139 research outputs found

    W/SiC x-ray multilayers optimized for use above 100 keV

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    We have developed a new depth-graded multilayer system comprising W and SiC layers, suitable for use as hard x-ray reflective coatings operating in the energy range 100-200 keV. Grazing-incidence x-ray reflectance at E = 8 keV was used to characterize the interface widths, as well as the temporal and thermal stability in both periodic and depth-graded W/SiC structures, whereas synchrotron radiation was used to measure the hard x-ray reflectance of a depth-graded multilayer designed specifically for use in the range E ~150-170 keV. We have modeled the hard x-ray reflectance using newly derived optical constants, which we determined from reflectance versus incidence angle measurements also made using synchrotron radiation, in the range E = 120-180 keV. We describe our experimental investigation in detail, compare the new W/SiC multilayers with both W/Si and W/B4C films that have been studied previously, and discuss the significance of these results with regard to the eventual development of a hard x-ray nuclear line telescope

    Native RNA Purification Method for Small RNA Molecules Based on Asymmetrical Flow Field-Flow Fractionation

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    RNA molecules provide promising new possibilities for the prevention and treatment of viral infections and diseases. The rapid development of RNA biology and medicine requires advanced methods for the purification of RNA molecules, which allow fast and efficient RNA processing, preferably under non-denaturing conditions. Asymmetrical flow field-flow fractionation (AF4) enables gentle separation and purification of macromolecules based on their diffusion coefficients. The aim of the study was to develop an AF4 method for efficient purification of enzymatically produced antiviral small interfering (si)RNA molecules and to evaluate the overall potential of AF4 in the separation of short single-stranded (ss) and double-stranded (ds) RNA molecules. We show that AF4 separates monomeric ssRNA from dsRNA molecules of the same size and monomeric ssRNA from multimeric forms of the same ssRNA. The developed AF4 method enabled the separation of enzymatically produced 27-nt siRNAs from partially digested substrate dsRNA, which is potentially toxic for mammalian cells. The recovery of AF4-purified enzymatically produced siRNA molecules was about 70%, which is about 20% higher than obtained using anion-exchange chromatography. The AF4-purified siRNAs were not toxic for mammalian cells and fully retained their biological activity as confirmed by efficient inhibition of herpes simplex virus 1 replication in cell culture. Our work is the first to develop AF4 methods for the separation of short RNA molecules

    Chromatin organization regulates viral egress dynamics

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    Various types of DNA viruses are known to elicit the formation of a large nuclear viral replication compartment and marginalization of the cell chromatin. We used three-dimensional soft x-ray tomography, confocal and electron microscopy, combined with numerical modelling of capsid diffusion to analyse the molecular organization of chromatin in herpes simplex virus 1 infection and its effect on the transport of progeny viral capsids to the nuclear envelope. Our data showed that the formation of the viral replication compartment at late infection resulted in the enrichment of heterochromatin in the nuclear periphery accompanied by the compaction of chromatin. Random walk modelling of herpes simplex virus 1-sized particles in a three-dimensional soft x-ray tomography reconstruction of an infected cell nucleus demonstrated that the peripheral, compacted chromatin restricts viral capsid diffusion, but due to interchromatin channels capsids are able to reach the nuclear envelope, the site of their nuclear egress.Peer reviewe

    Cormorants have negligible seascape-scale impacts on benthic vegetation communities

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    Cormorant populations in the Baltic Sea have rapidly expanded since the 1990s, raising concerns about their ecosystem impacts. Nutrient runoff from colonies, as well as cormorant predation on fish, can affect surrounding producer communities. Past studies have found cormorant impacts on producers in the immediate vicinity of colonies, but the importance of cormorants over a larger spatial scale is unknown, especially compared to other environmental variables. We used an extensive underwater vegetation inventory dataset (~18 000 data points along the Finnish coast) to determine the effects of cormorant colonies on macroalgae and plants. We compared community structure and species abundance/occurrence in near-colony (10 km from a colony) points, and determined the importance of cormorant influence (using an index incorporating colony size and distance from the colony) in near-colony sites. We found no significant differences in community structure between near-colony and control points in most habitats, and adding cormorant index only infinitesimally improved statistical models after incorporating other environmental factors. However, the abundance of several species did differ, in particular the foundation species bladderwrack (Fucus vesiculosus) and eelgrass (Zostera marina) were either less likely to occur in near-colony points or negatively correlated with cormorant index, possibly due to the effects of nutrient enrichment from colonies. Our findings confirm that cormorants can have effects on some producer species, but highlight that these effects are negligible when taking into account the scale and magnitude of other bottom up and top-down processes occurring in the Baltic Sea.</div

    Hole depletion and localization due to disorder in insulating PrBa2Cu3O7-d: a Compton scattering study

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    The (mostly) insulating behaviour of PrBa2Cu3O7-d is still unexplained and even more interesting since the occasional appearance of superconductivity in this material. Since YBa2Cu3O7-d is nominally iso-structural and always superconducting, we have measured the electron momentum density in these materials. We find that they differ in a striking way, the wavefunction coherence length in PrBa2Cu3O7-d being strongly suppressed. We conclude that Pr on Ba-site substitution disorder is responsible for the metal-insulator transition. Preliminary efforts at growth with a method to prevent disorder yield 90K superconducting PrBa2Cu3O7-d crystallites.Comment: 4 pages, 3 figures, revised version submitted to PR

    Native RNA Purification Method for Small RNA Molecules Based on Asymmetrical Flow Field-Flow Fractionation

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    RNA molecules provide promising new possibilities for the prevention and treatment of viral infections and diseases. The rapid development of RNA biology and medicine requires advanced methods for the purification of RNA molecules, which allow fast and efficient RNA processing, preferably under non-denaturing conditions. Asymmetrical flow field-flow fractionation (AF4) enables gentle separation and purification of macromolecules based on their diffusion coefficients. The aim of the study was to develop an AF4 method for efficient purification of enzymatically produced antiviral small interfering (si)RNA molecules and to evaluate the overall potential of AF4 in the separation of short single-stranded (ss) and double-stranded (ds) RNA molecules. We show that AF4 separates monomeric ssRNA from dsRNA molecules of the same size and monomeric ssRNA from multimeric forms of the same ssRNA. The developed AF4 method enabled the separation of enzymatically produced 27-nt siRNAs from partially digested substrate dsRNA, which is potentially toxic for mammalian cells. The recovery of AF4-purified enzymatically produced siRNA molecules was about 70%, which is about 20% higher than obtained using anion-exchange chromatography. The AF4-purified siRNAs were not toxic for mammalian cells and fully retained their biological activity as confirmed by efficient inhibition of herpes simplex virus 1 replication in cell culture. Our work is the first to develop AF4 methods for the separation of short RNA molecules

    Hard x-ray characterization of a HEFT single-reflection prototype

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    We have measured the hard X-ray reflectivity and imaging performance from depth graded W/Si multilayer coated mirror segments mounted in a single reflection cylindrical prototype for the hard X-ray telescopes to be flown on the High Energy Focusing Telescope (HEFT) balloon mission. Data have been obtained in the energy range from 18 - 170 keV at the European Synchrotron Radiation Facility and at the Danish Space Research Institute at 8 keV. The modeling of the reflectivity data demonstrate that the multilayer structure can be well described by the intended power law distribution of the bilayer thicknesses optimized for the telescope performance and we find that all the data is consistent with an interfacial width of 4.5 Å. We have also demonstrated that the required 5% uniformity of the coatings is obtained over the mirror surface and we have shown that it is feasible to use similar W/Si coatings for much higher energies than the nominal energy range of HEFT leading the way for designing Gamma-ray telescopes for future astronomical applications. Finally we have demonstrate 35 arcsecond Half Power Diameter imaging performance of the one bounce prototype throughout the energy range of the HEFT telescopes

    Global patterns in the impact of marine herbivores on benthic primary producers

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    Despite the importance of consumers in structuring communities, and the widespread assumption that consumption is strongest at low latitudes, empirical tests for global scale patterns in the magnitude of consumer impacts are limited. In marine systems, the long tradition of experimentally excluding herbivores in their natural environments allows consumer impacts to be quantified on global scales using consistent methodology. We present a quantitative synthesis of 613 marine herbivore exclusion experiments to test the influence of consumer traits, producer traits and the environment on the strength of herbivore impacts on benthic producers. Across the globe, marine herbivores profoundly reduced producer abundance (by 68% on average), with strongest effects in rocky intertidal habitats and the weakest effects on habitats dominated by vascular plants. Unexpectedly, we found little or no influence of latitude or mean annual water temperature. Instead, herbivore impacts differed most consistently among producer taxonomic and morphological groups. Our results show that grazing impacts on plant abundance are better predicted by producer traits than by large‐scale variation in habitat or mean temperature, and that there is a previously unrecognised degree of phylogenetic conservatism in producer susceptibility to consumption

    Herpes Simplex Virus Type 1 Clinical Isolates Respond to UL29-Targeted siRNA Swarm Treatment Independent of Their Acyclovir Sensitivity

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    Acyclovir is the drug of choice for the treatment of herpes simplex virus (HSV) infections. Acyclovir-resistant HSV strains may emerge, especially during long-term drug use, and subsequently cause difficult-to-treat exacerbations. Previously, we set up a novel treatment approach, based on enzymatically synthesized pools of siRNAs, or siRNA swarms. These swarms can cover kilobases-long target sequences, reducing the likelihood of resistance to treatment. Swarms targeting the UL29 essential gene of HSV-1 have demonstrated high efficacy against HSV-1 in vitro and in vivo. Here, we assessed the antiviral potential of a UL29 siRNA swarm against circulating strains of HSV-1, in comparison with acyclovir. All circulating strains were sensitive to both antivirals, with the half-maximal inhibitory concentrations (IC50) in the range of 350–1911 nM for acyclovir and 0.5–3 nM for the UL29 siRNA swarm. Additionally, we showed that an acyclovir-resistant HSV-1, devoid of thymidine kinase, is highly sensitive to UL29 siRNA treatment (IC50 1.0 nM; Imax 97%). Moreover, the detected minor variations in the RNAi target of the HSV strains had no effect on the potency or efficacy of UL29 siRNA swarm treatment. Our findings support the development of siRNA swarms for the treatment of HSV-1 infections, in order to circumvent any potential acyclovir resistance
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