278 research outputs found

    Characterization of a Clp Protease Gene Regulator and the Reaeration Response in Mycobacterium tuberculosis

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    Mycobacterium tuberculosis (MTB) enters a non-replicating state when exposed to low oxygen tension, a condition the bacillus encounters in granulomas during infection. Determining how mycobacteria enter and maintain this state is a major focus of research. However, from a public health standpoint the importance of latent TB is its ability to reactivate. The mechanism by which mycobacteria return to a replicating state upon re-exposure to favorable conditions is not understood. In this study, we utilized reaeration from a defined hypoxia model to characterize the adaptive response of MTB following a return to favorable growth conditions. Global transcriptional analysis identified the ∼100 gene Reaeration Response, induced relative to both log-phase and hypoxic MTB. This response includes chaperones and proteases, as well as the transcription factor Rv2745c, which we characterize as a Clp protease gene regulator (ClgR) orthologue. During reaeration, genes repressed during hypoxia are also upregulated in a wave of transcription that includes genes crucial to transcription, translation and oxidative phosphorylation and culminates in bacterial replication. In sum, this study defines a new transcriptional response of MTB with potential relevance to disease, and implicates ClgR as a regulator involved in resumption of replication following hypoxia

    Soil surface temperatures reveal moderation of the urban heat island effect by trees and shrubs

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    Urban areas are major contributors to air pollution and climate change, causing impacts on human health that are amplified by the microclimatological effects of buildings and grey infrastructure through the urban heat island (UHI) effect. Urban greenspaces may be important in reducing surface temperature extremes, but their effects have not been investigated at a city-wide scale. Across a midsized UK city we buried temperature loggers at the surface of greenspace soils at 100 sites, stratified by proximity to city centre, vegetation cover and land-use. Mean daily soil surface temperature over 11 months increased by 0.6 °C over the 5 km from the city outskirts to the centre. Trees and shrubs in non-domestic greenspace reduced mean maximum daily soil surface temperatures in the summer by 5.7 °C compared to herbaceous vegetation, but tended to maintain slightly higher temperatures in winter. Trees in domestic gardens, which tend to be smaller, were less effective at reducing summer soil surface temperatures. Our findings reveal that the UHI effects soil temperatures at a city-wide scale, and that in their moderating urban soil surface temperature extremes, trees and shrubs may help to reduce the adverse impacts of urbanization on microclimate, soil processes and human health

    Comprehensive Functional Analysis of Mycobacterium tuberculosis Toxin-Antitoxin Systems: Implications for Pathogenesis, Stress Responses, and Evolution

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    Toxin-antitoxin (TA) systems, stress-responsive genetic elements ubiquitous in microbial genomes, are unusually abundant in the major human pathogen Mycobacterium tuberculosis. Why M. tuberculosis has so many TA systems and what role they play in the unique biology of the pathogen is unknown. To address these questions, we have taken a comprehensive approach to identify and functionally characterize all the TA systems encoded in the M. tuberculosis genome. Here we show that 88 putative TA system candidates are present in M. tuberculosis, considerably more than previously thought. Comparative genomic analysis revealed that the vast majority of these systems are conserved in the M. tuberculosis complex (MTBC), but largely absent from other mycobacteria, including close relatives of M. tuberculosis. We found that many of the M. tuberculosis TA systems are located within discernable genomic islands and were thus likely acquired recently via horizontal gene transfer. We discovered a novel TA system located in the core genome that is conserved across the genus, suggesting that it may fulfill a role common to all mycobacteria. By expressing each of the putative TA systems in M. smegmatis, we demonstrate that 30 encode a functional toxin and its cognate antitoxin. We show that the toxins of the largest family of TA systems, VapBC, act by inhibiting translation via mRNA cleavage. Expression profiling demonstrated that four systems are specifically activated during stresses likely encountered in vivo, including hypoxia and phagocytosis by macrophages. The expansion and maintenance of TA genes in the MTBC, coupled with the finding that a subset is transcriptionally activated by stress, suggests that TA systems are important for M. tuberculosis pathogenesis

    Transcriptional Reprogramming in Nonhuman Primate (Rhesus Macaque) Tuberculosis Granulomas

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    In response to Mtb infection, the host remodels the infection foci into a dense mass of cells known as the granuloma. The key objective of the granuloma is to contain the spread of Mtb into uninfected regions of the lung. However, it appears that Mtb has evolved mechanisms to resist killing in the granuloma. Profiling granuloma transcriptome will identify key immune signaling pathways active during TB infection. Such studies are not possible in human granulomas, due to various confounding factors. Nonhuman Primates (NHPs) infected with Mtb accurately reflect human TB in clinical and pathological contexts.We studied transcriptomics of granuloma lesions in the lungs of NHPs exhibiting active TB, during early and late stages of infection. Early TB lesions were characterized by a highly pro-inflammatory environment, expressing high levels of immune signaling pathways involving IFNgamma, TNFalpha, JAK, STAT and C-C/C-X-C chemokines. Late TB lesions, while morphologically similar to the early ones, exhibited an overwhelming silencing of the inflammatory response. Reprogramming of the granuloma transcriptome was highly significant. The expression of approximately two-thirds of all genes induced in early lesions was later repressed.The transcriptional characteristics of TB granulomas undergo drastic changes during the course of infection. The overwhelming reprogramming of the initial pro-inflammatory surge in late lesions may be a host strategy to limit immunopathology. We propose that these host profiles can predict changes in bacterial replication and physiology, perhaps serving as markers for latency and reactivation

    Influence of freeze-thaw events on carbon dioxide emission from soils at different moisture and land use

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    BACKGROUND: The repeated freeze-thaw events during cold season, freezing of soils in autumn and thawing in spring are typical for the tundra, boreal, and temperate soils. The thawing of soils during winter-summer transitions induces the release of decomposable organic carbon and acceleration of soil respiration. The winter-spring fluxes of CO(2 )from permanently and seasonally frozen soils are essential part of annual carbon budget varying from 5 to 50%. The mechanisms of the freeze-thaw activation are not absolutely clear and need clarifying. We investigated the effect of repeated freezing-thawing events on CO(2 )emission from intact arable and forest soils (Luvisols, loamy silt; Central Germany) at different moisture (65% and 100% of WHC). RESULTS: Due to the measurement of the CO(2 )flux in two hours intervals, the dynamics of CO(2 )emission during freezing-thawing events was described in a detailed way. At +10°C (initial level) in soils investigated, carbon dioxide emission varied between 7.4 to 43.8 mg C m(-2)h(-1 )depending on land use and moisture. CO(2 )flux from the totally frozen soil never reached zero and amounted to 5 to 20% of the initial level, indicating that microbial community was still active at -5°C. Significant burst of CO(2 )emission (1.2–1.7-fold increase depending on moisture and land use) was observed during thawing. There was close linear correlation between CO(2 )emission and soil temperature (R(2 )= 0.86–0.97, P < 0.001). CONCLUSION: Our investigations showed that soil moisture and land use governed the initial rate of soil respiration, duration of freezing and thawing of soil, pattern of CO(2 )dynamics and extra CO(2 )fluxes. As a rule, the emissions of CO(2 )induced by freezing-thawing were more significant in dry soils and during the first freezing-thawing cycle (FTC). The acceleration of CO(2 )emission was caused by different processes: the liberation of nutrients upon the soil freezing, biological activity occurring in unfrozen water films, and respiration of cold-adapted microflora

    Is the northern high-latitude land-based CO2 sink weakening?

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    Author Posting. © American Geophysical Union, 2011. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Global Biogeochemical Cycles 25 (2011): GB3018, doi:10.1029/2010GB003813.Studies indicate that, historically, terrestrial ecosystems of the northern high-latitude region may have been responsible for up to 60% of the global net land-based sink for atmospheric CO2. However, these regions have recently experienced remarkable modification of the major driving forces of the carbon cycle, including surface air temperature warming that is significantly greater than the global average and associated increases in the frequency and severity of disturbances. Whether Arctic tundra and boreal forest ecosystems will continue to sequester atmospheric CO2 in the face of these dramatic changes is unknown. Here we show the results of model simulations that estimate a 41 Tg C yr−1 sink in the boreal land regions from 1997 to 2006, which represents a 73% reduction in the strength of the sink estimated for previous decades in the late 20th century. Our results suggest that CO2 uptake by the region in previous decades may not be as strong as previously estimated. The recent decline in sink strength is the combined result of (1) weakening sinks due to warming-induced increases in soil organic matter decomposition and (2) strengthening sources from pyrogenic CO2 emissions as a result of the substantial area of boreal forest burned in wildfires across the region in recent years. Such changes create positive feedbacks to the climate system that accelerate global warming, putting further pressure on emission reductions to achieve atmospheric stabilization targets.This study was supported through grants provided as part of the Arctic System Science Program (NSF OPP‐ 0531047), the North American Carbon Program (NASA NNG05GD25G), and the Bonanza Creek Long‐Term Ecological Program (funded jointly by NSF grant DEB‐0423442 and USDA Forest Service, Pacific Northwest Research Station grant PNW01‐JV11261952‐231)
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