610 research outputs found

    Numerical modelling of methyl iodide in the eastern tropical Atlantic

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    Methyl iodide (CH3I) is a volatile organic halogen compound that contributes significantly to the transport of iodine from the ocean to the atmosphere, where it plays an important role in tropospheric chemistry. CH3I is naturally produced and occurs in the global ocean. The processes involved in the formation of CH3I, however, are not fully understood. In fact, there is an ongoing debate whether production by phytoplankton or photochemical degradation of organic matter is the main source term. Here, both the biological and photochemical production mechanisms are considered in a biogeochemical module that is coupled to a one-dimensional water column model for the eastern tropical Atlantic. The model is able to reproduce observed subsurface maxima of CH3I concentrations. But, the dominating source process cannot be clearly identified as subsurface maxima can occur due to both direct biological and photochemical production. However, good agreement between the observed and simulated difference between surface and subsurface methyl iodide concentrations is achieved only when direct biological production is taken into account. Production rates for the biological CH3I source that were derived from published laboratory studies are shown to be inappropriate for explaining CH3I concentrations in the eastern tropical Atlantic

    Use of a dual reporter plasmid to demonstrate bactofection with an attenuated aroa- derivative of Pasteurella multocida b:2

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    A reporter plasmid pSRG has been developed which expresses red fluorescent protein (RFP) from a constitutive prokaryotic promoter within Pasteurella multocida B:2 and green fluorescent protein (GFP) from a constitutive eukaryotic promoter within mammalian cells. This construct has been used to determine the location and viability of the bacteria when moving from the extracellular environment into the intracellular compartment of mammalian cells. Invasion assays with embryonic bovine lung (EBL) cells and an attenuated AroA- derivative of Pasteurella multocida B:2 (strain JRMT12), harbouring the plasmid pSRG, showed that RFP-expressing bacteria could be detected intracellularly at 3 h post-invasion. At this stage, some EBL cells harbouring RFP-expressing bacteria were observed to express GFP simultaneously, indicating release of the plasmid into the intracellular environment. At 5 h post-invasion, more EBL cells were expressing GFP, while still harbouring RFP-expressing bacteria. Concurrently, some EBL cells were shown to express only GFP, indicating loss of viable bacteria within these cells. These experiments proved the functionality of the pSRG dual reporter system and the potential of P. multocida B:2 JRMT12 for bactofection and delivery of a DNA vaccine

    A simple viability analysis for unicellular cyanobacteria using a new autofluorescence assay, automated microscopy, and ImageJ

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    <p>Abstract</p> <p>Background</p> <p>Currently established methods to identify viable and non-viable cells of cyanobacteria are either time-consuming (eg. plating) or preparation-intensive (eg. fluorescent staining). In this paper we present a new and fast viability assay for unicellular cyanobacteria, which uses red chlorophyll fluorescence and an unspecific green autofluorescence for the differentiation of viable and non-viable cells without the need of sample preparation.</p> <p>Results</p> <p>The viability assay for unicellular cyanobacteria using red and green autofluorescence was established and validated for the model organism <it>Synechocystis </it>sp. PCC 6803. Both autofluorescence signals could be observed simultaneously allowing a direct classification of viable and non-viable cells. The results were confirmed by plating/colony count, absorption spectra and chlorophyll measurements. The use of an automated fluorescence microscope and a novel ImageJ based image analysis plugin allow a semi-automated analysis.</p> <p>Conclusions</p> <p>The new method simplifies the process of viability analysis and allows a quick and accurate analysis. Furthermore results indicate that a combination of the new assay with absorption spectra or chlorophyll concentration measurements allows the estimation of the vitality of cells.</p

    Combinatorial quorum sensing allows bacteria to resolve their social and physical environment

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    Quorum sensing (QS) is a cell–cell communication system that controls gene expression in many bacterial species, mediated by diffusible signal molecules. Although the intracellular regulatory mechanisms of QS are often well-understood, the functional roles of QS remain controversial. In particular, the use of multiple signals by many bacterial species poses a serious challenge to current functional theories. Here, we address this challenge by showing that bacteria can use multiple QS signals to infer both their social (density) and physical (mass-transfer) environment. Analytical and evolutionary simulation models show that the detection of, and response to, complex social/physical contrasts requires multiple signals with distinct half-lives and combinatorial (nonadditive) responses to signal concentrations. We test these predictions using the opportunistic pathogen Pseudomonas aeruginosa and demonstrate significant differences in signal decay betweeallyn its two primary signal molecules, as well as diverse combinatorial responses to dual-signal inputs. QS is associated with the control of secreted factors, and we show that secretome genes are preferentially controlled by synergistic “AND-gate” responses to multiple signal inputs, ensuring the effective expression of secreted factors in high-density and low mass-transfer environments. Our results support a new functional hypothesis for the use of multiple signals and, more generally, show that bacteria are capable of combinatorial communication
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