Identification and characterisation of enteroaggregative Escherichia coli subtypes associated with human disease

Enteroaggregative E. coli (EAEC) are a major cause of diarrhoea worldwide. Due to their heterogeneity and carriage in healthy individuals, identification of diagnostic virulence markers for pathogenic strains has been difficult. In this study, we have determined phenotypic and genotypic differences between EAEC strains of sequence types (STs) epidemiologically associated with asymptomatic carriage (ST31) and diarrhoeal disease (ST40). ST40 strains demonstrated significantly enhanced intestinal adherence, biofilm formation, and pro-inflammatory interleukin-8 secretion compared with ST31 isolates. This was independent of whether strains were derived from diarrhoea patients or healthy controls. Whole genome sequencing revealed differences in putative virulence genes encoding aggregative adherence fimbriae, E. coli common pilus, flagellin and EAEC heat-stable enterotoxin 1. Our results indicate that ST40 strains have a higher intrinsic potential of human pathogenesis due to a specific combination of virulence-related factors which promote host cell colonization and inflammation. These findings may contribute to the development of genotypic and/or phenotypic markers for EAEC strains of high virulence

A novel mouse model of in situ stenting

AIMS: Animal models of stenting are mostly limited to larger animals or involve substantial abdominal surgery in rodents. We aimed to develop a simple, direct model of murine stenting. METHODS AND RESULTS: We designed a miniature, self-expanding, nitinol wire coil stent that was pre-loaded into a metal stent sheath. This was advanced into the abdominal aorta of the mouse, via femoral access, and the stent deployed. In-stent restenosis was investigated at 1, 3, 7, and 28 days post-stenting. The model was validated by investigation of neointima formation in mice deficient in signalling via the interleukin-1 receptor (IL-1R1), compared with other injury models. Ninety-two per cent of mice undergoing the procedure were successfully stented. All stented vessels were patent. Inflammatory cells were seen in the adventitia and around the stent strut up to 3 days post-stenting. At 3 days, an early neointima was present, building to a mature neointima at 28 days. In mice lacking IL-1R1, the neointima was 64% smaller than that in wild-type controls at the 28-day timepoint, in agreement with other models. CONCLUSION: This is the first description of a successful model of murine in situ stenting, using a stent specifically tailored for use in small thin-walled arteries. The procedure can be undertaken by a single operator without the need for an advanced level of microsurgical skill and is reliable and reproducible. The utility of this model is demonstrated by a reduction in in-stent restenosis in IL-1R1-deficient mice

Drought: Understanding and reducing vulnerability through monitoring and early warning systems. Report of the DrIVER workshop, 17 March 2015.

As part of the Belmont Forum funded international DRIVER research project on linking indicators to impacts to improve drought monitoring and early warning systems (MEWs), a stakeholder workshop was held on 17th March 2015 in Wallingford, UK. The workshop was attended by representatives of various UK organisations with an interest in drought and MEWs and DRIVER researchers from the Centre for Ecology and Hydrology (UK), Open University (UK), University of Freiburg (Germany), National Drought Mitigation Center (USA) and CSIRO (Australia). The aims of the workshop were to introduce participants to the DRIVER project and recent RCUK drought projects; engage with stakeholders’ experiences, understandings and needs in relation to droughts; and identify needs for future MEWs. The design of the workshop was based on a commitment to social learning. It comprised a mix of presentations and interactive sessions using innovative techniques to develop collective insights, enabling participants to learn from others and contribute their experiences and ideas and concerns in relation to drought and MEWs. Themes emerging findings from the workshop were wide-ranging, but point to a range of issues, concerns and suggestions for improving drought management and MEWs centred on: recognition of different types of drought; uncertainties and risks relating to indicators; forecasting; impacts; politics of drought; public communication; role of stakeholders; and resilience strategies. The workshop suggests different ways of thinking and acting are required about drought and MEWs in particular. The discussions and presentations also suggest the complexity of droughts requires a more systemic understanding of drought policy, processes and practices in order to determine the role of MEWs and how these can be improved by linking indicators to impacts. The output of the workshop constitute a key source for informing and shaping ongoing DRIVER research activities and other events convened by RCUK drought research projects

A Freshwater Hydrobiid, cf. the Squat Duskysnail, Lyogyrus granum (Mollusca), Widespread in the Hampton Marsh, New Brunswick

In Canada, the Squat Duskysnail, Lyogyrus granum, is an apparently rare freshwater hydrobiid. We document putative L. granum to be widespread throughout the Hampton Marsh, New Brunswick, with densities sometimes exceeding 150 snails/m2 on substrates ranging from vegetated organic detritus over mud, to sparsely-vegetated cobble/sand lakeshore. That our identification of L. granum from Hampton Marsh remains tentative, in spite of large samples, emphasizes the need for detailed taxonomic study of any putative L. granum populations in Canada. Such study will be necessary before the true conservation status can be determined for this and any related taxa, as yet unrecognized in Canadian material

Water source dynamics influence macroinvertebrate communities across groundwater-fed streams in a glacierized catchment

Groundwater contributions to streamflow significantly influence the structure and function of riverine ecosystems, particularly in glacierized catchments where there are marked differences in water sources and subsurface flow paths. Here, we investigated spatial and temporal variation in relationships between water sources, flow paths, physical and chemical processes, organic matter, microbial biofilms, and macroinvertebrates across groundwater-fed streams in the glacierized Toklat River catchment of Denali National Park, Alaska. Streams fed predominantly by seepage from the valley sides were perennial, whereas streams sustained by glacial meltwater seepage were ephemeral. Differences in environmental conditions between flow regimes appeared to influence spatial and temporal patterns of organic matter, linking to macroinvertebrate community dynamics. Macroinvertebrates in perennial streams were supported by fine particulate organic matter from subsurface flow paths during summer, transitioning to a combination of fine particulate matter and leaf litter in autumn. In comparison, macroinvertebrates inhabiting ephemeral streams, which only flowed during autumn, were supported by leaf litter. Some macroinvertebrate taxa were unaffected by turnover in organic matter, indicating potential plasticity in organic matter resource use. Findings highlight the importance of considering spatial and temporal variation in groundwater-fed streams, considering that projected hydrological changes under a changing climate may have significant implications for these systems

Charged black holes: Wave equations for gravitational and electromagnetic perturbations

A pair of wave equations for the electromagnetic and gravitational perturbations of the charged Kerr black hole are derived. The perturbed Einstein-Maxwell equations in a new gauge are employed in the derivation. The wave equations refer to the perturbed Maxwell spinor $\Phi_0$ and to the shear $\sigma$ of a principal null direction of the Weyl curvature. The whole construction rests on the tripod of three distinct derivatives of the first curvature $\kappa$ of a principal null direction.Comment: 12 pages, to appear in Ap.

Demonstration of Universal Parametric Entangling Gates on a Multi-Qubit Lattice

We show that parametric coupling techniques can be used to generate selective entangling interactions for multi-qubit processors. By inducing coherent population exchange between adjacent qubits under frequency modulation, we implement a universal gateset for a linear array of four superconducting qubits. An average process fidelity of $\mathcal{F}=93\%$ is estimated for three two-qubit gates via quantum process tomography. We establish the suitability of these techniques for computation by preparing a four-qubit maximally entangled state and comparing the estimated state fidelity against the expected performance of the individual entangling gates. In addition, we prepare an eight-qubit register in all possible bitstring permutations and monitor the fidelity of a two-qubit gate across one pair of these qubits. Across all such permutations, an average fidelity of $\mathcal{F}=91.6\pm2.6\%$ is observed. These results thus offer a path to a scalable architecture with high selectivity and low crosstalk

RNA sequencing and proteomics approaches reveal novel deficits in the cortex of Mecp2-deficient mice, a model for Rett syndrome

BACKGROUND: Rett syndrome (RTT) is an X-linked neurodevelopmental disorder caused by mutations in the transcriptional regulator MeCP2. Much of our understanding of MeCP2 function is derived from transcriptomic studies with the general assumption that alterations in the transcriptome correlate with proteomic changes. Advances in mass spectrometry-based proteomics have facilitated recent interest in the examination of global protein expression to better understand the biology between transcriptional and translational regulation. METHODS: We therefore performed the first comprehensive transcriptome-proteome comparison in a RTT mouse model to elucidate RTT pathophysiology, identify potential therapeutic targets, and further our understanding of MeCP2 function. The whole cortex of wild-type and symptomatic RTT male littermates (n = 4 per genotype) were analyzed using RNA-sequencing and data-independent acquisition liquid chromatography tandem mass spectrometry. Ingenuity(R) Pathway Analysis was used to identify significantly affected pathways in the transcriptomic and proteomic data sets. RESULTS: Our results indicate these two omics data sets supplement one another. In addition to confirming previous works regarding mRNA expression in Mecp2-deficient animals, the current study identified hundreds of novel protein targets. Several selected protein targets were validated by Western blot analysis. These data indicate RNA metabolism, proteostasis, monoamine metabolism, and cholesterol synthesis are disrupted in the RTT proteome. Hits common to both data sets indicate disrupted cellular metabolism, calcium signaling, protein stability, DNA binding, and cytoskeletal cell structure. Finally, in addition to confirming disrupted pathways and identifying novel hits in neuronal structure and synaptic transmission, our data indicate aberrant myelination, inflammation, and vascular disruption. Intriguingly, there is no evidence of reactive gliosis, but instead, gene, protein, and pathway analysis suggest astrocytic maturation and morphological deficits. CONCLUSIONS: This comparative omics analysis supports previous works indicating widespread CNS dysfunction and may serve as a valuable resource for those interested in cellular dysfunction in RTT

Interleukin-1 regulates multiple atherogenic mechanisms in response to fat feeding

Background: Atherosclerosis is an inflammatory process that develops in individuals with known risk factors that include hypertension and hyperlipidaemia, influenced by diet. However, the interplay between diet, inflammatory mechanisms and vascular risk factors requires further research. We hypothesised that interleukin-1 (IL-1) signaling in the vessel wall would raise arterial blood pressure and promote atheroma. Methodology/Principal Findings: Apoe(-/-) and Apoe(-/-)/IL-1R1(-/-) mice were fed high fat diets for 8 weeks, and their blood pressure and atherosclerosis development measured. Apoe(-/-)/IL-R1(-/-) mice had a reduced blood pressure and significantly less atheroma than Apoe(-/-) mice. Selective loss of IL-1 signaling in the vessel wall by bone marrow transplantation also reduced plaque burden (p<0.05). This was associated with an IL-1 mediated loss of endothelium-dependent relaxation and an increase in vessel wall Nox 4. Inhibition of IL-1 restored endothelium-dependent vasodilatation and reduced levels of arterial oxidative stress. Conclusions/Significance: The IL-1 cytokine system links atherogenic environmental stimuli with arterial inflammation, oxidative stress, increased blood pressure and atherosclerosis. This is the first demonstration that inhibition of a single cytokine can block the rise in blood pressure in response to an environmental stimulus. IL-1 inhibition may have profound beneficial effects on atherogenesis in man

miR-486 is Essential for Muscle Function and Suppresses a Dystrophic Transcriptome

miR-486 is a muscle-enriched microRNA, or “myomiR,” that has reduced expression correlated with Duchenne muscular dystrophy (DMD). To determine the function of miR-486 in normal and dystrophin-deficient muscles and elucidate miR-486 target transcripts in skeletal muscle, we characterized mir-486 knockout mice (mir-486 KO). mir-486 KO mice developed disrupted myofiber architecture, decreased myofiber size, decreased locomotor activity, increased cardiac fibrosis, and metabolic defects were exacerbated in mir-486 KO:mdx5cv (DKO) mice. To identify direct in vivo miR-486 muscle target transcripts, we integrated RNA sequencing and chimeric miRNA eCLIP sequencing to identify key transcripts and pathways that contribute towards mir-486 KO and dystrophic disease pathologies. These targets included known and novel muscle metabolic and dystrophic structural remodeling factors of muscle and skeletal muscle contractile transcript targets. Together, our studies identify miR-486 as essential for normal muscle function, a driver of pathological remodeling in dystrophin-deficient muscle, a useful biomarker for dystrophic disease progression, and highlight the use of multiple omic platforms to identify in vivo microRNA target transcripts