Relating microstructure to shear strength of thin calcium phosphate coatings

The changes in shear strength and crystallinity along with microscopy of the thin calcium phosphate coating surface and sample fracture surfaces were analyzed. Samples of stainless steel were made the cathode in the reaction while platinum was made the anode. Coated samples and fractured surfaces from lapshear tests were carbon coated and viewed in a Jeol 840F FEG SEM at accelerating shear strength. It was found that increase in maturation time led to an increase in coating thickness and a reduction in surface porosity

Non-competitive cyclic peptides for targeting enzyme-substrate complexes

Affinity reagents are of central importance for selectively identifying proteins and investigating their interactions. We report on the development and use of cyclic peptides, identified by mRNA display-based RaPID methodology, that are selective for, and tight binders of, the human hypoxia inducible factor prolyl hydroxylases (PHDs) – enzymes crucial in hypoxia sensing. Biophysical analyses reveal the cyclic peptides to bind in a distinct site, away from the enzyme active site pocket, enabling conservation of substrate binding and catalysis. A biotinylated cyclic peptide captures not only the PHDs, but also their primary substrate hypoxia inducible factor HIF1-α. Our work highlights the potential for tight, non-active site binding cyclic peptides to act as promising affinity reagents for studying protein–protein interaction

Non-competitive cyclic peptides for targeting enzyme-substrate complexes

Affinity reagents are of central importance for selectively identifying proteins and investigating their interactions. We report on the development and use of cyclic peptides, identified by mRNA display-based RaPID methodology, that are selective for, and tight binders of, the human hypoxia inducible factor prolyl hydroxylases (PHDs) – enzymes crucial in hypoxia sensing. Biophysical analyses reveal the cyclic peptides to bind in a distinct site, away from the enzyme active site pocket, enabling conservation of substrate binding and catalysis. A biotinylated cyclic peptide captures not only the PHDs, but also their primary substrate hypoxia inducible factor HIF1-α. Our work highlights the potential for tight, non-active site binding cyclic peptides to act as promising affinity reagents for studying protein–protein interaction

Beamline K11 DIAD:a new instrument for dual imaging and diffraction at Diamond Light Source

The Dual Imaging and Diffraction (DIAD) beamline at Diamond Light Source is a new dual-beam instrument for full-field imaging/tomography and powder diffraction. This instrument provides the user community with the capability to dynamically image 2D and 3D complex structures and perform phase identification and/or strain mapping using micro-diffraction. The aim is to enable in situ and in operando experiments that require spatially correlated results from both techniques, by providing measurements from the same specimen location quasi-simultaneously. Using an unusual optical layout, DIAD has two independent beams originating from one source that operate in the medium energy range (7–38 keV) and are combined at one sample position. Here, either radiography or tomography can be performed using monochromatic or pink beam, with a 1.4 mm × 1.2 mm field of view and a feature resolution of 1.2 µm. Micro-diffraction is possible with a variable beam size between 13 µm × 4 µm and 50 µm × 50 µm. One key functionality of the beamline is image-guided diffraction, a setup in which the micro-diffraction beam can be scanned over the complete area of the imaging field-of-view. This moving beam setup enables the collection of location-specific information about the phase composition and/or strains at any given position within the image/tomography field of view. The dual beam design allows fast switching between imaging and diffraction mode without the need of complicated and time-consuming mode switches. Real-time selection of areas of interest for diffraction measurements as well as the simultaneous collection of both imaging and diffraction data of (irreversible) in situ and in operando experiments are possible

5-Carboxy-8-hydroxyquinoline is a broad spectrum 2-oxoglutarate oxygenase inhibitor which causes iron translocation

2-Oxoglutarate and iron dependent oxygenases are therapeutic targets for human diseases. Using a representative 2OG oxygenase panel, we compare the inhibitory activities of 5-carboxy-8-hydroxyquinoline (IOX1) and 4-carboxy-8-hydroxyquinoline (4C8HQ) with that of two other commonly used 2OG oxygenase inhibitors, N-oxalylglycine (NOG) and 2,4-pyridinedicarboxylic acid (2,4-PDCA). The results reveal that IOX1 has a broad spectrum of activity, as demonstrated by the inhibition of transcription factor hydroxylases, representatives of all 2OG dependent histone demethylase subfamilies, nucleic acid demethylases and γ-butyrobetaine hydroxylase. Cellular assays show that, unlike NOG and 2,4-PDCA, IOX1 is active against both cytosolic and nuclear 2OG oxygenases without ester derivatisation. Unexpectedly, crystallographic studies on these oxygenases demonstrate that IOX1, but not 4C8HQ, can cause translocation of the active site metal, revealing a rare example of protein ligand-induced metal movement. © 2013 The Royal Society of Chemistry

Biomass and nutrients in tree root systems-sustainable harvesting of an intensively managed Pinus pinaster

International audienceTo develop sources of renewable energy and to reduce greenhouse gas emissions, increasing attention has been given to the extraction of forest biomass, especially in the form of harvest residues. However, increasing the removal of biomass, and hence nutrients, has raised concerns about the sustainability of site fertility and forest productivity. The environmental cost of harvesting belowground biomass is still not fully understood. The objectives of this study were to (i) estimate the stocks of belowground biomass that potentially can be collected; (ii) measure the nutrient (N, P, K, Ca, Mg) concentrations of the different root compartments (stumps, coarse and thin roots); and to (iii) quantify the biomass and nutrient exports under different scenarios, including harvests of above and belowground compartments. The study was carried out on Pinus pinaster stands located in southwestern France. Results showed that roots could be a significant fuelwood resource, particularly at forest clear cutting. Negative relationships between root diameter and root nutrient concentration were observed, independently of root function or tree age. Such relationships can be used to accurately simulate nutrient concentrations in roots as well as nutrient exports. Combining our original results on roots with previously published data on the aboveground compartments showed that nutrient losses were higher in canopy harvest scenarios than in root harvest scenarios. This was mainly due to high nutrient concentrations of needles. We concluded that stump and root harvest could be sustainable in our study context, conversely to foliage harvest. Because thin roots have higher nutrient concentrations than coarse roots and the proportion of thin roots increased with an increase in the distance from the tree, collecting roots only in the close vicinity of the stumps should limit nutrient exports (particularly N) without unnecessarily reducing fuelwood biomass