Conservation of structure and mechanism in primary and secondary transporters exemplified by SiaP, a sialic acid binding virulence factor from Haemophilus influenzae

Extracytoplasmic solute receptors (ESRs) are important components of solute uptake systems in bacteria, having been studied extensively as parts of ATP binding cassette transporters. Herein we report the first crystal structure of an ESR protein from a functionally characterized electrochemical ion gradient-dependent secondary transporter. This protein, SiaP, forms part of a tripartite ATP-independent periplasmic transporter specific for sialic acid in Haemophilus influenzae. Surprisingly, the structure reveals an overall topology similar to ATP binding cassette ESR proteins, which is not apparent from the sequence, demonstrating that primary and secondary transporters can share a common structural component. The structure of SiaP in the presence of the sialic acid analogue 2,3-didehydro-2-deoxyN-acetylneuraminic acid reveals the ligand bound in a deep cavity with its carboxylate group forming a salt bridge with a highly conserved Arg residue. Sialic acid binding, which obeys simple bimolecular association kinetics as determined by stopped-flow fluorescence spectroscopy, is accompanied by domain closure about a hinge region and the kinking of an alpha-helix hinge component. The structure provides insight into the evolution, mechanism, and substrate specificity of ESR-dependent secondary transporters that are widespread in prokaryotes

The role of post-Variscan extensional tectonics and mantle melting in the generation of the Lower Permian granites and the giant W-As-Sn-Cu-Zn-Pb orefield of SW England

Copyright © 2003 Maney Publishin

Structure of purine nucleoside phosphorylase (DeoD) from Bacillus anthracis

Protein structures from the causative agent of anthrax (Bacillus anthracis) are being determined as part of a structural genomics programme. Amongst initial candidates for crystallographic analysis are enzymes involved in nucleotide biosynthesis, since these are recognized as potential targets in antibacterial therapy. Purine nucleoside phosphorylase is a key enzyme in the purine-salvage pathway. The crystal structure of purine nucleoside phosphorylase (DeoD) from B. anthracis has been solved by molecular replacement at 2.24 Å resolution and refined to an R factor of 18.4%. This is the first report of a DeoD structure from a Gram-positive bacterium

Psychological interventions for individuals with a diagnosis of borderline personality disorder in forensic settings:A systematic review

Borderline Personality Disorder (BPD) is a common diagnosis in forensic settings. Associated difficulties include impulsivity and emotional dysregulation, which can create a vulnerability to impulsive acts. A BPD diagnosis is also associated with significant distress and poor physical health, making it important to understand how to tailor interventions in forensic settings. This paper reviews evidence for the effectiveness of psychological interventions designed for individuals with a diagnosis of BPD when implemented in forensic settings. A systematic search found 3485 papers, of which 13 met the inclusion criteria. The papers reported nine separate studies that implemented four distinct interventions, often adapted for particular forensic settings. Six of the nine studies included control groups. Improvements in overall ‘BPD symptomatology’ and specific ‘BPD symptoms’ were reported for all intervention types, although few differences in outcome between intervention and control groups were found. There were also reported improvements in ‘BPD-related behaviours’, but data on offending behaviour were absent. Heterogeneity in study quality and design makes it challenging to draw any firm conclusions about the effectiveness of any one form of intervention over another, nor about which intervention may best suit a particular setting. Research of a higher quality is needed to answer these questions.</p

Gene-flow between populations of cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) is highly variable between years

Both large and small scale migrations of Helicoverpa armigera Hübner in Australia were investigated using AMOVA analysis and genetic assignment tests. Five microsatellite loci were screened across 3142 individuals from 16 localities in eight major cotton and grain growing regions within Australia, over a 38-month period (November 1999 to January 2003). From November 1999 to March 2001 relatively low levels of migration were characterized between growing regions. Substantially higher than average gene-flow rates and limited differentiation between cropping regions characterized the period from April 2001 to March 2002. A reduced migration rate in the year from April 2002 to March 2003 resulted in significant genetic structuring between cropping regions. This differentiation was established within two or three generations. Genetic drift alone is unlikely to drive genetic differentiation over such a small number of generations, unless it is accompanied by extreme bottlenecks and/or selection. Helicoverpa armigera in Australia demonstrated isolation by distance, so immigration into cropping regions is more likely to come from nearby regions than from afar. This effect was most pronounced in years with limited migration. However, there is evidence of long distance dispersal events in periods of high migration (April 2001–March 2002). The implications of highly variable migration patterns for resistance management are considered.K.D. Scott, K.S. Wilkinson, N. Lawrence, C.L. Lange, L.J. Scott, M.A. Merritt, A.J. Lowe and G.C Graha

Measurement and modelling of moisture—electrical resistivity relationship of fine-grained unsaturated soils and electrical anisotropy

A methodology for developing resistivity-moisture content relationships of materials associated with a clayey landslide is presented. Key elements of the methodology include sample selection and preparation, laboratory measurement of resistivity with changing moisture content, and the derivation of models describing the relationship between resistivity and moisture content. Laboratory resistivity measurements show that the techniques utilised (samples and square array) have considerable potential as a means of electropetrophysical calibration of engineering soils and weak rock. Experimental electrical resistivity results show a hierarchy of values dependent on sample lithology, with silty clay exhibiting the lowest resistivities, followed by siltstones and sands, which return the highest resistivities. In addition, finer grained samples show a greater degree of anisotropy between measurement orientations than coarser grained samples. However, suitability of results in light of issues such as sample cracking and electrical conduction must be identified and accounted for if the results are to be accurately up-scaled to inverted model resistivity results. The existence of directional anisotropy makes model calibration curve selection more difficult due to variability in the range of measured laboratory resistances. The use of larger measurement array size means that experimental data will be more representative of bulk lithological properties. In addition, use of electrodes with a relatively high surface area (wide diameter) help maintain low contact resistances and repeat measurement error, relative to narrow electrodes. Variation exists between the fit of experimental data and petrophysical models. Model fit is best for clay-dominated samples but fits less well for sand-dominated samples. Waxman–Smits equation is appropriately applied in this investigation as all samples have considerable clay mineral content, as is shown in non-negligible CEC results. The incorporation of pressure plate suction measurements on samples, allows suction dissipation to be quantified and evaluated alongside moisture content and electrical resistivity

3D ground model development for an active landslide in Lias mudrocks using geophysical, remote sensing and geotechnical methods

A ground model of an active and complex landslide system in instability prone Lias mudrocks of North Yorkshire, UK is developed through an integrated approach, utilising geophysical, geotechnical and remote sensing investigative methods. Surface geomorphology is mapped and interpreted using immersive 3D visualisation software to interpret airborne light detection and ranging data and aerial photographs. Subsurface structure is determined by core logging and 3D electrical resistivity tomography (ERT), which is deployed at two scales of resolution to provide a means of volumetrically characterising the subsurface expression of both site scale (tens of metres) geological structure, and finer (metre to sub-metre) scale earth-flow related structures. Petrophysical analysis of the borehole core samples is used to develop relationships between the electrical and physical formation properties, to aid calibration and interpretation of 3D ERT images. Results of the landslide investigation reveal that an integrated approach centred on volumetric geophysical imaging successfully achieves a detailed understanding of structure and lithology of a complex landslide system, which cannot be achieved through the use of remotely sensed data or discrete intrusive sampling alone

Cross-correlation based high resolution electron backscatter diffraction and electron channelling contrast imaging for strain mapping and dislocation distributions in InAlN thin films

We describe the development of cross-correlation based high resolution electron backscatter diffraction (HR-EBSD) and electron channelling contrast imaging (ECCI), in the scanning electron microscope (SEM), to quantitatively map the strain variation and lattice rotation and determine the density and identify dislocations in nitride semiconductor thin films. These techniques can provide quantitative, rapid, non-destructive analysis of the structural properties of materials with a spatial resolution of order of tens of nanometers. HR-EBSD has a sensitivity to changes of strain and rotation of the order of 10−4 and 0.01° respectively, while ECCI can be used to image single dislocations up to a dislocation density of order 1010 cm−2. In the present work, we report the application of the cross-correlation based HR-EBSD approach to determine the tilt, twist, elastic strain and the distribution and type of threading dislocations in InAlN/AlN/GaN high electron mobility transistor (HEMT) structures grown on two different substrates, namely SiC and sapphire. We describe our procedure to estimate the distribution of geometrically necessary dislocations (GND) based on Nye-Kroner analysis and compare them with the direct imaging of threading dislocations (TDs) by ECCI. Combining data from HR-EBSD and ECCI observations allowed the densities of pure edge, mixed and pure screw threading dislocations to be fully separated

Managing complex respiratory patients in the community: an evaluation of a pilot integrated respiratory care service

Introduction: In the UK, there is significant variation in respiratory care and outcomes. An integrated approach to the management of high-risk respiratory patients, incorporating specialist and primary care teams' expertise, is the basis for new integrated respiratory services designed to reduce this variation; however, this model needs evaluating.Methods: To evaluate an integrated service managing high-risk respiratory patients, electronic searches for patients with asthma and chronic obstructive pulmonary disease at risk of poor outcomes were performed in two general practitioner (GP) practices in a local service-development initiative. Patients were reviewed at joint clinics by primary and secondary care professionals. GPs also nominated patients for inclusion. Reviews were delivered to best standards of care including assessments of diagnosis, control, spirometry, self-management, education, medication, inhaler technique and smoking cessation support. Follow-up of routine clinical data collected at 9-months postclinic were compared with seasonally matched 9-months prior to integrated review.Results: 82 patients were identified, 55 attended. 13 (23.6%) had their primary diagnosis changed. In comparison with the seasonally adjusted baseline period, in the 9-month follow-up there was an increase in inhaled corticosteroid prescriptions of 23.3%, a reduction in short-acting β2-agonist prescription of 33.3%, a reduction in acute respiratory exacerbations of 67.6%, in unscheduled GP surgery visits of 53.3% and acute respiratory hospital admissions reduced from 3 to 0. Only 4 patients (7.3%) required referral to secondary care. Health economic evaluation showed respiratory-related costs per patient reduced by £231.86.Conclusions: Patients with respiratory disease in this region at risk of suboptimal outcomes identified proactively and managed by an integrated team improved outcomes without the need for hospital referral

Expression of soluble, active fragments of the morphogenetic protein SpoIIE from Bacillus subtilis using a library-based construct screen

SpoIIE is a dual function protein that plays important roles during sporulation in Bacillus subtilis. It binds to the tubulin-like protein FtsZ causing the cell division septum to relocate from mid-cell to the cell pole, and it dephosphorylates SpoIIAA phosphate leading to establishment of differential gene expression in the two compartments following the asymmetric septation. Its 872 residue polypeptide contains a multiple-membrane spanning sequence at the N-terminus and a PP2C phosphatase domain at the C-terminus. The central segment that binds to FtsZ is unlike domains of known structure or function, moreover the domain boundaries are poorly defined and this has hampered the expression of soluble fragments of SpoIIE at the levels required for structural studies. Here we have screened over 9000 genetic constructs of spoIIE using a random incremental truncation library approach, ESPRIT, to identify a number of soluble C-terminal fragments of SpoIIE that were aligned with the protein sequence to map putative domains and domain boundaries. The expression and purification of three fragments were optimised, yielding multimilligram quantities of the PP2C phosphatase domain, the putative FtsZ-binding domain and a larger fragment encompassing both these domains. All three fragments are monomeric and the PP2C domain-containing fragments have phosphatase activity