Potential Role for p53 in the Permissive Life Cycle of Human Cytomegalovirus

Infection of primary fibroblasts with human cytomegalovirus (HCMV) causes a rapid stabilization of the cellular protein p53. p53 is a major effector of the cellular damage response, and activation of this transcription factor can lead either to cell cycle arrest or to apoptosis. Viruses employ many tactics to avoid p53-mediated effects. One method HCMV uses to counteract p53 is sequestration into its viral replication centers. In order to determine whether or not HCMV benefits from this sequestration, we infected a p53 −/− fibroblast line. We find that although these cells are permissive for viral infection, several parameters are substantially altered compared to wild-type (wt) fibroblasts. p53 −/− cells show delayed and decreased accumulation of infectious viral particles compared to control fibroblasts, with the largest difference of 100-fold at 72 h post infection (p.i.) and peak titers decreased by approximately 10- to 20-fold at 144 h p.i. Viral DNA accumulation is also delayed and somewhat decreased in p53 −/− cells; however, on average, levels of DNA are not more than fivefold lower than wt at any time p.i. and thus cannot account entirely for the observed differences in titers. In addition, there are delays in the expression of several key viral proteins, including the early replication protein UL44 and some of the late structural proteins, pp28 (UL99) and MCP (UL86). UL44 localization also indicates delayed formation and maturation of the replication centers throughout the course of infection. Localization of the major tegument protein pp65 (UL83) is also altered in these p53 −/− cells. Partial reconstitution of the p53 −/− cells with a wt copy of p53 returns all parameters toward wt, while reconstitution with mutant p53 does not. Taken together, our data suggest that wt p53 enhances the ability of HCMV to replicate and produce high concentrations of infectious virions in permissive cells

Waste Isolation Safety Assessment Program scenario analysis methods for use in assessing the safety of the geologic isolation of nuclear waste.

The relative utility of the various safety analysis methods to scenario analysis for a repository system was evaluated by judging the degree to which certain criteria are satisfied by use of the method. Six safety analysis methods were reviewed in this report for possible use in scenario analysis of nuclear waste repositories: expert opinion, perspectives analysis, fault trees/event trees, Monte Carlo simulation, Markov chains, and classical systems analysis. Four criteria have been selected. The criteria suggest that the methods: (1) be quantitative and scientifically based; (2) model the potential disruptive events and processes, (3) model the system before and after failure (sufficiently detailed to provide for subsequent consequence analysis); and (4) be compatible with the level of available system knowledge and data. Expert opinion, fault trees/event trees, Monte Carlo simulation and classical systems analysis were judged to have the greatest potential appliation to the problem of scenario analysis. The methods were found to be constrained by limited data and by knowledge of the processes governing the system. It was determined that no single method is clearly superior to others when measured against all the criteria. Therefore, to get the best understanding of system behavior, a combination of the methods is recommended. Monte Carlo simulation was judged to be the most suitable matrix in which to incorporate a combination of methods

On the alarming state of freshwater biodiversity in Canada

Little is known about the current state of freshwater biodiversity in Canada, one of the countries with the greatest amount of surface waters in the world. To address this knowledge gap, we compiled a list of all available assessments of conservation status for freshwater species (over 3,000 taxa) and further evaluated the overall status of six distinct taxonomic groups, focusing on organisms reliant on freshwaters [i.e., aquatic plants, invertebrates (with a focus on freshwater mussels), fishes, herpetofauna (reptiles and amphibians), birds, and mammals]. Overall, 11.7% of all freshwater species of plants and animals assessed were found to be ‘at risk’ (i.e., listed as ‘Threatened’, ‘Endangered’, or ‘Extirpated’) and 17.9% identified as ‘Special Concern’. We found that 37.9% of species lacked sufficient data to enable their status to be assessed. Data gaps in Canada’s assessment of its freshwater species were most prevalent in invertebrates (excluding freshwater mussels). Given the alarming state of freshwater biodiversity in Canada, we conclude by providing recommendations that focus on evaluating temporal trends and informing conservation actions.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Evaluating the congruence between DNA ‐based and morphological taxonomic approaches in water and sediment trap samples: Analyses of a 36‐month time series from a temperate monomictic lake

International audiencePaleolimnological studies are central for identifying long-term changes, yet many studies rely on bioindicators that deposit detectable subfossils in sediments, such as diatoms and cladocerans. Emerging DNA-based approaches are expanding the taxonomic diversity that can be investigated. However, as sedimentary DNA-based approaches are expanding rapidly, calibration work is required to determine the advantages and limitations of these techniques. In this study, we assessed the congruence between morphological and DNA-based approaches applied to sediment trap samples for diatoms and crustaceans using both intracellular and extracellular DNA. We also evaluated which taxa are deposited in sediment traps from the water column to identify potential paleolimnological bioindicators of environmental variations. Based on 18S rRNA gene amplicons, we developed and analyzed a micro-eukaryotic, monthly time series that spanned 3 years and was comprised of paired water column and sediment trap samples from Cultus Lake, British Columbia, Canada. Comparisons of assemblages derived from our genetic and morphological analyses using RV coefficients revealed significant correlations for diatoms, but weaker correlations for crustaceans. Intracellular DNA reads correlated more strongly with diatom morphology, while extracellular DNA reads correlated more strongly with crustacean morphology. Additional analyses of amplicon sequence variants shared between water and sediment trap samples revealed a wide diversity of taxa to study in paleolimnology, including Ciliophora, Dinoflagellata, Chytridiomycota, Chrysophyceae, and Cryptophyceae. Partial RDAs identified significant environmental predictors of these shared assemblages. Overall, our study demonstrates the effectiveness of DNA-based approaches to track community dynamics from sediment samples, an essential step for successful paleolimnological studies