1,011 research outputs found

    Live Attenuated Influenza Vaccine contains Substantial and Unexpected Amounts of Defective Viral Genomic RNA

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    The live attenuated influenza vaccine FluMist® was withdrawn in the USA by the Centers for Disease Control and Prevention after its failure to provide adequate protective immunity during 2013–2016. The vaccine uses attenuated core type A and type B viruses, reconfigured each year to express the two major surface antigens of the currently circulating viruses. Here Fluenz™ Tetra, the European version of this vaccine, was examined directly for defective-interfering (DI) viral RNAs. DI RNAs are deleted versions of the infectious virus genome, and have powerful biological properties including attenuation of infection, reduction of infectious virus yield, and stimulation of some immune responses. Reverse transcription polymerase chain reaction followed by cloning and sequencing showed that Fluenz™ vaccine contains unexpected and substantial amounts of DI RNA arising from both its influenza A and influenza B components, with 87 different DI RNA sequences identified. Flu A DI RNAs from segment 3 replaced the majority of the genomic full-length segment 3, thus compromising its infectivity. DI RNAs arise during vaccine production and non-infectious DI virus replaces infectious virus pro rata so that fewer doses of the vaccine can be made. Instead the vaccine carries a large amount of non-infectious but biologically active DI virus. The presence of DI RNAs could significantly reduce the multiplication in the respiratory tract of the vaccine leading to reduced immunizing efficacy and could also stimulate the host antiviral responses, further depressing vaccine multiplication. The role of DI viruses in the performance of this and other vaccines requires further investigation

    Molecular evolution of the insect-specific flaviviruses

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    There has been an explosion in the discovery of ‘insect-specific’ flaviviruses and/or their related sequences in natural mosquito populations. Herein we review all ‘insect-specific’ flavivirus sequences currently available and conduct phylogenetic analyses of both the ‘insect-specific’ flaviviruses and available sequences of the entire genus Flavivirus. We show that there is no statistical support for virus–mosquito co-divergence, suggesting that the ‘insect-specific’ flaviviruses may have undergone multiple introductions with frequent host switching. We discuss potential implications for the evolution of vectoring within the family Flaviviridae. We also provide preliminary evidence for potential recombination events in the history of cell fusing agent virus. Finally, we consider priorities and guidelines for future research on ‘insect-specific’ flaviviruses, including the vast potential that exists for the study of biodiversity within a range of potential hosts and vectors, and its effect on the emergence and maintenance of the flaviviruses

    The Distances of the Magellanic Clouds

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    The present status of our knowledge of the distances to the Magellanic Clouds is evaluated from a post-Hipparcos perspective. After a brief summary of the effects of structure, reddening, age and metallicity, the primary distance indicators for the Large Magellanic Cloud are reviewed: The SN 1987A ring, Cepheids, RR Lyraes, Mira variables, and Eclipsing Binaries. Distances derived via these methods are weighted and combined to produce final "best" estimates for the Magellanic Clouds distance moduli.Comment: Invited review article to appear in ``Post Hipparcos Cosmic Candles'', F. Caputo & A. Heck (Eds.), Kluwer Academic Publ., Dordrecht, in pres

    Diversity and Abundance of Arbuscular–Mycorrhizal Fungus Spores in Tropical Forest and Pasture 1

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    As Neotropical forests are increasingly converted to agriculture (especially pastures), little is known about the impacts on microbial biodiversity. To assess such impacts, I compared spore abundance and diversity of arbuscular mycorrhizal fungi (AMF) in soils from lowland evergreen forests and pastures in Nicaragua and Costa Rica. Species composition, dominance–diversity curves, and Simpson's diversity indices were similar for both forest and pasture soils. Of 28 distinct fungus morphospecies, 11 produced more spores in pasture, while only 1 produced more spores in forest. According to species–accumulation curves, local AMF species richness did not significantly decline following conversion of forest to pasture. Because pastures contained a surprising abundance and diversity of AMF spores compared to native forest, a lack of mycorrhizal fungi is unlikely to limit plant succession, restoration, or reforestation in the pastures studied. At the regional scale, however, species-accumulation curves showed significantly greater gamma (G) diversity of spores in forest. In addition to these trends in diversity, species that sporulated more in pasture tended to have small spores, while the one species that sporulated more in forest had the largest spores. Similarly, only largespored fungi (>300 Μm) showed any seasonal variation in spore abundance, being more common in the wet season.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/71602/1/j.1744-7429.2000.tb00522.x.pd

    Analysis of Sulfur Poisoning on a PEM Fuel Cell Electrode

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    The extent of irreversible deactivation of Pt towards hydrogen oxidation reaction (HOR) due to sulfur adsorption and subsequent electrochemical oxidation is quantified in a functional PEM fuel cell. At 70 {\deg}C, sequential hydrogen sulfide (H2S) exposure and electrochemical oxidation experiments indicate that as much as 6% of total Pt sites are deactivated per monolayer sulfur adsorption at open circuit potential of a PEM fuel cell followed by its removal. The extent of such deactivation is much higher when the electrode is exposed to H2S when the fuel cell is operating at a finite load, and is dependent on the local overpotential and the duration of exposure. Regardless of this deactivation, the H2/O2 polarization curves obtained on post-recovery electrodes do not show performance losses suggesting that such performance curves alone cannot be used to assess the extent of recovery due to sulfur poisoning. A concise mechanism for the adsorption and electro-oxidation of H2S on Pt anode is presented. H2S dissociatively adsorbs onto Pt as two different sulfur species and at intermediate oxidation potentials, undergoes electro-oxidation to sulfur and then to sulfur dioxide (SO2). This mechanism is validated by charge balances between hydrogen desorption and sulfur electro-oxidation on Pt. The ignition potential for sulfur oxidation decreases with increase in temperature, which coupled with faster electro-oxidation kinetics result in the easier removal of adsorbed sulfur at higher temperatures. Furthermore, the adsorption potential is found to influence sulfur coverage of an electrode exposed to H2S. As an implication, the local potential of a PEM fuel cell anode exposed to H2S contaminated fuel should be kept below the equilibrium potential for sulfur oxidation to prevent irreversible loss of Pt sites.Comment: 28 pages, 15 figure

    Optimization of Cell Morphology Measurement via Single-Molecule Tracking PALM

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    In neurons, the shape of dendritic spines relates to synapse function, which is rapidly altered during experience-dependent neural plasticity. The small size of spines makes detailed measurement of their morphology in living cells best suited to super-resolution imaging techniques. The distribution of molecular positions mapped via live-cell Photoactivated Localization Microscopy (PALM) is a powerful approach, but molecular motion complicates this analysis and can degrade overall resolution of the morphological reconstruction. Nevertheless, the motion is of additional interest because tracking single molecules provides diffusion coefficients, bound fraction, and other key functional parameters. We used Monte Carlo simulations to examine features of single-molecule tracking of practical utility for the simultaneous determination of cell morphology. We find that the accuracy of determining both distance and angle of motion depend heavily on the precision with which molecules are localized. Strikingly, diffusion within a bounded region resulted in an inward bias of localizations away from the edges, inaccurately reflecting the region structure. This inward bias additionally resulted in a counterintuitive reduction of measured diffusion coefficient for fast-moving molecules; this effect was accentuated by the long camera exposures typically used in single-molecule tracking. Thus, accurate determination of cell morphology from rapidly moving molecules requires the use of short integration times within each image to minimize artifacts caused by motion during image acquisition. Sequential imaging of neuronal processes using excitation pulses of either 2 ms or 10 ms within imaging frames confirmed this: processes appeared erroneously thinner when imaged using the longer excitation pulse. Using this pulsed excitation approach, we show that PALM can be used to image spine and spine neck morphology in living neurons. These results clarify a number of issues involved in interpretation of single-molecule data in living cells and provide a method to minimize artifacts in single-molecule experiments
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