193 research outputs found

    Cell Envelope Associations Of Bacterial Flagella

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    Spatial and temporal changes in seasonal range attributes in a declining barren-ground caribou herd

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    From 1996 to 2015 the Bathurst caribou herd has declined from approximately 349,000 to 20,000 animals. Aboriginal traditional knowledge (TK) has recently observed the later arrival of the herd below the treeline, an attribute of the autumn range. Science also predicts that seasonal range attributes (e.g., area, location) likely vary with population size, and perhaps climate. We used Aboriginal TK and science to identify several seasonal range attributes that were ex­amined for changes through time (decreasing population abundance). Attributes of seasonal ranges for female Bathurst caribou were calculated using satellite radio-collar data from January 1996 through October 2013. Climate data from CircumArctic Rangifer Monitoring and Assessment Network were analyzed for trends from 1979 to 2009. Analyses showed a significant decrease in the area of post-calving and autumn ranges, but no changes in winter and spring ranges. Results supported Aboriginal TK that female caribou have shifted the autumn range farther from the treeline and moved into the forest later in the year. Analysis of climate variables found no trends at the spatio-temporal scale of the post-calving to autumn ranges. Working hypotheses to explain these patterns, which are not mutually exclusive, include reduced predation risk, increased use of core areas at lower population density, and greater utilization of areas of taiga where arboreal and ground lichen availability and accessibility are relatively higher than in the forest. This analysis demonstrates how including Aboriginal TK can lead to stronger connections and results, with potential to provide new and different insights for further investigations

    Combining physiological, environmental and locational sensors for citizen-oriented health applications

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    This work investigates the potential of combining the outputs of multiple low-cost sensor technologies for the direct measurement of spatio-temporal variations in phenomena that exist at the interface between our bodies and the environment. The example used herein is the measurement of personal exposure to traffic pollution, which may be considered as a function of the concentration of pollutants in the air and the frequency and volume of that air which enters our lungs. The sensor-based approach described in this paper removes the ‘traditional’ requirements either to model or interpolate pollution levels or to make assumptions about the physiology of an individual. Rather, a wholly empirical analysis into pollution exposure is possible, based upon high-resolution spatio-temporal data drawn from sensors for NO2, nasal airflow and location (GPS). Data are collected via a custom smartphone application and mapped to give an unprecedented insight into exposure to traffic pollution at the individual level. Whilst the quality of data from low-cost miniaturised sensors is not suitable for all applications, there certainly are many applications for which these data would be well suited, particularly those in the field of citizen science. This paper demonstrates both the potential and limitations of sensor-based approaches and discusses the wider relevance of these technologies for the advancement of citizen science

    A Socio-Spatial Approach to Enable Inclusive Well-Being in Cities: A Case Study of Birmingham, UK

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    This article examines density and deprivation, the two important parameters that define health and well-being in cities. Discussions are drawn from a case study conducted in Birmingham in four neighborhoods characterized by their different population density and deprivation levels. Data were collected through questionnaires developed from a set of subjective well-being measures and built environment audits, based on the Irvine Minnesota Inventory that evaluates the quality of streets and walkability in neighborhoods. The inferences from the study support the need for linking health, planning, policy and design research and decision-making to the socio-spatial practices of people, impacting well-being at the everyday level. The findings provide a holistic approach health and well-being research and suggests a conceptual framework for inclusive well-being in cities, which signifies the role of social and spatial parameters in determining peoples’ health and well-being. The study also highlights the lack of interdisciplinary research in understanding the association between well-being and social and behavioral practices in diverse communities

    [Re]-imagining vision and values:design as a driver for value creation in the Internet of Things

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    In digital age in which a myriad of products is becoming connected through the Internet of Things (IoT), new business opportunities and challenges arise for organisations regardless of size and product/service offerings. The traits of digital technology and digital economy results in not only profound changes in the ways organisations create value and develop their IoT products and services, but also emergent and dynamic business models. It also opens new opportunities in terms of how organisations increase turnover, thus conventional theories and practices of how value is created in the design and development process is constantly being challenged. Although, academics and practitioners often critically debate these emergent opportunities and challenges to the adoption of the ‘Internet of Things’, there is a paucity of established academic theories and industry practices to support and re-think traditional processes of product design and development to meet current needs and potential commercial opportunities in the era of IoT. This paper will offer a critical examination on how design processes have evolved with regard to the differences in value creation between goods-dominant logic and the service-dominant logic in order to identify an emergent design process feasible for IoT product and service development. In addition, factors that affect value creation, and design and development process in the IoT are reviewed. It concludes by offering key insights and observations as to where design can contribute to value creation in the internet of things

    FLYWCH1, a Multi-Functional Zinc Finger Protein Contributes to the DNA Repair Pathway

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    Over recent years, several Cys2-His2 (C2H2) domain-containing proteins have emerged as critical players in repairing DNA-double strand breaks. Human FLYWCH1 is a newly characterised nuclear transcription factor with (C2H2)-type zinc-finger DNA-binding domains. Yet, our knowledge about FLYWCH1 is still in its infancy. This study explores the expression, role and regulation of FLYWCH1 in the context of DNA damage and repair. We provide evidence suggesting a potential contribution of FLYWCH1 in facilitating the recruitment of DNA-damage response proteins (DDRPs). We found that FLYWCH1 colocalises with γH2AX in normal fibroblasts and colorectal cancer (CRC) cell lines. Importantly, our results showed that enforced expression of FLYWCH1 induces the expression of γH2AX, ATM and P53 proteins. Using an ATM-knockout (ATMKO) model, we indicated that FLYWCH1 mediates the phosphorylation of H2AX (Ser139) independently to ATM expression. On the other hand, the induction of DNA damage using UV-light induces the endogenous expression of FLYWCH1. Conversely, cisplatin treatment reduces the endogenous level of FLYWCH1 in CRC cell lines. Together, our findings uncover a novel FLYWCH1/H2AX phosphorylation axis in steady-state conditions and during the induction of the DNA-damage response (DDR). Although the role of FLYWCH1 within the DDR machinery remains largely uncharacterised and poorly understood, we here report for the first-time findings that implicate FLYWCH1 as a potential participant in the DNA damage response signaling pathways

    Coordinated Rearrangements between Cytoplasmic and Periplasmic Domains of the Membrane Protein Complex ExbB-ExbD of Escherichia coli

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    SummaryGram-negative bacteria rely on the ExbB-ExbD-TonB system for the import of essential nutrients. Despite decades of research, the stoichiometry, subunit organization, and mechanism of action of the membrane proteins of the Ton system remain unclear. We copurified ExbB with ExbD as an ∼240 kDa protein-detergent complex, measured by light scattering and by native gels. Quantitative Coomassie staining revealed a stoichiometry of ExbB4-ExbD2. Negative stain electron microscopy and 2D analysis showed particles of ∼10 nm diameter in multiple structural states. Nanogold labeling identified the position of the ExbD periplasmic domain. Random conical tilt was used to reconstruct the particles in three structural states followed by sorting of the single particles and refinement of each state. The different states are interpreted by coordinated structural rearrangements between the cytoplasmic domain and the periplasmic domain, concordant with in vivo predictions

    Siderophore Transport through Escherichia coli Outer Membrane Receptor FhuA with Disulfide-tethered Cork and Barrel Domains

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    The hydroxamate siderophore receptor FhuA is a TonB-dependent outer membrane protein of Escherichia coli composed of a C-terminal 22-stranded -barrel occluded by an N-terminal globular cork domain. During siderophore transport into the periplasm, the FhuA cork domain has been proposed to undergo conformational changes that allow transport through the barrel lumen; alternatively, the cork may be completely displaced from the barrel. To probe such changes, site-directed cysteine mutants in the cork domain (L109C and Q112C) and in the barrel domain (S356C and M383C) were created within the putative siderophore transport pathway. Molecular modeling predicted that the double cysteine mutants L109C/S356C and Q112C/M383C would form disulfide bonds, thereby tethering the cork and barrel domains. The double cysteine FhuA mutants were denatured under nonreducing conditions and fluorescently labeled with thiol-specific Oregon Green maleimide. Subsequent SDS-PAGE analysis revealed two distinct species: FhuA containing a disulfide bond and FhuA with free sulfhydryl groups. To address the role of the putative siderophore transport pathway and to evaluate possible rearrangements of the cork domain during ferricrocin transport, disulfide bond formation was enhanced by an oxidative catalyst. Cells containing double cysteine FhuA mutants that were subjected to oxidation during ferricrocin transport exhibited disulfide bond formation to near completion. After disulfide tethering of the cork to the barrel, ferricrocin transport was equivalent to transport by untreated cells. These results demonstrate that blocking the putative siderophore transport pathway does not abrogate ferricrocin uptake. We propose that, during siderophore transport through FhuA, the cork domain remains within the barrel rather than being displaced

    Enhanced Binding of TonB to a Ligand-loaded Outer Membrane Receptor: ROLE OF THE OLIGOMERIC STATE OF TonB IN FORMATION OF A FUNCTIONAL FhuA·TonB COMPLEX

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    The ferric hydroxymate uptake (FhuA) receptor from Escherichia coli facilitates transport of siderophores ferricrocin and ferrichrome and siderophore-antibiotic conjugates such as albomycin and rifamycin CGP 4832. FhuA is also the receptor for phages T5, T1, 80, UC-1, for colicin M and for the antimicrobial peptide microcin MccJ21. Energy for transport is provided by the cytoplasmic membrane complex TonB·ExbB·ExbD, which uses the proton motive force of the cytoplasmic membrane to transduce energy to the outer membrane. To accomplish energy transfer, TonB contacts outer membrane receptors. However, the stoichiometry of TonB· receptor complexes and their sites of interaction remain uncertain. In this study, analyses of FhuA interactions with two recombinant TonB proteins by analytical ultracentrifugation revealed that TonB forms a 2:1 complex with FhuA. The presence of the FhuA-specific ligand ferricrocin enhanced the amounts of complex but is not essential for its formation. Surface plasmon resonance experiments demonstrated that FhuA·TonB interactions are multiple and have apparent affinities in the nanomolar range. TonB also possesses two distinct binding regions: one in the C terminus of the protein, for which binding to FhuA is ferricrocin-independent, and a higher affinity region outside the C terminus, for which ferricrocin enhances interactions with FhuA. Together these experiments establish that FhuA·TonB interactions are more intricate than originally predicted, that the TonB·FhuA stoichiometry is 2:1, and that ferricrocin modulates binding of FhuA to TonB at regions outside the C-terminal domain of TonB

    Microarray-based comparative genomic profiling of reference strains and selected Canadian field isolates of Actinobacillus pleuropneumoniae

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    <p>Abstract</p> <p>Background</p> <p><it>Actinobacillus pleuropneumoniae</it>, the causative agent of porcine pleuropneumonia, is a highly contagious respiratory pathogen that causes severe losses to the swine industry worldwide. Current commercially-available vaccines are of limited value because they do not induce cross-serovar immunity and do not prevent development of the carrier state. Microarray-based comparative genomic hybridizations (M-CGH) were used to estimate whole genomic diversity of representative <it>Actinobacillus pleuropneumoniae </it>strains. Our goal was to identify conserved genes, especially those predicted to encode outer membrane proteins and lipoproteins because of their potential for the development of more effective vaccines.</p> <p>Results</p> <p>Using hierarchical clustering, our M-CGH results showed that the majority of the genes in the genome of the serovar 5 <it>A. pleuropneumoniae </it>L20 strain were conserved in the reference strains of all 15 serovars and in representative field isolates. Fifty-eight conserved genes predicted to encode for outer membrane proteins or lipoproteins were identified. As well, there were several clusters of diverged or absent genes including those associated with capsule biosynthesis, toxin production as well as genes typically associated with mobile elements.</p> <p>Conclusion</p> <p>Although <it>A. pleuropneumoniae </it>strains are essentially clonal, M-CGH analysis of the reference strains of the fifteen serovars and representative field isolates revealed several classes of genes that were divergent or absent. Not surprisingly, these included genes associated with capsule biosynthesis as the capsule is associated with sero-specificity. Several of the conserved genes were identified as candidates for vaccine development, and we conclude that M-CGH is a valuable tool for reverse vaccinology.</p
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