Flexibility in the receptor-binding domain of the enzymatic colicin E9 is required for toxicity against Escherichia coli cells

The events that occur after the binding of the enzymatic E colicins to Escherichia coli BtuB receptors that lead to translocation of the cytotoxic domain into the periplasmic space and, ultimately, cell killing are poorly understood. It has been suggested that unfolding of the coiled-coil Mull receptor binding domain of the E colicins may be an essential step that leads to the loss of immunity protein from the colicin and immunity protein complex and then triggers the events of translocation. We introduced pairs of cysteine mutations into the receptor binding domain of colicin E9 (ColE9) that resulted in the formation of a disulfide bond located near the middle or the top of the R domain. After dithiothreitol reduction, the ColE9 protein with the mutations L359C and F412C (ColE9 L359C-F412C) and the ColE9 protein with the mutations Y324C and L447C (ColE9 Y324C-L447C) were slightly less active than equivalent concentrations of ColE9. On oxidation with diamide, no significant biological activity was seen with the ColE9 L359C-F412C and the ColE9 Y324C-L447C mutant proteins; however diamide had no effect on the activity of ColE9. The presence of a disulfide bond was confirmed in both of the oxidized, mutant proteins by matrix-assisted laser desorption ionization-time of flight mass spectrometry. The loss of biological activity of the disulfide-containing mutant proteins was not due to an indirect effect on the properties of the translocation or DNase domains of the mutant colicins. The data are consistent with a requirement for the flexibility of the coiled-coil R domain after binding to BtuB

Effects of diabetes family history and exercise training on the expression of adiponectin and leptin and their receptors

The daughters of patients with diabetes have reduced insulin sensitivity index (ISI) scores compared with women with no family history of diabetes, but their ISI increase more in response to exercise training(1). The present study aimed to determine whether differences between these groups in exercise-induced changes in circulating adiponectin and leptin concentrations and expression of their genes and receptors in subcutaneous adipose tissue (SAT), could explain differences in the exercise-induced changes in ISI between women with and without a family history of diabetes

PMN J1632-0033: A new gravitationally lensed quasar

We report the discovery of a gravitationally lensed quasar resulting from our survey for lenses in the southern sky. Radio images of PMN J1632-0033 with the VLA and ATCA exhibit two compact, flat-spectrum components with separation 1.47" and flux density ratio 13.2. Images with the HST reveal the optical counterparts to the radio components and also the lens galaxy. An optical spectrum of the bright component, obtained with the first Magellan telescope, reveals quasar emission lines at redshift 3.42. Deeper radio images with MERLIN and the VLBA reveal a faint third radio component located near the center of the lens galaxy, which is either a third image of the background quasar or faint emission from the lens galaxy.Comment: 21 pp., including 4 figures; thoroughly revised in light of new MERLIN/HST data; accepted for publication in A

The role of discharge variability in determining alluvial stratigraphy

We illustrate the potential for using physics-based modeling to link alluvial stratigraphy to large river morphology and dynamics. Model simulations, validated using ground penetrating radar data from the Río Paraná, Argentina, demonstrate a strong relationship between bar-scale set thickness and channel depth, which applies across a wide range of river patterns and bar types. We show that hydrologic regime, indexed by discharge variability and flood duration, exerts a first-order influence on morphodynamics and hence bar set thickness, and that planform morphology alone may be a misleading variable for interpreting deposits. Indeed, our results illustrate that rivers evolving under contrasting hydrologic regimes may have very similar morphology, yet be characterized by marked differences in stratigraphy. This realization represents an important limitation on the application of established theory that links river topography to alluvial deposits, and highlights the need to obtain field evidence of discharge variability when developing paleoenvironmental reconstructions. Model simulations demonstrate the potential for deriving such evidence using metrics of paleocurrent variance

Generation and Culture of Blood Outgrowth Endothelial Cells from Human Peripheral Blood.

Historically, the limited availability of primary endothelial cells from patients with vascular disorders has hindered the study of the molecular mechanisms underlying endothelial dysfunction in these individuals. However, the recent identification of blood outgrowth endothelial cells (BOECs), generated from circulating endothelial progenitors in adult peripheral blood, may circumvent this limitation by offering an endothelial-like, primary cell surrogate for patient-derived endothelial cells. Beyond their value to understanding endothelial biology and disease modeling, BOECs have potential uses in endothelial cell transplantation therapies. They are also a suitable cellular substrate for the generation of induced pluripotent stem cells (iPSCs) via nuclear reprogramming, offering a number of advantages over other cell types. We describe a method for the reliable generation, culture and characterization of BOECs from adult peripheral blood for use in these and other applications. This approach (i) allows for the generation of patient-specific endothelial cells from a relatively small volume of adult peripheral blood and (ii) produces cells that are highly similar to primary endothelial cells in morphology, cell signaling and gene expression

Influence of Dunes on Channel‐Scale Flow and Sediment Transport in a Sand Bed Braided River

This is the final version. Available on open access from the American Geophysical Union via the DOI in this recordData availability: Project data is stored in, and available from, the UK Centre for Ecology & Hydrology (http://eidc.ceh.ac.uk).Current understanding of the role that dunes play in controlling bar and channel-scale processes and river morphodynamics is incomplete. We present results from a combined numerical modeling and field monitoring study that isolates the impact of dunes on depth-averaged and near-bed flow structure, with implications for morphodynamic modeling. Numerical simulations were conducted using the three-dimensional Computational Fluid Dynamics code OpenFOAM to quantify the time-averaged flow structure within a 400 m x 100 m channel using DEMs for which: (i) dunes and bars were present within the model; and (ii) only bar43 scale topographic features were resolved (dunes were removed). Comparison of these two simulations shows that dunes enhance lateral flows and reduce velocities over bar tops by as much as 30%. Dunes influence the direction of modeled sediment transport at spatial scales larger than individual bedforms due to their effect on topographic steering of the near-bed flow structure. We show that dunes can amplify, dampen or even reverse the deflection of sediment down lateral bar slopes, and this is closely associated with 3D and obliquely orientated dunes. Sediment transport patterns calculated using theory implemented in depth-averaged morphodynamic models suggests that gravitational deflection of sediment is still controlled by bar-scale topography, even in the presence of dunes. However, improved parameterizations of flow and sediment transport in depth-averaged morphodynamic models are needed that account for the effects of both dune- and bar- scale morphology on near-bed flow and sediment transport.Natural Environment Research Council (NERC

Quantification of bedform dynamics and bedload sediment flux in sandy braided rivers from airborne and satellite imagery

Images from specially‐commissioned aeroplane sorties (manned aerial vehicle, MAV), repeat unmanned aerial vehicle (UAV) surveys, and Planet CubeSat satellites are used to quantify dune and bar dynamics in the sandy braided South Saskatchewan River, Canada. Structure‐from‐Motion (SfM) techniques and application of a depth‐brightness model are used to produce a series of Digital Surface Models (DSMs) at low and near‐bankfull flows. A number of technical and image processing challenges are described that arise from the application of SfM in dry and submerged environments. A model for best practice is presented and analysis suggests a depth‐brightness model approach can represent the different scales of bedforms present in sandy braided rivers with low‐turbidity and shallow (< 2 m deep) water. The aerial imagery is used to quantify the spatial distribution of unit bar and dune migration rate in an 18 km reach and three ~1 km long reaches respectively. Dune and unit bar migration rates are highly variable in response to local variations in planform morphology. Sediment transport rates for dunes and unit bars, obtained by integrating migration rates (from UAV) with the volume of sediment moved (from DSMs using MAV imagery) show near‐equivalence in sediment flux. Hence, reach‐based sediment transport rate estimates can be derived from unit bar data alone. Moreover, it is shown that reasonable estimates of sediment transport rate can be made using just unit bar migration rates as measured from 2D imagery, including from satellite images, so long as informed assumptions are made regarding average bar shape and height. With recent availability of frequent, repeat satellite imagery, and the ease of undertaking repeat MAV and UAV surveys, for the first time, it may be possible to provide global estimates of bedload sediment flux for large or inaccessible low‐turbidity rivers that currently have sparse information on bedload sediment transport rates

Entamoeba histolytica Phagocytosis of Human Erythrocytes Involves PATMK, a Member of the Transmembrane Kinase Family

Entamoeba histolytica is the cause of amebic colitis and liver abscess. This parasite induces apoptosis in host cells and utilizes exposed ligands such as phosphatidylserine to ingest the apoptotic corpses and invade deeper into host tissue. The purpose of this work was to identify amebic proteins involved in the recognition and ingestion of dead cells. A member of the transmembrane kinase family, phagosome-associated TMK96 (PATMK), was identified in a proteomic screen for early phagosomal proteins. Anti-peptide affinity-purified antibody produced against PATMK demonstrated that it was a type I integral membrane protein that was expressed on the trophozoite surface, and that co-localized with human erythrocytes at the site of contact. The role of PATMK in erythrophagocytosis in vitro was demonstrated by: (i) incubation of ameba with anti-PATMK antibodies; (ii) PATMK mRNA knock-down using a novel shRNA expression system; and (iii) expression of a carboxy-truncation of PATMK (PATMKΔ932). Expression of the carboxy-truncation of PATMKΔ932 also caused a specific reduction in the ability of E. histolytica to establish infection in the intestinal model of amebiasis, however these amebae retained the ability to cause hepatic abscesses when directly injected in the liver. In conclusion, PATMK was identified as a member of the TMK family that participates in erythrophagocytosis and is uniquely required for intestinal infection