerardo; Torres, Carmen; Poeta, Patrícia

    2017-03-01

    The clonal diversity of extended-spectrum-β-lactamase (ESBL)-producing Escherichia coli isolates from nine different species of wild animals from distinct regions of Portugal and Spain and their content in replicon plasmids were analyzed. Among the initial 53 ESBL-producing E. coli isolates that were studied (from previous studies), 28 were selected, corresponding to different animal origins with distinct ESBL types and pulsed-field gel electrophoresis (PFGE) patterns. These 28 isolates produced different ESBLs ascribed to the following families: CTX-M, SHV and TEM. The isolates were classified into three phylogenetic groups: B1 (n = 11), A (n = 10) and D (n = 7). The seven E. coli of phylogroup D were then typed by multilocus sequence typing and ascribed to four distinct sequence types: ST117, ST115, ST2001 and ST69. The clonal diversity and relationship between isolates was studied by PFGE. Lastly, the plasmids were analyzed according to their incompatibility group using the PCR-based-replicon-typing scheme. A great diversity of replicon types was identified, with up to five per isolate. Most of the CTX-M-1 and SHV-12 producing E. coli isolates carried IncI1 or IncN replicons. The diversity of ESBL-producing E. coli isolates in wild animals, which can be disseminated in the environment, emphasizes the environmental and health problems that we face nowadays. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Fragmentation based

    Directory of Open Access Journals (Sweden)

    Shashank Srivastava

    2014-01-01

    Gaining the understanding of mobile agent architecture and the security concerns, in this paper, we proposed a security protocol which addresses security with mitigated computational cost. The protocol is a combination of self decryption, co-operation and obfuscation technique. To circumvent the risk of malicious code execution in attacking environment, we have proposed fragmentation based encryption technique. Our encryption technique suits the general mobile agent size and provides hard and thorny obfuscation increasing attacker’s challenge on the same plane providing better performance with respect to computational cost as compared to existing AES encryption.

  20. Diversity of plasmid replicons encoding the bla(CMY-2) gene in broad-spectrum cephalosporin-resistant Escherichia coli from livestock animals in Japan.

    Science.gov (United States)

    Hiki, Mototaka; Usui, Masaru; Kojima, Akemi; Ozawa, Manao; Ishii, Yoshikazu; Asai, Tetsuo

    2013-03-01

    Broad-spectrum cephalosporin (BSC) resistance has increased in Escherichia coli isolates from broiler chickens in Japan since 2004. The purpose of this study was to understand the epidemiology of BSC-resistant E. coli in livestock animals. Among 3274 E. coli isolates from 1767 feces of apparently healthy animals on 1767 farms between 2004 and 2009, 118 ceftiofur (CTF)-resistant isolates (CTF MIC ≥4 μg/mL) were identified on 74 farms. After elimination of apparently clonal isolates from a single animal, 75 selected CTF-resistant isolates (62 isolates from 61 broiler chickens, 10 isolates from 10 layer chickens, two isolates from two cows, and one isolate from a pig) were characterized. The bla(CMY-2) gene was most frequently detected in 50 isolates, followed by bla(CTX-M) (CTX-M-2: six isolates; CTX-M-14: four isolates; CTX-M-25: two isolates; CTX-M-1: one isolate) and bla(SHV) (SHV-12: seven isolates; SHV-2, SHV-2a, SHV-5: one isolate each). In particular, 42 of 62 broiler chicken isolates harbored bla(CMY-2). Pulsed-field gel electrophoresis analyses using XbaI revealed divergent profiles among the BSC-resistant isolates. The incompatibility groups of bla(CMY-2) plasmids from 34 of the 42 broiler chicken isolates belonged to IncIγ (10 isolates), IncA/C (nine isolates), IncB/O (seven isolates) and IncI1 (six isolates), or were nontypeable (two isolates). Co-transmission of resistance to non-β-lactam antibiotics was observed in transconjugants with IncA/C plasmids, but not with IncI1, IncIγ, and IncB/O plasmids except for one isolate with IncB/O. Our findings suggest that the bla(CMY-2) gene is a key player in BSC-resistant E. coli isolates and that coselection is unlikely to be associated with the abundance of bla(CMY-2) plasmids, except for IncA/C plasmids.

  1. Proteome analysis of liver cells expressing a full-length hepatitis C virus (HCV) replicon and biopsy specimens of posttransplantation liver from HCV-infected patients

    Czech Academy of Sciences Publication Activity Database

    Jacobs, J. M.; Diamond, D. L.; Chan, E. Y.; Gritsenko, M. A.; Qian, W.; Šťastná, Miroslava; Baas, T.; Camp II, D. G .H.; Carithers Jr., R. L.; Smith, R. D.; Katze, M. G.

    2005-01-01

    Roč. 79, č. 12 (2005), s. 7558-7569 ISSN 0022-538X Institutional research plan: CEZ:AV0Z40310501 Keywords : proteome analysis * hepatitis C Virus * mass spectrometry Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 5.178, year: 2005

  2. Foundation: Transforming data bases into knowledge bases

    Science.gov (United States)

    Purves, R. B.; Carnes, James R.; Cutts, Dannie E.

    1987-01-01

    One approach to transforming information stored in relational data bases into knowledge based representations and back again is described. This system, called Foundation, allows knowledge bases to take advantage of vast amounts of pre-existing data. A benefit of this approach is inspection, and even population, of data bases through an intelligent knowledge-based front-end.

  3. Geminiviral vectors based on bean yellow dwarf virus for production of vaccine antigens and monoclonal antibodies in plants.

    Science.gov (United States)

    Chen, Qiang; He, Junyun; Phoolcharoen, Waranyoo; Mason, Hugh S

    2011-03-01

    Expression of recombinant vaccine antigens and monoclonal antibodies using plant viral vectors has developed extensively during the past several years. The approach benefits from high yields of recombinant protein obtained within days after transient delivery of viral vectors to leaves of Nicotiana benthamiana, a tobacco relative. Modified viral genomes of both RNA and DNA viruses have been created. Geminiviruses such as bean yellow dwarf virus (BeYDV) have a small, single stranded DNA genome that replicates in the nucleus of an infected plant cell, using the cellular DNA synthesis apparatus and a virus-encoded replication initiator protein (Rep). BeYDV-derived expression vectors contain deletions of the viral genes encoding coat and movement proteins and insertion of an expression cassette for a protein of interest. Delivery of the geminiviral vector to leaf cells via Agrobacterium-mediated delivery produces very high levels of recombinant DNA that can act as a transcription template, yielding high levels of mRNA for the protein of interest. Several vaccine antigens, including Norwalk virus capsid protein and hepatitis B core antigen, were expressed using the BeYDV vector at levels up to 1 mg per g of leaf mass. BeYDV replicons can be stacked in the same vector molecule by linking them in tandem, which enables production of multi-subunit proteins like monoclonal antibody (mAb) heavy and light chains. The protective mAb 6D8 against Ebola virus was produced at 0.5 mg per g of leaf mass. Multi-replicon vectors could be conveniently used to produce protein complexes, e.g. virus-like particles that require two or more subunits.

  4. Expansion of the IncX plasmid family for improved identification and typing of novel plasmids in drug-resistant Enterobacteriaceae

    DEFF Research Database (Denmark)

    Johnson, Timothy J.; Bielak, Eliza Maria; Fortini, Daniela

    2012-01-01

    IncX plasmids are narrow host range plasmids of Enterobactericeae that have been isolated for over 50years. They are known to encode type IV fimbriae enabling their own conjugative transfer, and to provide accessory functions to their host bacteria such as resistance towards antimicrobial agents...... and biofilm formation. Previous plasmid-based replicon typing procedures have indicated that the prevalence of IncX plasmids is low among members of the Enterobacteriaceae. However, examination of a number of IncX-like plasmid sequences and their occurrence in various organisms suggests that IncX plasmid...... backbone, but that they are quite divergent with respect to nucleotide and amino acid similarity. Based on phylogenetic comparisons of the sequenced IncX plasmids, the IncX plasmid group has been expanded to include at least four subtypes, IncX1-IncX4. A revised IncX plasmid replicon typing procedure...

  5. Drug Targets in Infections With Ebola and Marburg Viruses

    Science.gov (United States)

    2009-01-01

    Venezuelan equine encephalitis (VEE) virus -like replicon (VRP), adenovirus 5 (AdS), vesicular stomatitis virus ( VSV )-based vaccines, and virus -like... virus itself may prove to be a very effective way to clear or prevent virus infection, an inherent flaw in this method does exist for several viruses ...immunofluorescence and ELISA. Rab9 siRNA also decreased replication of HIV and measles virus . This was not observed in non~enveloped viruses , which

  6. In vitro antiviral activity of SCH446211 (SCH6), a novel inhibitor of the hepatitis C virus NS3 serine protease.

    Science.gov (United States)

    Liu, Rong; Abid, Karim; Pichardo, John; Pazienza, Valerio; Ingravallo, Paul; Kong, Rong; Agrawal, Sony; Bogen, Stephane; Saksena, Anil; Cheng, Kuo-Chi; Prongay, Andrew; Njoroge, F George; Baroudy, Bahige M; Negro, Francesco

    2007-01-01

    Current hepatitis C virus (HCV) therapies may cure approximately 60% of infections. They are often contraindicated or poorly tolerated, underscoring the need for safer and more effective drugs. A novel, alpha-ketoamide-derived, substrate-based inhibitor of the HCV serine protease (SCH446211) was developed. Compared with earlier reported inhibitors of similar chemical class, it has a P1'-P2' extension which provides extended interaction with the protease active site. The aim of this study was to evaluate the in vitro antiviral activity of SCH446211. Binding constant of SCH446211 to HCV NS3 protease was measured with the chromogenic substrate in vitro cleavage assay. Cell-based activity of SCH446211 was evaluated in replicon cells, which are Huh-7 hepatoma cells stably transfected with a subgenomic HCV RNA as reported previously. After 72 h of incubation with SCH446211, viral transcription and protein expression were measured by real-time RT-PCR (TaqMan), quantitative in situ hybridization, immunoblot and indirect immunofluorescence. The binding constant of SCH446211 to HCV NS3 protease was 3.8 +/- 0.4 nM. HCV replication and protein expression were inhibited by SCH446211 in replicon cells as consistently shown by four techniques. In particular, based on quantitative real-time RT-PCR measurements, the IC50 and IC90 of SCH446211 were estimated to be 40 +/- 20 and 100 +/- 20 nM (n = 17), respectively. Long-term culture of replicon cells with SCH446211 reduced replicon RNA to <0.1 copy per cell. SCH446211 did not show cellular toxicity at concentrations up to 50 microM. SCH446211 is a potent inhibitor of HCV protease in vitro. Its extended interaction with the HCV NS3 protease active site is associated with potent in vitro antiviral activity. This observation is potentially a useful guide for development of future potent inhibitors against HCV NS3 protease.

  7. Mobile Inquiry Based Learning

    NARCIS (Netherlands)

    Specht, Marcus

    2012-01-01

    Specht, M. (2012, 8 November). Mobile Inquiry Based Learning. Presentation given at the Workshop "Mobile inquiry-based learning" at the Mobile Learning Day 2012 at the Fernuniversität Hagen, Hagen, Germany.

  8. Condition Based Maintenance

    National Research Council Canada - National Science Library

    Rabeno, Eric

    2008-01-01

    CONDITION BASED MAINTENANCE (CBM): A plan of maintenance for a system based not upon time intervals but rather upon the actual condition of the system as enabled by the application of usage, diagnostic, and prognostic processes...

  9. VectorBase

    Data.gov (United States)

    U.S. Department of Health & Human Services — VectorBase is a Bioinformatics Resource Center for invertebrate vectors. It is one of four Bioinformatics Resource Centers funded by NIAID to provide web-based...

  10. Community-Based Care

    Science.gov (United States)

    ... our e-newsletter! Aging & Health A to Z Community-Based Care Basic Facts & Information A variety of ... Adult Day Care Adult day care is a community-based option that has become more common. It ...

  11. Beyond Zero Based Budgeting.

    Science.gov (United States)

    Ogden, Daniel M., Jr.

    1978-01-01

    Suggests that the most practical budgeting system for most managers is a formalized combination of incremental and zero-based analysis because little can be learned about most programs from an annual zero-based budget. (Author/IRT)

  12. Convergent Filter Bases

    Directory of Open Access Journals (Sweden)

    Coghetto Roland

    2015-09-01

    Full Text Available We are inspired by the work of Henri Cartan [16], Bourbaki [10] (TG. I Filtres and Claude Wagschal [34]. We define the base of filter, image filter, convergent filter bases, limit filter and the filter base of tails (fr: filtre des sections.

  13. Stolen Base Physics

    Science.gov (United States)

    Kagan, David

    2013-01-01

    Few plays in baseball are as consistently close and exciting as the stolen base. While there are several studies of sprinting, the art of base stealing is much more nuanced. This article describes the motion of the base-stealing runner using a very basic kinematic model. The model will be compared to some data from a Major League game. The…

  14. Outcome-based education.

    Science.gov (United States)

    Davis, Margery H

    2003-01-01

    The move to outcome-based education has been one of the most important trends in health-profession education in recent years. This paper defines outcomes and outcome-based education, describes the development of outcome-based education, identifies several different ways that outcomes have been presented, and discusses the advantages and disadvantages of the outcome-based educational approach. The implementation of outcome-based education at the University of Dundee medical school, Scotland, UK is described as a case study for curriculum planners in veterinary medical education. The lessons learned in Dundee from six years' experience with outcome-based education are identified to aid veterinary medical educators wishing to implement the approach.

  15. Information based speech transduction

    OpenAIRE

    Juel Henrichsen, Peter

    2011-01-01

    Modern hearing aids use a variety of advanced digital signal processing methods in order to improve speech intelligibility. These methods are based on knowledge about the acoustics outside the ear as well as psychoacoustics. We present a novel observation based on the fact that acoustic prominence is not equal to information prominence for time intervals at the syllabic and sub-syllabic levels. The idea is that speech elements with a high degree of information can be robustly identified based...

  16. Base Oils from Petroleum

    Science.gov (United States)

    Prince, R. J.

    The source, composition and suitability of crude oils for base oil production are reviewed. The physical and chemical properties of alkanes, naphthenes and aromatics and their characteristics for lubricant applications are examined. Properties and applications of various base oils are defined and specified. Production of conventional mineral oils is described, including the various processes to remove wax and other deleterious substances, followed by increasingly severe hydrogenation to produce base oils of increased quality and performance. The API categorization of mineral base oils, either direct from the refinery or after hydrotreatment of increasing severity, is described, together with sub-categories.

  17. Imagery Data Base Facility

    Data.gov (United States)

    Federal Laboratory Consortium — The Imagery Data Base Facility supports AFRL and other government organizations by providing imagery interpretation and analysis to users for data selection, imagery...

  18. Monitoring Knowledge Base (MKB)

    Data.gov (United States)

    U.S. Environmental Protection Agency — The Monitoring Knowledge Base (MKB) is a compilation of emissions measurement and monitoring techniques associated with air pollution control devices, industrial...

  19. Plasma Based Devices

    National Research Council Canada - National Science Library

    Gundersen, M

    2001-01-01

    .... Energy-efficient plasma-based technologies, supported through this grant, are now under commercial investigation for pollution abatement, and have potential for reduced emissions, higher efficiencies...

  20. Strengths-based Learning

    DEFF Research Database (Denmark)

    Ledertoug, Mette Marie

    Strength-based learning - Children͛s Character Strengths as Means to their Learning Potential͛ is a Ph.D.-project aiming to create a strength-based mindset in school settings and at the same time introducing strength-based interventions as specific tools to improve both learning and well......-being. The Ph.D.-project in Strength-based learning took place in a Danish school with 750 pupils age 6-16 and a similar school was functioning as a control group. The presentation will focus on both the aware-explore-apply processes and the practical implications for the schools involved, and on measurable...

  1. Case-based reasoning

    CERN Document Server

    Kolodner, Janet

    1993-01-01

    Case-based reasoning is one of the fastest growing areas in the field of knowledge-based systems and this book, authored by a leader in the field, is the first comprehensive text on the subject. Case-based reasoning systems are systems that store information about situations in their memory. As new problems arise, similar situations are searched out to help solve these problems. Problems are understood and inferences are made by finding the closest cases in memory, comparing and contrasting the problem with those cases, making inferences based on those comparisons, and asking questions whe

  2. Development of a broad-host-range sacB-based vector for unmarked allelic exchange

    Directory of Open Access Journals (Sweden)

    Marx Christopher J

    2008-02-01

    Full Text Available Abstract Background Although genome sequences are available for an ever-increasing number of bacterial species, the availability of facile genetic tools for physiological analysis have generally lagged substantially behind traditional genetic models. Results Here I describe the development of an improved, broad-host-range "in-out" allelic exchange vector, pCM433, which permits the generation of clean, marker-free genetic manipulations. Wild-type and mutant alleles were reciprocally exchanged at three loci in Methylobacterium extorquens AM1 in order to demonstrate the utility of pCM433. Conclusion The broad-host-range vector for marker-free allelic exchange described here, pCM433, has the advantages of a high copy, general Escherichia coli replicon for easy cloning, an IncP oriT enabling conjugal transfer, an extensive set of restriction sites in its polylinker, three antibiotic markers, and sacB (encoding levansucrase for negative selection upon sucrose plates. These traits should permit pCM433 to be broadly applied across many bacterial taxa for marker-free allelic exchange, which is particularly important if multiple manipulations or more subtle genetic manipulations such as point mutations are desired.

  3. Secure base stations

    NARCIS (Netherlands)

    Bosch, Peter; Brusilovsky, Alec; McLellan, Rae; Mullender, Sape J.; Polakos, Paul

    2009-01-01

    With the introduction of the third generation (3G) Universal Mobile Telecommunications System (UMTS) base station router (BSR) and fourth generation (4G) base stations, such as the 3rd Generation Partnership Project (3GPP) Long Term Evolution (LTE) Evolved Node B (eNB), it has become important to

  4. Hydrogel based occlusion systems

    NARCIS (Netherlands)

    Stam, F.A.; Jackson, N.; Dubruel, P.; Adesanya, K.; Embrechts, A.; Mendes, E.; Neves, H.P.; Herijgers, P.; Verbrugghe, Y.; Shacham, Y.; Engel, L.; Krylov, V.

    2013-01-01

    A hydrogel based occlusion system, a method for occluding vessels, appendages or aneurysms, and a method for hydrogel synthesis are disclosed. The hydrogel based occlusion system includes a hydrogel having a shrunken and a swollen state and a delivery tool configured to deliver the hydrogel to a

  5. Diffusion Based Photon Mapping

    DEFF Research Database (Denmark)

    Schjøth, Lars; Fogh Olsen, Ole; Sporring, Jon

    2007-01-01

    . To address this problem we introduce a novel photon mapping algorithm based on nonlinear anisotropic diffusion. Our algorithm adapts according to the structure of the photon map such that smoothing occurs along edges and structures and not across. In this way we preserve the important illumination features......, while eliminating noise. We call our method diffusion based photon mapping....

  6. Evidence based practice

    DEFF Research Database (Denmark)

    Hjørland, Birger

    2011-01-01

    Evidence-based practice (EBP) is an influential interdisciplinary movement that originated in medicine as evidence-based medicine (EBM) about 1992. EBP is of considerable interest to library and information science (LIS) because it focuses on a thorough documentation of the basis for the decision...

  7. Work based learning

    OpenAIRE

    Adams, David M.

    2014-01-01

    Entry submitted for publication in The SAGE Encyclopedia of Action Research. Provides a full definition of Work based learning, including origins, historical context, significant characteristics, the relationship between work based learning and action research and their shared foundations, and future prospects.

  8. Zero-Based Budgeting.

    Science.gov (United States)

    Wichowski, Chester

    1979-01-01

    The zero-based budgeting approach is designed to achieve the greatest benefit with the fewest undesirable consequences. Seven basic steps make up the zero-based decision-making process: (1) identifying program goals, (2) classifying goals, (3) identifying resources, (4) reviewing consequences, (5) developing decision packages, (6) implementing a…

  9. Diffusion Based Photon Mapping

    DEFF Research Database (Denmark)

    Schjøth, Lars; Olsen, Ole Fogh; Sporring, Jon

    2006-01-01

    . To address this problem we introduce a novel photon mapping algorithm based on nonlinear anisotropic diffusion. Our algorithm adapts according to the structure of the photon map such that smoothing occurs along edges and structures and not across. In this way we preserve the important illumination features......, while eliminating noise. We call our method diffusion based photon mapping....

  10. Evidence-based radiography

    Energy Technology Data Exchange (ETDEWEB)

    Hafslund, Bjorg [Institute of Radiography, Faculty of Health and Social Sciences, Bergen University College, P.O. Box 7030, N-5020 Bergen (Norway)], E-mail: bhaf@hib.no; Clare, Judith; Graverholt, Birgitte; Wammen Nortvedt, Monica [Centre for Evidence Based Practice, Faculty of Health and Social Sciences, Bergen University College, Bergen (Norway)

    2008-11-15

    Evidence-based practice (EBP) offers the integration of the best research evidence with clinical knowledge and expertise and patient values. EBP is a well known term in health care. This paper discusses the implementation of EBP into radiography and introduces the term evidence-based radiography. Evidence-based radiography is radiography informed and based on the combination of clinical expertise and the best available research-based evidence, patient preferences and resources available. In Norway, EBP in radiography is being debated and radiographers are discussing the challenges of implementing EBP in both academic and clinical practice. This discussion paper explains why EBP needs to be a basis for a radiography curriculum and a part of radiographers' practice. We argue that Norwegian radiographers must increase participation in research and developing practice within their specific radiographic domain.

  11. QuickBase

    CERN Document Server

    Conner, Nancy

    2007-01-01

    Ready to put Intuit's QuickBase to work? Our new Missing Manual shows you how to capture, modify, share, and manage data and documents with this web-based data-sharing program quickly and easily. No longer do you have to coordinate your team through a blizzard of emails or play frustrating games of "guess which document is the right one."QuickBase saves your organization time and money, letting you manage and share the information that makes your business tick: sales figures, project timelines, drafts of documents, purchase or work requests--whatever information you need to keep business flowi

  12. Nature-based integration

    DEFF Research Database (Denmark)

    Pitkänen, Kati; Oratuomi, Joose; Hellgren, Daniela

    based integration by case study descriptions from Denmark, Sweden Norway and Finland. Across Nordic countries several practical projects and initiatives have been launched to promote the benefits of nature in integration and there is also growing academic interest in the topic. Nordic countries have......Increased attention to, and careful planning of the integration of migrants into Nordic societies is ever more important. Nature based integration is a new solution to respond to this need. This report presents the results of a Nordic survey and workshop and illustrates current practices of nature...... the potential of becoming real forerunners in nature based integration even at the global scale....

  13. Data base management study

    Science.gov (United States)

    1976-01-01

    Data base management techniques and applicable equipment are described. Recommendations which will assist potential NASA data users in selecting and using appropriate data base management tools and techniques are presented. Classes of currently available data processing equipment ranging from basic terminals to large minicomputer systems were surveyed as they apply to the needs of potential SEASAT data users. Cost and capabilities projections for this equipment through 1985 were presented. A test of a typical data base management system was described, as well as the results of this test and recommendations to assist potential users in determining when such a system is appropriate for their needs. The representative system tested was UNIVAC's DMS 1100.

  14. Evidence-based dermatology

    Directory of Open Access Journals (Sweden)

    Ching-Chi Chi

    2013-03-01

    Full Text Available Evidence-based medicine (EBM has become a hot topic in medical practice, education, and research. However, a large number of senior doctors did not have an opportunity to learn EBM in medical schools. Firstly, this article addresses the history of EBM and the principle of practicing EBM, i.e., asking, acquiring, appraisal, application, and auditing. Secondly, this article also provides a brief introduction to evidence-based dermatology and compares the introduction of clinical practice guidelines between Europe, the UK, and the US. Finally, this article addresses the present condition and future perspective of evidence-based dermatology in Taiwan.

  15. Value-based pricing

    Directory of Open Access Journals (Sweden)

    Netseva-Porcheva Tatyana

    2010-01-01

    Full Text Available The main aim of the paper is to present the value-based pricing. Therefore, the comparison between two approaches of pricing is made - cost-based pricing and value-based pricing. The 'Price sensitively meter' is presented. The other topic of the paper is the perceived value - meaning of the perceived value, the components of perceived value, the determination of perceived value and the increasing of perceived value. In addition, the best company strategies in matrix 'value-cost' are outlined. .

  16. Network-Based Effectiveness

    National Research Council Canada - National Science Library

    Friman, Henrik

    2006-01-01

    ... (extended from Leavitt, 1965). This text identifies aspects of network-based effectiveness that can benefit from a better understanding of leadership and management development of people, procedures, technology, and organizations...

  17. Hanscom Air Force Base

    Data.gov (United States)

    Federal Laboratory Consortium — MIT Lincoln Laboratory occupies 75 acres (20 acres of which are MIT property) on the eastern perimeter of Hanscom Air Force Base, which is at the nexus of Lexington,...

  18. Biomimetics: nature based innovation

    National Research Council Canada - National Science Library

    Bar-Cohen, Yoseph

    2012-01-01

    "Based on the concept that nature offers numerous sources of inspiration for inventions related to mechanisms, materials, processes, and algorithms, this book covers the topic of biomimetics and the inspired innovation...

  19. Kelomehele preemia Baseli festivalil

    Index Scriptorium Estoniae

    2000-01-01

    Baselis festivalil "VIPER - International Festival for Film Video and New Media" tunnistati parimaks CD-ROMiks Gustav Deutschi/Anna Schimeki "Odysee today", netiprojektiks itaallaste "01.ORG", äramärkimispreemia - Raivo Kelomehe "Videoweaver"

  20. WormBase

    Data.gov (United States)

    U.S. Department of Health & Human Services — WormBase is an international consortium of biologists and computer scientists dedicated to providing the research community with accurate, current, accessible...

  1. Mindfulness-Based Movement.

    Science.gov (United States)

    Lucas, Alexander R; Klepin, Heidi D; Porges, Stephen W; Rejeski, W Jack

    2016-12-01

    Compelling evidence suggests that physical activity is an effective intervention for cancer survivors, including for those undergoing active cancer treatments. However, to date most evidence has emerged from interventions that have promoted moderate to vigorous physical activity. In this conceptual review, we argue that attention should be given to the entire continuum of physical activity from reducing sedentary behavior to increasing higher levels of physical activity when possible. In addition, considerable evidence in the cancer literature supports the value of mindfulness-based interventions as a means of helping patients and survivors cope with the variety of threats that accompany this disease. Based on the success of these two areas of research, we argue for conceptualizing and promoting physical activity as Mindfulness-Based Movement, using Polyvagal Theory as a theoretical framework to understand the role and value of Mindfulness-Based Movement as a potential intervention for cancer care and control.

  2. Cheboygan Vessel Base

    Data.gov (United States)

    Federal Laboratory Consortium — Cheboygan Vessel Base (CVB), located in Cheboygan, Michigan, is a field station of the USGS Great Lakes Science Center (GLSC). CVB was established by congressional...

  3. Benzimidazole Based Aerogel Materials

    Science.gov (United States)

    Rhine, Wendell E. (Inventor); Mihalcik, David (Inventor)

    2016-01-01

    The present invention provides aerogel materials based on imidazoles and polyimidazoles. The polyimidazole based aerogel materials can be thermally stable up to 500 C or more, and can be carbonized to produce a carbon aerogel having a char yield of 60% or more, specifically 70% or more. The present invention also provides methods of producing polyimidazole based aerogel materials by reacting at least one monomer in a suitable solvent to form a polybenzimidazole gel precursor solution, casting the polybenzimidazole gel precursor solution into a fiber reinforcement phase, allowing the at least one gel precursor in the precursor solution to transition into a gel material, and drying the gel materials to remove at least a portion of the solvent, to obtain an polybenzimidazole-based aerogel material.

  4. REST based mobile applications

    Science.gov (United States)

    Rambow, Mark; Preuss, Thomas; Berdux, Jörg; Conrad, Marc

    2008-02-01

    Simplicity is the major advantage of REST based webservices. Whereas SOAP is widespread in complex, security sensitive business-to-business aplications, REST is widely used for mashups and end-user centric applicatons. In that context we give an overview of REST and compare it to SOAP. Furthermore we apply the GeoDrawing application as an example for REST based mobile applications and emphasize on pros and cons for the use of REST in mobile application scenarios.

  5. Hydrogel based occlusion systems

    OpenAIRE

    Stam, F.A.; Jackson, N.; Dubruel, P.; Adesanya, K.; Embrechts, A.; Mendes, E.; Neves, H.P.; Herijgers, P.; Verbrugghe, Y.; Shacham, Y.; Engel, L; Krylov, V.

    2013-01-01

    A hydrogel based occlusion system, a method for occluding vessels, appendages or aneurysms, and a method for hydrogel synthesis are disclosed. The hydrogel based occlusion system includes a hydrogel having a shrunken and a swollen state and a delivery tool configured to deliver the hydrogel to a target occlusion location. The hydrogel is configured to permanently occlude the target occlusion location in the swollen state. The hydrogel may be an electro-activated hydrogel (EAH) which could be ...

  6. Object Based Systems Engineering

    Science.gov (United States)

    2011-10-17

    Based Systems Engineering ( MBSE ) has shifted the emphasis of the Systems Engineering community away from documents towards view-based artifacts. These...Engineering lies primarily in these objects, not the containers that deliver them. FIGURE 1: Evolution of Systems Engineering Practice MBSE ...capture minority viewpoints and discussion threads associated with each object of interest. If the majority view doesn’t lead to success, this data may

  7. Evidence-Based Development

    DEFF Research Database (Denmark)

    Hertzum, Morten; Simonsen, Jesper

    2004-01-01

    Systems development is replete with projects that represent substantial resource investments but result in systems that fail to meet users’ needs. Evidence-based development is an emerging idea intended to provide means for managing customer-vendor relationships and working systematically toward...... and electronic patient records for diabetes patients, this paper reports research in progress regarding the prospects and pitfalls of evidence-based development....

  8. Participatory design based research

    DEFF Research Database (Denmark)

    Dau, Susanne; Falk, Lars; Jensen, Louise Bach

    This poster reveal how participatory design based research by the use of a CoED inspired creative process can be used for designing solutions to problems regarding students study activities outside campus.......This poster reveal how participatory design based research by the use of a CoED inspired creative process can be used for designing solutions to problems regarding students study activities outside campus....

  9. ACTIVITY - BASED COSTING DESIGNING

    OpenAIRE

    Wioletta Skibiñska; Marta Kad³ubek

    2010-01-01

    The traditional costing system sometimes does not give accurate information about the consumption of different resources and the activities of the organisation. The activity-based costing system is an information-rich costing system which is more and more necessary for the success of many European companies. Base of designing and implementation of an ABC system in the enterprises are presented in the article.

  10. Knowledge Based Text Generation

    Science.gov (United States)

    1989-08-01

    from data bases, so Kukich [1984] developed a system, ANA , which generates stock reports from a knowledge base of daily trading on the Dow Jones stock...MACHIAVELLI (topic organization and phraseology), CICERO (realization), FREUD (monitoring the origins of rhetorical plans), and LEIBNITZ (a "concept...68 Bossie and Mani 8 Alla Fiera dell’est 37 brain 2 frame 29 Alshawi 49 Brown and Yule 51 amplification 38 Cambridge University 40 ANA 15 canned text 7

  11. Evidence-Based Toxicology.

    Science.gov (United States)

    Hoffmann, Sebastian; Hartung, Thomas; Stephens, Martin

    Evidence-based toxicology (EBT) was introduced independently by two groups in 2005, in the context of toxicological risk assessment and causation as well as based on parallels between the evaluation of test methods in toxicology and evidence-based assessment of diagnostics tests in medicine. The role model of evidence-based medicine (EBM) motivated both proposals and guided the evolution of EBT, whereas especially systematic reviews and evidence quality assessment attract considerable attention in toxicology.Regarding test assessment, in the search of solutions for various problems related to validation, such as the imperfectness of the reference standard or the challenge to comprehensively evaluate tests, the field of Diagnostic Test Assessment (DTA) was identified as a potential resource. DTA being an EBM discipline, test method assessment/validation therefore became one of the main drivers spurring the development of EBT.In the context of pathway-based toxicology, EBT approaches, given their objectivity, transparency and consistency, have been proposed to be used for carrying out a (retrospective) mechanistic validation.In summary, implementation of more evidence-based approaches may provide the tools necessary to adapt the assessment/validation of toxicological test methods and testing strategies to face the challenges of toxicology in the twenty first century.

  12. LDEF materials data bases

    Science.gov (United States)

    Funk, Joan G.; Strickland, John W.; Davis, John M.

    1993-01-01

    The Long Duration Exposure Facility (LDEF) and the accompanying experiments were composed of and contained a wide variety of materials representing the largest collection of materials flown in low Earth orbit (LEO) and retrieved for ground based analysis to date. The results and implications of the mechanical, thermal, optical, and electrical data from these materials are the foundation on which future LEO space missions will be built. The LDEF Materials Special Investigation Group (MSIG) has been charged with establishing and developing data bases to document these materials and their performance to assure not only that the data are archived for future generations but also that the data are available to the spacecraft user community in an easily accessed, user-friendly form. This paper discusses the format and content of the three data bases developed or being developed to accomplish this task. The hardware and software requirements for each of these three data bases are discussed along with current availability of the data bases. This paper also serves as a user's guide to the MAPTIS LDEF Materials Data Base.

  13. Paper based electronics platform

    KAUST Repository

    Nassar, Joanna Mohammad

    2017-07-20

    A flexible and non-functionalized low cost paper-based electronic system platform fabricated from common paper, such as paper based sensors, and methods of producing paper based sensors, and methods of sensing using the paper based sensors are provided. A method of producing a paper based sensor can include the steps of: a) providing a conventional paper product to serve as a substrate for the sensor or as an active material for the sensor or both, the paper product not further treated or functionalized; and b) applying a sensing element to the paper substrate, the sensing element selected from the group consisting of a conductive material, the conductive material providing contacts and interconnects, sensitive material film that exhibits sensitivity to pH levels, a compressible and/or porous material disposed between a pair of opposed conductive elements, or a combination of two of more said sensing elements. The method of sensing can further include measuring, using the sensing element, a change in resistance, a change in voltage, a change in current, a change in capacitance, or a combination of any two or more thereof.

  14. Architecture and functions of a multipartite genome of the methylotrophic bacterium Paracoccus aminophilus JCM 7686, containing primary and secondary chromids.

    Science.gov (United States)

    Dziewit, Lukasz; Czarnecki, Jakub; Wibberg, Daniel; Radlinska, Monika; Mrozek, Paulina; Szymczak, Michal; Schlüter, Andreas; Pühler, Alfred; Bartosik, Dariusz

    2014-02-12

    Paracoccus aminophilus JCM 7686 is a methylotrophic α-Proteobacterium capable of utilizing reduced one-carbon compounds as sole carbon and energy source for growth, including toxic N,N-dimethylformamide, formamide, methanol, and methylamines, which are widely used in the industry. P. aminophilus JCM 7686, as many other Paracoccus spp., possesses a genome representing a multipartite structure, in which the genomic information is split between various replicons, including chromids, essential plasmid-like replicons, with properties of both chromosomes and plasmids. In this study, whole-genome sequencing and functional genomics approaches were applied to investigate P. aminophilus genome information. The P. aminophilus JCM 7686 genome has a multipartite structure, composed of a single circular chromosome and eight additional replicons ranging in size between 5.6 and 438.1 kb. Functional analyses revealed that two of the replicons, pAMI5 and pAMI6, are essential for host viability, therefore they should be considered as chromids. Both replicons carry housekeeping genes, e.g. responsible for de novo NAD biosynthesis and ammonium transport. Other mobile genetic elements have also been identified, including 20 insertion sequences, 4 transposons and 10 prophage regions, one of which represents a novel, functional serine recombinase-encoding bacteriophage, ϕPam-6. Moreover, in silico analyses allowed us to predict the transcription regulatory network of the JCM 7686 strain, as well as components of the stress response, recombination, repair and methylation machineries. Finally, comparative genomic analyses revealed that P. aminophilus JCM 7686 has a relatively distant relationship to other representatives of the genus Paracoccus. P. aminophilus genome exploration provided insights into the overall structure and functions of the genome, with a special focus on the chromids. Based on the obtained results we propose the classification of bacterial chromids into two types

  15. Gossip-Based Dissemination

    Science.gov (United States)

    Friedman, Roy; Kermarrec, Anne-Marie; Miranda, Hugo; Rodrigues, Luís

    Gossip-based networking has emerged as a viable approach to disseminate information reliably and efficiently in large-scale systems. Initially introduced for database replication [222], the applicability of the approach extends much further now. For example, it has been applied for data aggregation [415], peer sampling [416] and publish/subscribe systems [845]. Gossip-based protocols rely on a periodic peer-wise exchange of information in wired systems. By changing the way each peer is selected for the gossip communication, and which data are exchanged and processed [451], gossip systems can be used to perform different distributed tasks, such as, among others: overlay maintenance, distributed computation, and information dissemination (a collection of papers on gossip can be found in [451]). In a wired setting, the peer sampling service, allowing for a random or specific peer selection, is often provided as an independent service, able to operate independently from other gossip-based services [416].

  16. Problem Based Learning

    DEFF Research Database (Denmark)

    de Graaff, Erik; Guerra, Aida

    Problem-Based Learning (PBL) is an innovative method to organize the learning process in such a way that the students actively engage in finding answers by themselves. During the past 40 years PBL has evolved and diversified resulting in a multitude in variations in models and practices. However......, the key principles remain the same everywhere. Graaff & Kolmos (2003) identify the main PBL principles as follows: 1. Problem orientation 2. Project organization through teams or group work 3. Participant-directed 4. Experiental learning 5. Activity-based learning 6. Interdisciplinary learning and 7...... in Engineering Education. In answer to the requests for visits the Aalborg Centre for Problem Based Learning in Engineering Science and Sustainability under the auspices of UNESCO (UCPBL) a two days programme for visitors is offered two times a year. The workshop is an introduction workshop to the Aalborg PBL...

  17. Design-Based Research

    DEFF Research Database (Denmark)

    Gynther, Karsten; Christensen, Ove; Petersen, Trine Brun

    2012-01-01

    I denne artikel introduceres Design Based Research for første gang på dansk i et videnskabeligt tidsskrift. Artiklen præsenterer de grundlæggende antagelser, som ligger til grund for Design Based Research-traditionen, og artiklen diskuterer de principper, som ligger til grund for gennemførelse af...... et DBR-forskningsprojekt. Med udgangspunkt i forsknings- og udviklingsprojektet ELYK: E-læring, Yderområder og Klyngedannelse, præsenteres den innovationsmodel, som projektet har udviklet med udgangspunkt i Design Based Research traditionen. ELYKs DBR innovationsmodel har vist sig effektiv i forhold...... til projektfremdrift, brugerinvolvering og vidensgenerering, og den vil kunne inspirere andre med interesse i forskningsbaseret udvikling af didaktisk design, der er medieret af digitale teknologier....

  18. Inkjet-based micromanufacturing

    CERN Document Server

    Korvink, Jan G; Shin, Dong-Youn; Brand, Oliver; Fedder, Gary K; Hierold, Christofer; Tabata, Osamu

    2012-01-01

    Inkjet-based Micromanufacturing Inkjet technology goes way beyond putting ink on paper: it enables simpler, faster and more reliable manufacturing processes in the fields of micro- and nanotechnology. Modern inkjet heads are per se precision instruments that deposit droplets of fluids on a variety of surfaces in programmable, repeating patterns, allowing, after suitable modifications and adaptations, the manufacturing of devices such as thin-film transistors, polymer-based displays and photovoltaic elements. Moreover, inkjet technology facilitates the large-scale production of flexible RFID tr

  19. Carbon-based spintronics

    Science.gov (United States)

    Chen, Peng; Zhang, GuangYu

    2013-01-01

    Carbon-based spintronics refers mainly to the spin injection and transport in carbon materials including carbon nanotubes, graphene, fullerene, and organic materials. In the last decade, extraordinary development has been achieved for carbon-based spintronics, and the spin transport has been studied in both local and nonlocal spin valve devices. A series of theoretical and experimental studies have been done to reveal the spin relaxation mechanisms and spin transport properties in carbon materials, mostly for graphene and carbon nanotubes. In this article, we provide a brief review on spin injection and transport in graphene, carbon nanotubes, fullerene and organic thin films.

  20. Game-Based Teaching

    DEFF Research Database (Denmark)

    Hanghøj, Thorkild

    2013-01-01

    , pedagogical knowledge practices, and everyday knowledge practices. Secondly, the chapter emphasizes how teachers must be able to shift back and forth between various interactional roles in order to facilitate game scenarios. Finally, a discussion is presented on how teachers choose different pedagogical......This chapter outlines theoretical and empirical perspectives on how Game-Based Teaching can be integrated within the context of formal schooling. Initially, this is done by describing game scenarios as models for possible actions that need to be translated into curricular knowledge practices...... approaches to game-based teaching, which may or may not correspond with the pedagogical models of particular games....

  1. Iron-based superconductivity

    CERN Document Server

    Johnson, Peter D; Yin, Wei-Guo

    2015-01-01

    This volume presents an in-depth review of experimental and theoretical studies on the newly discovered Fe-based superconductors.  Following the Introduction, which places iron-based superconductors in the context of other unconventional superconductors, the book is divided into three sections covering sample growth, experimental characterization, and theoretical understanding.  To understand the complex structure-property relationships of these materials, results from a wide range of experimental techniques and theoretical approaches are described that probe the electronic and magnetic proper

  2. Process-based costing.

    Science.gov (United States)

    Lee, Robert H; Bott, Marjorie J; Forbes, Sarah; Redford, Linda; Swagerty, Daniel L; Taunton, Roma Lee

    2003-01-01

    Understanding how quality improvement affects costs is important. Unfortunately, low-cost, reliable ways of measuring direct costs are scarce. This article builds on the principles of process improvement to develop a costing strategy that meets both criteria. Process-based costing has 4 steps: developing a flowchart, estimating resource use, valuing resources, and calculating direct costs. To illustrate the technique, this article uses it to cost the care planning process in 3 long-term care facilities. We conclude that process-based costing is easy to implement; generates reliable, valid data; and allows nursing managers to assess the costs of new or modified processes.

  3. Bases de dades

    OpenAIRE

    del Valle López, Alfredo Eugenio

    2013-01-01

    Este proyecto final de carrera ha consistido principalmente en la planificación y ejecución del diseño e implementación de una base de datos (BD) relacional para la gestión sanitaria. El sistema resultante permite tanto la gestión de las consultas externas de los centros hospitalarios como la de los pacientes atendidos. Toda la gestión y acceso a los datos de las tablas se ha hecho mediante procedimientos almacenados creados en la propia base de datos. También se ha definido un almacén de dat...

  4. Video-based rendering

    CERN Document Server

    Magnor, Marcus A

    2005-01-01

    Driven by consumer-market applications that enjoy steadily increasing economic importance, graphics hardware and rendering algorithms are a central focus of computer graphics research. Video-based rendering is an approach that aims to overcome the current bottleneck in the time-consuming modeling process and has applications in areas such as computer games, special effects, and interactive TV. This book offers an in-depth introduction to video-based rendering, a rapidly developing new interdisciplinary topic employing techniques from computer graphics, computer vision, and telecommunication en

  5. Identity-based encryption

    CERN Document Server

    Chatterjee, Sanjit

    2011-01-01

    Identity Based Encryption (IBE) is a type of public key encryption and has been intensely researched in the past decade. Identity-Based Encryption summarizes the available research for IBE and the main ideas that would enable users to pursue further work in this area. This book will also cover a brief background on Elliptic Curves and Pairings, security against chosen Cipher text Attacks, standards and more. Advanced-level students in computer science and mathematics who specialize in cryptology, and the general community of researchers in the area of cryptology and data security will find Ide

  6. Model-Based Testing

    NARCIS (Netherlands)

    Timmer, Mark; Brinksma, Hendrik; Stoelinga, Mariëlle Ida Antoinette; Broy, M.; Leuxner, C.; Hoare, C.A.R.

    This paper provides a comprehensive introduction to a framework for formal testing using labelled transition systems, based on an extension and reformulation of the ioco theory introduced by Tretmans. We introduce the underlying models needed to specify the requirements, and formalise the notion of

  7. Dictionary Based Image Segmentation

    DEFF Research Database (Denmark)

    Dahl, Anders Bjorholm; Dahl, Vedrana Andersen

    2015-01-01

    We propose a method for weakly supervised segmentation of natural images, which may contain both textured or non-textured regions. Our texture representation is based on a dictionary of image patches. To divide an image into separated regions with similar texture we use an implicit level sets...

  8. Skull Base Tumors

    Science.gov (United States)

    Schulz-Ertner, Daniela

    In skull base tumors associated with a low radiosensitivity for conventional radiotherapy (RT), irradiation with proton or carbon ion beams facilitates a safe and accurate application of high tumor doses due to the favorable beam localization properties of these particle beams. Cranial nerves, the brain stem and normal brain tissue can at the same time be optimally spared.

  9. Office-based anaesthesia

    African Journals Online (AJOL)

    setting. Data for South Africa are lacking, but there seems to be consistent growth in this area, ... inter-patient variation in response to a given dose of a given drug or drug ... Successful office-based anaesthesia requires a particular set of skills.

  10. Sea Basing Logistiek

    Science.gov (United States)

    2007-11-01

    flexibiliteit, dynamiek en configureerbaarheid afhankelijk van de missie behoefte is wat hen aanspreekt: ’Joint Sea Base is a frame of mind, not a...en dat tijdens de sustain fase selectief gelost kan worden, is bet van belang orn voorafgaand aan de close fase stil te staan bij de manier waarop

  11. Schiff base ligand

    Indian Academy of Sciences (India)

    Unknown

    Schiff base; dicopper (II/II) complexes; imidazolidine-bridged; molecular mechanics; cy- ... nuclear complex of copper(II)–zinc(II) held by an ... ineffectiveness of the imidazolidine bridges in im- porting any magnetic communication between the two copper(II) centres has also been here established through EPR study. 2.

  12. 80537 based distance relay

    DEFF Research Database (Denmark)

    Pedersen, Knud Ole Helgesen

    1999-01-01

    A method for implementing a digital distance relay in the power system is described.Instructions are given on how to program this relay on a 80537 based microcomputer system.The problem is used as a practical case study in the course 53113: Micocomputer applications in the power system.The relay...

  13. schiff base complexes

    African Journals Online (AJOL)

    DR. AMINU

    ABSTRACT. The complexes of iron (II) and nickel (II) with schiff base derived from benzoin and 2-amino benzoic acid have been prepared. Solubility, melting point, decomposition temperature, conductance measurement, infrared (IR) and UV – Visible spectrophotometric studies were used in characterizing the compounds.

  14. Cotton-based nonwovens

    Science.gov (United States)

    This article is an abbreviated description of a new cotton-based nonwovens research program at the Southern Regional Research Center, which is one of the four regional research centers of the Agricultural Research Service, U.S. Department of Agriculture. Since cotton is a significant cash crop inte...

  15. Fundamental research data base

    Science.gov (United States)

    1983-01-01

    A fundamental research data base containing ground truth, image, and Badhwar profile feature data for 17 North Dakota, South Dakota, and Minnesota agricultural sites is described. Image data was provided for a minimum of four acquisition dates for each site and all four images were registered to one another.

  16. Trajectory Based Traffic Analysis

    DEFF Research Database (Denmark)

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

    2013-01-01

    We present the INTRA system for interactive path-based traffic analysis. The analyses are developed in collaboration with traffic researchers and provide novel insights into conditions such as congestion, travel-time, choice of route, and traffic-flow. INTRA supports interactive point...

  17. Diffusion Based Photon Mapping

    DEFF Research Database (Denmark)

    Schjøth, Lars; Sporring, Jon; Fogh Olsen, Ole

    2008-01-01

    . To address this problem, we introduce a photon mapping algorithm based on nonlinear anisotropic diffusion. Our algorithm adapts according to the structure of the photon map such that smoothing occurs along edges and structures and not across. In this way, we preserve important illumination features, while...

  18. Status-Based Identity.

    Science.gov (United States)

    Destin, Mesmin; Rheinschmidt-Same, Michelle; Richeson, Jennifer A

    2017-03-01

    Psychological research on socioeconomic status (SES) has grown significantly over the past decade. In this article, we build upon and integrate existing approaches to direct greater attention toward investigating the subjective meaning and value that people attach to understanding their own SES as an identity. We use the term status-based identity to organize relevant research and examine how people understand and make meaning of their SES from moment to moment in real time. Drawing from multiple areas of research on identity, we suggest that even temporary shifts in how people construe their status-based identities predict changes in thought, affect, motivation, and behavior. This novel focus is positioned to examine the psychological effects of status transitions (e.g., upward or downward mobility). Further, in initial empirical work, we introduce a new measure to assess uncertainty regarding one's SES (i.e., status-based identity uncertainty) and offer evidence that greater uncertainty regarding one's status-based identity is associated with lower individual well-being. In sum, we argue that insight from the literature on identity will both expand and serve to organize the burgeoning literature on the psychology of SES and, in so doing, reveal promising new directions for research.

  19. Internet based benchmarking

    DEFF Research Database (Denmark)

    Bogetoft, Peter; Nielsen, Kurt

    2005-01-01

    We discuss the design of interactive, internet based benchmarking using parametric (statistical) as well as nonparametric (DEA) models. The user receives benchmarks and improvement potentials. The user is also given the possibility to search different efficiency frontiers and hereby to explore...

  20. Animation-based Sketching

    DEFF Research Database (Denmark)

    Vistisen, Peter

    This thesis is based on the results of a three-year long PhD-study at the Department of Communication and Psychology at Aalborg University. The thesis consist of five original papers, a book manuscript, as well as a linking text with the thesis’ research questions, research design, and summary...

  1. SCHIFF-BASE

    African Journals Online (AJOL)

    Preferred Customer

    Schiff bases due to structural varieties and unique characteristics are the most versatile studied ligands in coordination chemistry [1, 2] and their metal complexes play an important role in the development of inorganic chemistry [3]. A variety of applications, including biological [4, 5], analytical [6, 7] and industrial use as.

  2. Performance based fault diagnosis

    DEFF Research Database (Denmark)

    Niemann, Hans Henrik

    2002-01-01

    Different aspects of fault detection and fault isolation in closed-loop systems are considered. It is shown that using the standard setup known from feedback control, it is possible to formulate fault diagnosis problems based on a performance index in this general standard setup. It is also shown...

  3. ISFET based enzyme sensors

    NARCIS (Netherlands)

    van der Schoot, Bart H.; Bergveld, Piet

    1987-01-01

    This paper reviews the results that have been reported on ISFET based enzyme sensors. The most important improvement that results from the application of ISFETs instead of glass membrane electrodes is in the method of fabrication. Problems with regard to the pH dependence of the response and the

  4. Problem-based learning

    NARCIS (Netherlands)

    Loyens, Sofie; Kirschner, Paul A.; Paas, Fred

    2010-01-01

    Loyens, S. M. M., Kirschner, P. A., & Paas, F. (2011). Problem-based learning. In S. Graham (Editor-in-Chief), A. Bus, S. Major, & L. Swanson (Associate Editors), APA educational psychology handbook: Vol. 3. Application to learning and teaching (pp. 403-425). Washington, DC: American Psychological

  5. Project-Based Science

    Science.gov (United States)

    Krajcik, Joe

    2015-01-01

    Project-based science is an exciting way to teach science that aligns with the "Next Generation Science Standards" ("NGSS"). By focusing on core ideas along with practices and crosscutting concepts, classrooms become learning environments where teachers and students engage in science by designing and carrying out…

  6. Refractive index based measurements

    DEFF Research Database (Denmark)

    2014-01-01

    In a method for performing a refractive index based measurement of a property of a fluid such as chemical composition or temperature, a chirp in the local spatial frequency of interference fringes of an interference pattern is reduced by mathematical manipulation of the recorded light intensity...

  7. Refractive index based measurements

    DEFF Research Database (Denmark)

    2014-01-01

    A refractive index based measurement of a property of a fluid is measured in an apparatus comprising a variable wavelength coherent light source (16), a sample chamber (12), a wavelength controller (24), a light sensor (20), a data recorder (26) and a computation apparatus (28), by - directing...

  8. Refractive index based measurements

    DEFF Research Database (Denmark)

    2014-01-01

    In a method for performing a refractive index based measurement of a property of a fluid such as chemical composition or temperature by observing an apparent angular shift in an interference fringe pattern produced by back or forward scattering interferometry, ambiguities in the measurement caused...

  9. Science Based Stockpile Stewardship

    Science.gov (United States)

    1994-11-01

    low level of radioactivity produced are comparable to those in TFTR presently operating routinely on the Forrestal Campus of Princeton University and...to nu- merous astronomical objects and processes (e.g., primordial nucleosynthesis, stellar evoiution, and hydrodynamic instabilities in supernova ... radioactive decay heat, reactivity excursions beyond the subcriticality margin etc. A specific APS system based on the thorium fuel cycle has recently

  10. Lidar base specification

    Science.gov (United States)

    Heidemann, Hans Karl

    2012-01-01

    In late 2009, a $14.3 million allocation from the “American Recovery and Reinvestment Act” for new light detection and ranging (lidar) elevation data prompted the U.S. Geological Survey (USGS) National Geospatial Program (NGP) to develop a common base specification for all lidar data acquired for The National Map. Released as a draft in 2010 and formally published in 2012, the USGS–NGP “Lidar Base Specification Version 1.0” (now Lidar Base Specification) was quickly embraced as the foundation for numerous state, county, and foreign country lidar specifications. Prompted by a growing appreciation for the wide applicability and inherent value of lidar, a USGS-led consortium of Federal agencies commissioned a National Enhanced Elevation Assessment (NEEA) study in 2010 to quantify the costs and benefits of a national lidar program. A 2012 NEEA report documented a substantial return on such an investment, defined five Quality Levels (QL) for elevation data, and recommended an 8-year collection cycle of Quality Level 2 (QL2) lidar data as the optimum balance of benefit and affordability. In response to the study, the USGS–NGP established the 3D Elevation Program (3DEP) in 2013 as the interagency vehicle through which the NEEA recommendations could be realized. Lidar is a fast evolving technology, and much has changed in the industry since the final draft of the “Lidar Base Specification Version 1.0” was written. Lidar data have improved in accuracy and spatial resolution, geospatial accuracy standards have been revised by the American Society for Photogrammetry and Remote Sensing (ASPRS), industry standard file formats have been expanded, additional applications for lidar have become accepted, and the need for interoperable data across collections has been realized. This revision to the “Lidar Base Specification Version 1.0” publication addresses those changes and provides continued guidance towards a nationally consistent lidar dataset.

  11. Venezuelan Equine Encephalitis Replicon Immunization Overcomes Intrinsic Tolerance and Elicits Effective Anti-Tumor Immunity to the ’Self’ Tumor-Associated Antigen, neu in a Rat Mammary Tumor Model

    Science.gov (United States)

    2003-01-01

    Immunol Immunother 50: 615–624, 2002 75. Rongcun Y, Salazar- Onfray F, Charo J, Malmberg KJ, Evrin K, Maes H, Kono K, Hising C, Petersson M, Larsson O...cultured cells infected by Semliki Forest virus. J Gen Virol 78: 1559–1563, 1997 100. Grandgirard D, Studer E, Monney L, Belser T, Fellay I, Borner C, Michel

  12. Evidence-based policy

    DEFF Research Database (Denmark)

    Vohnsen, Nina Holm

    2013-01-01

    A current ambition in welfare states as diverse as Denmark, the UK, and in the USA is to base political decision making on rigorous research (Cartwright et al 2009; Mulgan 2009; Bason 2010). Sound as this might seem the ambition has nevertheless been problematized by both policy-makers and the re......A current ambition in welfare states as diverse as Denmark, the UK, and in the USA is to base political decision making on rigorous research (Cartwright et al 2009; Mulgan 2009; Bason 2010). Sound as this might seem the ambition has nevertheless been problematized by both policy...... a full account, see Vohnsen 2011). These insights will be relevant for the anthropological researcher of legislative processes who wishes to move beyond a merely discursive approach to the study of policy and politics....

  13. Ontology Based Access Control

    Directory of Open Access Journals (Sweden)

    Özgü CAN

    2010-02-01

    Full Text Available As computer technologies become pervasive, the need for access control mechanisms grow. The purpose of an access control is to limit the operations that a computer system user can perform. Thus, access control ensures to prevent an activity which can lead to a security breach. For the success of Semantic Web, that allows machines to share and reuse the information by using formal semantics for machines to communicate with other machines, access control mechanisms are needed. Access control mechanism indicates certain constraints which must be achieved by the user before performing an operation to provide a secure Semantic Web. In this work, unlike traditional access control mechanisms, an "Ontology Based Access Control" mechanism has been developed by using Semantic Web based policies. In this mechanism, ontologies are used to model the access control knowledge and domain knowledge is used to create policy ontologies.

  14. Maintaining Relationship Based Procurement

    Directory of Open Access Journals (Sweden)

    Peter Davis

    2012-11-01

    Full Text Available Alliance and relationship projects are increasingin number and represent a large pool of work. Tobe successful relationship style contracts dependon soft-dollar factors, particularly the participants'ability to work together within an agreedframework, generally they are not based on lowbid tendering. Participants should be prepared todo business in an open environment based ontrust and mutually agreed governance. Theresearch evaluates relationship maintenance inthe implementation phase of constructionalliances - a particular derivative of relationshipstyle contracts. To determine the factors thatcontribute to relationship maintenance forty-nineexperienced Australian alliance projectmanagers were interviewed. The main findingswere; the development of relationships early inthe project form building blocks of success fromwhich relationships are maintained and projectvalue added; quality facilitation plays animportant part in relationship maintenance and ahybrid organisation created as a result of alliancedevelopment overcomes destructiveorganisational boundaries. Relationshipmaintenance is integral to alliance project controland failure to formalise it and pay attention toprocess and past outcomes will undermine analliance project's potential for success.

  15. Sparse approximation with bases

    CERN Document Server

    2015-01-01

    This book systematically presents recent fundamental results on greedy approximation with respect to bases. Motivated by numerous applications, the last decade has seen great successes in studying nonlinear sparse approximation. Recent findings have established that greedy-type algorithms are suitable methods of nonlinear approximation in both sparse approximation with respect to bases and sparse approximation with respect to redundant systems. These insights, combined with some previous fundamental results, form the basis for constructing the theory of greedy approximation. Taking into account the theoretical and practical demand for this kind of theory, the book systematically elaborates a theoretical framework for greedy approximation and its applications.  The book addresses the needs of researchers working in numerical mathematics, harmonic analysis, and functional analysis. It quickly takes the reader from classical results to the latest frontier, but is written at the level of a graduate course and do...

  16. Problem Based Learning

    Directory of Open Access Journals (Sweden)

    Paola Cappola

    2014-01-01

    Full Text Available In this current work, I am proposing a general close examination of Problem Based Learning as a student centered educational method in which a problem constitutes  the starting point of the learning process. Such a method provides students with the suitable knowledge for problem solving and presents numerous and significant differences compared to traditional education.In particular, I analyze the theoretic aspects of problem learning by tracing a history and presenting its structure, clarifying the role of the tutor in the various phases of the learning process. The method has found a wide diffusion since the beginning of the 70s and numerous studies have confirmed the advantages. The effectiveness of PBL is construable and is based on principles of constructivism and cognitivism.

  17. Bases de datos Multimedia

    OpenAIRE

    Blanco Andrés, Pedro José

    2005-01-01

    Aquest treball mira d'aproximar-se a la situació actual de les bases de dades multimèdia. Per a fer-ho, s'ha optat per un enfocament múltiple que té en compte diferents aspectes del producte multimèdia: els aspectes econòmics, és a dir, la situació del mercat (nombre de bases de dades, volum de negoci, creixement, etc.), els aspectes tècnics, és a dir, l'eina emprada, la seva evolució, les seves limitacions i problemes, i les seves aplicacions i serveis en diferents àrees (amb un interès espe...

  18. Conducting Polymer Based Nanobiosensors

    Directory of Open Access Journals (Sweden)

    Chul Soon Park

    2016-06-01

    Full Text Available In recent years, conducting polymer (CP nanomaterials have been used in a variety of fields, such as in energy, environmental, and biomedical applications, owing to their outstanding chemical and physical properties compared to conventional metal materials. In particular, nanobiosensors based on CP nanomaterials exhibit excellent performance sensing target molecules. The performance of CP nanobiosensors varies based on their size, shape, conductivity, and morphology, among other characteristics. Therefore, in this review, we provide an overview of the techniques commonly used to fabricate novel CP nanomaterials and their biosensor applications, including aptasensors, field-effect transistor (FET biosensors, human sense mimicking biosensors, and immunoassays. We also discuss prospects for state-of-the-art nanobiosensors using CP nanomaterials by focusing on strategies to overcome the current limitations.

  19. Knowledge based maintenance

    Energy Technology Data Exchange (ETDEWEB)

    Sturm, A. [Hamburgische Electacitaets-Werke AG Hamburg (Germany)

    1997-12-31

    The establishment of maintenance strategies is of crucial significance for the reliability of a plant and the economic efficiency of maintenance measures. Knowledge about the condition of components and plants from the technical and business management point of view therefore becomes one of the fundamental questions and the key to efficient management and maintenance. A new way to determine the maintenance strategy can be called: Knowledge Based Maintenance. A simple method for determining strategies while taking the technical condition of the components of the production process into account to the greatest possible degree which can be shown. A software with an algorithm for Knowledge Based Maintenance leads the user during complex work to the determination of maintenance strategies for this complex plant components. (orig.)

  20. Spintronics-based computing

    CERN Document Server

    Prenat, Guillaume

    2015-01-01

    This book provides a comprehensive introduction to spintronics-based computing for the next generation of ultra-low power/highly reliable logic, which is widely considered a promising candidate to replace conventional, pure CMOS-based logic. It will cover aspects from device to system-level, including magnetic memory cells, device modeling, hybrid circuit structure, design methodology, CAD tools, and technological integration methods. This book is accessible to a variety of readers and little or no background in magnetism and spin electronics are required to understand its content.  The multidisciplinary team of expert authors from circuits, devices, computer architecture, CAD and system design reveal to readers the potential of spintronics nanodevices to reduce power consumption, improve reliability and enable new functionality.  .

  1. Problem Based Game Design

    DEFF Research Database (Denmark)

    Reng, Lars; Schoenau-Fog, Henrik

    2011-01-01

    At Aalborg University’s department of Medialogy, we are utilizing the Problem Based Learning method to encourage students to solve game design problems by pushing the boundaries and designing innovative games. This paper is concerned with describing this method, how students employ it in various ...... projects and how they learn to analyse, design, and develop for innovation by using it. We will present various cases to exemplify the approach and focus on how the method engages students and aspires for innovation in digital entertainment and games.......At Aalborg University’s department of Medialogy, we are utilizing the Problem Based Learning method to encourage students to solve game design problems by pushing the boundaries and designing innovative games. This paper is concerned with describing this method, how students employ it in various...

  2. Terrorism Knowledge Base (TKB)

    Science.gov (United States)

    2008-04-01

    These sorts of inferences are trivial for humans, but they can’t be performed by machines that lack the commonsense knowledge that makes them valid...inference abilities in conjunction with its commonsense geospatial knowledge enable it to conclude the fact that Rafik was not in Los Angeles at T, from...comprehensive knowledge base ontology and schema, formulated by and agreed upon by panels of internationally recognized terrorism experts; produced a

  3. REST based service composition

    DEFF Research Database (Denmark)

    Grönvall, Erik; Ingstrup, Mads; Pløger, Morten

    2011-01-01

    This paper presents an ongoing work developing and testing a Service Composition framework based upon the REST architecture named SECREST. A minimalistic approach have been favored instead of a creating a complete infrastructure. One focus has been on the system's interaction model. Indeed, an aim...... is to allow users in different healthcare scenarios to experiment with service composition to support highly individual and changing needs....

  4. Geovisualization based upon KML

    OpenAIRE

    Marianna Zichar

    2012-01-01

    The extension kml refers to a human readable file format that is based on a markup language deriving from XML and became rather popular mainly due to Google Earth. This type of files uses not only the classical geometric data, but coordinates of longitude and latitude (in case of 3D system also coordinate of altitude) to visualize the different geographically referenced objects, phenomena and processes. Nowadays more than 10 millions of kml files can be accessed via the internet, and this num...

  5. Air Force Smart Bases

    Science.gov (United States)

    2017-10-19

    bases and missions toward an enterprise solution for incorporating smart technologies in the future (defined in the Information Environment Mission... technology might enrich and protect our nation, businesses, and lives. As a human-centric design center, we seek out unique ways to connect Air Force...warfighters with current and future technology in meaningful ways. We look to transfer, license, and share promising prototypes, solutions, and knowledge

  6. Luxury-based Growth

    OpenAIRE

    Shiro Kuwahara

    2006-01-01

    Assuming that there exists a preference for luxury goods and a knowledge spillover from luxury goods production to goods production, this paper constructs an endogenous economic growth model. The model predicts two steady states: one is a steady positive growth state with regard to luxury goods production, and the other is a zero growth state in the absence of luxury goods production. Thus, this study examines the polarization of economies based on luxury goods consumption

  7. Spiritual-based Leadership

    DEFF Research Database (Denmark)

    Pruzan, Peter

    2015-01-01

    Although far from mainstream, the concept of spiritual-based leadership is emerging as an inclusive and yet highly personal approach to leadership that integrates a leader’s inner perspectives on identity, purpose, responsibility and success with her or his decisions and actions in the outer world...... of business—and therefore it is also emerging as a significant framework for understanding, practicing, communicating and teaching the art and profession of leadership....

  8. Boid Based Timetable Generation

    Science.gov (United States)

    Fulmański, Piotr

    2008-09-01

    The general timetabling problem consists of scheduling a number of events into a finite number of periods so that no individual entity is required to attend two or more events simultaneously. In most cases there will also be restrictions on the amount of resources, time etc. In this document we try to investigate a new method of timetable generation based not on popular evolutionary approach but on boids concept.

  9. As bases do petismo

    Directory of Open Access Journals (Sweden)

    David Samuels

    2004-10-01

    Full Text Available A partir dos dados do ESEB de 2002 o autor realiza um estudo das bases eleitorais do PT e de hipóteses sobre a natureza do petismo. Através de técnicas estatísticas multivariadas, são testadas relações do petismo com variáveis demográficas, socioeconômicas e variáveis relativas a questões políticas específicas. Os resultados apontam que apenas a escolaridade tem uma associação específica com o petismo, com implicações para o seu comportamento sócio-político.Based on the results of the 2002 Brazilian Electoral Study, the author analyses the electoral bases of the Worker´s Party and the factors associated with the "petismo". The relationships between the "petismo"and the socioeconomic, demographic and political variables are tested using multivariate analysis. The results indicate that the only "social category"associated with "petismo"is level of education, and it has clear implications to their social and political behavior.

  10. Fluorescence-based biosensors.

    Science.gov (United States)

    Strianese, Maria; Staiano, Maria; Ruggiero, Giuseppe; Labella, Tullio; Pellecchia, Claudio; D'Auria, Sabato

    2012-01-01

    The field of optical sensors has been a growing research area over the last three decades. A wide range of books and review articles has been published by experts in the field who have highlighted the advantages of optical sensing over other transduction methods. Fluorescence is by far the method most often applied and comes in a variety of schemes. Nowadays, one of the most common approaches in the field of optical biosensors is to combine the high sensitivity of fluorescence detection in combination with the high selectivity provided by ligand-binding proteins. In this chapter we deal with reviewing our recent results on the implementation of fluorescence-based sensors for monitoring environmentally hazardous gas molecules (e.g. nitric oxide, hydrogen sulfide). Reflectivity-based sensors, fluorescence correlation spectroscopy-based (FCS) systems, and sensors relying on the enhanced fluorescence emission on silver island films (SIFs) coupled to the total internal reflection fluorescence mode (TIRF) for the detection of gliadin and other prolamines considered toxic for celiac patients are also discussed herein.

  11. Nanowire-based thermoelectrics

    Science.gov (United States)

    Ali, Azhar; Chen, Yixi; Vasiraju, Venkata; Vaddiraju, Sreeram

    2017-07-01

    Research on thermoelectrics has seen a huge resurgence since the early 1990s. The ability of tuning a material’s electrical and thermal transport behavior upon nanostructuring has led to this revival. Nevertheless, thermoelectric performances of nanowires and related materials lag far behind those achieved with thin-film superlattices and quantum dot-based materials. This is despite the fact that nanowires offer many distinct advantages in enhancing the thermoelectric performances of materials. The simplicity of the strategy is the first and foremost advantage. For example, control of the nanowire diameters and their surface roughnesses will aid in enhancing their thermoelectric performances. Another major advantage is the possibility of obtaining high thermoelectric performances using simpler nanowire chemistries (e.g., elemental and binary compound semiconductors), paving the way for the fabrication of thermoelectric modules inexpensively from non-toxic elements. In this context, the topical review provides an overview of the current state of nanowire-based thermoelectrics. It concludes with a discussion of the future vision of nanowire-based thermoelectrics, including the need for developing strategies aimed at the mass production of nanowires and their interface-engineered assembly into devices. This eliminates the need for trial-and-error strategies and complex chemistries for enhancing the thermoelectric performances of materials.

  12. Microbead agglutination based assays

    KAUST Repository

    Kodzius, Rimantas

    2013-01-21

    We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

  13. Aperiodic-metamaterial-based absorber

    OpenAIRE

    Quanlong Yang; Xieyu Chen; Yanfeng Li; Xueqian Zhang; Yuehong Xu; Zhen Tian; Chunmei Ouyang; Jianqiang Gu; Jiaguang Han; Weili Zhang

    2017-01-01

    The periodic-metamaterial-based perfect absorber has been studied broadly. Conversely, if the unit cell in the metamaterial-based absorber is arranged aperiodically (aperiodic-metamaterial-based absorber), how does it perform? Inspired by this, here we present a systematic study of the aperiodic-metamaterial-based absorber. By investigating the response of metamaterial absorbers based on periodic, Fibonacci, Thue-Morse, and quasicrystal lattices, we found that aperiodic-metamaterial-based abs...

  14. Nanoplatform-based molecular imaging

    National Research Council Canada - National Science Library

    Chen, Xiaoyuan

    2011-01-01

    "Nanoplathform-Based Molecular Imaging provides rationale for using nanoparticle-based probes for molecular imaging, then discusses general strategies for this underutilized, yet promising, technology...

  15. Polyolefin-Based Aerogels

    Science.gov (United States)

    Lee, Je Kyun; Gould, George

    2012-01-01

    An organic polybutadiene (PB) rubberbased aerogel insulation material was developed that will provide superior thermal insulation and inherent radiation protection, exhibiting the flexibility, resiliency, toughness, and durability typical of the parent polymer, yet with the low density and superior insulation properties associated with the aerogels. The rubbery behaviors of the PB rubber-based aerogels are able to overcome the weak and brittle nature of conventional inorganic and organic aerogel insulation materials. Additionally, with higher content of hydrogen in their structure, the PB rubber aerogels will also provide inherently better radiation protection than those of inorganic and carbon aerogels. Since PB rubber aerogels also exhibit good hydrophobicity due to their hydrocarbon molecular structure, they will provide better performance reliability and durability as well as simpler, more economic, and environmentally friendly production over the conventional silica or other inorganic-based aerogels, which require chemical treatment to make them hydrophobic. Inorganic aerogels such as silica aerogels demonstrate many unusual and useful properties. There are several strategies to overcoming the drawbacks associated with the weakness and brittleness of silica aerogels. Development of the flexible fiber-reinforced silica aerogel composite blanket has proven one promising approach, providing a conveniently fielded form factor that is relatively robust toward handling in industrial environments compared to silica aerogel monoliths. However, the flexible silica aerogel composites still have a brittle, dusty character that may be undesirable, or even intolerable, in certain applications. Although the cross-linked organic aerogels such as resorcinol-formaldehyde (RF), polyisocyanurate, and cellulose aerogels show very high impact strength, they are also very brittle with little elongation (i.e., less rubbery). Also, silica and carbon aerogels are less efficient

  16. Knowledge based Entrepreneurship

    DEFF Research Database (Denmark)

    Heebøll, John

    This book is dedicated enterprising people with a technical or a scientific background who consider commercializing ideas and inventions within their field of expertise via a new business activity or a new company. It aims at distilling experiences from many successful and not so successful start......-up ventures from the Technical University of Denmark, 1988 – 2008 into practical, portable knowledge that can be used by future knowledge-based entrepreneurs to set up new companies efficiently or to stay away from it; to do what’s needed and avoid the pitfalls....

  17. Vision-based interaction

    CERN Document Server

    Turk, Matthew

    2013-01-01

    In its early years, the field of computer vision was largely motivated by researchers seeking computational models of biological vision and solutions to practical problems in manufacturing, defense, and medicine. For the past two decades or so, there has been an increasing interest in computer vision as an input modality in the context of human-computer interaction. Such vision-based interaction can endow interactive systems with visual capabilities similar to those important to human-human interaction, in order to perceive non-verbal cues and incorporate this information in applications such

  18. Linear optics based nanoscopy.

    Science.gov (United States)

    Gur, Aviram; Fixler, Dror; Micó, Vicente; Garcia, Javier; Zalevsky, Zeev

    2010-10-11

    Classically, optical systems are considered to have a fundamental resolution limit due to wave nature of light. This article presents a novel method for observing sub-wavelength features in a conventional optical microscope using linear optics. The operation principle is based on a random and time varying flow of nanoparticles moving in proximity to the inspected sample. Those particles excite the evanescent waves and couple them into harmonic waves. The sub-wavelength features are encoded and later on digitally decoded by proper image processing of a sequence of images. The achievable final resolution limit corresponds to the size of the nanoparticles. Experimental proof of principle validation of the technique is reported.

  19. Sustainability Base Construction Update

    Science.gov (United States)

    Mewhinney, Michael

    2012-01-01

    Construction of the new Sustainability Base Collaborative support facility, expected to become the highest performing building in the federal government continues at NASA's Ames Research Center, Moffet Field, Calif. The new building is designed to achieve a platinum rating under the leadership in Energy and Environment Design (LEED) new construction standards for environmentally sustainable construction developed by the U. S. Green Building Council, Washington, D. C. When completed by the end of 2011, the $20.6 million building will feature near zero net energy consumption, use 90 percent less potable water than conventionally build buildings of equivalent size, and will result in reduced building maintenance costs.

  20. LIGHTWEIGHT CONCRETE BASED GRANSHLAK

    Directory of Open Access Journals (Sweden)

    NETESA M. I.

    2016-02-01

    Full Text Available Raising of problem. Concrete advisable to obtain a low strength with local secondary resources for recycling and reduce the environmental burden on the environment. But it is important to design such concrete compositions with a reduced flow of cement. It is known that the coefficient of efficiency of use of cement in the concrete of the heavy and B10 is less than about 0.5, which is almost two times smaller than in class B15 concrete and above. Even lower coefficient of efficiency in light concrete cement low strength. Therefore, it is important to find patterns determining the composition of lightweight concrete based on local-products industry with more efficient use of cement in them. Purpose.. Based on the analysis of earlier research results, including with the use of methods of mathematical planning of experiments to determine the concrete contents, which can provide the requirements for the underlying layers of the floor, the compressive strength of which should correspond to the class B5. It is important to provide the required strength at minimum flow of the cement, which is the most expensive and energy-intensive part of concrete. Conclusion. Analysis of the test results of control samples of concrete in 28-day-old, the following laws. The required tensile strength of concrete compressive strength of 7.0 MPa can be obtained in the test range when used in formulations as a filler as the Dnieper hydroelectric power station fly ash and tailings Krivoy Rog iron ore YuGOK. To ensure providing the required characteristic strength of the concrete in the underlying layers of the floor is advisable to use a nominal composition per cubic meter of concrete: cement 160 kg granshlaka Plant named after Petrovsky, 675 kg of fly ash Dnieper HPP 390 kg, 400 kg of sand, 230 liters of water. Thus, while ensuring rational grain composition components can obtain the desired strength lightweight concrete based granshlaka plant Petrovsky, using as fillers

  1. Location-based Scheduling

    DEFF Research Database (Denmark)

    Andersson, Niclas; Christensen, Knud

    The coordination of activities and resources in order to establish an effective production flow is central to the management of construction projects. The traditional technique for coordination of activities and resources in construction projects is the CPM-scheduling, which has been...... on the market. However, CPM is primarily an activity based method that takes the activity as the unit of focus and there is criticism raised, specifically in the case of construction projects, on the method for deficient management of construction work and continuous flow of resources. To seek solutions...

  2. [Evidence-based physiotherapy].

    Science.gov (United States)

    Bender, Tamás

    2013-12-01

    This article on physiotherapy presents some current evidence stating the strengths and weaknesses of the physiotherapeutic procedures. In the area of physiotherapy empirical data obtained during decades were overtaken by evidence from current studies. The author points out the great problem of physiotherapy, namely the heterogeneity of the applied parameters. Knowledge of current evidence may be very important and helpful for the physicians, but the author proposes, from the practical point of view, that physiotherapeutical procedures based on exprience and used for many years should not be entirely neglected. Nowadays physiotherapy plays an important role in the treament of locomotor diseases but its use is increasing in other fields of medicine, as well.

  3. Evidence-based guidelines

    DEFF Research Database (Denmark)

    Rovira, Àlex; Wattjes, Mike P; Tintoré, Mar

    2015-01-01

    The clinical use of MRI in patients with multiple sclerosis (MS) has advanced markedly over the past few years. Technical improvements and continuously emerging data from clinical trials and observational studies have contributed to the enhanced performance of this tool for achieving a prompt...... diagnosis in patients with MS. The aim of this article is to provide guidelines for the implementation of MRI of the brain and spinal cord in the diagnosis of patients who are suspected of having MS. These guidelines are based on an extensive review of the recent literature, as well as on the personal...... of MRI in clinical practice for the diagnosis of MS....

  4. Knowledge based Entrepreneurship

    DEFF Research Database (Denmark)

    Heebøll, John

    This book is dedicated enterprising people with a technical or a scientific background who consider commercializing ideas and inventions within their field of expertise via a new business activity or a new company. It aims at distilling experiences from many successful and not so successful start-up...... ventures from the Technical University of Denmark, 1988 – 2008 into practical, portable knowledge that can be used by future knowledge-based entrepreneurs to set up new companies efficiently or to stay away from it; to do what’s needed and avoid the pitfalls....

  5. Polymerization Using Phosphazene Bases

    KAUST Repository

    Zhao, Junpeng

    2015-09-01

    In the recent rise of metal-free polymerization techniques, organic phosphazene superbases have shown their remarkable strength as promoter/catalyst for the anionic polymerization of various types of monomers. Generally, the complexation of phosphazene base with the counterion (proton or lithium cation) significantly improves the nucleophilicity of the initiator/chain end resulting in highly enhanced polymerization rates, as compared with conventional metalbased initiating systems. In this chapter, the general features of phosphazenepromoted/catalyzed polymerizations and the applications in macromolecular engineering (synthesis of functionalized polymers, block copolymers, and macromolecular architectures) are discussed with challenges and perspectives being pointed out.

  6. Mars base buildup scenarios

    Energy Technology Data Exchange (ETDEWEB)

    Blacic, J.D.

    1985-01-01

    Two surface base build-up scenarios are presented in order to help visualize the mission and to serve as a basis for trade studies. In the first scenario, direct manned landings on the Martian surface occur early in the missions and scientific investigation is the main driver and rationale. In the second scenario, early development of an infrastructure to exploite the volatile resources of the Martian moons for economic purposes is emphasized. Scientific exploration of the surface is delayed at first, but once begun develops rapidly aided by the presence of a permanently manned orbital station.

  7. Unification-Based Glossing

    CERN Document Server

    Hatzivassiloglou, V; Hatzivassiloglou, Vasileios; Knight, Kevin

    1995-01-01

    We present an approach to syntax-based machine translation that combines unification-style interpretation with statistical processing. This approach enables us to translate any Japanese newspaper article into English, with quality far better than a word-for-word translation. Novel ideas include the use of feature structures to encode word lattices and the use of unification to compose and manipulate lattices. Unification also allows us to specify abstract features that delay target-language synthesis until enough source-language information is assembled. Our statistical component enables us to search efficiently among competing translations and locate those with high English fluency.

  8. Agent-Based Optimization

    CERN Document Server

    Jędrzejowicz, Piotr; Kacprzyk, Janusz

    2013-01-01

    This volume presents a collection of original research works by leading specialists focusing on novel and promising approaches in which the multi-agent system paradigm is used to support, enhance or replace traditional approaches to solving difficult optimization problems. The editors have invited several well-known specialists to present their solutions, tools, and models falling under the common denominator of the agent-based optimization. The book consists of eight chapters covering examples of application of the multi-agent paradigm and respective customized tools to solve  difficult optimization problems arising in different areas such as machine learning, scheduling, transportation and, more generally, distributed and cooperative problem solving.

  9. Bi-based superconductor

    Directory of Open Access Journals (Sweden)

    S E Mousavi

    2009-08-01

    Full Text Available   In this paper, Bi-Sr-Ca-Cu-O (BCSCCO system superconductor is made by the solid state reaction method. The effect of doping Pb, Cd, Sb, Cu and annealing time on the critical temperature and critical current density have been investigated. The microstructure and morphology of the samples have been studied by X-ray diffraction, scanning electron microscope and energy dispersive X-ray. The results show that the fraction of Bi-2223 phase in the Bi- based superconductor, critical temperature and critical current density depend on the annealing temperature, annealing time and the kind and amount of doping .

  10. Cellular based cancer vaccines

    DEFF Research Database (Denmark)

    Hansen, Morten; Met, O; Svane, I M

    2012-01-01

    Cancer vaccines designed to re-calibrate the existing host-tumour interaction, tipping the balance from tumor acceptance towards tumor control holds huge potential to complement traditional cancer therapies. In general, limited success has been achieved with vaccines composed of tumor...... in vitro migration via autocrine receptor-mediated endocytosis of CCR7. In the current review, we discuss optimal design of DC maturation focused on pre-clinical as well as clinical results from standard and polarized dendritic cell based cancer vaccines....

  11. Cellular based cancer vaccines

    DEFF Research Database (Denmark)

    Hansen, M; Met, Ö; Svane, I M

    2012-01-01

    Cancer vaccines designed to re-calibrate the existing host-tumour interaction, tipping the balance from tumor acceptance towards tumor control holds huge potential to complement traditional cancer therapies. In general, limited success has been achieved with vaccines composed of tumor...... to transiently affect in vitro migration via autocrine receptor-mediated endocytosis of CCR7. In the current review, we discuss optimal design of DC maturation focused on pre-clinical as well as clinical results from standard and polarized dendritic cell based cancer vaccines....

  12. In Silico Detection and Typing of Plasmids using PlasmidFinder and Plasmid Multilocus Sequence Typing

    DEFF Research Database (Denmark)

    Carattoli, Alessandra; Zankari, Ea; García-Fernández, Aurora

    2014-01-01

    into a Web tool called PlasmidFinder that can be used for replicon sequence analysis of raw, contig group, or completely assembled and closed plasmid sequencing data. The PlasmidFinder database currently consists of 116 replicon sequences that match with at least at 80% nucleotide identity all replicon...... of Salmonella enterica serovar Typhimurium isolates. PlasmidFinder identified a total of 103 replicons and between zero and five different plasmid replicons within each of 49 S. Typhimurium draft genomes tested. The pMLST Web tool was able to subtype genomic sequencing data of plasmids, revealing both known...

  13. SPACE BASED INTERCEPTOR SCALING

    Energy Technology Data Exchange (ETDEWEB)

    G. CANAVAN

    2001-02-01

    Space Based Interceptor (SBI) have ranges that are adequate to address rogue ICBMs. They are not overly sensitive to 30-60 s delay times. Current technologies would support boost phase intercept with about 150 interceptors. Higher acceleration and velocity could reduce than number by about a factor of 3 at the cost of heavier and more expensive Kinetic Kill Vehicles (KKVs). 6g SBI would reduce optimal constellation costs by about 35%; 8g SBI would reduce them another 20%. Interceptor ranges fall rapidly with theater missile range. Constellations increase significantly for ranges under 3,000 km, even with advanced interceptor technology. For distributed launches, these estimates recover earlier strategic scalings, which demonstrate the improved absentee ratio for larger or multiple launch areas. Constellations increase with the number of missiles and the number of interceptors launched at each. The economic estimates above suggest that two SBI per missile with a modest midcourse underlay is appropriate. The SBI KKV technology would appear to be common for space- and surface-based boost phase systems, and could have synergisms with improved midcourse intercept and discrimination systems. While advanced technology could be helpful in reducing costs, particularly for short range theater missiles, current technology appears adequate for pressing rogue ICBM, accidental, and unauthorized launches.

  14. Design Based Wilderness Education

    Directory of Open Access Journals (Sweden)

    Christopher R. Saulnier

    2015-02-01

    Full Text Available The Massachusetts Institute of Technology (MIT has been collaborating since 2010 with the Singapore Ministry of Education to help develop the Singapore University of Technology and Design (SUTD. One element of this collaboration, the Global Leadership Program (GLP, aims to provide SUTD students with the opportunity to interact with the MIT community and experience MIT’s academic culture. During GLP students participate in a program designed to develop leadership ability while also increasing their understanding of engineering science and design thinking. This paper introduces a curriculum combining the pedagogies of design-based learning and wilderness education that was implemented in the summer of 2014 to holistically address the development of these three competencies. Through design-based learning activities, both for and in a natural environment, students were encouraged to develop competencies in engineering science and engineering design while exploring the diverse attributes essential for success as an engineer. This paper examines the results of a retrospective post-then-pre survey administered to the participants upon completion of the program to explore the effects of the program on the development of professional engineering competencies. We find a statistically significant increase in items associated with Individual Leadership Skill, Group Leadership Skill and the role of Society and the Economy. These results are triangulated with student exit interviews and instructor observations.

  15. Microlaser-based displays

    Science.gov (United States)

    Bergstedt, Robert; Fink, Charles G.; Flint, Graham W.; Hargis, David E.; Peppler, Philipp W.

    1997-07-01

    Laser Power Corporation has developed a new type of projection display, based upon microlaser technology and a novel scan architecture, which provides the foundation for bright, extremely high resolution images. A review of projection technologies is presented along with the limitations of each and the difficulties they experience in trying to generate high resolution imagery. The design of the microlaser based projector is discussed along with the advantage of this technology. High power red, green, and blue microlasers have been designed and developed specifically for use in projection displays. These sources, in combination with high resolution, high contrast modulator, produce a 24 bit color gamut, capable of supporting the full range of real world colors. The new scan architecture, which reduces the modulation rate and scan speeds required, is described. This scan architecture, along with the inherent brightness of the laser provides the fundamentals necessary to produce a 5120 by 4096 resolution display. The brightness and color uniformity of the display is excellent, allowing for tiling of the displays with far fewer artifacts than those in a traditionally tiled display. Applications for the display include simulators, command and control centers, and electronic cinema.

  16. Model Based Definition

    Science.gov (United States)

    Rowe, Sidney E.

    2010-01-01

    In September 2007, the Engineering Directorate at the Marshall Space Flight Center (MSFC) created the Design System Focus Team (DSFT). MSFC was responsible for the in-house design and development of the Ares 1 Upper Stage and the Engineering Directorate was preparing to deploy a new electronic Configuration Management and Data Management System with the Design Data Management System (DDMS) based upon a Commercial Off The Shelf (COTS) Product Data Management (PDM) System. The DSFT was to establish standardized CAD practices and a new data life cycle for design data. Of special interest here, the design teams were to implement Model Based Definition (MBD) in support of the Upper Stage manufacturing contract. It is noted that this MBD does use partially dimensioned drawings for auxiliary information to the model. The design data lifecycle implemented several new release states to be used prior to formal release that allowed the models to move through a flow of progressive maturity. The DSFT identified some 17 Lessons Learned as outcomes of the standards development, pathfinder deployments and initial application to the Upper Stage design completion. Some of the high value examples are reviewed.

  17. Challenge Based Innovation gala

    CERN Multimedia

    CERN. Geneva; Utriainen, Tuuli Maria; Toivonen, Harri; Nordberg, Markus

    2014-01-01

    Challenge Based Innovation gala   There’s a new experiment starting in CERN called IdeaLab where we work together with detector R&D researchers to help them to bridge their knowledge into a more human, societally oriented context. Currently we are located in B153, but will move our activities to a new facility next to the Globe in May 2014. One of our first pilot projects is a 5 month course CBI (Challenge Based Innovation) where two multidisciplinary student teams join forces with Edusafe & TALENT projects at CERN. Their goal is to discover what kind of tools for learning could be created in collaboration with the two groups. After months of user interviews and low resolution prototyping they are ready to share the results with us in the form of an afternoon gala. We warmly welcome you to join us to see the students' results and experience the prototypes they have conceived. The event is in three parts, you are welcome to visit all of them,...

  18. Constraint-based reachability

    Directory of Open Access Journals (Sweden)

    Arnaud Gotlieb

    2013-02-01

    Full Text Available Iterative imperative programs can be considered as infinite-state systems computing over possibly unbounded domains. Studying reachability in these systems is challenging as it requires to deal with an infinite number of states with standard backward or forward exploration strategies. An approach that we call Constraint-based reachability, is proposed to address reachability problems by exploring program states using a constraint model of the whole program. The keypoint of the approach is to interpret imperative constructions such as conditionals, loops, array and memory manipulations with the fundamental notion of constraint over a computational domain. By combining constraint filtering and abstraction techniques, Constraint-based reachability is able to solve reachability problems which are usually outside the scope of backward or forward exploration strategies. This paper proposes an interpretation of classical filtering consistencies used in Constraint Programming as abstract domain computations, and shows how this approach can be used to produce a constraint solver that efficiently generates solutions for reachability problems that are unsolvable by other approaches.

  19. Skull base tumor model.

    Science.gov (United States)

    Gragnaniello, Cristian; Nader, Remi; van Doormaal, Tristan; Kamel, Mahmoud; Voormolen, Eduard H J; Lasio, Giovanni; Aboud, Emad; Regli, Luca; Tulleken, Cornelius A F; Al-Mefty, Ossama

    2010-11-01

    Resident duty-hours restrictions have now been instituted in many countries worldwide. Shortened training times and increased public scrutiny of surgical competency have led to a move away from the traditional apprenticeship model of training. The development of educational models for brain anatomy is a fascinating innovation allowing neurosurgeons to train without the need to practice on real patients and it may be a solution to achieve competency within a shortened training period. The authors describe the use of Stratathane resin ST-504 polymer (SRSP), which is inserted at different intracranial locations to closely mimic meningiomas and other pathological entities of the skull base, in a cadaveric model, for use in neurosurgical training. Silicone-injected and pressurized cadaveric heads were used for studying the SRSP model. The SRSP presents unique intrinsic metamorphic characteristics: liquid at first, it expands and foams when injected into the desired area of the brain, forming a solid tumorlike structure. The authors injected SRSP via different passages that did not influence routes used for the surgical approach for resection of the simulated lesion. For example, SRSP injection routes included endonasal transsphenoidal or transoral approaches if lesions were to be removed through standard skull base approach, or, alternatively, SRSP was injected via a cranial approach if the removal was planned to be via the transsphenoidal or transoral route. The model was set in place in 3 countries (US, Italy, and The Netherlands), and a pool of 13 physicians from 4 different institutions (all surgeons and surgeons in training) participated in evaluating it and provided feedback. All 13 evaluating physicians had overall positive impressions of the model. The overall score on 9 components evaluated--including comparison between the tumor model and real tumor cases, perioperative requirements, general impression, and applicability--was 88% (100% being the best possible

  20. MS Based Metabonomics

    Energy Technology Data Exchange (ETDEWEB)

    Want, Elizabeth J.; Metz, Thomas O.

    2010-03-01

    Metabonomics is the latest and least mature of the systems biology triad, which also includes genomics and proteomics, and has its origins in the early orthomolecular medicine work pioneered by Linus Pauling and Arthur Robinson. It was defined by Nicholson and colleagues in 1999 as the quantitative measurement of perturbations in the metabolite complement of an integrated biological system in response to internal or external stimuli, and is often used today to describe many non-global types of metabolite analyses. Applications of metabonomics are extensive and include toxicology, nutrition, pharmaceutical research and development, physiological monitoring and disease diagnosis. For example, blood samples from millions of neonates are tested routinely by mass spectrometry (MS) as a diagnostic tool for inborn errors of metabolism. The metabonome encompasses a wide range of structurally diverse metabolites; therefore, no single analytical platform will be sufficient. Specialized sample preparation and detection techniques are required, and advances in NMR and MS technologies have led to enhanced metabonome coverage, which in turn demands improved data analysis approaches. The role of MS in metabonomics is still evolving as instrumentation and software becomes more sophisticated and as researchers realize the strengths and limitations of current technology. MS offers a wide dynamic range, high sensitivity, and reproducible, quantitative analysis. These attributes are essential for addressing the challenges of metabonomics, as the range of metabolite concentrations easily exceeds nine orders of magnitude in biofluids, and the diversity of molecular species ranges from simple amino and organic acids to lipids and complex carbohydrates. Additional challenges arise in generating a comprehensive metabolite profile, downstream data processing and analysis, and structural characterization of important metabolites. A typical workflow of MS-based metabonomics is shown in Figure

  1. Transaction based approach

    Science.gov (United States)

    Hunka, Frantisek; Matula, Jiri

    2017-07-01

    Transaction based approach is utilized in some methodologies in business process modeling. Essential parts of these transactions are human beings. The notion of agent or actor role is usually used for them. The paper on a particular example describes possibilities of Design Engineering Methodology for Organizations (DEMO) and Resource-Event-Agent (REA) methodology. Whereas the DEMO methodology can be regarded as a generic methodology having its foundation in the theory of Enterprise Ontology the REA methodology is regarded as the domain specific methodology and has its origin in accountancy systems. The results of these approaches is that the DEMO methodology captures everything that happens in the reality with a good empirical evidence whereas the REA methodology captures only changes connected with economic events. Economic events represent either change of the property rights to economic resource or consumption or production of economic resources. This results from the essence of economic events and their connection to economic resources.

  2. Gossip-Based Broadcast

    Science.gov (United States)

    Leitão, João; Pereira, José; Rodrigues, Luís

    Gossip, or epidemic, protocols have emerged as a powerful strategy to implement highly scalable and resilient reliable broadcast primitives on large scale peer-to-peer networks. Epidemic protocols are scalable because they distribute the load among all nodes in the system and resilient because they have an intrinsic level of redundancy that masks node and network failures. This chapter provides an introduction to gossip-based broadcast on large-scale unstructured peer-to-peer overlay networks: it surveys the main results in the field, discusses techniques to build and maintain the overlays that support efficient dissemination strategies, and provides an in-depth discussion and experimental evaluation of two concrete protocols, named HyParView and Plumtree.

  3. Droplet-based microfluidics.

    Science.gov (United States)

    Sharma, Sanjiv; Srisa-Art, Monpichar; Scott, Steven; Asthana, Amit; Cass, Anthony

    2013-01-01

    Droplet-based microfluidics or digital microfluidics is a subclass of microfluidic devices, wherein droplets are generated using active or passive methods. The active method for generation of droplets involves the use of an external factor such as an electric field for droplet generation. Two techniques that fall in this category are dielectrophoresis (DEP) and electrowetting on dielectric (EWOD). In passive methods, the droplet generation depends on the geometry and dimensions of the device. T-junction and flow focusing methods are examples of passive methods used for generation of droplets. In this chapter the methods used for droplet generation, mixing of contents of droplets, and the manipulation of droplets are described in brief. A review of the applications of digital microfluidics with emphasis on the last decade is presented.

  4. Graphene based biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Gürel, Hikmet Hakan, E-mail: hhakan.gurel@kocaeli.edu.tr [Kocaeli University, Kocaeli (Turkey); Salmankurt, Bahadır [Sakarya University, Sakarya (Turkey)

    2016-03-25

    Nanometer-sized graphene as a 2D material has unique chemical and electronic properties. Because of its unique physical, chemical, and electronic properties, its interesting shape and size make it a promising nanomaterial in many biological applications. It is expected that biomaterials incorporating graphene will be developed for the graphene-based drug delivery systems and biomedical devices. The interactions of biomolecules and graphene are long-ranged and very weak. Development of new techniques is very desirable for design of bioelectronics sensors and devices. In this work, we present first-principles calculations within density functional theory to calculate effects of charging on nucleobases on graphene. It is shown that how modify structural and electronic properties of nucleobases on graphene by applied charging.

  5. Graphene based biosensors

    Science.gov (United States)

    Gürel, Hikmet Hakan; Salmankurt, Bahadır

    2016-03-01

    Nanometer-sized graphene as a 2D material has unique chemical and electronic properties. Because of its unique physical, chemical, and electronic properties, its interesting shape and size make it a promising nanomaterial in many biological applications. It is expected that biomaterials incorporating graphene will be developed for the graphene-based drug delivery systems and biomedical devices. The interactions of biomolecules and graphene are long-ranged and very weak. Development of new techniques is very desirable for design of bioelectronics sensors and devices. In this work, we present first-principles calculations within density functional theory to calculate effects of charging on nucleobases on graphene. It is shown that how modify structural and electronic properties of nucleobases on graphene by applied charging.

  6. Pitch Based Sound Classification

    DEFF Research Database (Denmark)

    Nielsen, Andreas Brinch; Hansen, Lars Kai; Kjems, U

    2006-01-01

    -max output function. Both linear and quadratic inputs are used. The model is trained on 2 hours of sound and tested on publicly available data. A test classification error below 0.05 with 1 s classification windows is achieved. Further more it is shown that linear input performs as well as a quadratic......A sound classification model is presented that can classify signals into music, noise and speech. The model extracts the pitch of the signal using the harmonic product spectrum. Based on the pitch estimate and a pitch error measure, features are created and used in a probabilistic model with soft......, and that even though classification gets marginally better, not much is achieved by increasing the window size beyond 1 s....

  7. Base isolation: Fresh insight

    Energy Technology Data Exchange (ETDEWEB)

    Shustov, V.

    1993-07-15

    The objective of the research is a further development of the engineering concept of seismic isolation. Neglecting the transient stage of seismic loading results in a widespread misjudgement: The force of resistance associated with velocity is mostly conceived as a source of damping vibrations, though it is an active force at the same time, during an earthquake type excitation. For very pliant systems such as base isolated structures with relatively low bearing stiffness and with artificially added heavy damping mechanism, the so called `damping`` force may occur even the main pushing force at an earthquake. Thus, one of the two basic pillars of the common seismic isolation philosophy, namely, the doctrine of usefulness and necessity of a strong damping mechanism, is turning out to be a self-deception, sometimes even jeopardizing the safety of structures and discrediting the very idea of seismic isolation. There is a way out: breaking with damping dependancy.

  8. Surface Plasmon Based Spectrometer

    Science.gov (United States)

    Wig, Andrew; Passian, Ali; Boudreaux, Philip; Ferrell, Tom

    2008-03-01

    A spectrometer that uses surface plasmon excitation in thin metal films to separate light into its component wavelengths is described. The use of surface plasmons as a dispersive medium sets this spectrometer apart from prism, grating, and interference based variants and allows for the miniaturization of this device. Theoretical and experimental results are presented for two different operation models. In the first case surface plasmon tunneling in the near field is used to provide transmission spectra of different broad band-pass, glass filters across the visible wavelength range with high stray-light rejection at low resolution as well as absorption spectra of chlorophyll extracted from a spinach leaf. The second model looks at the far field components of surface plasmon scattering.

  9. Touching base with OPERA

    CERN Multimedia

    CERN Bulletin

    2011-01-01

    Three seminars – at CERN, at Gran Sasso and in Japan – and an article calling for the scrutiny of the scientific community: the OPERA Collaboration opened its research publicly. In addition to huge press coverage, this triggered welcome reactions from colleagues around the world, many of whom will attempt to independently interpret and reproduce the measurement. OPERA’s Spokesperson touches base with the Bulletin.   The CERN Main Auditorium was crowded as OPERA Physics co-ordinator Dario Autiero presented the results of their research (23 September 2011). According to the OPERA strategy, the results of the measurements are in the hands of the scientific community and, as for any other scientific result, several months will be needed before other groups will be able to perform an independent measurement. In the meantime, the OPERA Collaboration is dealing with an avalanche of emails from the scientific community, members of the general public, and the press. &...

  10. PROBLEM-BASED LEARNING

    Directory of Open Access Journals (Sweden)

    David Esema

    2012-12-01

    Full Text Available Problem-Based Learning (PBL, adalah model pembelajaran yang awalnya dikembangkan di dunia pendidikan medis, namun kini telah digunakan secara luas di berbagai jenjang dan bidang pendidikan. Basis teortits dari PBL adalah kolaborativisme dan konstruktivisme. Kemitraan, keterbukaan dan kejujuran, rasa hormat, dan kepercayaan menjadi nilai-nilai yang mendasari dan sekaligus menjadi prasyarat bagi keberhasilan PBL Sintaks PBL meliputi: pengenalan dan pemahaman konsep dasar, eksplorasi fakta dan informasi relevan secara mandiri, bertukar pemahaman dalam kelompok/kelas, kesimpulan dan evaluasi. Beberapa riset mengenai PBL baik di luar maupun di dalam negeri yang dipaparkan dan dibandingkan dalam tulisan ini menunjukkan data tentang efektifitas PBL dalam kegiatan pembelajaran.

  11. Surfel Based Geometry Resonstruction

    DEFF Research Database (Denmark)

    Andersen, Vedrana; Aanæs, Henrik; Bærentzen, Jakob Andreas

    2010-01-01

    We propose a method for retrieving a piecewise smooth surface from noisy data. In data acquired by a scanning process sampled points are almost never on the discontinuities making reconstruction of surfaces with sharp features difficult. Our method is based on a Markov Random Field (MRF......) formulation of a surface prior, with the surface represented as a collection of small planar patches, the surfels, associated with each data point. The main advantage of using surfels is that we avoid treating data points as vertices. MRF formulation of the surface prior allows us to separately model...... the likelihood (related to the mesh formation process) and the local surface properties. We chose to model the smoothness by considering two terms: the parallelism between neighboring surfels, and their overlap. We have demonstrated the feasibility of this approach on both synthetical and scanned data. In both...

  12. Lunar based massdriver applications

    Science.gov (United States)

    Ehresmann, Manfred; Gabrielli, Roland Atonius; Herdrich, Georg; Laufer, René

    2017-05-01

    The results of a lunar massdriver mission and system analysis are discussed and show a strong case for a permanent lunar settlement with a site near the lunar equator. A modular massdriver concept is introduced, which uses multiple acceleration modules to be able to launch large masses into a trajectory that is able to reach Earth. An orbital mechanics analysis concludes that the launch site will be in the Oceanus Procellarum a flat, Titanium rich lunar mare area. It is further shown that the bulk of massdriver components can be manufactured by collecting lunar minerals, which are broken down into its constituting elements. The mass to orbit transfer rates of massdriver case study are significant and can vary between 1.8 kt and 3.3 megatons per year depending on the available power. Thus a lunar massdriver would act as a catalyst for any space based activities and a game changer for the scale of feasible space projects.

  13. Technology based Education System

    DEFF Research Database (Denmark)

    Kant Hiran, Kamal; Doshi, Ruchi; Henten, Anders

    2016-01-01

    Abstract - Education plays a very important role for the development of the country. Education has multiple dimensions from schooling to higher education and research. In all these domains, there is invariably a need for technology based teaching and learning tools are highly demanded...... in the academic institutions. Thus, there is a need of comprehensive technology support system to cater the demands of all educational actors. Cloud Computing is one such comprehensive and user-friendly technology support environment that is the need of an hour. Cloud computing is the emerging technology that has...... and operational data that is used within a university for daily routine work. This paper presents a hybrid cloud computing model for higher education institutions to share intellectual data. Moreover, it proposes, the strategies for the implementation of the cloud computing in the academic institutions. Keywords...

  14. Problem Based Learning Online

    DEFF Research Database (Denmark)

    Kolbæk, Ditte

    2018-01-01

    Problem- and project- based learning (PBL) was originally developed for collaboration between physically present students and teachers, but political decisions at many universities also require that collaboration, dialogues, and other PBL activities take place online. Teachers design learning...... “How do two online learning designs affect student engagement in the PBL online modules?” The empirical data were collected and analyzed using a netnographic approach. The study finds that concepts such as self-directed learning and active involvement may be perceived very differently from the students...... situations with the intention of supporting the students. This study investigates how technology influenced the interaction between teachers and students and between students in two different online modules for master’s students at Aalborg University, Denmark. The study aims to answer the research question...

  15. Rate based failure detection

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, Brett Emery Trabun; Gamage, Thoshitha Thanushka; Bakken, David Edward

    2018-01-02

    This disclosure describes, in part, a system management component and failure detection component for use in a power grid data network to identify anomalies within the network and systematically adjust the quality of service of data published by publishers and subscribed to by subscribers within the network. In one implementation, subscribers may identify a desired data rate, a minimum acceptable data rate, desired latency, minimum acceptable latency and a priority for each subscription. The failure detection component may identify an anomaly within the network and a source of the anomaly. Based on the identified anomaly, data rates and or data paths may be adjusted in real-time to ensure that the power grid data network does not become overloaded and/or fail.

  16. Bases para proyectiles dirigidos

    Directory of Open Access Journals (Sweden)

    Editorial, Equipo

    1959-03-01

    Full Text Available Aunque actualmente no se ha llegado a una línea general de métodos o sistemas que gobiernen un tipo característico de rampa y servicios auxiliares necesarios para el lanzamiento al espacio de proyectiles dirigidos a grandes alturas y distancias, las experiencias obtenidas en diferentes ensayos, utilizando distintos tipos de proyectiles y trayectorias balísticas, han sentado toda una serie de procedimientos, datos y conclusiones de gran valor balístico que, aun teniendo en cuenta la continua evolución del proyectil, sus formas, combustibles y alcances, se conocen ya, con bastante aproximación, las condiciones mínimas que ha de reunir una base dedicada a este tipo de lanzamientos.

  17. Organic Biochar Based Fertilization

    Science.gov (United States)

    Schmidt, Hans-Peter; Pandit, Bishnu Hari; Cornelissen, Gerard; Kammann, Claudia

    2017-04-01

    Biochar produced in cost-efficient flame curtain kilns (Kon-Tiki) was nutrient enriched either with cow urine or with dissolved mineral (NPK) fertilizer to produce biochar-based fertilizers containing between 60-100 kg N, 5-60 kg P2O5 and 60-100 kg K2O, respectively, per ton of biochar. In 21 field trials nutrient-enriched biochars were applied at rates of 0.5 to 2 t ha-1 into the root zone of 13 different annual and perennial crops. Treatments combining biochar, compost and organic or chemical fertilizer were evaluated; control treatments contained the same amounts of nutrients but without biochar. All nutrient-enriched biochar substrates improved yields compared to their respective no-biochar controls. Biochar enriched with dissolved NPK produced on average 20% ± 5.1% (N=4) higher yields than standard NPK fertilization without biochar. Cow urine-enriched biochar blended with compost resulted on average in 123% ± 76.7% (N=13) higher yields compared to the organic farmer practice with cow urine-blended compost and outcompeted NPK-enriched biochar (same nutrient dose) by 103% ± 12.4% (N=4) on average. 21 field trials robustly revealed that low-dosage root zone application of organic biochar-based fertilizers caused substantial yield increases in rather fertile silt loam soils compared to traditional organic fertilization and to mineral NPK- or NPK-biochar fertilization. This can likely be explained by the nutrient carrier effect of biochar causing a slow nutrient release behavior, more balanced nutrient fluxes and reduced nutrient losses especially when liquid organic nutrients are used for the biochar enrichment. The results promise new pathways for optimizing organic farming and improving on-farm nutrient cycling.

  18. PROcess Based Diagnostics PROBE

    Science.gov (United States)

    Clune, T.; Schmidt, G.; Kuo, K.; Bauer, M.; Oloso, H.

    2013-01-01

    Many of the aspects of the climate system that are of the greatest interest (e.g., the sensitivity of the system to external forcings) are emergent properties that arise via the complex interplay between disparate processes. This is also true for climate models most diagnostics are not a function of an isolated portion of source code, but rather are affected by multiple components and procedures. Thus any model-observation mismatch is hard to attribute to any specific piece of code or imperfection in a specific model assumption. An alternative approach is to identify diagnostics that are more closely tied to specific processes -- implying that if a mismatch is found, it should be much easier to identify and address specific algorithmic choices that will improve the simulation. However, this approach requires looking at model output and observational data in a more sophisticated way than the more traditional production of monthly or annual mean quantities. The data must instead be filtered in time and space for examples of the specific process being targeted.We are developing a data analysis environment called PROcess-Based Explorer (PROBE) that seeks to enable efficient and systematic computation of process-based diagnostics on very large sets of data. In this environment, investigators can define arbitrarily complex filters and then seamlessly perform computations in parallel on the filtered output from their model. The same analysis can be performed on additional related data sets (e.g., reanalyses) thereby enabling routine comparisons between model and observational data. PROBE also incorporates workflow technology to automatically update computed diagnostics for subsequent executions of a model. In this presentation, we will discuss the design and current status of PROBE as well as share results from some preliminary use cases.

  19. Mainstreaming gesture based interfaces

    Directory of Open Access Journals (Sweden)

    David Procházka

    2013-01-01

    Full Text Available Gestures are a common way of interaction with mobile devices. They emerged especially with the iPhone production. Gestures in currently used devices are usually based on the original gestures presented by Apple in its iOS (iPhone Operating System. Therefore, there is a wide agreement on the mobile gesture design. In last years, it is possible to see experiments with gesture usage also in the other areas of consumer electronics and computers. The examples can include televisions, large projections etc. These gestures can be marked as spatial or 3D gestures. They are connected with a natural 3D environment rather than with a flat 2D screen. Nevertheless, it is hard to find a comparable design agreement within the spatial gestures. Various projects are based on completely different gesture sets. This situation is confusing for their users and slows down spatial gesture adoption.This paper is focused on the standardization of spatial gestures. The review of projects focused on spatial gesture usage is provided in the first part. The main emphasis is placed on the usability point-of-view. On the basis of our analysis, we argue that the usability is the key issue enabling the wide adoption. The mobile gesture emergence was possible easily because the iPhone gestures were natural. Therefore, it was not necessary to learn them.The design and implementation of our presentation software, which is controlled by gestures, is outlined in the second part of the paper. Furthermore, the usability testing results are provided as well. We have tested our application on a group of users not instructed in the implemented gestures design. These results were compared with the other ones, obtained with our original implementation. The evaluation can be used as the basis for implementation of similar projects.

  20. Base Research Program

    Energy Technology Data Exchange (ETDEWEB)

    Everett Sondreal; John Hendrikson

    2009-03-31

    In June 2009, the Energy & Environmental Research Center (EERC) completed 11 years of research under the U.S. Department of Energy (DOE) Base Cooperative Agreement No. DE-FC26-98FT40320 funded through the Office of Fossil Energy (OFE) and administered at the National Energy Technology Laboratory (NETL). A wide range of diverse research activities were performed under annual program plans approved by NETL in seven major task areas: (1) resource characterization and waste management, (2) air quality assessment and control, (3) advanced power systems, (4) advanced fuel forms, (5) value-added coproducts, (6) advanced materials, and (7) strategic studies. This report summarizes results of the 67 research subtasks and an additional 50 strategic studies. Selected highlights in the executive summary illustrate the contribution of the research to the energy industry in areas not adequately addressed by the private sector alone. During the period of performance of the agreement, concerns have mounted over the impact of carbon emissions on climate change, and new programs have been initiated by DOE to ensure that fossil fuel resources along with renewable resources can continue to supply the nation's transportation fuel and electric power. The agreement has addressed DOE goals for reductions in CO{sub 2} emissions through efficiency, capture, and sequestration while expanding the supply and use of domestic energy resources for energy security. It has further contributed to goals for near-zero emissions from highly efficient coal-fired power plants; environmental control capabilities for SO{sub 2}, NO{sub x}, fine respirable particulate (PM{sub 2.5}), and mercury; alternative transportation fuels including liquid synfuels and hydrogen; and synergistic integration of fossil and renewable resources (e.g., wind-, biomass-, and coal-based electrical generation).

  1. Loyalty-based management.

    Science.gov (United States)

    Reichheld, F F

    1993-01-01

    Despite a flurry of activities aimed at serving customers better, few companies have systematically revamped their operations with customer loyalty in mind. Instead, most have adopted improvement programs ad hoc, and paybacks haven't materialized. Building a highly loyal customer base must be integral to a company's basic business strategy. Loyalty leaders like MBNA credit cards are successful because they have designed their entire business systems around customer loyalty--a self-reinforcing system in which the company delivers superior value consistently and reinvents cash flows to find and keep high-quality customers and employees. The economic benefits of high customer loyalty are measurable. When a company consistently delivers superior value and wins customer loyalty, market share and revenues go up, and the cost of acquiring new customers goes down. The better economics mean the company can pay workers better, which sets off a whole chain of events. Increased pay boosts employee moral and commitment; as employees stay longer, their productivity goes up and training costs fall; employees' overall job satisfaction, combined with their experience, helps them serve customers better; and customers are then more inclined to stay loyal to the company. Finally, as the best customers and employees become part of the loyalty-based system, competitors are left to survive with less desirable customers and less talented employees. To compete on loyalty, a company must understand the relationships between customer retention and the other parts of the business--and be able to quantify the linkages between loyalty and profits. It involves rethinking and aligning four important aspects of the business: customers, product/service offering, employees, and measurement systems.

  2. The office based CHIVA

    Directory of Open Access Journals (Sweden)

    Passariello F

    2013-09-01

    Full Text Available Fausto Passariello,1 Stefano Ermini,2 Massimo Cappelli,3 Roberto Delfrate,4 Claude Franceschi5 1Centro Diagnostico Aquarius, Napoli, Italy; 2Private Practice, Grassina, Italy; 3Private Practice, Firenze, Italy; 4Casa di Cure Figlie di Maria, Cremona, Italy; 5Hospital St Joseph, Service d'Explorations Vasculaires, Paris, France Abstract: The cure Conservatrice Hémodynamique de l'Insuffisance Veineuse en Ambulatoire (CHIVA can be office based (OB. The OB-CHIVA protocol is aimed at transferring CHIVA procedures to specialists rooms. The protocol will check the feasibility of OB-CHIVA, data pertaining to recurrence, and will offer the opportunity to study saphenous femoral junction (SFJ stump evolution, the role of the washing vessels and the arch recanalization rate, and gather new data about the effect of the length of the treated saphenous vein. A simplified diagnostic procedure will allow an essential ultrasound examination of the venous net while a schematic and easily readable algorithm guides therapeutic choices. The Riobamba draining crossotomy (RDC tactic is composed of a set of OB procedures. While some of these procedures are, at the moment, only proposals, others are already applied. Devices generally used in ablative procedures such as Light Amplification by Stimulated Emission of Radiation (LASER, radio frequency, steam, and mechanical devices are used in this context to serve to conservative interventions for CHIVA. New techniques have also been proposed for devalvulation and tributary disconnection. Detailed follow-up is necessary in order to determine the effects of therapy and possible disease evolution. Finally, information is added about the informed consent and the ethical considerations of OB-CHIVA research. Keywords: CHIVA, office based procedures, LASER, RF, steam

  3. Telephone-Based Coaching.

    Science.gov (United States)

    Boccio, Mindy; Sanna, Rashel S; Adams, Sara R; Goler, Nancy C; Brown, Susan D; Neugebauer, Romain S; Ferrara, Assiamira; Wiley, Deanne M; Bellamy, David J; Schmittdiel, Julie A

    2017-03-01

    Many Americans continue to smoke, increasing their risk of disease and premature death. Both telephone-based counseling and in-person tobacco cessation classes may improve access for smokers seeking convenient support to quit. Little research has assessed whether such programs are effective in real-world clinical populations. Retrospective cohort study comparing wellness coaching participants with two groups of controls. Kaiser Permanente Northern California, a large integrated health care delivery system. Two hundred forty-one patients who participated in telephonic tobacco cessation coaching from January 1, 2011, to March 31, 2012, and two control groups: propensity-score-matched controls, and controls who participated in a tobacco cessation class during the same period. Wellness coaching participants received an average of two motivational interviewing-based coaching sessions that engaged the patient, evoked their reason to consider quitting, and helped them establish a quit plan. Self-reported quitting of tobacco and fills of tobacco cessation medications within 12 months of follow-up. Logistic regressions adjusting for age, gender, race/ethnicity, and primary language. After adjusting for confounders, tobacco quit rates were higher among coaching participants vs. matched controls (31% vs. 23%, p Coaching participants and class attendees filled tobacco-cessation prescriptions at a higher rate (47% for both) than matched controls (6%, p coaching was as effective as in-person classes and was associated with higher rates of quitting compared to no treatment. The telephonic modality may increase convenience and scalability for health care systems looking to reduce tobacco use and improve health.

  4. A new and improved host-independent plasmid system for RK2-based conjugal transfer.

    Directory of Open Access Journals (Sweden)

    Trine Aakvik Strand

    Full Text Available Bacterial conjugation is a process that is mediated either by a direct cell-to-cell junction or by formation of a bridge between the cells. It is often used to transfer DNA constructs designed in Escherichia coli to recipient bacteria, yeast, plants and mammalian cells. Plasmids bearing the RK2/RP4 origin of transfer (oriT are mostly mobilized using the E. coli S17-1/SM10 donor strains, in which transfer helper functions are provided from a chromosomally integrated RP4::Mu. We have observed that large plasmids were occasionally modified after conjugal transfer when using E. coli S17-1 as a donor. All modified plasmids had increased in size, which most probably was a result of co-transfer of DNA from the chromosomally located oriT. It has earlier also been demonstrated that the bacteriophage Mu is silently transferred to recipient cells by these donor strains, and both occurrences are very likely to lead to mutations within the recipient DNA. Here we report the construction of a new biological system addressing both the above mentioned problems in which the transfer helper functions are provided by a plasmid lacking a functional oriT. This system is compatible with all other replicons commonly used in conjugation experiments and further enables the use of diverse bacterial strains as donors. Plasmids containing large inserts were successfully conjugated and the plasmid modifications observed when E. coli S17-1 was used as donor were eliminated by the use of the new host-independent vector system.

  5. Implementation of Task-based and problem-based ...

    African Journals Online (AJOL)

    This paper describes implementation of new task-based and problem-based pharmacotherapy course for 3rd year pre-clinical medical students at Faculty of Medicine and Health Sciences, Omdurman Islamic University. Keywords: Pharmacotherapy, Task-based learning, P -drug. Sudan Journal of Medical Science Vol.

  6. Flow-Based Provenance

    Directory of Open Access Journals (Sweden)

    Sabah Al-Fedaghi

    2017-02-01

    Full Text Available Aim/Purpose: With information almost effortlessly created and spontaneously available, current progress in Information and Communication Technology (ICT has led to the complication that information must be scrutinized for trustworthiness and provenance. Information systems must become provenance-aware to be satisfactory in accountability, reproducibility, and trustworthiness of data. Background:\tMultiple models for abstract representation of provenance have been proposed to describe entities, people, and activities involved in producing a piece of data, including the Open Provenance Model (OPM and the World Wide Web Consortium. These models lack certain concepts necessary for specifying workflows and encoding the provenance of data products used and generated. Methodology: Without loss of generality, the focus of this paper is on OPM depiction of provenance in terms of a directed graph. We have redrawn several case studies in the framework of our proposed model in order to compare and evaluate it against OPM for representing these cases. Contribution: This paper offers an alternative flow-based diagrammatic language that can form a foundation for modeling of provenance. The model described here provides an (abstract machine-like representation of provenance. Findings: The results suggest a viable alternative in the area of diagrammatic representation for provenance applications. Future Research: Future work will seek to achieve more accurate comparisons with current models in the field.

  7. Mentalization-Based Treatment.

    Science.gov (United States)

    Bateman, Anthony; Fonagy, Peter

    2013-11-01

    The concept of mentalizing has captured the interest and imagination of an astonishing range of people-from psychoanalysts to neuroscientists, from child development researchers to geneticists, from existential philosophers to phenomenologists-all of whom seem to have found it useful. According to the Thompson Reuter maintained Web of Science, the use of the term in titles and abstracts of scientific papers increased from 10 to 2,750 between 1991 and 2011. Clinicians in particular have enthusiastically embraced the idea, and have put it to innovative use in their practices. Mentalization-based treatment (MBT)-making mentalizing a core focus of therapy-was initially developed for the treatment of borderline personality disorder (BPD) in routine clinical services delivered in group and individual modalities. Therapy with mentalizing as a central component is currently being developed for treatment of numerous groups, including people with antisocial personality disorder, substance abuse, eating disorders, and at-risk mothers with infants and children (A. Bateman & Fonagy, 2011). It is also being used with families and adolescents, in schools, and in managing social groups (Asen & Fonagy, 2011; Fonagy et al., 2009; Twemlow, Fonagy, & Sacco, 2005a, 2005b). In this article, we focus on MBT in the treatment of BPD.

  8. Mixture Based Outlier Filtration

    Directory of Open Access Journals (Sweden)

    P. Pecherková

    2006-01-01

    Full Text Available Success/failure of adaptive control algorithms – especially those designed using the Linear Quadratic Gaussian criterion – depends on the quality of the process data used for model identification. One of the most harmful types of process data corruptions are outliers, i.e. ‘wrong data’ lying far away from the range of real data. The presence of outliers in the data negatively affects an estimation of the dynamics of the system. This effect is magnified when the outliers are grouped into blocks. In this paper, we propose an algorithm for outlier detection and removal. It is based on modelling the corrupted data by a two-component probabilistic mixture. The first component of the mixture models uncorrupted process data, while the second models outliers. When the outlier component is detected to be active, a prediction from the uncorrupted data component is computed and used as a reconstruction of the observed data. The resulting reconstruction filter is compared to standard methods on simulated and real data. The filter exhibits excellent properties, especially in the case of blocks of outliers. 

  9. Fluorescence lifetime based bioassays

    Science.gov (United States)

    Meyer-Almes, Franz-Josef

    2017-12-01

    Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.

  10. Fiber Based Dispersion Compensation

    CERN Document Server

    Ramachandran, Siddharth

    2007-01-01

    Dispersion management is a critical design criterion that characterizes the performance of an optical network, and has impacted almost every aspect of the physical layer of an optical transmission line. The past 10 years have seen an explosion in the variety of device effects exploited to obtain optimal performance from dispersion compensators, and this is the first book that deals exclusively with this technology. It starts with an exposition on the fundamental physics underlying dispersion and its effects on optical pulses, followed by at least one chapter devoted to each of the fiber-based dispersion-compensating devices that have either been deployed or are considered as serious candidates for future networks. The final section of this book then describes the systems-level impact of these devices, hence providing a one-stop reference for all aspects of optical-communication-network design pertaining to dispersion-management. Each chapter is written by the leading experts in the field, drawn from both acad...

  11. Nanowire-based detector

    Science.gov (United States)

    Berggren, Karl K; Hu, Xiaolong; Masciarelli, Daniele

    2014-06-24

    Systems, articles, and methods are provided related to nanowire-based detectors, which can be used for light detection in, for example, single-photon detectors. In one aspect, a variety of detectors are provided, for example one including an electrically superconductive nanowire or nanowires constructed and arranged to interact with photons to produce a detectable signal. In another aspect, fabrication methods are provided, including techniques to precisely reproduce patterns in subsequently formed layers of material using a relatively small number of fabrication steps. By precisely reproducing patterns in multiple material layers, one can form electrically insulating materials and electrically conductive materials in shapes such that incoming photons are redirected toward a nearby electrically superconductive materials (e.g., electrically superconductive nanowire(s)). For example, one or more resonance structures (e.g., comprising an electrically insulating material), which can trap electromagnetic radiation within its boundaries, can be positioned proximate the nanowire(s). The resonance structure can include, at its boundaries, electrically conductive material positioned proximate the electrically superconductive nanowire such that light that would otherwise be transmitted through the sensor is redirected toward the nanowire(s) and detected. In addition, electrically conductive material can be positioned proximate the electrically superconductive nanowire (e.g. at the aperture of the resonant structure), such that light is directed by scattering from this structure into the nanowire.

  12. Base Camp Architecture

    Directory of Open Access Journals (Sweden)

    Warebi Gabriel Brisibe

    2016-03-01

    Full Text Available Longitudinal or time line studies of change in the architecture of a particular culture are common, but an area still open to further research is change across space or place. In particular, there is need for studies on architectural change of cultures stemming from the same ethnic source split between their homeland and other Diasporas. This change may range from minor deviations to drastic shifts away from an architectural norm and the accumulation of these shifts within a time frame constitutes variations. This article focuses on identifying variations in the architecture of the Ijo fishing group that migrates along the coastline of West Africa. It examines the causes of cross-cultural variation between base camp dwellings of Ijo migrant fishermen in the Bakassi Peninsula in Cameroon and Bayelsa State in Nigeria. The study draws on the idea of the inevitability of cultural and social change over time as proposed in the theories of cultural dynamism and evolution. It tests aspects of cultural transmission theory using the principal coordinates analysis to ascertain the possible causes of variation. From the findings, this research argues that migration has enhanced the forces of cultural dynamism, which have resulted in significant variations in the architecture of this fishing group.

  13. Managing the Gap between Curriculum Based and Problem Based Learning

    DEFF Research Database (Denmark)

    Bygholm, Ann; Buus, Lillian

    2009-01-01

    /or but rather both/and. In this paper we describe an approach to design and delivery of online courses in computer science which on the one hand is based on a specified curriculum and on the other hand gives room for different learning strategies, problem based learning being one of them. We discuss...... the challenges in applying problem based learning strategies in a context where several universities, with different cultures of teaching, collaboratively develop and deliver online courses. We present a pedagogical framework embracing both problem based and curriculum based strategies and show how we used...... this as a basis for trying out various online learning strategies....

  14. Mindfulness-Based Stress Reduction

    Science.gov (United States)

    ... R S T U V W X Y Z Mindfulness-Based Stress Reduction (MBSR) Information 6 Things You ... Disease and Dementia (12/20/13) Research Spotlights Mindfulness-Based Stress Reduction, Cognitive-Behavioral Therapy Shown To ...

  15. Organizational Semantic Web based Portals

    OpenAIRE

    Necula, Sabina-Cristiana

    2011-01-01

    This paper tries to treat organizational semantic web based portals. The first part of the paper focuses on concepts regarding semantic web based portals. After discussing some concepts we treat the basic functionalities that a semantic web based portal must have and we finish by presenting these functionalities by actual examples. We present semantic web based portals after studying the necessary implementations from literature and practice. We develop some examples that use semantic web ...

  16. Model-based distance sampling

    OpenAIRE

    Buckland, Stephen Terrence; Oedekoven, Cornelia Sabrina; Borchers, David Louis

    2015-01-01

    CSO was part-funded by EPSRC/NERC Grant EP/1000917/1. Conventional distance sampling adopts a mixed approach, using model-based methods for the detection process, and design-based methods to estimate animal abundance in the study region, given estimated probabilities of detection. In recent years, there has been increasing interest in fully model-based methods. Model-based methods are less robust for estimating animal abundance than conventional methods, but offer several advantages: they ...

  17. Web-based collaboration tools.

    Science.gov (United States)

    Wink, Diane M

    2009-01-01

    In this bimonthly series, the author examines how nurse educators can use Internet and Web-based computer technologies such as search, communication, and collaborative writing tools; social networking and social bookmarking sites; virtual worlds; and Web-based teaching and learning programs. This article describes Web-based collaboration tools and techniques to increase their effectiveness.

  18. On Pimsner–Popa bases

    Indian Academy of Sciences (India)

    Home; Journals; Proceedings – Mathematical Sciences; Volume 127; Issue 1. On Pimsner-Popa bases ... Keywords. Subfactor; basic construction; connected inclusion; Pimsner-Popa bases. ... of basic constructions. Finally we use these bases to describe the phenomenon of multistep basic constructions (in both the cases).

  19. Different perspectives on economic base.

    Science.gov (United States)

    Lisa K. Crone; Richard W. Haynes; Nicholas E. Reyna

    1999-01-01

    Two general approaches for measuring the economic base are discussed. Each method is used to define the economic base for each of the counties included in the Interior Columbia Basin Ecosystem Management Project area. A more detailed look at four selected counties results in similar findings from different approaches. Limitations of economic base analysis also are...

  20. Securing web-based exams

    NARCIS (Netherlands)

    Sessink, O.D.T.; Beeftink, H.H.; Tramper, J.; Hartog, R.J.M.

    2004-01-01

    Learning management systems may offer web-based exam facilities. Such facilities entail a higher risk to exams fraud than traditional paper-based exams. The article discusses security issues with web-based exams, and proposes precautionary measures to reduce the risks. A security model is presented

  1. Ferrocene-Based Nanoelectronics

    Energy Technology Data Exchange (ETDEWEB)

    Lawrence R. Sita

    2006-02-15

    Ferrocene-based molecular components for nanoelectronics offer a number of distinct advantages relative to all carbon frameworks due to metal-centered molecular states that should be closer in energy to the Fermi levels of the metal electrodes in metal / molecule / metal heterojunctions. Given this, the overall goal of the project was to investigate the conduction physics of a variety of proposed ferrocene diode / transistor designs in order to address the fundamental question; can electron transport within nm-length scale structures be modulated in a controlled fashion? During the funded period, substantial progress towards achieving this goal was made by surmounting a number of scientific and technical obstacles. More specifically, a concise and general synthetic route to several mono- and diferrocene dithiols and monothiols was achieved that now allows for the directed and controlled assembly of a variety of metal / molecule /metal test structures for the single molecule conductance measurements and the fabrication of self-assembled monolayers (SAMs) on Au(111) that are amenable to quantitative electrochemical characterization of electron-transfer rates. Most importantly, by using an electromigrated test structure, reproducible I/V data for one of the ferrocene dithiol molecules have been collected which exhibit surprisingly high conductance. Exceptional agreement of this result with theory serves to substantiate the original hypothesis that metal-centered states within a molecular bridge can indeed serve to establish higher conductance relative to all-organic molecular bridges. Overall, the successful demonstration of the ability of ferrocene-molecular frameworks to serve as exceptional molecular conductors will play an important role in the continued evolution in design of molecular components for nanoelectronic devices, which in turn, will have a positive impact on the science and potential technologies associated with these systems.

  2. Accelerator based fusion reactor

    Science.gov (United States)

    Liu, Keh-Fei; Chao, Alexander Wu

    2017-08-01

    A feasibility study of fusion reactors based on accelerators is carried out. We consider a novel scheme where a beam from the accelerator hits the target plasma on the resonance of the fusion reaction and establish characteristic criteria for a workable reactor. We consider the reactions d+t\\to n+α,d+{{}3}{{H}\\text{e}}\\to p+α , and p+{{}11}B\\to 3α in this study. The critical temperature of the plasma is determined from overcoming the stopping power of the beam with the fusion energy gain. The needed plasma lifetime is determined from the width of the resonance, the beam velocity and the plasma density. We estimate the critical beam flux by balancing the energy of fusion production against the plasma thermo-energy and the loss due to stopping power for the case of an inert plasma. The product of critical flux and plasma lifetime is independent of plasma density and has a weak dependence on temperature. Even though the critical temperatures for these reactions are lower than those for the thermonuclear reactors, the critical flux is in the range of {{10}22}-{{10}24}~\\text{c}{{\\text{m}}-2}~{{\\text{s}}-1} for the plasma density {ρt}={{10}15}~\\text{c}{{\\text{m}}-3} in the case of an inert plasma. Several approaches to control the growth of the two-stream instability are discussed. We have also considered several scenarios for practical implementation which will require further studies. Finally, we consider the case where the injected beam at the resonance energy maintains the plasma temperature and prolongs its lifetime to reach a steady state. The equations for power balance and particle number conservation are given for this case.

  3. Space-based detectors

    Science.gov (United States)

    Sesana, A.; Weber, W. J.; Killow, C. J.; Perreur-Lloyd, M.; Robertson, D. I.; Ward, H.; Fitzsimons, E. D.; Bryant, J.; Cruise, A. M.; Dixon, G.; Hoyland, D.; Smith, D.; Bogenstahl, J.; McNamara, P. W.; Gerndt, R.; Flatscher, R.; Hechenblaikner, G.; Hewitson, M.; Gerberding, O.; Barke, S.; Brause, N.; Bykov, I.; Danzmann, K.; Enggaard, A.; Gianolio, A.; Vendt Hansen, T.; Heinzel, G.; Hornstrup, A.; Jennrich, O.; Kullmann, J.; Møller-Pedersen, S.; Rasmussen, T.; Reiche, J.; Sodnik, Z.; Suess, M.; Armano, M.; Sumner, T.; Bender, P. L.; Akutsu, T.; Sathyaprakash, B. S.

    2014-12-01

    The parallel session C5 on Space-Based Detectors gave a broad overview over the planned space missions related to gravitational wave detection. Overviews of the revolutionary science to be expected from LISA was given by Alberto Sesana and Sasha Buchman. The launch of LISA Pathfinder (LPF) is planned for 2015. This mission and its payload "LISA Technology Package" will demonstrate key technologies for LISA. In this context, reference masses in free fall for LISA, and gravitational physics in general, was described by William Weber, laser interferometry at the pico-metre level and the optical bench of LPF was presented by Christian Killow and the performance of the LPF optical metrology system by Paul McNamara. While LPF will not yet be sensitive to gravitational waves, it may nevertheless be used to explore fundamental physics questions, which was discussed by Michele Armano. Some parts of the LISA technology that are not going to be demonstrated by LPF, but under intensive development at the moment, were presented by Oliver Jennrich and Oliver Gerberding. Looking into the future, Japan is studying the design of a mid-frequency detector called DECIGO, which was discussed by Tomotada Akutsu. Using atom interferometry for gravitational wave detection has also been recently proposed, and it was critically reviewed by Peter Bender. In the nearer future, the launch of GRACE Follow-On (for Earth gravity observation) is scheduled for 2017, and it will include a Laser Ranging Interferometer as technology demonstrator. This will be the first inter-spacecraft laser interferometer and has many aspects in common with the LISA long arm, as discussed by Andrew Sutton.

  4. Lunar Base Heat Pump

    Science.gov (United States)

    Walker, D.; Fischbach, D.; Tetreault, R.

    1996-01-01

    The objective of this project was to investigate the feasibility of constructing a heat pump suitable for use as a heat rejection device in applications such as a lunar base. In this situation, direct heat rejection through the use of radiators is not possible at a temperature suitable for lde support systems. Initial analysis of a heat pump of this type called for a temperature lift of approximately 378 deg. K, which is considerably higher than is commonly called for in HVAC and refrigeration applications where heat pumps are most often employed. Also because of the variation of the rejection temperature (from 100 to 381 deg. K), extreme flexibility in the configuration and operation of the heat pump is required. A three-stage compression cycle using a refrigerant such as CFC-11 or HCFC-123 was formulated with operation possible with one, two or three stages of compression. Also, to meet the redundancy requirements, compression was divided up over multiple compressors in each stage. A control scheme was devised that allowed these multiple compressors to be operated as required so that the heat pump could perform with variable heat loads and rejection conditions. A prototype heat pump was designed and constructed to investigate the key elements of the high-lift heat pump concept. Control software was written and implemented in the prototype to allow fully automatic operation. The heat pump was capable of operation over a wide range of rejection temperatures and cooling loads, while maintaining cooling water temperature well within the required specification of 40 deg. C +/- 1.7 deg. C. This performance was verified through testing.

  5. Military Bases and Conservation Markets

    Energy Technology Data Exchange (ETDEWEB)

    Mark Ankeny

    2007-09-01

    Over time, DoD is likely to be one of the largest buyers and sellers in a water quality trading market. The Department of Defense (DoD) operates military bases that resemble small cities in infrastructure. As units redeploy, bases are likely to find themselves well within their environmental limits at the originating base and potentially bumping against limits such as nitrate and phosphate loading at the destination base. Stricter rules and heavier loadings in growing watersheds also present challenges to local bases and municipalities as regulators clamp down on loadings from existing Waste Water Treatment Plants (WWTPs) to meet water quality standards.

  6. Workplace Based Assessment in Psychiatry

    Directory of Open Access Journals (Sweden)

    Ayse Devrim Basterzi

    2009-11-01

    Full Text Available Workplace based assessment refers to the assessment of working practices based on what doctors actually do in the workplace, and is predominantly carried out in the workplace itself. Assessment drives learning and it is therefore essential that workplace-based assessment focuses on important attributes rather than what is easiest to assess. Workplacebased assessment is usually competency based. Workplace based assesments may well facilitate and enhance various aspects of educational supervisions, including its structure, frequency and duration etc. The structure and content of workplace based assesments should be monitored to ensure that its benefits are maximised by remaining tailored to individual trainees' needs. Workplace based assesment should be used for formative and summative assessments. Several formative assessment methods have been developed for use in the workplace such as mini clinical evaluation exercise (mini-cex, evidence based journal club assesment and case based discussion, multi source feedback etc. This review discusses the need of workplace based assesments in psychiatry graduate education and introduces some of the work place based assesment methods.

  7. Leveraging Evidence-Based Practice through Partnerships Based on Practice-Based Evidence

    Science.gov (United States)

    Cook, Bryan G.; Cook, Lysandra

    2016-01-01

    Evidence-based practice is among the most influential and compelling reforms in contemporary education. Despite their potential to improve the outcomes of students with disabilities, adoption and implementation of evidence-based reforms have been disappointing, with the gap between research and practice remaining wide. Practice-based evidence…

  8. Case-based reasoning: The marriage of knowledge base and data base

    Science.gov (United States)

    Pulaski, Kirt; Casadaban, Cyprian

    1988-01-01

    The coupling of data and knowledge has a synergistic effect when building an intelligent data base. The goal is to integrate the data and knowledge almost to the point of indistinguishability, permitting them to be used interchangeably. Examples given in this paper suggest that Case-Based Reasoning is a more integrated way to link data and knowledge than pure rule-based reasoning.

  9. Knowledge base verification based on enhanced colored petri net

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jong Hyun; Seong, Poong Hyun [Korea Advanced Institute of Science and Technology, Taejon (Korea, Republic of)

    1997-12-31

    Verification is a process aimed at demonstrating whether a system meets it`s specified requirements. As expert systems are used in various applications, the knowledge base verification of systems takes an important position. The conventional Petri net approach that has been studied recently in order to verify the knowledge base is found that it is inadequate to verify the knowledge base of large and complex system, such as alarm processing system of nuclear power plant. Thus, we propose an improved method that models the knowledge base as enhanced colored Petri net. In this study, we analyze the reachability and the error characteristics of the knowledge base and apply the method to verification of simple knowledge base. 8 refs., 4 figs. (Author)

  10. XML-Based SHINE Knowledge Base Interchange Language

    Science.gov (United States)

    James, Mark; Mackey, Ryan; Tikidjian, Raffi

    2008-01-01

    The SHINE Knowledge Base Interchange Language software has been designed to more efficiently send new knowledge bases to spacecraft that have been embedded with the Spacecraft Health Inference Engine (SHINE) tool. The intention of the behavioral model is to capture most of the information generally associated with a spacecraft functional model, while specifically addressing the needs of execution within SHINE and Livingstone. As such, it has some constructs that are based on one or the other.

  11. Improved Biogeography-Based Optimization Based on Affinity Propagation

    OpenAIRE

    Zhihao Wang; Peiyu Liu; Min Ren; Yuzhen Yang; Xiaoyan Tian

    2016-01-01

    To improve the search ability of biogeography-based optimization (BBO), this work proposed an improved biogeography-based optimization based on Affinity Propagation. We introduced the Memetic framework to the BBO algorithm, and used the simulated annealing algorithm as the local search strategy. MBBO enhanced the exploration with the Affinity Propagation strategy to improve the transfer operation of the BBO algorithm. In this work, the MBBO algorithm was applied to IEEE Congress on Evolutiona...

  12. Complete Genome Sequence of the Type Strain Cupriavidus necator N-1 ▿ †

    Science.gov (United States)

    Poehlein, Anja; Kusian, Bernhard; Friedrich, Bärbel; Daniel, Rolf; Bowien, Botho

    2011-01-01

    Here we announce the complete genome sequence of the copper-resistant bacterium Cupriavidus necator N-1, the type strain of the genus Cupriavidus. The genome consists of two chromosomes and two circular plasmids. Based on genome comparison, the chromosomes of C. necator N-1 share a high degree of similarity with the two chromosomal replicons of the bioplastic-producing hydrogen bacterium Ralstonia eutropha H16. The two strains differ in their plasmids and the presence of hydrogenase genes, which are absent in strain N-1. PMID:21742890

  13. Computer-based and web-based radiation safety training

    Energy Technology Data Exchange (ETDEWEB)

    Owen, C., LLNL

    1998-03-01

    The traditional approach to delivering radiation safety training has been to provide a stand-up lecture of the topic, with the possible aid of video, and to repeat the same material periodically. New approaches to meeting training requirements are needed to address the advent of flexible work hours and telecommuting, and to better accommodate individuals learning at their own pace. Computer- based and web-based radiation safety training can provide this alternative. Computer-based and web- based training is an interactive form of learning that the student controls, resulting in enhanced and focused learning at a time most often chosen by the student.

  14. Acids and bases solvent effects on acid-base strenght

    CERN Document Server

    Cox, Brian G

    2013-01-01

    Acids and bases are ubiquitous in chemistry. Our understanding of them, however, is dominated by their behaviour in water. Transfer to non-aqueous solvents leads to profound changes in acid-base strengths and to the rates and equilibria of many processes: for example, synthetic reactions involving acids, bases and nucleophiles; isolation of pharmaceutical actives through salt formation; formation of zwitter- ions in amino acids; and chromatographic separation of substrates. This book seeks to enhance our understanding of acids and bases by reviewing and analysing their behaviour in non-aqueous solvents. The behaviour is related where possible to that in water, but correlations and contrasts between solvents are also presented.

  15. Aperiodic-metamaterial-based absorber

    Directory of Open Access Journals (Sweden)

    Quanlong Yang

    2017-09-01

    Full Text Available The periodic-metamaterial-based perfect absorber has been studied broadly. Conversely, if the unit cell in the metamaterial-based absorber is arranged aperiodically (aperiodic-metamaterial-based absorber, how does it perform? Inspired by this, here we present a systematic study of the aperiodic-metamaterial-based absorber. By investigating the response of metamaterial absorbers based on periodic, Fibonacci, Thue-Morse, and quasicrystal lattices, we found that aperiodic-metamaterial-based absorbers could display similar absorption behaviors as the periodic one in one hand. However, their absorption behaviors show different tendency depending on the thicknesses of the spacer. Further studies on the angle and polarization dependence of the absorption behavior are also presented.

  16. The knowledge base of journalism

    DEFF Research Database (Denmark)

    Svith, Flemming

    In this paper I propose the knowledge base as a fruitful way to apprehend journalism. With the claim that the majority of practice is anchored in knowledge – understood as 9 categories of rationales, forms and levels – this knowledge base appears as a contextual look at journalists’ knowledge......, and place. As an analytical framework, the knowledge base is limited to understand the practice of newspaper journalists, but, conversely, the knowledge base encompasses more general beginnings through the inclusion of overall structural relationships in the media and journalism and general theories...... on practice and knowledge. As the result of an abductive reasoning is a theory proposal, there is a need for more deductive approaches to test the validity of this knowledge base claim. It is thus relevant to investigate which rationales are included in the knowledge base of journalism, as the dimension does...

  17. Aperiodic-metamaterial-based absorber

    Science.gov (United States)

    Yang, Quanlong; Chen, Xieyu; Li, Yanfeng; Zhang, Xueqian; Xu, Yuehong; Tian, Zhen; Ouyang, Chunmei; Gu, Jianqiang; Han, Jiaguang; Zhang, Weili

    2017-09-01

    The periodic-metamaterial-based perfect absorber has been studied broadly. Conversely, if the unit cell in the metamaterial-based absorber is arranged aperiodically (aperiodic-metamaterial-based absorber), how does it perform? Inspired by this, here we present a systematic study of the aperiodic-metamaterial-based absorber. By investigating the response of metamaterial absorbers based on periodic, Fibonacci, Thue-Morse, and quasicrystal lattices, we found that aperiodic-metamaterial-based absorbers could display similar absorption behaviors as the periodic one in one hand. However, their absorption behaviors show different tendency depending on the thicknesses of the spacer. Further studies on the angle and polarization dependence of the absorption behavior are also presented.

  18. Concurrent array-based queue

    Science.gov (United States)

    Heidelberger, Philip; Steinmacher-Burow, Burkhard

    2015-01-06

    According to one embodiment, a method for implementing an array-based queue in memory of a memory system that includes a controller includes configuring, in the memory, metadata of the array-based queue. The configuring comprises defining, in metadata, an array start location in the memory for the array-based queue, defining, in the metadata, an array size for the array-based queue, defining, in the metadata, a queue top for the array-based queue and defining, in the metadata, a queue bottom for the array-based queue. The method also includes the controller serving a request for an operation on the queue, the request providing the location in the memory of the metadata of the queue.

  19. Veto-based Malware Detection

    OpenAIRE

    Shahzad, Raja Khurram; Lavesson, Niklas

    2012-01-01

    Malicious software (malware) represents a threat to the security and privacy of computer users. Traditional signature-based and heuristic-based methods are unsuccessful in detecting some forms of malware. This paper presents a malware detection approach based on supervised learning. The main contributions of the paper are an ensemble learning algorithm, two pre-processing techniques, and an empirical evaluation of the proposed algorithm. Sequences of operational codes are extracted as feature...

  20. Principles of models based engineering

    Energy Technology Data Exchange (ETDEWEB)

    Dolin, R.M.; Hefele, J.

    1996-11-01

    This report describes a Models Based Engineering (MBE) philosophy and implementation strategy that has been developed at Los Alamos National Laboratory`s Center for Advanced Engineering Technology. A major theme in this discussion is that models based engineering is an information management technology enabling the development of information driven engineering. Unlike other information management technologies, models based engineering encompasses the breadth of engineering information, from design intent through product definition to consumer application.

  1. Trace-Based Code Generation for Model-Based Testing

    NARCIS (Netherlands)

    Kanstrén, T.; Piel, E.; Gross, H.G.

    2009-01-01

    Paper Submitted for review at the Eighth International Conference on Generative Programming and Component Engineering. Model-based testing can be a powerful means to generate test cases for the system under test. However, creating a useful model for model-based testing requires expertise in the

  2. Brain-Based Learning and Standards-Based Elementary Science.

    Science.gov (United States)

    Konecki, Loretta R.; Schiller, Ellen

    This paper explains how brain-based learning has become an area of interest to elementary school science teachers, focusing on the possible relationships between, and implications of, research on brain-based learning to the teaching of science education standards. After describing research on the brain, the paper looks at three implications from…

  3. solar-based boost differen based boost differen ased boost

    African Journals Online (AJOL)

    eobe

    This paper depicts about a power conversion circu known as a Solar known as a Solar-based boost differential single ph based boost differential single ph an open loop Solar ... aximum power point tracker, Boost differential inverter, PID Voltage Con al device that ..... as compared to capacitor, and still under no feedback.

  4. Capability-based computer systems

    CERN Document Server

    Levy, Henry M

    2014-01-01

    Capability-Based Computer Systems focuses on computer programs and their capabilities. The text first elaborates capability- and object-based system concepts, including capability-based systems, object-based approach, and summary. The book then describes early descriptor architectures and explains the Burroughs B5000, Rice University Computer, and Basic Language Machine. The text also focuses on early capability architectures. Dennis and Van Horn's Supervisor; CAL-TSS System; MIT PDP-1 Timesharing System; and Chicago Magic Number Machine are discussed. The book then describes Plessey System 25

  5. Forest biomass-based energy

    Science.gov (United States)

    Janaki R. R. Alavalapati; Pankaj Lal; Andres Susaeta; Robert C. Abt; David N. Wear

    2013-01-01

    Key FindingsHarvesting woody biomass for use as bioenergy is projected to range from 170 million to 336 million green tons by 2050, an increase of 54 to 113 percent over current levels.Consumption projections for forest biomass-based energy, which are based on Energy Information Administration projections, have a high level of...

  6. Knowledge-Based Asynchronous Programming

    NARCIS (Netherlands)

    Haan, Hendrik Wietze de; Hesselink, Wim H.; Renardel de Lavalette, Gerard R.

    2004-01-01

    A knowledge-based program is a high-level description of the behaviour of agents in terms of knowledge that an agent must have before (s)he may perform an action. The definition of the semantics of knowledge-based programs is problematic, since it involves a vicious circle; the knowledge of an agent

  7. Trust-based belief change

    OpenAIRE

    Lorini, Emiliano; Jiang, Guifei; Perrussel, Laurent

    2014-01-01

    International audience; We propose a modal logic that supports reasoning about trust-based belief change. The term trust-based belief change refers to belief change that depends on the degree of trust the receiver has in the source of information.

  8. Evidence-based clinical practice

    DEFF Research Database (Denmark)

    Gluud, Christian

    2002-01-01

    Evidence-based medicine combines the patient's preferences with clinical experience and the best research evidence. Randomized clinical trials are considered the most valid research design for evaluating health-care interventions. However, empirical research shows that intervention effects may...... practice. By investments in education, applied research, and The Cochrane Collaboration, evidence-based medicine may form a stronger basis for clinical practice....

  9. Defining Nature-Based Preschools

    Science.gov (United States)

    Larimore, Rachel

    2016-01-01

    Nature-based early childhood education. Nature-based preschool. Nature preschool. Forest kindergarten. Nature kindergarten. Waldkindergarten. Forest school. These are a few of the program terms currently being discussed among early childhood environmental education professionals in the United States. Why is there so much discussion about the names…

  10. Cloud-Based Data Storage

    Science.gov (United States)

    Waters, John K.

    2011-01-01

    The vulnerability and inefficiency of backing up data on-site is prompting school districts to switch to more secure, less troublesome cloud-based options. District auditors are pushing for a better way to back up their data than the on-site, tape-based system that had been used for years. About three years ago, Hendrick School District in…

  11. Methods in Logic Based Control

    DEFF Research Database (Denmark)

    Christensen, Georg Kronborg

    1999-01-01

    Desing and theory of Logic Based Control systems.Boolean Algebra, Karnaugh Map, Quine McClusky's algorithm. Sequential control design. Logic Based Control Method, Cascade Control Method. Implementation techniques: relay, pneumatic, TTL/CMOS,PAL and PLC- and Soft_PLC implementation. PLC...

  12. Plasma-based accelerator structures

    Energy Technology Data Exchange (ETDEWEB)

    Schroeder, Carl B. [Univ. of California, Berkeley, CA (United States)

    1999-12-01

    Plasma-based accelerators have the ability to sustain extremely large accelerating gradients, with possible high-energy physics applications. This dissertation further develops the theory of plasma-based accelerators by addressing three topics: the performance of a hollow plasma channel as an accelerating structure, the generation of ultrashort electron bunches, and the propagation of laser pulses is underdense plasmas.

  13. Dog Mathematics: Exploring Base-4

    Science.gov (United States)

    Kurz, Terri L.; Yanik, H. Bahadir; Lee, Mi Yeon

    2016-01-01

    Using a dog's paw as a basis for numerical representation, sixth grade students explored how to count and regroup using the dog's four digital pads. Teachers can connect these base-4 explorations to the conceptual meaning of place value and regrouping using base-10.

  14. Constraint-Based Scheduling System

    Science.gov (United States)

    Zweben, Monte; Eskey, Megan; Stock, Todd; Taylor, Will; Kanefsky, Bob; Drascher, Ellen; Deale, Michael; Daun, Brian; Davis, Gene

    1995-01-01

    Report describes continuing development of software for constraint-based scheduling system implemented eventually on massively parallel computer. Based on machine learning as means of improving scheduling. Designed to learn when to change search strategy by analyzing search progress and learning general conditions under which resource bottleneck occurs.

  15. Team-based global organizations

    DEFF Research Database (Denmark)

    Zander, Lena; Butler, Christina; Mockaitis, Audra

    2015-01-01

    of challenges and opportunities of future global organizing as being team-based, we presented new advancements in the study of global teams, leadership, process and outcomes divided into four themes. Two specifically address processes in global teams: 1) the benefits of openness towards linguistic and value......This chapter draws on a panel discussion of the future of global organizing as a team-based organization at EIBA 2014 in Uppsala, Sweden. We began by discussing contemporary developments of hybrid forms of hierarchy and teams-based organizing, but we venture to propose that as organizations become...... characterized by decreased importance of hierarchal structures, more fluidity across borders, even a possible dissolution of firm boundaries, we move towards team-based organizing as an alternative to more traditional forms of hierarchical-based organizing in global firms. To provide input for a discussion...

  16. Content-Based Image Retrial Based on Hadoop

    Directory of Open Access Journals (Sweden)

    DongSheng Yin

    2013-01-01

    Full Text Available Generally, time complexity of algorithms for content-based image retrial is extremely high. In order to retrieve images on large-scale databases efficiently, a new way for retrieving based on Hadoop distributed framework is proposed. Firstly, a database of images features is built by using Speeded Up Robust Features algorithm and Locality-Sensitive Hashing and then perform the search on Hadoop platform in a parallel way specially designed. Considerable experimental results show that it is able to retrieve images based on content on large-scale cluster and image sets effectively.

  17. Skull base chondrosarcoma: evidence-based treatment paradigms.

    Science.gov (United States)

    Bloch, Orin; Parsa, Andrew T

    2013-01-01

    Chondrosarcomas are indolent but invasive chondroid malignancies that can form in the skull base. Standard management of chondrosarcoma involves surgical resection and adjuvant radiation therapy. This review evaluates evidence from the literature to assess the importance of the surgical approach and extent of resection on outcomes for patients with skull base chondrosarcoma. Also evaluated is the ability of the multiple modalities of radiation therapy, such as conventional fractionated radiotherapy, proton beam, and stereotactic radiosurgery, to control tumor growth. Finally, emerging therapies for the treatment of skull-base chondrosarcoma are discussed. Copyright © 2013. Published by Elsevier Inc.

  18. Analysis of ParB-centromere interactions by multiplex SPR imaging reveals specific patterns for binding ParB in six centromeres of Burkholderiales chromosomes and plasmids.

    Directory of Open Access Journals (Sweden)

    Flavien Pillet

    Full Text Available Bacterial centromeres-also called parS, are cis-acting DNA sequences which, together with the proteins ParA and ParB, are involved in the segregation of chromosomes and plasmids. The specific binding of ParB to parS nucleates the assembly of a large ParB/DNA complex from which ParA-the motor protein, segregates the sister replicons. Closely related families of partition systems, called Bsr, were identified on the chromosomes and large plasmids of the multi-chromosomal bacterium Burkholderia cenocepacia and other species from the order Burkholeriales. The centromeres of the Bsr partition families are 16 bp palindromes, displaying similar base compositions, notably a central CG dinucleotide. Despite centromeres bind the cognate ParB with a narrow specificity, weak ParB-parS non cognate interactions were nevertheless detected between few Bsr partition systems of replicons not belonging to the same genome. These observations suggested that Bsr partition systems could have a common ancestry but that evolution mostly erased the possibilities of cross-reactions between them, in particular to prevent replicon incompatibility. To detect novel similarities between Bsr partition systems, we have analyzed the binding of six Bsr parS sequences and a wide collection of modified derivatives, to their cognate ParB. The study was carried out by Surface Plasmon Resonance imaging (SPRi mulitplex analysis enabling a systematic survey of each nucleotide position within the centromere. We found that in each parS some positions could be changed while maintaining binding to ParB. Each centromere displays its own pattern of changes, but some positions are shared more or less widely. In addition from these changes we could speculate evolutionary links between these centromeres.

  19. Molecular characterisation of blaESBL-harbouring conjugative plasmids identified in multi-drug resistant Escherichia coli isolated from food-producing animals and healthy humans

    Directory of Open Access Journals (Sweden)

    Juan eWang

    2013-07-01

    Full Text Available Background: Extended-spectrum β-lactamse (ESBL-encoding genes are frequently mapped to plasmids, yet few of these structures have been characterized at the molecular level, to date.Methods: Eighty-seven ESBL-producing E. coli were isolated from fecal samples of food-producing animals and healthy humans in Switzerland from 2009 to 2011. Plasmid DNA of all isolates was purified. Broth mating assays were carried out individually for 32 isolates to determine if the ESBL marker could be transferred by conjugation. The plasmid sizes were determined by S1 nuclease pulsed-field gel electrophoresis (PFGE and the plasmids were typed by PCR-based replicon typing. Susceptibility tests by disk diffusion followed with a re-analysis S1-nuclease PFGE and PCR reactions were performed to confirm plasmid transfer. Microarray was performed to detect additional antibiotic resistance markers and multi-locus sequence typing (MLST was also performed in selected donor strains. The phylotypes were identified by triplex PCR.Results: About half (n=46 of the 87 isolates carried small (< 20-kb plasmids. All selected 32 isolates contained large plasmids (ranging in sizes from 20- to 600-kb. Eleven plasmid replicon types were detected. Of these, IncFIA (n=5, IncFIB (n=9 and IncK/B (n=4 were common. Nine isolates demonstrated the ability to transfer their cefotaxime resistance marker at high transfer rates. Plasmid profile re-analysis of these transconjugants identified 16 plasmids. IncFIB and IncI1 were the most prevalent replicon types. Phylogenetic grouping showed that five of the nine donor strains belonged to phylogroup B1. Nine different STs were identified in nine tested donor strains.Conclusions: Characterization of these ESBL-encoding conjugative plasmids extends our understanding on these resistance markers in multi-drug resistant E. coli cultured from healthy human and animal sources.

  20. Free fatty acids induce ER stress and block antiviral activity of interferon alpha against hepatitis C virus in cell culture

    Directory of Open Access Journals (Sweden)

    Gunduz Feyza

    2012-08-01

    Full Text Available Abstract Background Hepatic steatosis is recognized as a major risk factor for liver disease progression and impaired response to interferon based therapy in chronic hepatitis C (CHC patients. The mechanism of response to interferon-alpha (IFN-α therapy under the condition of hepatic steatosis is unexplored. We investigated the effect of hepatocellular steatosis on hepatitis C virus (HCV replication and IFN-α antiviral response in a cell culture model. Methods Sub-genomic replicon (S3-GFP and HCV infected Huh-7.5 cells were cultured with a mixture of saturated (palmitate and unsaturated (oleate long-chain free fatty acids (FFA. Intracytoplasmic fat accumulation in these cells was visualized by Nile red staining and electron microscopy then quantified by microfluorometry. The effect of FFA treatment on HCV replication and IFN-α antiviral response was measured by flow cytometric analysis, Renilla luciferase activity, and real-time RT-PCR. Results FFA treatment induced dose dependent hepatocellular steatosis and lipid droplet accumulation in the HCV replicon cells was confirmed by Nile red staining, microfluorometry, and by electron microscopy. Intracellular fat accumulation supports replication more in the persistently HCV infected culture than in the sub-genomic replicon (S3-GFP cell line. FFA treatment also partially blocked IFN-α response and viral clearance by reducing the phosphorylation of Stat1 and Stat2 dependent IFN-β promoter activation. We show that FFA treatment induces endoplasmic reticulum (ER stress response and down regulates the IFNAR1 chain of the type I IFN receptor leading to defective Jak-Stat signaling and impaired antiviral response. Conclusion These results suggest that intracellular fat accumulation in HCV cell culture induces ER stress, defective Jak-Stat signaling, and attenuates the antiviral response, thus providing an explanation to the clinical observation regarding how hepatocellular steatosis influences IFN

  1. Analysis of ParB-centromere interactions by multiplex SPR imaging reveals specific patterns for binding ParB in six centromeres of Burkholderiales chromosomes and plasmids

    Science.gov (United States)

    Pillet, Flavien; Passot, Fanny Marie

    2017-01-01

    Bacterial centromeres–also called parS, are cis-acting DNA sequences which, together with the proteins ParA and ParB, are involved in the segregation of chromosomes and plasmids. The specific binding of ParB to parS nucleates the assembly of a large ParB/DNA complex from which ParA—the motor protein, segregates the sister replicons. Closely related families of partition systems, called Bsr, were identified on the chromosomes and large plasmids of the multi-chromosomal bacterium Burkholderia cenocepacia and other species from the order Burkholeriales. The centromeres of the Bsr partition families are 16 bp palindromes, displaying similar base compositions, notably a central CG dinucleotide. Despite centromeres bind the cognate ParB with a narrow specificity, weak ParB-parS non cognate interactions were nevertheless detected between few Bsr partition systems of replicons not belonging to the same genome. These observations suggested that Bsr partition systems could have a common ancestry but that evolution mostly erased the possibilities of cross-reactions between them, in particular to prevent replicon incompatibility. To detect novel similarities between Bsr partition systems, we have analyzed the binding of six Bsr parS sequences and a wide collection of modified derivatives, to their cognate ParB. The study was carried out by Surface Plasmon Resonance imaging (SPRi) mulitplex analysis enabling a systematic survey of each nucleotide position within the centromere. We found that in each parS some positions could be changed while maintaining binding to ParB. Each centromere displays its own pattern of changes, but some positions are shared more or less widely. In addition from these changes we could speculate evolutionary links between these centromeres. PMID:28562673

  2. Characterization of Extended-Spectrum β-Lactamase-Carrying Plasmids in Clinical Isolates of Klebsiella pneumoniae from Taiwan.

    Science.gov (United States)

    Chang, Chung-Yu; Lin, Heng-Jia; Chang, Lin-Li; Ma, Ling; Siu, L Kristopher; Tung, Yi-Ching; Lu, Po-Liang

    2017-01-01

    We analyzed the replicon types, sizes, and restriction fragment length polymorphism (RFLP) typing of plasmids carrying extended-spectrum β-lactamase (ESBL) genes in Klebsiella pneumoniae isolates from Taiwan. Fifty-one Escherichia coli transconjugant strains with plasmids from ESBL-producing K. pneumoniae from the Taiwan Surveillance of Antimicrobial Resistance III Program in 2002 were included. All the 51 plasmids carried a bla CTX-M gene, the majority of which were bla CTX-M-3 (28/51 [54.9%]). Plasmids ranged in size from 126 to 241 kb by S1 nuclease digestion and subsequent pulsed-field gel electrophoresis, and the most common plasmid size (37.3%) was 161-170 kb. The most common replicon type of plasmids was incompatibility group (Inc)A/C (60.8%). The IncA/C plasmids all carried bla CTX-M (bla CTX-M-3, -14, -15 ), and some also carried bla SHV (bla SHV-5, -12 ) genes. All 51 plasmids could be typed with PstI, and 27 (52.9%) belonged to 10 clusters. Thirty-eight of the 51 plasmids were typable with BamHI, and 21 plasmids (55.3%) fell into 7 clusters. Plasmids in the same cluster belonged to the same incompatibility group, with the exception of cluster C6. In conclusion, IncA/C plasmids are the main plasmid type responsible for the dissemination of ESBL genes of K. pneumoniae from Taiwan. RFLP with PstI possessed better discriminatory power than that with BamHI and PCR-based replicon typing for ESBL-carrying plasmids in K. pneumoniae in this study. Greater than 50% of plasmids fell into clusters, and >60% of cluster-classified plasmids were present in clonally unrelated isolates, indicating that horizontal transfer of plasmids plays an important role in the spread of ESBL genes.

  3. SB certification for mixture-based specification for flexible base.

    Science.gov (United States)

    2012-10-01

    Presentation topics: : Establish List of Qualified Producers; : Producers Responsible for Process Control/Quality Control; : Reduce TxDOT Sampling and Testing; : Expedite Aggregate Base Acceptance; : Share Responsibility (Producer/TxDOT) for Quality ...

  4. Value-based metrics and Internet-based enterprises

    Science.gov (United States)

    Gupta, Krishan M.

    2001-10-01

    Within the last few years, a host of value-based metrics like EVA, MVA, TBR, CFORI, and TSR have evolved. This paper attempts to analyze the validity and applicability of EVA and Balanced Scorecard for Internet based organizations. Despite the collapse of the dot-com model, the firms engaged in e- commerce continue to struggle to find new ways to account for customer-base, technology, employees, knowledge, etc, as part of the value of the firm. While some metrics, like the Balance Scorecard are geared towards internal use, others like EVA are for external use. Value-based metrics are used for performing internal audits as well as comparing firms against one another; and can also be effectively utilized by individuals outside the firm looking to determine if the firm is creating value for its stakeholders.

  5. Survey paper on Sketch Based and Content Based Image Retrieval

    OpenAIRE

    Gaidhani, Prachi A.; Bagal, S.B.

    2015-01-01

    International audience; This survey paper presents an overview of development of Sketch Based Image Retrieval (SBIR) and Content based image retrieval (CBIR) in the past few years. There is awful growth in bulk of images as well as the far-flung application in too many fields. The main attributes to represent as well index the images are color, shape, texture, spatial layout. These features of images are extracted to check similarity among the images. Generation of special query is the main p...

  6. Coronavirus phylogeny based on triplets of nucleic acids bases

    Science.gov (United States)

    Liao, Bo; Liu, Yanshu; Li, Renfa; Zhu, Wen

    2006-04-01

    We considered the fully overlapping triplets of nucleotide bases and proposed a 2D graphical representation of protein sequences consisting of 20 amino acids and a stop code. Based on this 2D graphical representation, we outlined a new approach to analyze the phylogenetic relationships of coronaviruses by constructing a covariance matrix. The evolutionary distances are obtained through measuring the differences among the two-dimensional curves.

  7. Insect acid-base physiology.

    Science.gov (United States)

    Harrison, J F

    2001-01-01

    Acid-base status influences many aspects of insect biology, including insect distributions in aquatic systems, insect-plant and insect-pathogen interactions, membrane transport phenomena, and the mode of action of pesticides. Acid-base status in the hemolymph and gut lumen of insects is generally well regulated but varies somewhat within individuals owing to effects of temperature, activity, discontinuous ventilation, and diet. The pH of the midgut lumen varies with the phylogeny and feeding ecology. Insect fluids have buffer values similar to those of vertebrates. The respiratory system participates in acid-base homeostasis primarily by regulating the internal carbon dioxide (partial) pressure via changes in spiracular opening and convective ventilation. The epithelia of the renal system and gut participate in hemolymph acid-base regulation by varying acid-base transport in response to organismal acid-base status. Evidence to date suggests that the dominant mechanisms for control of renal acid-base excretion involve hormonal regulation of H+-V-ATPase activity.

  8. Execution-Based Model Checking of Interrupt-Based Systems

    Science.gov (United States)

    Drusinsky, Doron; Havelund, Klaus

    2003-01-01

    Execution-based model checking (EMC) is a verification technique based on executing a multi-threaded/multiprocess program repeatedly in a systematic manner in order to explore the different interleavings of the program. This is in contrast to traditional model checking, where a model of a system is analyzed Several execution-based model-checking tools exist at this point, such as for example Verisoft and Java PathFinder. The most common formal specification languages used by EMC tools are un- timed, either just assertions, or linear-time temporal logic (LTL). An alternative verification technique is Runtime Execution Monitoring (REM), which is based on monitor- ing the execution of a program, checking that the execution trace conforms to a requirement specification. The Temporal Rover and DBRover are such tools. They provide a very rich specification language, being an extension of LTL with real-time constraints and time-series. We show how execution-based model checking, combined with runtime execution monitoring, can be used for the verification of a large class of safety critical systems commonly known as interrupt-based systems. The proposed approach is novel in that: (i) it supports model checking of a large class of applications not practically verifiable using conventional EMC tools, (ii) it supports verification of LTL assertions extended with real-time and time-series constraints, and (iii) it supports the verification of custom schedulers.

  9. Solar based hydrogen production systems

    CERN Document Server

    Dincer, Ibrahim

    2013-01-01

    This book provides a comprehensive analysis of various solar based hydrogen production systems. The book covers first-law (energy based) and second-law (exergy based) efficiencies and provides a comprehensive understanding of their implications. It will help minimize the widespread misuse of efficiencies among students and researchers in energy field by using an intuitive and unified approach for defining efficiencies. The book gives a clear understanding of the sustainability and environmental impact analysis of the above systems. The book will be particularly useful for a clear understanding

  10. Wavelength conversion based spectral imaging

    DEFF Research Database (Denmark)

    Dam, Jeppe Seidelin

    There has been a strong, application driven development of Si-based cameras and spectrometers for imaging and spectral analysis of light in the visible and near infrared spectral range. This has resulted in very efficient devices, with high quantum efficiency, good signal to noise ratio and high...... resolution for this spectral region. Today, an increasing number of applications exists outside the spectral region covered by Si-based devices, e.g. within cleantech, medical or food imaging. We present a technology based on wavelength conversion which will extend the spectral coverage of state of the art...... visible or near infrared cameras and spectrometers to include other spectral regions of interest....

  11. Mitochondrial base excision repair assays

    DEFF Research Database (Denmark)

    Maynard, Scott; de Souza-Pinto, Nadja C; Scheibye-Knudsen, Morten

    2010-01-01

    The main source of mitochondrial DNA (mtDNA) damage is reactive oxygen species (ROS) generated during normal cellular metabolism. The main mtDNA lesions generated by ROS are base modifications, such as the ubiquitous 8-oxoguanine (8-oxoG) lesion; however, base loss and strand breaks may also occur....... Many human diseases are associated with mtDNA mutations and thus maintaining mtDNA integrity is critical. All of these lesions are repaired primarily by the base excision repair (BER) pathway. It is now known that mammalian mitochondria have BER, which, similarly to nuclear BER, is catalyzed by DNA...

  12. Formal Component-Based Semantics

    Directory of Open Access Journals (Sweden)

    Ken Madlener

    2011-08-01

    Full Text Available One of the proposed solutions for improving the scalability of semantics of programming languages is Component-Based Semantics, introduced by Peter D. Mosses. It is expected that this framework can also be used effectively for modular meta theoretic reasoning. This paper presents a formalization of Component-Based Semantics in the theorem prover Coq. It is based on Modular SOS, a variant of SOS, and makes essential use of dependent types, while profiting from type classes. This formalization constitutes a contribution towards modular meta theoretic formalizations in theorem provers. As a small example, a modular proof of determinism of a mini-language is developed.

  13. Coal data base - thesaurus 1980

    Energy Technology Data Exchange (ETDEWEB)

    1980-01-01

    The thesaurus contains the vocabulary used to index the Coal Data Base maintained by IEA Coal Research Technical Information Service. The Data Base contains indexed and abstracted references to publicly-available literature covering all aspects of the coal industry. The subject areas covered include: economics and management, reserves and exploration, mining, preparation, transport and handling, coal properties and constitution, processing and conversion, combustion, waste management, environ mental aspects, coal products, and health and safety. The indexing terms are used in the preparation of the annual subject index to Coal Abstracts and should be useful in searching other data bases for material relevant to the coal industry. (Available from IEA Coal Research)

  14. Evidence-Based Medicine: Rhinoplasty.

    Science.gov (United States)

    Lee, Matthew K; Most, Sam P

    2015-08-01

    Evidence-based medicine has become increasingly prominent in the climate of modern day healthcare. The practice of evidence-based medicine involves the integration of the best available evidence with clinical experience and expertise to help guide clinical decision-making. The essential tenets of evidence-based medicine can be applied to both functional and aesthetic rhinoplasty. Current outcome measures in functional and aesthetic rhinoplasty, including objective, subjective, and clinician-reported measures, is summarized and the current data is reviewed. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Home-based Healthcare Technology

    DEFF Research Database (Denmark)

    Verdezoto, Nervo

    of these systems target a specific treatment or condition and might not be sufficient to support the care management work at home. Based on a case study approach, my research investigates home-based healthcare practices and how they can inform future design of home-based healthcare technology that better account......Sustaining daily, unsupervised healthcare activities in non-clinical settings such as the private home can challenge, among others, older adults. To support such unsupervised care activities, an increasingly number of reminders and monitoring systems are being designed. However, most...... for the home setting and people’s everyday activities....

  16. Evidence-based playground design

    DEFF Research Database (Denmark)

    Refshauge, Anne Dahl; Stigsdotter, Ulrika K.; Lamm, Bettina

    2015-01-01

    This paper develops, explores and evaluates an evidence-based approach to playground design, with a public park playground (playlab Cph) in Copenhagen as a case study. In the increasingly urbanised world, park playgrounds are valuable places that support healthy child development by providing...... opportunities for play, nature exploration and sensory stimulation. As it is increasingly expected that designers base their decisions on research evidence, there is a need to develop approaches to facilitate this, which also applies to playground design. The design of PlayLab Cph was based on relevant evidence...

  17. Querying Natural Logic Knowledge Bases

    DEFF Research Database (Denmark)

    Andreasen, Troels; Bulskov, Henrik; Jensen, Per Anker

    2017-01-01

    This paper describes the principles of a system applying natural logic as a knowledge base language. Natural logics are regimented fragments of natural language employing high level inference rules. We advocate the use of natural logic for knowledge bases dealing with querying of classes...... in ontologies and class-relationships such as are common in life-science descriptions. The paper adopts a version of natural logic with recursive restrictive clauses such as relative clauses and adnominal prepositional phrases. It includes passive as well as active voice sentences. We outline a prototype...... for partial translation of natural language into natural logic, featuring further querying and conceptual path finding in natural logic knowledge bases....

  18. Surface stress-based biosensors.

    Science.gov (United States)

    Sang, Shengbo; Zhao, Yuan; Zhang, Wendong; Li, Pengwei; Hu, Jie; Li, Gang

    2014-01-15

    Surface stress-based biosensors, as one kind of label-free biosensors, have attracted lots of attention in the process of information gathering and measurement for the biological, chemical and medical application with the development of technology and society. This kind of biosensors offers many advantages such as short response time (less than milliseconds) and a typical sensitivity at nanogram, picoliter, femtojoule and attomolar level. Furthermore, it simplifies sample preparation and testing procedures. In this work, progress made towards the use of surface stress-based biosensors for achieving better performance is critically reviewed, including our recent achievement, the optimally circular membrane-based biosensors and biosensor array. The further scientific and technological challenges in this field are also summarized. Critical remark and future steps towards the ultimate surface stress-based biosensors are addressed. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Base Flow Model Validation Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The program focuses on turbulence modeling enhancements for predicting high-speed rocket base flows. A key component of the effort is the collection of high-fidelity...

  20. Chip-based droplet sorting

    Science.gov (United States)

    Beer, Neil Reginald; Lee, Abraham; Hatch, Andrew

    2017-11-21

    A non-contact system for sorting monodisperse water-in-oil emulsion droplets in a microfluidic device based on the droplet's contents and their interaction with an applied electromagnetic field or by identification and sorting.

  1. Reliability-Based Code Calibration

    DEFF Research Database (Denmark)

    Faber, M.H.; Sørensen, John Dalsgaard

    2003-01-01

    The present paper addresses fundamental concepts of reliability based code calibration. First basic principles of structural reliability theory are introduced and it is shown how the results of FORM based reliability analysis may be related to partial safety factors and characteristic values....... Thereafter the code calibration problem is presented in its principal decision theoretical form and it is discussed how acceptable levels of failure probability (or target reliabilities) may be established. Furthermore suggested values for acceptable annual failure probabilities are given for ultimate...... and serviceability limit states. Finally the paper describes the Joint Committee on Structural Safety (JCSS) recommended procedure - CodeCal - for the practical implementation of reliability based code calibration of LRFD based design codes....

  2. Industry Based Survey (IBS) Yellowtail

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The "Southern New England Yellowtail Flounder Industry-Based Survey" was a collaboration between the Rhode Island Division of Fish and Wildlife and the fishing...

  3. FEMA DFIRM Base Flood Elevations

    Data.gov (United States)

    Minnesota Department of Natural Resources — The Base Flood Elevation (BFE) table is required for any digital data where BFE lines will be shown on the corresponding Flood Insurance Rate Map (FIRM). Normally,...

  4. Cage-based performance capture

    CERN Document Server

    Savoye, Yann

    2014-01-01

    Nowadays, highly-detailed animations of live-actor performances are increasingly easier to acquire and 3D Video has reached considerable attentions in visual media production. In this book, we address the problem of extracting or acquiring and then reusing non-rigid parametrization for video-based animations. At first sight, a crucial challenge is to reproduce plausible boneless deformations while preserving global and local captured properties of dynamic surfaces with a limited number of controllable, flexible and reusable parameters. To solve this challenge, we directly rely on a skin-detached dimension reduction thanks to the well-known cage-based paradigm. First, we achieve Scalable Inverse Cage-based Modeling by transposing the inverse kinematics paradigm on surfaces. Thus, we introduce a cage inversion process with user-specified screen-space constraints. Secondly, we convert non-rigid animated surfaces into a sequence of optimal cage parameters via Cage-based Animation Conversion. Building upon this re...

  5. Chip-based droplet sorting

    Energy Technology Data Exchange (ETDEWEB)

    Beer, Neil Reginald; Lee, Abraham; Hatch, Andrew

    2017-11-21

    A non-contact system for sorting monodisperse water-in-oil emulsion droplets in a microfluidic device based on the droplet's contents and their interaction with an applied electromagnetic field or by identification and sorting.

  6. PHYSIOLOGY OF ACID BASE BALANCE

    Directory of Open Access Journals (Sweden)

    Awati

    2014-12-01

    Full Text Available Acid-base, electrolyte, and metabolic disturbances are common in the intensive care unit. Almost all critically ill patients often suffer from compound acid-base and electrolyte disorders. Successful evaluation and management of such patients requires recognition of common patterns (e.g., metabolic acidosis and the ability to dissect one disorder from another. The intensivists needs to identify and correct these condition with the easiest available tools as they are the associated with multiorgan failure. Understanding the elements of normal physiology in these areas is very important so as to diagnose the pathological condition and take adequate measures as early as possible. Arterial blood gas analysis is one such tool for early detection of acid base disorder. Physiology of acid base is complex and here is the attempt to simplify it in our day to day application for the benefit of critically ill patients.

  7. Imprinted silicon-based nanophotonics

    DEFF Research Database (Denmark)

    Borel, Peter Ingo; Olsen, Brian Bilenberg; Frandsen, Lars Hagedorn

    2007-01-01

    We demonstrate and optically characterize silicon-on-insulator based nanophotonic devices fabricated by nanoimprint lithography. In our demonstration, we have realized ordinary and topology-optimized photonic crystal waveguide structures. The topology-optimized structures require lateral pattern ...

  8. Cell phone based balance trainer

    National Research Council Canada - National Science Library

    Lee, Beom-Chan; Kim, Jeonghee; Chen, Shu; Sienko, Kathleen H

    2012-01-01

    ..., weight, complexity, calibration procedures, cost, and fragility. We have designed and developed a cell phone based vibrotactile feedback system for potential use in balance rehabilitation training in clinical and home environments...

  9. Hospital Value-Based Purchasing

    Data.gov (United States)

    U.S. Department of Health & Human Services — Hospital Value-Based Purchasing (VBP) is part of the Centers for Medicare and Medicaid Services (CMS) long-standing effort to link Medicares payment system to a...

  10. Chip-based droplet sorting

    Science.gov (United States)

    Beer, Neil Reginald; Lee, Abraham; Hatch, Andrew

    2014-07-01

    A non-contact system for sorting monodisperse water-in-oil emulsion droplets in a microfluidic device based on the droplet's contents and their interaction with an applied electromagnetic field or by identification and sorting.

  11. NORTHWOODS Wildlife Habitat Data Base

    Science.gov (United States)

    Mark D. Nelson; Janine M. Benyus; Richard R. Buech

    1992-01-01

    Wildlife habitat data from seven Great Lakes National Forests were combined into a wildlife-habitat matrix named NORTHWOODS. Several electronic file formats of NORTHWOODS data base and documentation are available on floppy disks for microcomputers.

  12. Graph Model Based Indoor Tracking

    DEFF Research Database (Denmark)

    Jensen, Christian Søndergaard; Lu, Hua; Yang, Bin

    2009-01-01

    The tracking of the locations of moving objects in large indoor spaces is important, as it enables a range of applications related to, e.g., security and indoor navigation and guidance. This paper presents a graph model based approach to indoor tracking that offers a uniform data management...... infrastructure for different symbolic positioning technologies, e.g., Bluetooth and RFID. More specifically, the paper proposes a model of indoor space that comprises a base graph and mappings that represent the topology of indoor space at different levels. The resulting model can be used for one or several...... indoor positioning technologies. Focusing on RFID-based positioning, an RFID specific reader deployment graph model is built from the base graph model. This model is then used in several algorithms for constructing and refining trajectories from raw RFID readings. Empirical studies with implementations...

  13. Consent Based Verification System (CBSV)

    Data.gov (United States)

    Social Security Administration — CBSV is a fee-based service offered by SSA's Business Services Online (BSO). It is used by private companies to verify the SSNs of their customers and clients that...

  14. Industry Based Survey (IBS) Cod

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The "Gulf of Maine Atlantic Cod Industry-Based Survey" was a collaboration of the Massachusetts Division of Marine Fisheries and the fishing industry, with support...

  15. Dimensions of problem based learning

    DEFF Research Database (Denmark)

    Nielsen, Jørgen Lerche; Andreasen, Lars Birch

    2013-01-01

    The article contributes to the literature on problem based learning and problem-oriented project work, building on and reflecting the experiences of the authors through decades of work with problem-oriented project pedagogy. The article explores different dimensions of problem based learning such...... and Learning (MIL). We discuss changes in the roles of the teachers as supervisors within this learning environment, and we explore the involvement of students as active participants and co-designers of how course and project activities unfold.......The article contributes to the literature on problem based learning and problem-oriented project work, building on and reflecting the experiences of the authors through decades of work with problem-oriented project pedagogy. The article explores different dimensions of problem based learning...

  16. Studying Sensing-Based Systems

    DEFF Research Database (Denmark)

    Kjærgaard, Mikkel Baun

    2013-01-01

    Recent sensing-based systems involve a multitude of users, devices, and places. These types of systems challenge existing approaches for conducting valid system evaluations. Here, the author discusses such evaluation challenges and revisits existing system evaluation methodologies....

  17. Base Flood Elevation (BFE) Lines

    Data.gov (United States)

    Department of Homeland Security — The Base Flood Elevation (BFE) table is required for any digital data where BFE lines will be shown on the corresponding Flood Insurance Rate Map (FIRM). Normally if...

  18. US Air Force Base Observations

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Hourly observations taken by U.S. Air Force personnel at bases in the United States and around the world. Foreign observations concentrated in the Middle East and...

  19. Consumption-based Equity Valuation

    DEFF Research Database (Denmark)

    Bach, Christian; Christensen, Peter O.

    2016-01-01

    implementations of standard valuation models, both in terms of median absolute valuation errors (MAVE) and in terms of excess returns on simple investment strategies based on the differences between model and market prices. The CCAPM-based valuation model yields a significantly lower MAVE than the best performing...... standard valuation model. Both types of models can identify investment strategies with subsequent excess returns. The CCAPM-based valuation model yields time-series of realized hedge returns with more and higher positive returns and fewer and less negative returns compared with the time-series of realized...... through a risk-adjusted cost of equity in the denominator. The risk adjustments are derived based on assumptions about the time-series properties of residual income returns and aggregate consumption rather than on historical stock returns. We compare the performance of the model with several...

  20. Virtual Reality based Learning Systems

    OpenAIRE

    Cheng, Yang

    2016-01-01

    This article is based on studies of the existing literature, focusing on the states-of-the-arts on virtual reality (VR) and its potential uses in learning. Different platforms have been used to improve the learning effects of VR that offers exciting opportunities in various fields. As more and more students want in a distance, part-time, or want to continue their education, VR has attracted considerable attention in learning, training, and traditional education. VR based learning enables oper...

  1. Carbon Nanostructure-Based Sensors

    OpenAIRE

    Sarkar, Tapan

    2012-01-01

    One-dimensional (1-D) nanostructure-based sensors provide better sensitivity as compared to conventional thin film-based sensors due to their comparable dimensions with respect to Debye length. Single-walled carbon nanotubes (SWNTs) are 1-D nanostructures having high electrical mobility, high mechanical strength and high specific surface area that facilitate building of low-power, ultrahigh density sensors within limited space. However, pristine SWNTs posses limited sensitivity and selectivit...

  2. Evidence-based cancer imaging

    Energy Technology Data Exchange (ETDEWEB)

    Shinagare, Atul B.; Khorasani, Ramin [Dept. of Radiology, Brigham and Women' s Hospital, Boston (Korea, Republic of)

    2017-01-15

    With the advances in the field of oncology, imaging is increasingly used in the follow-up of cancer patients, leading to concerns about over-utilization. Therefore, it has become imperative to make imaging more evidence-based, efficient, cost-effective and equitable. This review explores the strategies and tools to make diagnostic imaging more evidence-based, mainly in the context of follow-up of cancer patients.

  3. Method for gesture based modeling

    DEFF Research Database (Denmark)

    2006-01-01

    A computer program based method is described for creating models using gestures. On an input device, such as an electronic whiteboard, a user draws a gesture which is recognized by a computer program and interpreted relative to a predetermined meta-model. Based on the interpretation, an algorithm...... is assigned to the gesture drawn by the user. The executed algorithm may, for example, consist in creating a new model element, modifying an existing model element, or deleting an existing model element....

  4. A comprehensive physiologically based pharmacokinetic ...

    Science.gov (United States)

    Published physiologically based pharmacokinetic (PBPK) models from peer-reviewed articles are often well-parameterized, thoroughly-vetted, and can be utilized as excellent resources for the construction of models pertaining to related chemicals. Specifically, chemical-specific parameters and in vivo pharmacokinetic data used to calibrate these published models can act as valuable starting points for model development of new chemicals with similar molecular structures. A knowledgebase for published PBPK-related articles was compiled to support PBPK model construction for new chemicals based on their close analogues within the knowledgebase, and a web-based interface was developed to allow users to query those close analogues. A list of 689 unique chemicals and their corresponding 1751 articles was created after analysis of 2,245 PBPK-related articles. For each model, the PMID, chemical name, major metabolites, species, gender, life stages and tissue compartments were extracted from the published articles. PaDEL-Descriptor, a Chemistry Development Kit based software, was used to calculate molecular fingerprints. Tanimoto index was implemented in the user interface as measurement of structural similarity. The utility of the PBPK knowledgebase and web-based user interface was demonstrated using two case studies with ethylbenzene and gefitinib. Our PBPK knowledgebase is a novel tool for ranking chemicals based on similarities to other chemicals associated with existi

  5. Evidence based medical imaging (EBMI)

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Tony [Senior Lecturer in Medical Radiation Science, University Department of Rural Health, School of Health Sciences, Faculty of Health, University of Newcastle, Locked Bag 9783 NEMSC, Tamworth, NSW 2348 (Australia)], E-mail: tony.smith@hnehealth.nsw.gov.au

    2008-08-15

    Background: The evidence based paradigm was first described about a decade ago. Previous authors have described a framework for the application of evidence based medicine which can be readily adapted to medical imaging practice. Purpose: This paper promotes the application of the evidence based framework in both the justification of the choice of examination type and the optimisation of the imaging technique used. Methods: The framework includes five integrated steps: framing a concise clinical question; searching for evidence to answer that question; critically appraising the evidence; applying the evidence in clinical practice; and, evaluating the use of revised practices. Results: This paper illustrates the use of the evidence based framework in medical imaging (that is, evidence based medical imaging) using the examples of two clinically relevant case studies. In doing so, a range of information technology and other resources available to medical imaging practitioners are identified with the intention of encouraging the application of the evidence based paradigm in radiography and radiology. Conclusion: There is a perceived need for radiographers and radiologists to make greater use of valid research evidence from the literature to inform their clinical practice and thus provide better quality services.

  6. Chicken Swarm Optimization Based on Elite Opposition-Based Learning

    Directory of Open Access Journals (Sweden)

    Chiwen Qu

    2017-01-01

    Full Text Available Chicken swarm optimization is a new intelligent bionic algorithm, simulating the chicken swarm searching for food in nature. Basic algorithm is likely to fall into a local optimum and has a slow convergence rate. Aiming at these deficiencies, an improved chicken swarm optimization algorithm based on elite opposition-based learning is proposed. In cock swarm, random search based on adaptive t distribution is adopted to replace that based on Gaussian distribution so as to balance the global exploitation ability and local development ability of the algorithm. In hen swarm, elite opposition-based learning is introduced to promote the population diversity. Dimension-by-dimension greedy search mode is used to do local search for individual of optimal chicken swarm in order to improve optimization precision. According to the test results of 18 standard test functions and 2 engineering structure optimization problems, this algorithm has better effect on optimization precision and speed compared with basic chicken algorithm and other intelligent optimization algorithms.

  7. Graph Cuts based Image Segmentation using Fuzzy Rule Based System

    Directory of Open Access Journals (Sweden)

    M. R. Khokher

    2012-12-01

    Full Text Available This work deals with the segmentation of gray scale, color and texture images using graph cuts. From input image, a graph is constructed using intensity, color and texture profiles of the image simultaneously. Based on the nature of image, a fuzzy rule based system is designed to find the weight that should be given to a specific image feature during graph development. The graph obtained from the fuzzy rule based weighted average of different image features is further used in normalized graph cuts framework. Graph is iteratively bi-partitioned through the normalized graph cuts algorithm to get optimum partitions resulting in the segmented image. Berkeley segmentation database is used to test our algorithm and the segmentation results are evaluated through probabilistic rand index, global consistency error, sensitivity, positive predictive value and Dice similarity coefficient. It is shown that the presented segmentation method provides effective results for most types of images.

  8. Improved Biogeography-Based Optimization Based on Affinity Propagation

    Directory of Open Access Journals (Sweden)

    Zhihao Wang

    2016-07-01

    Full Text Available To improve the search ability of biogeography-based optimization (BBO, this work proposed an improved biogeography-based optimization based on Affinity Propagation. We introduced the Memetic framework to the BBO algorithm, and used the simulated annealing algorithm as the local search strategy. MBBO enhanced the exploration with the Affinity Propagation strategy to improve the transfer operation of the BBO algorithm. In this work, the MBBO algorithm was applied to IEEE Congress on Evolutionary Computation (CEC 2015 benchmarks optimization problems to conduct analytic comparison with the first three winners of the CEC 2015 competition. The results show that the MBBO algorithm enhances the exploration, exploitation, convergence speed and solution accuracy and can emerge as the best solution-providing algorithm among the competing algorithms.

  9. Generalized eigenvalue based spectrum sensing

    KAUST Repository

    Shakir, Muhammad

    2012-01-01

    Spectrum sensing is one of the fundamental components in cognitive radio networks. In this chapter, a generalized spectrum sensing framework which is referred to as Generalized Mean Detector (GMD) has been introduced. In this context, we generalize the detectors based on the eigenvalues of the received signal covariance matrix and transform the eigenvalue based spectrum sensing detectors namely: (i) the Eigenvalue Ratio Detector (ERD) and two newly proposed detectors which are referred to as (ii) the GEometric Mean Detector (GEMD) and (iii) the ARithmetic Mean Detector (ARMD) into an unified framework of generalize spectrum sensing. The foundation of the proposed framework is based on the calculation of exact analytical moments of the random variables of the decision threshold of the respective detectors. The decision threshold has been calculated in a closed form which is based on the approximation of Cumulative Distribution Functions (CDFs) of the respective test statistics. In this context, we exchange the analytical moments of the two random variables of the respective test statistics with the moments of the Gaussian (or Gamma) distribution function. The performance of the eigenvalue based detectors is compared with the several traditional detectors including the energy detector (ED) to validate the importance of the eigenvalue based detectors and the performance of the GEMD and the ARMD particularly in realistic wireless cognitive radio network. Analytical and simulation results show that the newly proposed detectors yields considerable performance advantage in realistic spectrum sensing scenarios. Moreover, the presented results based on proposed approximation approaches are in perfect agreement with the empirical results. © 2012 Springer Science+Business Media Dordrecht.

  10. Characteristic properties of Fibonacci-based mutually unbiased bases

    Energy Technology Data Exchange (ETDEWEB)

    Seyfarth, Ulrich; Alber, Gernot [Institut fuer Angewandte Physik, Technische Universitaet Darmstadt, 64289 Darmstadt (Germany); Ranade, Kedar [Institut fuer Quantenphysik, Universitaet Ulm, Albert-Einstein-Allee 11, 89069 Ulm (Germany)

    2012-07-01

    Complete sets of mutually unbiased bases (MUBs) offer interesting applications in quantum information processing ranging from quantum cryptography to quantum state tomography. Different construction schemes provide different perspectives on these bases which are typically also deeply connected to various mathematical research areas. In this talk we discuss characteristic properties resulting from a recently established connection between construction methods for cyclic MUBs and Fibonacci polynomials. As a remarkable fact this connection leads to construction methods which do not involve any relations to mathematical properties of finite fields.

  11. Lignin-Based Thermoplastic Materials.

    Science.gov (United States)

    Wang, Chao; Kelley, Stephen S; Venditti, Richard A

    2016-04-21

    Lignin-based thermoplastic materials have attracted increasing interest as sustainable, cost-effective, and biodegradable alternatives for petroleum-based thermoplastics. As an amorphous thermoplastic material, lignin has a relatively high glass-transition temperature and also undergoes radical-induced self-condensation at high temperatures, which limits its thermal processability. Additionally, lignin-based materials are usually brittle and exhibit poor mechanical properties. To improve the thermoplasticity and mechanical properties of technical lignin, polymers or plasticizers are usually integrated with lignin by blending or chemical modification. This Review attempts to cover the reported approaches towards the development of lignin-based thermoplastic materials on the basis of published information. Approaches reviewed include plasticization, blending with miscible polymers, and chemical modifications by esterification, etherification, polymer grafting, and copolymerization. Those lignin-based thermoplastic materials are expected to show applications as engineering plastics, polymeric foams, thermoplastic elastomers, and carbon-fiber precursors. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. A Knowledge Base for Knowledge-Based Multiagent System Construction

    Science.gov (United States)

    2000-10-01

    step_p.html [5] M. J. Raphael, “Knowledge Based Support for Design and Synthesis of Multiagent Systems.” MS Thesis AFIT/ENG/00M21. School of...Wooldridge, N. Jennings, and D. Kinny, “The Gaia Methodology for Agent-Oriented Analysis and Design,” Journal of Autonomous Agents and Multi-Agent

  13. Image based Monument Recognition using Graph based Visual Saliency

    DEFF Research Database (Denmark)

    Kalliatakis, Grigorios; Triantafyllidis, Georgios

    2013-01-01

    , the images have been previously processed according to the Graph Based Visual Saliency model in order to keep either SIFT or SURF features corresponding to the actual monuments while the background “noise” is minimized. The application is then able to classify these images, helping the user to better...

  14. gis-based hydrological model based hydrological model upstream

    African Journals Online (AJOL)

    eobe

    Hydrological. Hydrological modeling tools have been increasingl modeling tools have been increasingl watershed watershed level. The application of these tools hav. The application of these tools hav sensing and G sensing and Geographical Information System (GIS) eographical Information System (GIS) based models ...

  15. Centre-based versus home-based childcare

    NARCIS (Netherlands)

    Bauchmüller, R.

    2013-01-01

    Centre‐based childcare is seen as a public investment to facilitate maternal employment. Recent theoretical research proposes that such investments potentially lead to substantial gains in child development and thus to high returns for society as a whole. However, the empirical evidence is still

  16. Isotope-based quantum information

    CERN Document Server

    G Plekhanov, Vladimir

    2012-01-01

    The present book provides to the main ideas and techniques of the rapid progressing field of quantum information and quantum computation using isotope - mixed materials. It starts with an introduction to the isotope physics and then describes of the isotope - based quantum information and quantum computation. The ability to manipulate and control electron and/or nucleus spin in semiconductor devices provides a new route to expand the capabilities of inorganic semiconductor-based electronics and to design innovative devices with potential application in quantum computing. One of the major challenges towards these objectives is to develop semiconductor-based systems and architectures in which the spatial distribution of spins and their properties can be controlled. For instance, to eliminate electron spin decoherence resulting from hyperfine interaction due to nuclear spin background, isotopically controlled devices are needed (i.e., nuclear spin-depleted). In other emerging concepts, the control of the spatial...

  17. Agent Based Individual Traffic Guidance

    DEFF Research Database (Denmark)

    Wanscher, Jørgen

    This thesis investigates the possibilities in applying Operations Research (OR) to autonomous vehicular traffic. The explicit difference to most other research today is that we presume that an agent is present in every vehicle - hence Agent Based Individual Traffic guidance (ABIT). The next...... evolutionary step for the in-vehicle route planners is the introduction of two-way communication. We presume that the agent is capable of exactly this. Based on this presumption we discuss the possibilities and define a taxonomy and use this to discuss the ABIT system. Based on a set of scenarios we conclude...... of the project were not previously considered. We define a special inseparable cost function and develop a solution complex capable of using this cost function. In relation to calibration and estimation of statistical models used for dynamic route guidance we worked with generating random number sequences...

  18. Fluorous-based carbohydrate microarrays.

    Science.gov (United States)

    Ko, Kwang-Seuk; Jaipuri, Firoz A; Pohl, Nicola L

    2005-09-28

    The success of microarrays, such as DNA chips, for biosample screening with minimal sample usage has led to a variety of technologies for assays on glass slides. Unfortunately, for small molecules, such as carbohydrates, these methods usually rely on covalent bond formation, which requires unique functional handles and multiple chemical steps. A new simpler concept in microarray formation is based on noncovalent fluorous-based interactions. A fluorous tail is designed not only to aid in saccharide purification but also to allow direct formation of carbohydrate microarrays on fluorous-derivatized glass slides for biological screening with lectins, such as concanavalin A. The noncovalent interactions in the fluorous-based array are even strong enough to withstand the detergents used in assays with the Erythrina crystagalli lectin. Additionally, the utility of benzyl carbonate protecting groups on fucose building blocks for the formation of alpha-linkages is demonstrated.

  19. Eye-based head gestures

    DEFF Research Database (Denmark)

    Mardanbegi, Diako; Witzner Hansen, Dan; Pederson, Thomas

    2012-01-01

    A novel method for video-based head gesture recognition using eye information by an eye tracker has been proposed. The method uses a combination of gaze and eye movement to infer head gestures. Compared to other gesture-based methods a major advantage of the method is that the user keeps the gaze...... on the interaction object while interacting. This method has been implemented on a head-mounted eye tracker for detecting a set of predefined head gestures. The accuracy of the gesture classifier is evaluated and verified for gaze-based interaction in applications intended for both large public displays and small...... mobile phone screens. The user study shows that the method detects a set of defined gestures reliably....

  20. Brain plasticity-based therapeutics

    Science.gov (United States)

    Merzenich, Michael M.; Van Vleet, Thomas M.; Nahum, Mor

    2014-01-01

    The primary objective of this review article is to summarize how the neuroscience of brain plasticity, exploiting new findings in fundamental, integrative and cognitive neuroscience, is changing the therapeutic landscape for professional communities addressing brain-based disorders and disease. After considering the neurological bases of training-driven neuroplasticity, we shall describe how this neuroscience-guided perspective distinguishes this new approach from (a) the more-behavioral, traditional clinical strategies of professional therapy practitioners, and (b) an even more widely applied pharmaceutical treatment model for neurological and psychiatric treatment domains. With that background, we shall argue that neuroplasticity-based treatments will be an important part of future best-treatment practices in neurological and psychiatric medicine. PMID:25018719

  1. Event-Based Conceptual Modeling

    DEFF Research Database (Denmark)

    Bækgaard, Lars

    The paper demonstrates that a wide variety of event-based modeling approaches are based on special cases of the same general event concept, and that the general event concept can be used to unify the otherwise unrelated fields of information modeling and process modeling. A set of event......-based modeling approaches are analyzed and the results are used to formulate a general event concept that can be used for unifying the seemingly unrelated event concepts. Events are characterized as short-duration processes that have participants, consequences, and properties, and that may be modeled in terms...... of information structures. The general event concept can be used to guide systems analysis and design and to improve modeling approaches....

  2. Solar Panel based Milk Pasteurization

    DEFF Research Database (Denmark)

    Nielsen, Kirsten Mølgaard; Pedersen, Tom Søndergaard

    This paper treats the subject of analysis, design and development of the control system for a solar panel based milk pasteurization system to be used in small villages in Tanzania. The analysis deals with the demands for an acceptable pasteurization, the varying energy supply and the low cost, low...... complexity, simple user interface and high reliability demands. Based on these demands a concept for the pasteurization system is established and a control system is developed. A solar panel has been constructed and the energy absorption has been tested in Tanzania. Based on the test, the pasteurization...... system is dimensioned. A functional prototype of the pasteurization facility with a capacity of 200 l milk/hour has been developed and tested. The system is prepared for solar panels as the main energy source and is ready for a test in Tanzania....

  3. Solar Panel based Milk Pasteurization

    DEFF Research Database (Denmark)

    Nielsen, Kirsten Mølgaard; Pedersen, Tom Søndergaard

    2002-01-01

    This paper treats the subject of analysis, design and development of the control system for a solar panel based milk pasteurization system to be used in small villages in Tanzania. The analysis deals with the demands for an acceptable pasteurization, the varying energy supply and the low cost, low...... complexity, simple user interface and high reliability demands. Based on these demands a concept for the pasteurization system is established and a control system is developed. A solar panel has been constructed and the energy absorption has been tested in Tanzania. Based on the test, the pasteurization...... system is dimensioned. A functional prototype of the pasteurization facility with a capacity of 200 l milk/hour has been developed and tested. The system is prepared for solar panels as the main energy source and is ready for a test in Tanzania....

  4. Optical Fiber Grating based Sensors

    DEFF Research Database (Denmark)

    Michelsen, Susanne

    2003-01-01

    In this thesis differenct optical fiber gratings are used for sensor purposes. If a fiber with a core concentricity error (CCE) is used, a directional dependent bend sensor can be produced. The CCE direction can be determined by means of diffraction. This makes it possible to produce long......-period gratings in a fiber with a CCE direction parallel or perpendicular to the writing direction. The maximal bending sensitivity is independent on the writing direction, but the detailed bending response is different in the two cases. A temperature and strain sensor, based on a long-period grating and two...... sampled gratings, was produced and investigated. It is based on the different temperature and strain response of these gratings. Both a transfer matrix method and an overlap calculation is performed to explain the sensor response. Another type of sensor is based on tuning and modulation of a laser...

  5. Thiophene-Based Organic Semiconductors.

    Science.gov (United States)

    Turkoglu, Gulsen; Cinar, M Emin; Ozturk, Turan

    2017-10-24

    Thiophene-based π-conjugated organic small molecules and polymers are the research subject of significant current interest owing to their potential use as organic semiconductors in material chemistry. Despite simple and similar molecular structures, the hitherto reported properties of thiophene-based organic semiconductors are rather diverse. Design of high performance organic semiconducting materials requires a thorough understanding of inter- and intra-molecular interactions, solid-state packing, and the influence of both factors on the charge carrier transport. In this chapter, thiophene-based organic semiconductors, which are classified in terms of their chemical structures and their structure-property relationships, are addressed for the potential applications as organic photovoltaics (OPVs), organic field-effect transistors (OFETs) and organic light emitting diodes (OLEDs).

  6. Energy Data Base: Subject Thesaurus

    Energy Technology Data Exchange (ETDEWEB)

    Raridon, M.H. (ed.)

    1987-09-01

    This seventh edition of the subject thesaurus contains the standard vocabulary of indexing terms (descriptors) developed and structured by the technical staff of the Office of Scientific and Technical Information as part of its subject indexing activities for building and maintaining the Energy Data Base (EDB) and other energy information data bases for the Department of Energy (DOE). The purpose of this thesaurus is to enhance the efficiency of information retrieval from these data bases. This structured vocabulary reflects the scope of DOE's research, development, and technological programs and encompasses terminology derived not only from the basic sciences for also from areas of energy resources, conservation, safety, environmental impact, and regulation. There are 21,080 valid descriptors and 5683 forbidden terms in this edition of the Thesaurus. These descriptors are listed alphabetically.

  7. Brain Plasticity-Based Therapeutics

    Directory of Open Access Journals (Sweden)

    Michael eMerzenich

    2014-06-01

    Full Text Available The primary objective of this review article is to summarize how the neuroscience of brain plasticity, exploiting new findings in fundamental, integrative and cognitive neuroscience, is changing the therapeutic landscape for professional communities addressing brain-based disorders and disease. After considering the neurological bases of training-driven neuroplasticity, we shall describe how this neuroscience-guided perspective distinguishes this new approach from a the more-behavioral, traditional clinical strategies of professional therapy practitioners, and b an even more widely applied pharmaceutical treatment model for neurological and psychiatric treatment domains. With that background, we shall argue that neuroplasticity-based treatments will be an important part of future best-treatment practices in neurological and psychiatric medicine.

  8. Graphene-based smart materials

    Science.gov (United States)

    Yu, Xiaowen; Cheng, Huhu; Zhang, Miao; Zhao, Yang; Qu, Liangti; Shi, Gaoquan

    2017-09-01

    The high specific surface area and the excellent mechanical, electrical, optical and thermal properties of graphene make it an attractive component for high-performance stimuli-responsive or 'smart' materials. Complementary to these inherent properties, functionalization or hybridization can substantially improve the performance of these materials. Typical graphene-based smart materials include mechanically exfoliated perfect graphene, chemical vapour deposited high-quality graphene, chemically modified graphene (for example, graphene oxide and reduced graphene oxide) and their macroscopic assemblies or composites. These materials are sensitive to a range of stimuli, including gas molecules or biomolecules, pH value, mechanical strain, electrical field, and thermal or optical excitation. In this Review, we outline different graphene-based smart materials and their potential applications in actuators, chemical or strain sensors, self-healing materials, photothermal therapy and controlled drug delivery. We also introduce the working mechanisms of graphene-based smart materials and discuss the challenges facing the realization of their practical applications.

  9. Developing Constraint-based Recommenders

    Science.gov (United States)

    Felfernig, Alexander; Friedrich, Gerhard; Jannach, Dietmar; Zanker, Markus

    Traditional recommendation approaches (content-based filtering [48] and collaborative filtering[40]) are well-suited for the recommendation of quality&taste products such as books, movies, or news. However, especially in the context of products such as cars, computers, appartments, or financial services those approaches are not the best choice (see also Chapter 11). For example, apartments are not bought very frequently which makes it rather infeasible to collect numerous ratings for one specific item (exactly such ratings are required by collaborative recommendation algorithms). Furthermore, users of recommender applications would not be satisfied with recommendations based on years-old item preferences (exactly such preferences would be exploited in this context by content-based filtering algorithms).

  10. Photoelectrochemical based direct conversion systems

    Energy Technology Data Exchange (ETDEWEB)

    Kocha, S.; Arent, D.; Peterson, M. [National Renewable Energy Lab., Golden, CO (United States)] [and others

    1995-09-01

    The goal of this research is to develop a stable, cost effective, photoelectrochemical based system that will split water upon illumination, producing hydrogen and oxygen directly, using sunlight as the only energy input. This type of direct conversion system combines a photovoltaic material and an electrolyzer into a single monolithic device. We report on our studies of two multifunction multiphoton photoelectrochemical devices, one based on the ternary semiconductor gallium indium phosphide, (GaInP{sub 2}), and the other one based on amorphous silicon carbide. We also report on our studies of the solid state surface treatment of GaInP{sub 2} as well as our continuing effort to develop synthetic techniques for the attachment of transition metal complexes to the surface of semiconductor electrodes. All our surface studies are directed at controlling the interface energetics and forming stable catalytic surfaces.

  11. SILAC-Based Temporal Phosphoproteomics

    DEFF Research Database (Denmark)

    Francavilla, Chiara; Hekmat, Omid; Blagoev, Blagoy

    2014-01-01

    In recent years, thanks to advances in Mass Spectrometry (MS)-based quantitative proteomics, studies on signaling pathways have moved from a detailed description of individual components to system-wide analysis of entire signaling cascades, also providing spatio-temporal views of intracellular...... tyrosines by immunoaffinity and then further enriched for phosphorylated serine/threonine peptides by strong cation exchange in combination with titanium dioxide-beads chromatography. Analysis of enriched peptides on Orbitrap-based MS results in comprehensive and accurate reconstruction of temporal changes...... signaling events. Here we provide an optimized SILAC-based proteomic workflow to analyze temporal changes in phosphoproteomes, which involve a generic three step enrichment protocol for phosphopeptides. SILAC-labeled peptides from digested whole cell lysates are as a first step enriched for phosphorylated...

  12. Coal Reserves Data Base report

    Energy Technology Data Exchange (ETDEWEB)

    Jones, R.W.; Glass, G.B.

    1991-12-05

    The Coal Reserves Data Base (CRDB) Program is a cooperative data base development program sponsored by the Energy Information Administration (EIA). The objective of the CRDB Program is to involve knowledgeable coal resource authorities from the major coal-bearing regions in EIA's effort to update the Nation's coal reserves data. This report describes one of two prototype studies to update State-level reserve estimates. The CRDB data are intended for use in coal supply analyses and to support analyses of policy and legislative issues. They will be available to both Government and non-Government analysts. The data also will be part of the information used to supply United States energy data for international data bases and for inquiries from private industry and the public. (VC)

  13. Gold based bulk metallic glass

    OpenAIRE

    Schroers, Jan; Lohwongwatana, Boonrat; Johnson, William L.; Peker, Atakan

    2005-01-01

    Gold-based bulk metallic glass alloys based on Au-Cu-Si are introduced. The alloys exhibit a gold content comparable to 18-karat gold. They show very low liquidus temperature, large supercooled liquid region, and good processibility. The maximum casting thickness exceeds 5 mm in the best glassformer. Au49Ag5.5Pd2.3Cu26.9Si16.3 has a liquidus temperature of 644 K, a glass transition temperature of 401 K, and a supercooled liquid region of 58 K. The Vickers hardness of the alloys in this system...

  14. Texture-based correspondence display

    Science.gov (United States)

    Gerald-Yamasaki, Michael

    2005-03-01

    Texture-based correspondence display is a methodology to display corresponding data elements in visual representations of complex multidimensional, multivariate data. Texture is utilized as a persistent medium to contain a visual representation model and as a means to create multiple renditions of data where color is used to identify correspondence. Corresponding data elements are displayed over a variety of visual metaphors in a normal rendering process without adding extraneous linking metadata creation and maintenance. Texture-based correspondence display extends the effectiveness of visual representation for understanding data to the understanding and creation of visual representation models.

  15. Behavior-based evacuation planning

    KAUST Repository

    Rodriguez, Samuel

    2010-05-01

    In this work, we present a formulation of an evacuation planning problem that is inspired by motion planning and describe an integrated behavioral agent-based and roadmap-based motion planning approach to solve it. Our formulation allows users to test the effect on evacuation of a number of different environmental factors. One of our main focuses is to provide a mechanism to investigate how the interaction between agents influences the resulting evacuation plans. Specifically, we explore how various types of control provided by a set of directing agents effects the overall evacuation planning strategies of the evacuating agents. ©2010 IEEE.

  16. Laser-based coatings removal

    Energy Technology Data Exchange (ETDEWEB)

    Freiwald, J.G.; Freiwald, D.A. [F2 Associates, Inc., Albuquerque, NM (United States)

    1995-10-01

    Over the years as building and equipment surfaces became contaminated with low levels of uranium or plutonium dust, coats of paint were applied to stabilize the contaminants in place. Most of the earlier paint used was lead-based paint. More recently, various non-lead-based paints, such as two-part epoxy, are used. For D&D (decontamination and decommissioning), it is desirable to remove the paints or other coatings rather than having to tear down and dispose of the entire building. This report describes the use of pulse-repetetion laser systems for the removal of paints and coatings.

  17. Community-based recreational football

    DEFF Research Database (Denmark)

    Bruun, Ditte Marie; Bjerre, Eik; Krustrup, Peter

    2014-01-01

    is limited and the majority of prostate cancer survivors remain sedentary. Hence, novel approaches to evaluate and promote physical activity are warranted. This paper presents the rationale behind the delivery and evaluation of community-based recreational football offered in existing football clubs under...... the Danish Football Association to promote quality of life and physical activity adherence in prostate cancer survivors. The RE-AIM framework will be applied to evaluate the impact of the intervention including outcomes both at the individual and organizational level. By introducing community-based sport...

  18. Case-based medical informatics.

    Science.gov (United States)

    Pantazi, Stefan V; Arocha, José F; Moehr, Jochen R

    2004-11-08

    The "applied" nature distinguishes applied sciences from theoretical sciences. To emphasize this distinction, we begin with a general, meta-level overview of the scientific endeavor. We introduce the knowledge spectrum and four interconnected modalities of knowledge. In addition to the traditional differentiation between implicit and explicit knowledge we outline the concepts of general and individual knowledge. We connect general knowledge with the "frame problem," a fundamental issue of artificial intelligence, and individual knowledge with another important paradigm of artificial intelligence, case-based reasoning, a method of individual knowledge processing that aims at solving new problems based on the solutions to similar past problems. We outline the fundamental differences between Medical Informatics and theoretical sciences and propose that Medical Informatics research should advance individual knowledge processing (case-based reasoning) and that natural language processing research is an important step towards this goal that may have ethical implications for patient-centered health medicine. We focus on fundamental aspects of decision-making, which connect human expertise with individual knowledge processing. We continue with a knowledge spectrum perspective on biomedical knowledge and conclude that case-based reasoning is the paradigm that can advance towards personalized healthcare and that can enable the education of patients and providers. We center the discussion on formal methods of knowledge representation around the frame problem. We propose a context-dependent view on the notion of "meaning" and advocate the need for case-based reasoning research and natural language processing. In the context of memory based knowledge processing, pattern recognition, comparison and analogy-making, we conclude that while humans seem to naturally support the case-based reasoning paradigm (memory of past experiences of problem-solving and powerful case matching

  19. Towards a lunar base programme

    Science.gov (United States)

    Duke, M. B.; Mendell, W. W.; Roberts, B. B.

    1985-01-01

    When the requisite technlogy exists, the U.S. political process will inevitably include lunar surface activities as a major space objective. This article examines a manned lunar base in terms of three distinct functions: the scientific investigation of the moon and its environment; development of the capability to use lunar resources for beneficial purposes throughout the earth-moon system; and conduct of R and D leading to a self-sufficient and self-supporting manned lunar base. Three scenarios are outlined with respect to each possible function.

  20. Agent Based Individual Traffic guidance

    DEFF Research Database (Denmark)

    Wanscher, Jørgen Bundgaard

    2004-01-01

    When working with traffic planning or guidance it is common practice to view the vehicles as a combined mass. >From this models are employed to specify the vehicle supply and demand for each region. As the models are complex and the calculations are equally demanding the regions and the detail...... of the road network is aggregated. As a result the calculations reveal only what the mass of vehicles are doing and not what a single vehicle is doing. This is the crucial difference to ABIT (Agent Based Individual Trafficguidance). ABIT is based on the fact that information on the destination of each vehicle...

  1. Transistor-based interface circuitry

    Science.gov (United States)

    Taubman, Matthew S [Richland, WA

    2004-02-24

    Among the embodiments of the present invention is an apparatus that includes a transistor, a servo device, and a current source. The servo device is operable to provide a common base mode of operation of the transistor by maintaining an approximately constant voltage level at the transistor base. The current source is operable to provide a bias current to the transistor. A first device provides an input signal to an electrical node positioned between the emitter of the transistor and the current source. A second device receives an output signal from the collector of the transistor.

  2. Risk Based Optimal Fatigue Testing

    DEFF Research Database (Denmark)

    Sørensen, John Dalsgaard; Faber, M.H.; Kroon, I.B.

    1992-01-01

    Optimal fatigue life testing of materials is considered. Based on minimization of the total expected costs of a mechanical component a strategy is suggested to determine the optimal stress range levels for which additional experiments are to be performed together with an optimal value of the maxi......Optimal fatigue life testing of materials is considered. Based on minimization of the total expected costs of a mechanical component a strategy is suggested to determine the optimal stress range levels for which additional experiments are to be performed together with an optimal value...

  3. Flexible experimental FPGA based platform

    DEFF Research Database (Denmark)

    Andersen, Karsten Holm; Nymand, Morten

    2016-01-01

    This paper presents an experimental flexible Field Programmable Gate Array (FPGA) based platform for testing and verifying digital controlled dc-dc converters. The platform supports different types of control strategies, dc-dc converter topologies and switching frequencies. The controller platform...... interface supporting configuration and reading of setup parameters, controller status and the acquisition memory in a simple way. The FPGA based platform, provides an easy way within education or research to use different digital control strategies and different converter topologies controlled by an FPGA...

  4. Case-based medical informatics

    Directory of Open Access Journals (Sweden)

    Arocha José F

    2004-11-01

    Full Text Available Abstract Background The "applied" nature distinguishes applied sciences from theoretical sciences. To emphasize this distinction, we begin with a general, meta-level overview of the scientific endeavor. We introduce the knowledge spectrum and four interconnected modalities of knowledge. In addition to the traditional differentiation between implicit and explicit knowledge we outline the concepts of general and individual knowledge. We connect general knowledge with the "frame problem," a fundamental issue of artificial intelligence, and individual knowledge with another important paradigm of artificial intelligence, case-based reasoning, a method of individual knowledge processing that aims at solving new problems based on the solutions to similar past problems. We outline the fundamental differences between Medical Informatics and theoretical sciences and propose that Medical Informatics research should advance individual knowledge processing (case-based reasoning and that natural language processing research is an important step towards this goal that may have ethical implications for patient-centered health medicine. Discussion We focus on fundamental aspects of decision-making, which connect human expertise with individual knowledge processing. We continue with a knowledge spectrum perspective on biomedical knowledge and conclude that case-based reasoning is the paradigm that can advance towards personalized healthcare and that can enable the education of patients and providers. We center the discussion on formal methods of knowledge representation around the frame problem. We propose a context-dependent view on the notion of "meaning" and advocate the need for case-based reasoning research and natural language processing. In the context of memory based knowledge processing, pattern recognition, comparison and analogy-making, we conclude that while humans seem to naturally support the case-based reasoning paradigm (memory of past experiences

  5. Aptasensors Based on Stripping Voltammetry

    Directory of Open Access Journals (Sweden)

    Wenjing Qi

    2016-07-01

    Full Text Available Aptasensors based on stripping voltammetry exhibit several advantages, such as high sensitivity and multi-target detection from stripping voltammetric technology, and high selectivity from the specific binding of apamers with targets. This review comprehensively discusses the recent accomplishments in signal amplification strategies based on nanomaterials, such as metal nanoparticles, semiconductor nanoparticles, and nanocomposite materials, which are detected by stripping voltammetry after suitable dissolution. Focus will be put in discussing multiple amplification strategies that are widely applied in aptasensors for small biomolecules, proteins, disease markers, and cancer cells.

  6. Fidelity based measurement induced nonlocality

    Science.gov (United States)

    Muthuganesan, R.; Sankaranarayanan, R.

    2017-09-01

    In this paper, we propose measurement induced nonlocality (MIN) using a metric based on fidelity to capture global nonlocal effect of a quantum state due to locally invariant projective measurements. This quantity is a remedy for local ancilla problem in the original definition of MIN. We present an analytical expression of the proposed version of MIN for pure bipartite state and 2 × n dimensional mixed state. We also provide an upper bound of the MIN for general mixed state. Finally, we compare this quantity with MINs based on Hilbert-Schmidt norm and skew information for higher dimensional Werner and isotropic states.

  7. Cluster identification based on correlations.

    Science.gov (United States)

    Schulman, L S

    2012-04-01

    The problem addressed is the identification of cooperating agents based on correlations created as a result of the joint action of these and other agents. A systematic method for using correlations beyond second moments is developed. The technique is applied to a didactic example, the identification of alphabet letters based on correlations among the pixels used in an image of the letter. As in this example, agents can belong to more than one cluster. Moreover, the identification scheme does not require that the patterns be known ahead of time.

  8. Inquiry-based science education

    DEFF Research Database (Denmark)

    Østergaard, Lars Domino; Sillasen, Martin Krabbe; Hagelskjær, Jens

    2010-01-01

    Inquiry-based science education (IBSE) er en internationalt afprøvet naturfagsdidaktisk metode der har til formål at øge elevernes interesse for og udbytte af naturfag. I artiklen redegøres der for metoden, der kan betegnes som en elevstyret problem- og undersøgelsesbaseret naturfagsundervisnings......Inquiry-based science education (IBSE) er en internationalt afprøvet naturfagsdidaktisk metode der har til formål at øge elevernes interesse for og udbytte af naturfag. I artiklen redegøres der for metoden, der kan betegnes som en elevstyret problem- og undersøgelsesbaseret...

  9. Population Set based Optimization Method

    Science.gov (United States)

    Manekar, Y.; Verma, H. K.

    2013-09-01

    In this paper an population set based optimization method is proposed for solving some benchmark functions and also to solve optimal power flow problem like `combined economic and emission dispatch problem (CEED)' with multiple objective functions. This algorithm has taken into consideration all the equality and inequality constraints. The improvement in system performance is based on reduction in cost of power generation and active power loss. The proposed algorithms have been compared with the other methods like GA, PSO etc reported in the literature. The results are impressive and encouraging. The study results show that the proposed method holds better solutions in CEED problems.

  10. Technology base for microgravity horticulture

    Science.gov (United States)

    Sauer, R. L.; Magnuson, J. W.; Scruby, R. R.; Scheld, H. W.

    1987-01-01

    Advanced microgravity plant biology research and life support system development for the spacecraft environment are critically hampered by the lack of a technology base. This inadequacy stems primarily from the fact that microgravity results in a lack of convective currents and phase separation as compared to the one gravity environment. A program plan is being initiated to develop this technology base. This program will provide an iterative flight development effort that will be closely integrated with both basic science investigations and advanced life support system development efforts incorporating biological processes. The critical considerations include optimum illumination methods, root aeration, root and shoot support, and heat rejection and gas exchange in the plant canopy.

  11. Four segment piezo based micropump

    Science.gov (United States)

    Haldkar, Rakesh Kumar; Sheorey, Tanuja; Gupta, Vijay Kumar; Ansari, M. Zahid

    2017-06-01

    In recent years, micropumps have been investigated by various researchers as drug delivery and disease diagnostic devices. Many of these micropumps have been designed, considering available micro fabrication technologies rather than appropriate pump performance analysis. Piezoelectric based micro pumps are more popular as compared to other smart materials being explored. In this paper, four segment piezoelectric bimorph actuator (FSPB) are compared with circular disc piezoelectric bimorph actuator (CDPB) based pump. The static and transient behaviors under various electric fields have been analyzed by using ANSYS 12.1(R) finite element software. Simulation results show that dividing the actuator in segment can amplify the deflection and improve the performance of the pump.

  12. Transistor-based interface circuitry

    Science.gov (United States)

    Taubman, Matthew S.

    2007-02-13

    Among the embodiments of the present invention is an apparatus that includes a transistor, a servo device, and a current source. The servo device is operable to provide a common base mode of operation of the transistor by maintaining an approximately constant voltage level at the transistor base. The current source is operable to provide a bias current to the transistor. A first device provides an input signal to an electrical node positioned between the emitter of the transistor and the current source. A second device receives an output signal from the collector of the transistor.

  13. Pheromone based alternative route planning

    Directory of Open Access Journals (Sweden)

    Liangbing Feng

    2016-08-01

    Full Text Available In this work, we propose an improved alternative route calculation based on alternative figures, which is suitable for practical environments. The improvement is based on the fact that the main traffic route is the road network skeleton in a city. Our approach using nodes may generate a higher possibility of overlapping. We employ a bidirectional Dijkstra algorithm to search the route. To measure the quality of an Alternative Figures (AG, three quotas are proposed. The experiment results indicate that the improved algorithm proposed in this paper is more effective than others.

  14. Knowledge-Based Replanning System.

    Science.gov (United States)

    1987-05-01

    INTERFACE 2.0 INTRODUCTION 12 2.1 HISTORY OF APE-Il PARSER IMPLEMENTATION 12 2.1.1 ATN-Based Natural Language 13 2.1.2 APE-Il Development 14 2.2...understanding using a conceptual-dependency parser for question-answer- ing and control, and a constraint-based approach to planning. This complex architecture...eacri extension to the ssisem JJ! LL It. t’alxt and Increalses the cost of future exttnsion-, aind the lawk if dilmnis hinl. returns comenic lr ~i

  15. Why Internet-based Education?

    Directory of Open Access Journals (Sweden)

    Morton Ann Gernsbacher

    2015-01-01

    Full Text Available This essay illustrates five ways that Internet-based higher education can capitalize on fundamental principles of learning. Internet-based education can enable better mastery through distributed (shorter, more frequent practice rather than massed (longer, less frequent practice; it can optimize performance because it allows students to learn at their peak time of their day; it can deepen memory because it requires cheat-proof assignments and tests; it can promote critical thinking because it necessitates intellectual winnowing and sifting; and it can enhance writing skills by requiring students to write frequently and for a broad audience.

  16. Model-Based Requirements Management in Gear Systems Design Based On Graph-Based Design Languages

    Directory of Open Access Journals (Sweden)

    Kevin Holder

    2017-10-01

    Full Text Available For several decades, a wide-spread consensus concerning the enormous importance of an in-depth clarification of the specifications of a product has been observed. A weak clarification of specifications is repeatedly listed as a main cause for the failure of product development projects. Requirements, which can be defined as the purpose, goals, constraints, and criteria associated with a product development project, play a central role in the clarification of specifications. The collection of activities which ensure that requirements are identified, documented, maintained, communicated, and traced throughout the life cycle of a system, product, or service can be referred to as “requirements engineering”. These activities can be supported by a collection and combination of strategies, methods, and tools which are appropriate for the clarification of specifications. Numerous publications describe the strategy and the components of requirements management. Furthermore, recent research investigates its industrial application. Simultaneously, promising developments of graph-based design languages for a holistic digital representation of the product life cycle are presented. Current developments realize graph-based languages by the diagrams of the Unified Modelling Language (UML, and allow the automatic generation and evaluation of multiple product variants. The research presented in this paper seeks to present a method in order to combine the advantages of a conscious requirements management process and graph-based design languages. Consequently, the main objective of this paper is the investigation of a model-based integration of requirements in a product development process by means of graph-based design languages. The research method is based on an in-depth analysis of an exemplary industrial product development, a gear system for so-called “Electrical Multiple Units” (EMU. Important requirements were abstracted from a gear system

  17. Problem Based Learning and sustainability

    DEFF Research Database (Denmark)

    Pizzol, Massimo; Løkke, Søren; Schmidt, Jannick Højrup

    Problem-Based Learning (PBL) is a style of active learning based on problem solving. PBL aims at providing university students with flexible knowledge, capacity to self-learning, and skills in problem solving and collaboration. In this context, the present study explores the advantages and challe......Problem-Based Learning (PBL) is a style of active learning based on problem solving. PBL aims at providing university students with flexible knowledge, capacity to self-learning, and skills in problem solving and collaboration. In this context, the present study explores the advantages...... and challenges that the PBL model offers for developing five key competences in sustainability: (i) system thinking, (ii) interpersonal competence, (iii) anticipatory competence, (iv) strategic competence, (v) normative competences. The study draws on the experiences from PBL activities performed at Aalborg...... University (AAU), Denmark, and focuses on the teaching of Life Cycle Assessment as a method for sustainability assessment. The objective is providing recommendations for future LCA teaching and learning. PBL activites performed at AAU were evaluated critically to detemine to what extent they addressed...

  18. Converse PUF-based authentication

    NARCIS (Netherlands)

    Kocabas, U.; Peter, Andreas; Katzenbeisser, S.; Sadeghi, A.

    Physically Unclonable Functions (PUFs) are key tools in the construction of lightweight authentication and key exchange protocols. So far, all existing PUF-based authentication protocols follow the same paradigm: A resource-constrained prover, holding a PUF, wants to authenticate to a resource-rich

  19. Risk-based decisionmaking (Panel)

    Energy Technology Data Exchange (ETDEWEB)

    Smith, T.H.

    1995-12-31

    By means of a panel discussion and extensive audience interaction, explore the current challenges and progress to date in applying risk considerations to decisionmaking related to low-level waste. This topic is especially timely because of the proposed legislation pertaining to risk-based decisionmaking and because of the increased emphasis placed on radiological performance assessments of low-level waste disposal.

  20. India-based Neutrino Observatory

    Indian Academy of Sciences (India)

    2012-11-17

    Nov 17, 2012 ... Abstract. The current status of the India-based Neutrino Observatory (INO) is summarized. The main physics goals are described followed by the motivation for building a magnetized iron calori- metric (ICAL) detector. The charge identification capability of ICAL would make it complementary to large water ...

  1. TEE-Based Trusted Storage

    DEFF Research Database (Denmark)

    Gonzalez, Javier; Bonnet, Philippe

    Today, it is safe to assume that any program or data can be compromised, if they are not protected by hardware within a secure area. Systems based on crypto-processors (e.g., a trusted platform module, a smart card or a hardware security module) rely on the properties of tamper resistant hardware...

  2. Community-Based Assessment Pedagogy

    Science.gov (United States)

    Inoue, Asao B.

    2004-01-01

    This article attempts to structure student assessment practices in the classroom. Informed by fourth generation evaluation, it discusses a pedagogy based on a recursive framework of writing, assessment, and reflection activities that move students toward productive praxis. Implemented over three semesters at a land grant university in the U.S.,…

  3. The LADM based on INTERLIS

    NARCIS (Netherlands)

    Germann, M.; Kaufmann, J.; Lemmen, C.H.J.; Van Oosterom, P.J.M.; De Zeeuw, K.

    2015-01-01

    Both the conceptual schema language INTERLIS and the land administration domain model (LADM) share the same model driven architecture (MDA) principles. In this paper we explore how INTERLIS and LADM complement each other in actual implementation of land administration system based on the LADM using

  4. Consumption-based Equity Valuation

    DEFF Research Database (Denmark)

    Bach, Christian; Christensen, Peter Ove

    2013-01-01

    Using a CCAPM-based risk-adjustment model consistent with general asset pricing theory, we perform yearly valuations of a large sample of stocks listed on NYSE, AMEX and NASDAQ over a thirty-year period. The model differs from standard valuation models in the sense that it adjusts forecasted resi...

  5. Graphite-based photovoltaic cells

    Science.gov (United States)

    Lagally, Max; Liu, Feng

    2010-12-28

    The present invention uses lithographically patterned graphite stacks as the basic building elements of an efficient and economical photovoltaic cell. The basic design of the graphite-based photovoltaic cells includes a plurality of spatially separated graphite stacks, each comprising a plurality of vertically stacked, semiconducting graphene sheets (carbon nanoribbons) bridging electrically conductive contacts.

  6. Bandit-Based Genetic Programming

    OpenAIRE

    Hoock, Jean-Baptiste; Teytaud, Olivier

    2010-01-01

    International audience; We consider the validation of randomly generated patterns in a Monte-Carlo Tree Search program. Our bandit-based genetic programming (BGP) algorithm, with proved mathematical properties, outperformed a highly optimized handcrafted module of a well-known computer-Go program with several world records in the game of Go.

  7. Mesoporous Silicon-Based Anodes

    Science.gov (United States)

    Peramunage, Dharmasena

    2015-01-01

    For high-capacity, high-performance lithium-ion batteries. A new high-capacity anode composite based on mesoporous silicon is being developed. With a structure that resembles a pseudo one-dimensional phase, the active anode material will accommodate significant volume changes expected upon alloying and dealloying with lithium (Li).

  8. Model-based Software Engineering

    DEFF Research Database (Denmark)

    Kindler, Ekkart

    2010-01-01

    The vision of model-based software engineering is to make models the main focus of software development and to automatically generate software from these models. Part of that idea works already today. But, there are still difficulties when it comes to behaviour. Actually, there is no lack in models...

  9. Activity based costing (ABC Method

    Directory of Open Access Journals (Sweden)

    Prof. Ph.D. Saveta Tudorache

    2008-05-01

    Full Text Available In the present paper the need and advantages are presented of using the Activity BasedCosting method, need arising from the need of solving the information pertinence issue. This issue has occurreddue to the limitation of classic methods in this field, limitation also reflected by the disadvantages ofsuch classic methods in establishing complete costs.

  10. Context based multimedia information retrieval

    DEFF Research Database (Denmark)

    Mølgaard, Lasse Lohilahti

    with the help of contextual knowledge. Our approach to model the context of multimedia is based on unsupervised methods to automatically extract meaning. We investigate two paths of context modelling. The first part extracts context from the primary media, in this case broadcast news speech, by extracting...

  11. Interactive example-based hatching

    NARCIS (Netherlands)

    Gerl, Moritz; Isenberg, Tobias

    2013-01-01

    We present an approach for interactively generating pen-and-ink hatching renderings based on hand-drawn examples. We aim to overcome the regular and synthetic appearance of the results of existing methods by incorporating human virtuosity and illustration skills in the computer generation of such

  12. Exchanging Description Logic Knowledge Bases

    NARCIS (Netherlands)

    Arenas, M.; Botoeva, E.; Calvanese, D.; Ryzhikov, V.; Sherkhonov, E.

    2012-01-01

    In this paper, we study the problem of exchanging knowledge between a source and a target knowledge base (KB), connected through mappings. Differently from the traditional database exchange setting, which considers only the exchange of data, we are interested in exchanging implicit knowledge. As

  13. Evidence-based management reconsidered.

    Science.gov (United States)

    Kovner, Anthony R; Rundall, Thomas G

    2006-01-01

    Reports of medical mistakes have splashed across newspapers and magazines in the United States. At the same time, instances of overuse, underuse, and misuse of management tactics and strategies receive far less attention. The sense of urgency associated with improving the quality of medical care does not exist with respect to improving the quality of management decision making. A more evidence-based approach would improve the competence of the decision-makers and their motivation to use more scientific methods when making a decision. The authors of this article consider a study of 68 U.S. health services managers that found a low level of evidence-based management behaviors. From the findings, four strategies are suggested to increase health systems managers' use of research evidence to improve decision making: focusing evidence-based decision making on strategically important issues, developing committees and other structures to diffuse management research throughout the organization, building a management culture that values research, and training managers in the competencies required to apply research evidence to health services management decisions. To aid the manager in understanding and applying an evidenced-based approach to decision making, the article provides practical tools, techniques, and resources for immediate use.

  14. Web Based ATM PVC Management

    NARCIS (Netherlands)

    van der Waaij, B.D.; Sprenkels, Ron; van Beijnum, Bernhard J.F.; Pras, Aiko

    1998-01-01

    This paper discusses the design of a public domain web based ATM PVC Management tool for the Dutch SURFnet research ATM network. The aim of this tool is to assists in the creation and deletion of PVCs through local and remote ATM network domains. The tool includes security mechanisms to restrict the

  15. Colour Mixing Based on Daylight

    Science.gov (United States)

    Meyn, Jan-Peter

    2008-01-01

    Colour science is based on the sensation of monochromatic light. In contrast to that, surface colours are caused by reflection of wide sections of the daylight spectrum. Non-spectral colours like magenta and purple appear homologous to colours with spectral hue, if the approach of mixing monochromatic light is abandoned. It is shown that a large…

  16. Collaborative Inquiry-based Learning

    NARCIS (Netherlands)

    Suarez, Angel

    2017-01-01

    This thesis presents the results of the conducted research and development of applications to support collaborative inquiry-based learning, with a special focus on leveraging learners’ agency. The reported results are structured into three parts: the theoretical foundations, the design and

  17. Intelligence-based systems engineering

    CERN Document Server

    Tolk, Andreas

    2011-01-01

    The International Council on Systems Engineering (INCOSE) defines Systems Engineering as an interdisciplinary approach and means to enable the realization of successful systems. Researchers are using intelligence-based techniques to support the practices of systems engineering in an innovative way. This research volume includes a selection of contributions by subject experts to design better systems.

  18. Activity based costing (ABC Method)

    OpenAIRE

    Prof. Ph.D. Saveta Tudorache; University Lecturer Anca Bratu

    2008-01-01

    In the present paper the need and advantages are presented of using the Activity Based Costing method, need arising from the need of solving the information pertinence issue. This issue has occurred due to the limitation of classic methods in this field, limitation also reflected by the disadvantages of such classic methods in establishing complete costs.

  19. Ensemble based co-training

    NARCIS (Netherlands)

    Tanha, J.; van Someren, M.; Afsarmanesh, H.

    2011-01-01

    Recently Semi-Supervised learning algorithms such as co-training are used in many application domains. In co-training, two classifiers based on different views of data or on different learning algorithms are trained in parallel and then unlabeled data that are classified differently by the

  20. Accelerator-Based Neutrino Beams

    OpenAIRE

    Hübner, K

    2003-01-01

    The design principles of accelerator-based neutrino beams are outlined and the beams currently in operation or under construction are briefly described. The concepts and basic features of the different types of advanced neutrino beams which are under study are summarized.