Structures of an apo and a binary complex of an evolved archeal b family DNA polymerase capable of synthesising highly cy-dye labelled DNA

Thermophilic DNA polymerases of the polB family are of great importance in biotechnological applications including high-fidelity PCR. Of particular interest is the relative promiscuity of engineered versions of the exo- form of polymerases from the Thermo- and Pyrococcales families towards non-canonical substrates, which enables key advances in Next-generation sequencing. Despite this there is a paucity of structural information to guide further engineering of this group of polymerases. Here we report two structures, of the apo form and of a binary complex of a previously described variant (E10) of Pyrococcus furiosus (Pfu) polymerase with an ability to fully replace dCTP with Cyanine dye-labeled dCTP (Cy3-dCTP or Cy5-dCTP) in PCR and synthesise highly fluorescent “CyDNA” densely decorated with cyanine dye heterocycles. The apo form of Pfu-E10 closely matches reported apo form structures of wild-type Pfu. In contrast, the binary complex (in the replicative state with a duplex DNA oligonucleotide) reveals a closing movement of the thumb domain, increasing the contact surface with the nascent DNA duplex strand. Modelling based on the binary complex suggests how bulky fluorophores may be accommodated during processive synthesis and has aided the identification of residues important for the synthesis of unnatural nucleic acid polymers

Public Participation Organizations and Open Policy:A Constitutional Moment for British Democracy?

This article builds on work in Science and Technology Studies and cognate disciplines concerning the institutionalization of public engagement and participation practices. It describes and analyses ethnographic qualitative research into one “organization of participation,” the UK government–funded Sciencewise program. Sciencewise’s interactions with broader political developments are explored, including the emergence of “open policy” as a key policy object in the UK context. The article considers what the new imaginary of openness means for institutionalized forms of public participation in science policymaking, asking whether this is illustrative of a “constitutional moment” in relations between society and science policymaking

Development of microstructure and crystallographic texture in a double-sided friction stir welded microalloyed steel

The evolution of microstructure and crystallographic texture has been investigated in double-sided friction stir welded microalloyed steel, using electron backscatter diffraction (EBSD). The microstructure analyses show that the centre of stirred zone reached a temperature between Ac1 – Ac3 during FSW, resulting in a dual phase austenitic/ ferritic microstructure. The temperatures in the thermo-mechanically affected zone and the overlapped area between the first and second weld pass did not exceed the Ac1. The shear generated by the rotation probe occurs in austenitic/ferritic phase field where the austenite portion of the microstructure is transformed to a bainitic ferrite, on cooling. Analysis of crystallographic textures with regard to shear flow lines generated by the probe tool, show the dominance of simple shear components across the whole weld. The austenite texture at Ac1 – Ac3 is dominated by the B{11 ̅2} and ¯B {1 ̅12 ̅ } simple shear texture components, where the bainite phase textures formed on cooling were inherited from the shear textures of the austenite phase with relatively strong variant selection. The ferrite portion of the stirred zone and the ferrites in the thermo-mechanically affected zones and the overlapped area underwent shear deformation with textures dominated by the D1{1 ̅1 ̅2} and D2{112 ̅ } simple shear texture components. The formation of ultra-fine equiaxed ferrite with submicron grain size has been observed in the overlapped area between the first and second weld pass. This is due to continuous dynamic strain-induced recrystallisation as a result of simultaneous severe shear deformation and drastic undercooling

Industrial constructions of publics and public knowledge: a qualitative investigation of practice in the UK chemicals industry

This is a post print version of the article. The official published version can be obtained from the link below - © 2007 by SAGE PublicationsWhile the rhetoric of public engagement is increasingly commonplace within industry, there has been little research that examines how lay knowledge is conceptualized and whether it is really used within companies. Using the chemicals sector as an example, this paper explores how companies conceive of publics and "public knowledge," and how this relates to modes of engagement/communication with them. Drawing on qualitative empirical research in four companies, we demonstrate that the public for industry are primarily conceived as "consumers" and "neighbours," having concerns that should be allayed rather than as groups with knowledge meriting engagement. We conclude by highlighting the dissonance between current advocacy of engagement and the discourses and practices prevalent within industry, and highlight the need for more realistic strategies for industry/public engagement.Funding was received from the ESRC Science in Society Programme

Using machining force feedback to quantify grain size in beta titanium

The fluctuating forces on the cutting tool generated during machining of β processed Ti-17 alloy are shown to contain sufficient information to enable measurement of β grain size to an equivalent accuracy of standard etching methods. Three orthogonal forces were gathered, cutting force tangential to the rotation, the force in the feed (radial) direction, and the normal force in the longitudinal axis. Each individual force produced a microstructure image with a high level of contrast but in some cases did not fully highlight all features as shown in the optical image of the equivalent area. By normalising and combining the three forces into a vector, followed by noise reduction, a high-resolution image with sufficient detail to undertake grain size measurements using the linear intercept was produced. The measured grain size differed by no more than 5% with respect the grain size measured in the etched micrograph. It is believed that the forces which have a higher proportion of elastic response in their total values, i.e., the feed and normal forces, produced the higher contrast images, indicating that elastic stresses produce the highest contrast between grains and plastic strains smear out the grain to grain variation

Towards an analytical framework of science communication models

This chapter reviews the discussion in science communication circles of models for public communication of science and technology (PCST). It questions the claim that there has been a large-scale shift from a ‘deficit model’ of communication to a ‘dialogue model’, and it demonstrates the survival of the deficit model along with the ambiguities of that model. Similar discussions in related fields of communication, including the critique of dialogue, are briefly sketched. Outlining the complex circumstances governing approaches to PCST, the author argues that communications models often perceived to be opposed can, in fact, coexist when the choices are made explicit. To aid this process, the author proposes an analytical framework of communication models based on deficit, dialogue and participation, including variations on each

Quantitative characterization of machining-induced white layers in Ti–6Al–4V

Machining-induced white layers can affect the functional performance of engineered components, due to the resulting mechanical and microstructural properties. Destructive inspection methods such as cross-sectional microscopy are typically used to identify white layers, however, these methods are inherently costly and time-consuming. It is, therefore, desirable to detect this anomalous surface feature using non-destructive methods which requires improved knowledge around the characteristics of white layers. The present paper reports on the characterization of white layers formed during machining of Ti–6Al–4V, to aid future development of a reliable non-destructive assessment method. The microstructure of the material in the white layer was found to have a basal α-hexagonal close packed texture and there was no evidence of an α→β phase transformation during white layer formation. The white layer has a highly refined grain structure with an increased nanohardness of up to 15% compared with the bulk material. It is proposed that white layers in Ti–6Al–4V are formed by continuous dynamic recrystallization driven by severe plastic deformation during machining. According to the measured micro-mechanical properties of the white layer, suitable non-destructive testing methods are suggested for the detection of this surface feature

The use of Fe-30% Ni and Fe-30% Ni-Nb alloys as model systems for studying the microstructural evolution during the hot deformation of austenite

The development of physically-based models of microstructural evolution during thermomechanical processing of metallic materials requires knowledge of the internal state variable data, such as microstructure, texture, and dislocation substructure characteristics, over a range of processing conditions. This is a particular problem for steels, where transformation of the austenite to a variety of transformation products eradicates the hot deformed microstructure. This article reports on a model Fe-30wt% Ni-based alloy, which retains a stable austenitic structure at room temperature, and has, therefore, been used to model the development of austenite microstructure during hot deformation of conventional low carbon-manganese steels. It also provides an excellent model alloy system for microalloy additions. Evolution of the microstructure and crystallographic texture was characterized in detail using optical microscopy, X-ray diffraction (XRD), SEM, EBSD, and TEM. The dislocation substructure has been quantified as a function of crystallographic texture component for a variety of deformation conditions for the Fe-30% Ni-based alloy. An extension to this study, as the use of a microalloyed Fe-30% Ni-Nb alloy in which the strain induced precipitation mechanism was studied directly. The work has shown that precipitation can occur at a much finer scale and higher number density than hitherto considered, but that pipe diffusion leads to rapid coarsening. The implications of this for model development are discussed

Tool for automatic macrozone characterization from EBSD data sets of titanium alloys

Microtexture heterogeneities are commonly found in titanium forgings because of the thermomechanical processing. Also known as macrozones, these regions can reach millimetres in length, with grains sharing a similar crystallographic orientation leading to less resistance to crack propagation. Since the link between macrozones and the reduction of cold‐dwell‐fatigue performance on rotative components in gas turbine engines was established, efforts have been put into macrozone definition and characterization. The electron backscatter diffraction (EBSD) technique, widely used for texture analysis, allows for a qualitative macrozone characterization; however, further processing is required to define the boundaries and disorientation spread of each macrozone. Current approaches often use c‐axis misorientation criteria, but this can sometimes lead to a large disorientation spread within a macrozone. This article describes the development and application of a computational tool implemented in MATLAB for automatic macrozone identification from EBSD data sets on the basis of a more conservative approach where both the c‐axis tilting and rotation are considered. The tool allows for detection of macrozones according to the disorientation angle and density‐fraction criteria. The clustering efficiency is validated by pole‐figure plots, and the effects of the key parameters defining the macrozone clustering (disorientation and fraction) are discussed. In addition, this tool was successfully applied to both fully equiaxed and bimodal microstructures of titanium forgings

Quark-gluon plasma phenomenology from anisotropic lattice QCD

The FASTSUM collaboration has been carrying out simulations of N_f=2+1 QCD at nonzero temperature in the fixed-scale approach using anisotropic lattices. Here we present the status of these studies, including recent results for electrical conductivity and charge diffusion, and heavy quarkonium (charm and beauty) physics.Comment: Talk given at Quark Confinement and the Hadron Spectrum (Confinement XI), 8-12 September, St. Petersburg, Russia. 8 pages, 7 figure