90 research outputs found

    The Design and Assessment of Attention-Getting Rear Brake Light Signals

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    This paper summarizes work intended to further characterize and develop rear brake light signals likely to improve driver reaction to hard-braking lead-vehicle events, emphasizing unique and novel approaches not previously studied. The work developed optimized signal lighting configurations, including specifications for LED signal approaches (flash frequencies, brightness levels, patterns), and performed field evaluations to assess eye-drawing capability of candidate signals for drivers who were looking away from the forward view. Results indicate that newer rear signaling designs can be very effective at drawing drivers’ eyes back to the forward roadway, and that flashing and brightness are two important signal properties moderating effectiveness (attention-getting). Remarkable performance gains can be achieved via use of LED signal approaches which both flash and increase signal intensity or lamp brightness

    News discourse and readers’ comments: expanding the range of citizenship positions?

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    First Published May 15, 2017.Little attention has been paid to the relation between citizens’ representation in news media and citizen participation in readers’ comments, and to the roles both discourses may play in fostering public engagement in official consultation processes. This article offers a discursive analysis of these questions by focusing on how commenters, through their uses of language in connection with news texts, address the political ordering of news discourse and their positioning therein. Using Critical Discourse Analysis and other interaction-oriented forms of discourse analysis, we examine, first, the topics and the framing of voices in news coverage and, second, the interactional order, stance markers and style features of readers’ comments. Based on data regarding a policy plan on hydroelectric power in Portugal that was submitted to public consultation, we show that citizen positionings emerging from the interaction between news texts and comments change the balance of power within the discussion, but their participatory potential is restrained by traditional citizenship regimes.The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was developed for the project COMPOLIS: 'Communication and Political Engagement with Environmental Issues' and supported by the Fundacao para a Ciencia e a Tecnologia (Portugal) [grant number EXPL/IVC-COM/1717/2012] through national funds (PIDDAC) and co-funded by the European Fund of Regional Development (FEDER) through COMPETE - Operational Program Competitive Factors (POFC).info:eu-repo/semantics/publishedVersio

    Fluorescent carbon dioxide indicators

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    Over the last decade, fluorescence has become the dominant tool in biotechnology and medical imaging. These exciting advances have been underpinned by the advances in time-resolved techniques and instrumentation, probe design, chemical / biochemical sensing, coupled with our furthered knowledge in biology. Complementary volumes 9 and 10, Advanced Concepts of Fluorescence Sensing: Small Molecule Sensing and Advanced Concepts of Fluorescence Sensing: Macromolecular Sensing, aim to summarize the current state of the art in fluorescent sensing. For this reason, Drs. Geddes and Lakowicz have invited chapters, encompassing a broad range of fluorescence sensing techniques. Some chapters deal with small molecule sensors, such as for anions, cations, and CO2, while others summarize recent advances in protein-based and macromolecular sensors. The Editors have, however, not included DNA or RNA based sensing in this volume, as this were reviewed in Volume 7 and is to be the subject of a more detailed volume in the near future

    Neil3-dependent base excision repair regulates lipid metabolism and prevents atherosclerosis in Apoe-deficient mice

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    Increasing evidence suggests that oxidative DNA damage accumulates in atherosclerosis. Recently, we showed that a genetic variant in the human DNA repair enzyme NEIL3 was associated with increased risk of myocardial infarction. Here, we explored the role of Neil3/NEIL3 in atherogenesis by both clinical and experimental approaches. Human carotid plaques revealed increased NEIL3 mRNA expression which significantly correlated with mRNA levels of the macrophage marker CD68. Apoe−/−Neil3−/− mice on high-fat diet showed accelerated plaque formation as compared to Apoe−/− mice, reflecting an atherogenic lipid profile, increased hepatic triglyceride levels and attenuated macrophage cholesterol efflux capacity. Apoe−/−Neil3−/− mice showed marked alterations in several pathways affecting hepatic lipid metabolism, but no genotypic alterations in genome integrity or genome-wide accumulation of oxidative DNA damage. These results suggest a novel role for the DNA glycosylase Neil3 in atherogenesis in balancing lipid metabolism and macrophage function, potentially independently of genome-wide canonical base excision repair of oxidative DNA damage

    The human DNA glycosylases NEIL1 and NEIL3 excise psoralen-induced DNA-DNA cross-links in a four-stranded DNA structure

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    Interstrand cross-links (ICLs) are highly cytotoxic DNA lesions that block DNA replication and transcription by preventing strand separation. Previously, we demonstrated that the bacterial and human DNA glycosylases Nei and NEIL1 excise unhooked psoralen-derived ICLs in three-stranded DNA via hydrolysis of the glycosidic bond between the crosslinked base and deoxyribose sugar. Furthermore, NEIL3 from Xenopus laevis has been shown to cleave psoralen- and abasic site-induced ICLs in Xenopus egg extracts. Here we report that human NEIL3 cleaves psoralen-induced DNA-DNA cross-links in three-stranded and four-stranded DNA substrates to generate unhooked DNA fragments containing either an abasic site or a psoralen-thymine monoadduct. Furthermore, while Nei and NEIL1 also cleave a psoralen-induced four-stranded DNA substrate to generate two unhooked DNA duplexes with a nick, NEIL3 targets both DNA strands in the ICL without generating single-strand breaks. The DNA substrate specificities of these Nei-like enzymes imply the occurrence of long uninterrupted three- and four-stranded crosslinked DNA-DNA structures that may originate in vivo from DNA replication fork bypass of an ICL. In conclusion, the Nei-like DNA glycosylases unhook psoralen-derived ICLs in various DNA structures via a genuine repair mechanism in which complex DNA lesions can be removed without generation of highly toxic double-strand breaks

    Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging

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    This study investigates the temporal and spatial interchange of the aromatic amino acid phenylalanine (Phe) between human retinal pigment epithelial cell line (ARPE-19) and tachyzoites of the apicomplexan protozoan parasite Toxoplasma gondii (T. gondii). Stable isotope labelling by amino acids in cell culture (SILAC) is combined with Raman micro-spectroscopy to selectively monitor the incorporation of deuterium-labelled Phe into proteins in individual live tachyzoites. Our results show a very rapid uptake of L-Phe(D8) by the intracellular growing parasite. T. gondii tachyzoites are capable of extracting L-Phe(D8) from host cells as soon as it invades the cell. L-Phe(D8) from the host cell completely replaces the L-Phe within T. gondii tachyzoites 7–9 hours after infection. A quantitative model based on Raman spectra allowed an estimation of the exchange rate of Phe as 0.5–1.6 × 104 molecules/s. On the other hand, extracellular tachyzoites were not able to consume L-Phe(D8) after 24 hours of infection. These findings further our understanding of the amino acid trafficking between host cells and this strictly intracellular parasite. In particular, this study highlights new aspects of the metabolism of amino acid Phe operative during the interaction between T. gondii and its host cell

    Antibody-free magnetic cell sorting of genetically modified primary human CD4+ T cells by one-step streptavidin affinity purification.

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    Existing methods for phenotypic selection of genetically modified mammalian cells suffer disadvantages of time, cost and scalability and, where antibodies are used to bind exogenous cell surface markers for magnetic selection, typically yield cells coated with antibody-antigen complexes and beads. To overcome these limitations we have developed a method termed Antibody-Free Magnetic Cell Sorting in which the 38 amino acid Streptavidin Binding Peptide (SBP) is displayed at the cell surface by the truncated Low Affinity Nerve Growth Receptor (LNGFRF) and used as an affinity tag for one-step selection with streptavidin-conjugated magnetic beads. Cells are released through competition with the naturally occurring vitamin biotin, free of either beads or antibody-antigen complexes and ready for culture or use in downstream applications. Antibody-Free Magnetic Cell Sorting is a rapid, cost-effective, scalable method of magnetic selection applicable to either viral transduction or transient transfection of cell lines or primary cells. We have optimised the system for enrichment of primary human CD4+ T cells expressing shRNAs and exogenous genes of interest to purities of >99%, and used it to isolate cells following Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 genome editing

    Transforming derivational complexity of term rewriting to runtime complexity

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    Derivational complexity of term rewriting considers the length of the longest rewrite sequence for arbitrary start terms, whereas runtime complexity restricts start terms to basic terms. Recently, there has been notable progress in automatic inference of upper and lower bounds for runtime complexity. We propose a novel transformation that allows an off-the-shelf tool for inference of upper or lower bounds for runtime complexity to be used to determine upper or lower bounds for derivational complexity as well. Our approach is applicable to derivational complexity problems for innermost rewriting and for full rewriting. We have implemented the transformation in the tool AProVE and conducted an extensive experimental evaluation. Our results indicate that bounds for derivational complexity can now be inferred for rewrite systems that have been out of reach for automated analysis thus far
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