A simple device for multiplex ELISA made from melt-extruded plastic microcapillary film

We present a simple device for multiplex quantitative enzyme-linked immunosorbant assays (ELISA) made from a novel melt-extruded microcapillary film (MCF) containing a parallel array of 200µm capillaries along its length. To make ELISA devices different protein antigens or antibodies were immobilised inside individual microcapillaries within long reels of MCF extruded from fluorinated ethylene propylene (FEP). Short pieces of coated film were cut and interfaced with a pipette, allowing sequential uptake of samples and detection solutions into all capillaries from a reagent well. As well as being simple to produce, these FEP MCF devices have excellent light transmittance allowing direct optical interrogation of the capillaries for simple signal quantification. Proof of concept experiments demonstrate both quantitative and multiplex assays in FEP MCF devices using a standard direct ELISA procedure and read using a flatbed scanner. This new multiplex immunoassay platform should find applications ranging from lab detection to point-of-care and field diagnostics.

Historicizing counterurbanization: in-migration and the reconstruction of rural space in Berkshire (UK), 1901-51

This article uses census data for Berkshire to argue that large-scale counterurbanization began much earlier than is generally recognized in some parts of southern England. This was not just movement down the urban hierarchy, which as Pooley and Turnbull have demonstrated was a long-term feature of England’s settlement system, but in some cases at least amenity-driven migration to rural areas of the kind increasingly recognized as a core component of recent counterurbanization. Despite a reduction of acreage Berkshire’s rural districts saw a 54% rise in population between 1901 and 1951. The sub-regional pattern of growth is assessed to gauge whether ‘clean break’ migration to the remote west of the county (which remained effectively out of commuting range from London throughout the period) was taking place, or whether counterurbanization was confined to the more accessible eastern districts. However, whilst population did increase in both west and east, it was in fact the central districts that grew most impressively. Three case study parishes are investigated in order to gauge the nature and consequences of counterurbanization at a local level. Professional and business migrants figure prominently, seeking to preserve and promote the rural attributes of their new communities, without however cutting their ties to urban centres. It is argued that migration to rural Berkshire in the first half of the twentieth century cannot adequately be described either as a form of extended suburbanization or an anti-metropolitan ‘clean break’. Rather, early counterurbanization marks the first stage on the long road to a post-productivist countryside, in which countryside becomes detached from agriculture, there is socio-economic convergence between town and country, and the ‘rural’ increasingly becomes defined by landscape and identity rather than economic function.

Motivation in second language acquisition in Chinese research students in the UK

The present study sets out to examine motivation to learn English by Chinese research students in an informal setting. Data were collected, using semi-structured interviews, from four research students at two points in time during their first year in the UK. The main findings are: they believed that learning English was important; their main goal orientations were instrumental and extrinsic; they set learning goals and persisted to attain them; they valued their current learning environment in general and saw it as supportive of their goals; they held both positive and negative attitudes towards the British, which had differential effects on their motivation; their self-perceived support seemed to have a positive impact on their motivation and the development of self-confidence; they tended to attribute their success to stable causes such as the environment and failure to unstable but controllable causes such as effort. It is concluded that qualitative data of this kind may complement insights from quantitative research. Implications for target country institutions in the provision of support are discussed.

Transputer based real time robot control

A parallel processor architecture based on a communicating sequential processor chip, the transputer, is described. The architecture is easily linearly extensible to enable separate functions to be included in the controller. To demonstrate the power of the resulting controller some experimental results are presented comparing PID and full inverse dynamics on the first three joints of a Puma 560 robot. Also examined are some of the sample rate issues raised by the asynchronous updating of inertial parameters, and the need for full inverse dynamics at every sample interval is questioned.

“Ce n’est pas elle qu’on voit”: the photograph in the work of Annie Saumont

The incorporation of ekphrastic evocations of photographs into fictional works is a growing trend charted by (mostly) literary and (occasionally) art critics interested in the effect of their inclusion in a narrative. What has emerged as a veritable affinity of photography with literature has produced a fertile interdisciplinary critical discourse around areas of intersection between visual and verbal. With regard to short fiction, the photograph is often subject to investigation as analogy, the photograph and the short story being considered metonymically related with regard to form and effect. This notion of a structural equivalence between short story and photograph is one stressed by author/photographer Julio Cortàzar, concerned to highlight the quality of intensity he ascribes to both forms, which he saw as ‘cutting out a piece of reality’ in order to ‘breaking out’ into a wider one. Given Annie Saumont’s oft-cited admiration of Cortàzar’s work it is unsurprising that in her own writing – of stories themselves often classed, in their elliptical density, as verbal snapshots – she should take an interest in photographs and/or photographers. This article seeks to explore and analyse different values Saumont ascribes to what was paradoxically described by Barthes as ‘invisible’, in that what we see when viewing a photograph is, (often treacherously), ‘ pas elle qu’on voit’: never, or never solely, the actual object itself …

Identifying antimicrobial resistance genes of human clinical relevance within Salmonella isolated from food animals in Great Britain

To investigate the occurrence of antimicrobial resistance genes of human clinical relevance in Salmonella isolated from livestock in Great Britain. Two hundred and twenty-five Salmonella enterica isolates were characterized using an antimicrobial resistance gene chip and disc diffusion assays. Plasmid profiling, conjugation experiments and identification of Salmonella genomic island 1 (SGI1) were performed for selected isolates. Approximately 43% of Salmonella harboured single or multiple antimicrobial resistance genes with pig isolates showing the highest numbers where 96% of Salmonella Typhimurium harboured one or more resistance genes. Isolates harbouring multiple resistances divided into three groups. Group 1 isolates harboured ampicillin/streptomycin/sulphonamide/tetracycline resistance and similar phenotypes. This group contained isolates from pigs, cattle and poultry that were from several serovars including Typhimurium, 4,[5],12:i:-, Derby, Ohio and Indiana. All Group 2 isolates were from pigs and were Salmonella Typhimurium. They contained a non-sul-type class 1 integron and up to 13 transferrable resistances. All Group 3 isolates harboured a class 1 integron and were isolated from all animal species included in the study. Most isolates were Salmonella Typhimurium and harboured SGI1. Salmonella isolated from livestock was shown to harbour antimicrobial resistance genes although no or little resistance to third-generation cephalosporins or ciprofloxacin, respectively, was detected. The preponderance in pigs of multidrug-resistant Salmonella Typhimurium makes it important to introduce control measures such as improved biosecurity to ensure that they do not pass through the food chain and limit human therapeutic options.

Optimizing layer-by-layer deposition of interpolymer complexes on solid substrates using Biacore

The layer-by-layer deposition of polymers onto surfaces allows the fabrication of multilayered materials for a wide range of applications, from drug delivery to biosensors. This work describes the analysis of complex formation between poly(acrylic acid) and methylcellulose in aqueous solutions using Biacore, a surface plasmon resonance analytical technique, traditionally used to examine biological interactions. This technique characterized the layer-by-layer deposition of these polymers on the surface of a Biacore sensor chip. The results were subsequently used to optimize the experimental conditions for sequential layer deposition on glass slides. The role of the solution pH and poly(acrylic acid) molecular weight on the formation of interpolymer multilayered coatings was researched, and showed that the optimal deposition of the polymer complexes was achieved at pHs ≤2.5 with a poly(acrylic acid) molecular weight of 450 kDa.

Use of miniaturized protein arrays for Escherichia coli O serotyping

Serological typing of Escherichia coli O antigens is a well-established method used for differentiation and identification of O serotypes commonly associated with disease. In this feasibility study, we have developed a novel somatic antibody-based miniaturized microarray chip, using 17 antisera, which can be used to detect bound whole-cell E. coli antigen with its corresponding immobilized antibody, to assess the feasibility of this approach. The chip was tested using the related 17 control strains, and the O types found by the microarray chip showed 100% correlation with the O types found by conventional typing. A blind trial was performed in which 100 E. coli isolates that had been O serotyped previously by the conventional assay were tested by the array approach. Overall, the O serotypes of 88% of isolates were correctly identified by the microarray method. For several isolates, ambiguity of O-type designation by microarray arose due to increased sensitivity of this method, allowing signal intensities of cross-reactions to be quantified. Investigation of discrepancies between conventional and microarray O serotyping indicated that some isolates upon storage had become untypeable and, therefore, gave poor signal intensity when tested by the microarray or retested by conventional means. For all 20 serotype O26 and O157 isolates, the apparent discrepancy in O serotyping was analyzed further by a third independent test, which confirmed the microarray results. Therefore, the use of miniaturized protein arrays increases the speed and efficiency of O serotyping in a cost-effective manner, and these preliminary findings suggest the microarray approach may have a higher accuracy than those of traditional O-serotyping methods.

Robotic multiwell planar patch-clamp for native and primary mammalian cells

Robotic multiwell planar patch-clamp has become common in drug development and safety programs because it enables efficient and systematic testing of compounds against ion channels during voltage-clamp. It has not, however, been adopted significantly in other important areas of ion channel research, where conventional patch-clamp remains the favored method. Here, we show the wider potential of the multiwell approach with the ability for efficient intracellular solution exchange, describing protocols and success rates for recording from a range of native and primary mammalian cells derived from blood vessels, arthritic joints and the immune and central nervous systems. The protocol involves preparing a suspension of single cells to be dispensed robotically into 4-8 microfluidic chambers each containing a glass chip with a small aperture. Under automated control, giga-seals and whole-cell access are achieved followed by preprogrammed routines of voltage paradigms and fast extracellular or intracellular solution exchange. Recording from 48 chambers usually takes 1-6 h depending on the experimental design and yields 16-33 cell recordings.

Denial of change: the 'Military Revolution' as seen by contemporaries

Writers on military matters from the 14th century until the late 18th century either regretted the decadence of their times compared with Antiquity, or they saw no great change in military affairs since Antiquity. Few saw a revolutionary change ushered in by gunpowder, although this number increased since the great "querelle" about the Ancients and the Moderns under Louis XIV. In the early 19th century, the balance tipped, and few would have denied that technology had profoundly changed warfare. All this is a far cry, however, from any contemporary perception of a "Military Revolution" in the 16th and 17th centuries.

Victory, peace, and justice : the neglected trinity

The assumption that the most important aim of war is to create a better peace than existed before the war, i.e. a peace with justice, was self-evident for writers prior to Clausewitz. This does not mean that princes saw this as their priority, but theoreticians did. This changed dramatically with the Napoleonic Wars: Clausewitz initiated an era where writers on strategy paid no heed to what would come after military victory, now seen as the be-all and end-all of war. Terrible consequences flowed from this, and a series of ephemeral victories leading to new wars. It was only around the Second World War, to some in itself the consequence of this obsession with victory and not with peace, that it began to dawn on writers that peace, not military victory must be the ultimate aim to be kept in sight.

Concentrations of free amino acids and sugars in nine potato varieties: effects of storage and relationship with acrylamide formation

Acrylamide forms during cooking and processing predominately from the reaction of free asparagine and reducing sugars in the Maillard reaction. The identification of low free asparagine and reducing sugar varieties of crops is therefore an important target. In this study, nine varieties of potato (French fry varieties Maris Piper (from two suppliers), Pentland Dell, King Edward, Daisy, and Markies; and chipping varieties Lady Claire, Lady Rosetta, Saturna, and Hermes) grown in the United Kingdom in 2009 were analyzed at monthly intervals through storage from November 2009 to July 2010. Acrylamide formation was measured in heated flour and chips fried in oil. Analysis of variance revealed significant interactions between varieties nested within type (French fry and chipping) and storage time for most free amino acids, glucose, fructose, and acrylamide formation. Acrylamide formed in chips correlated significantly with acrylamide formed in flour and with chip color. There were significant correlations between glucose or total reducing sugar concentration and acrylamide formation in both variety types, but with fructose the correlation was much stronger for chipping than for French fry varieties. Conversely, there were significant correlations with acrylamide formation for both total free amino acid and free asparagine concentration in the French fry but not chipping varieties. The study showed the potential of variety selection for preventing unacceptable levels of acrylamide formation in potato products and the variety-dependent effect of long-term storage on acrylamide risk. It also highlighted the complex relationship between precursor concentration and acrylamide risk in potatoes.

Sub-surface terahertz imaging through uneven surfaces: visualizing Neolithic wall paintings in Çatalhöyük

Pulsed terahertz imaging is being developed as a technique to image obscured mural paintings. Due to significant advances in terahertz technology, portable systems are now capable of operating in unregulated environments and this has prompted their use on archaeological excavations. August 2011 saw the first use of pulsed terahertz imaging at the archaeological site of Çatalhöyük, Turkey, where mural paintings dating from the Neolithic period are continuously being uncovered by archaeologists. In these particular paintings the paint is applied onto an uneven surface, and then covered by an equally uneven surface. Traditional terahertz data analysis has proven unsuccessful at sub-surface imaging of these paintings due to the effect of these uneven surfaces. For the first time, an image processing technique is presented, based around Gaussian beam-mode coupling, which enables the visualization of the obscured painting.

Microcontroller-based peak current mode control using digital slope compensation

Microcontroller-based peak current mode control of a buck converter is investigated. The new solution uses a discrete time controller with digital slope compensation. This is implemented using only a single-chip microcontroller to achieve desirable cycle-by-cycle peak current limiting. The digital controller is implemented as a two-pole, two-zero linear difference equation designed using a continuous time model of the buck converter and a discrete time transform. Subharmonic oscillations are removed with digital slope compensation using a discrete staircase ramp. A 16 W hardware implementation directly compares analog and digital control. Frequency response measurements are taken and it is shown that the crossover frequency and expected phase margin of the digital control system match that of its analog counterpart.

Social and environmental report assurance: some interview evidence

The steady growth of social and environmental reporting (SER) is being accompanied by an increase in social and environmental reporting assurance (SERA). The existing literature on SERA suggests that it is necessary to build credibility and trust among corporate stakeholders. Prior work has also found evidence of managerial and professional capture of SERA. In this paper, we present empirical evidence from interviews with corporate social responsibility representatives from 20 UK listed companies on whether they consider SERA to be necessary. We believe this to be the first research into SERA that uses an interview method. Our interviews revealed mixed feelings. Half of the respondents believed that external SERA would enhance credibility and trust which confirmed the prior literature. However, the other half believed that external SERA was not necessary, believing that internal assurance was sufficient. This was because they saw SERA as predominantly a managerial tool, useful for checking the efficiency of internal management control systems, rather than as a mechanism for enhancing corporate accountability to stakeholders and building credibility and trust. The potential for SERA to be a mechanism whereby greater dialogue is created between companies and their stakeholders on social and environmental issues is not being harnessed. This paper thus demonstrates a fundamental difference between the external prior normative literature and the managerial motivation in the SERA area.