Development and Validation of a PulseNet Standardized Protocol for Subtyping Isolates of Cronobacter Species

Cronobacter (formerly known as Enterobacter sakazakii) is a genus comprising seven species regarded as opportunistic pathogens that can be found in a wide variety of environments and foods, including powdered infant formula (PIF). Cronobacter sakazakii, the major species of this genus, has been epidemiologically linked to cases of bacteremia, meningitis in neonates, and necrotizing enterocolitis, and contaminated PIF has been identified as an important source of infection. Robust and reproducible subtyping methods are required to aid in the detection and investigation, of foodborne outbreaks. In this study, a pulsed-field gel electrophoresis (PFGE) protocol was developed and validated for subtyping Cronobacter species. It was derived from an existing modified PulseNet protocol, wherein XbaI and SpeI were the primary and secondary restriction enzymes used, generating an average of 14.7 and 20.3 bands, respectively. The PFGE method developed was both reproducible and discriminatory for subtyping Cronobacter species.

Development and Validation of a Compact Thermal Model for an Aircraft Compartment

The development of accurate structural/thermal numerical models of complex systems, such as aircraft fuselage barrels, is often limited and determined by the smallest scales that need to be modelled. The development of reduced order models of the smallest scales and consequently their integration with higher level models can be a way to minimise the bottle neck present, while still having efficient, robust and accurate numerical models. In this paper a methodology on how to develop compact thermal fluid models (CTFMs) for compartments where mixed convection regimes are present is demonstrated. Detailed numerical simulations (CFD) have been developed for an aircraft crown compartment and validated against experimental data obtained from a 1:1 scale compartment rig. The crown compartment is defined as the confined area between the upper fuselage and the passenger cabin in a single aisle commercial aircraft. CFD results were utilised to extract average quantities (temperature and heat fluxes) and characteristic parameters (heat transfer coefficients) to generate CTFMs. The CTFMs have then been compared with the results obtained from the detailed models showing average errors for temperature predictions lower than 5%. This error can be deemed acceptable when compared to the nominal experimental error associated with the thermocouple measurements.

The CTFMs methodology developed allows to generate accurate reduced order models where accuracy is restricted to the region of Boundary Conditions applied. This limitation arises from the sensitivity of the internal flow structures to the applied boundary condition set. CTFMs thus generated can be then integrated in complex numerical modelling of whole fuselage sections.

Further steps in the development of an exhaustive methodology would be the implementation of a logic ruled based approach to extract directly from the CFD simulations numbers and positions of the nodes for the CTFM.

Psychic Distance, its Business Impact and Modes of Coping: A Study of British and Indian Partner SMEs

This paper reports one of the first investigations to analyze inter-partner perceptions of psychic distance between two countries. Its empirical focus is British and Indian SMEs engaged in business with each other. It examines different dimensions of psychic distance, their impact and modes of coping with them. Potential firm-level and individual influences are also taken into account. The paper aims to transcend some of the conceptual and methodological limitations of previous research on the subject and to identify the theoretical and practical implications that arise. A ‘mirror’ approach is applied, accessing both partners’ perceptions. These are assessed through a ‘mixed’ method combining quantitative measurement with qualitative interpretations. Psychic distance dimensions are found to vary in their impact on doing business with the other country, and there is also variation according to the firm’s sector. There is considerable asymmetry in British and Indian partners’ perceptions of psychic distance but the degree of difference between their psychic distance evaluations lacks predictive power. Culturally embedded psychic distance dimensions tend to have less impact and to be easier to cope with than institutionally embedded dimensions. Four categories of coping are identified. The principal theoretical implication of this study is that a contingency perspective needs to be adopted in the field of ‘distance’ research, taking account of factors such as a firm’s sector, and that this will require a more complex analytical framework that hitherto.

Development and validation of the first high performance-lateral flow immunoassay (HP-LFIA) for the rapid screening of domoic acid from shellfish extracts

A lateral flow immunoassay (LFIA) has been developed and fully validated to detect the primary amnesic shellfish poisoning (ASP) toxin, domoic acid (DA). The performance characteristics of two versions of the test were investigated using spiked and naturally contaminated shellfish (mussels, scallops, oysters, clams, and cockles). The tests provide a qualitative result, to indicate the absence or presence of DA in extracts of shellfish tissues, at concentrations that are relevant to regulatory limits. The new rapid assay (LFIA version 2) was designed to overcome the performance limitations identified in the first version of the assay. The improved test uses an electronic reader to remove the subjective nature of the generated results, and the positive cut-off for screening of DA in shellfish was increased from 10 ppm (version 1) to 17.5 ppm (version 2). A simple extraction and test procedure was employed, which required minimal equipment and materials; results were available 15 min after sample preparation. Stability of the aqueous extracts at room temperature (22 C) at four time points (up to 245 min after extraction) and across a range of DA concentrations was 100.3±1.3% and 98.8±2.4% for pre- and post-buffered extracts, respectively. The assay can be used both within laboratory settings and in remote locations. The accuracy of the new assay, to indicate negative results at or below 10 ppm DA, and positive results at or above 17.5 ppm, was 99.5% (n=216 tests). Validation data were obtained from a 2-day, randomised, blind study consisting of multiple LFIA lots (n=3), readers (n=3) and operators (n=3), carrying out multiple extractions of mussel tissue (n=3) at each concentration (0, 10, 17.5, and 20 ppm). No matrix effects were observed on the performance of the assay with different species (mussels, scallops, oysters, clams, and cockles). There was no impact on accuracy or interference from other phycotoxins, glutamic acid or glutamine with various strip incubations (8, 10, and 12 min). The accuracy of the assay, using naturally contaminated samples to indicate negative results at or below 12.5 ppm and positive results at or above 17.5 ppm, was 100%. Variability between three LFIA lots across a range of DA concentrations, expressed as coefficient of variation (% CV), was 1.1±0.4% (n=2 days) based on quantitative readings from the electronic reader. During an 8 week stability study, accuracy of the method with test strips stored at various temperatures (6, 22, 37 and 50 C) was 100%. Validation for both versions included comparisons with results obtained using reference LC-UV methods. © 2013 Elsevier B.V.

Development and validation of a rapid multiplex ELISA for pyrrolizidine alkaloids and their N-oxides in honey and feed Rapid Detection in Food and Feed

Pyrrolizidine alkaloids (PAs) are a group of plant secondary metabolites with carcinogenic and hepatotoxic properties. When PA-producing plants contaminate crops, toxins can be transferred through the food chain and cause illness in humans and animals, most notably hepatic veno-occlusive disease. Honey has been identified as a direct risk of human exposure. The European Food Safety Authority has recently identified four groups of PAs that are of particular importance for food and feed: senecionine-type, lycopsamine-type, heliotrine-type and monocrotaline-type. Liquid or gas chromatography methods are currently used to detect PAs but there are no rapid screening assays available commercially. Therefore, the aim of this study was to develop a rapid multiplex ELISA test for the representatives of three groups of alkaloids (senecionine, lycopsamine and heliotrine types) that would be used as a risk-management tool for the screening of these toxic compounds in food and feed. The method was validated for honey and feed matrices and was demonstrated to have a detection capability less than 25 µg/kg for jacobine, lycopsamine, heliotrine and senecionine. The zinc reduction step introduced to the extraction procedure allows for the additional detection of the presence of N-oxides of PAs. This first multiplex immunoassay for PA detection with N-oxide reduction can be used for the simultaneous screening of 21 samples for >12 PA analytes. Honey samples (n?=?146) from various origins were analysed for PA determination. Six samples were determined to contain measurable PAs >25 µg/kg by ELISA which correlated to >10 µg/kg by LC-MS/MS.

Development and assessment of a robust optimization metric for WAMPAC modeling

A PMU based WAMS is to be placed on a weakly coupled section of distribution grid, with high levels of distributed generation. In anticipation of PMU data a Siemens PSS/E model of the electrical environment has been used to return similar data to that expected from the WAMS. This data is then used to create a metric that reflects optimization, control and protection in the region. System states are iterated through with the most desirable one returning the lowest optimization metric, this state is assessed against the one returned by PSS/E under normal circumstances. This paper investigates the circumstances that trigger SPS in the region, through varying generation between 0 and 110% and compromising the network through line loss under summer minimum and winter maximum conditions. It is found that the optimized state can generally tolerate an additional 2 MW of generation (3% of total) before encroaching the same thresholds and in one instance moves the triggering to 100% of generation output.

Microstructural analysis of non-woven fabrics using scanning electron microscopy and image processing. Part 1:Development and verification of the methods

This paper reports image analysis methods that have been developed to study the microstructural changes of non-wovens made by the hydroentanglement process. The validity of the image processing techniques has been ascertained by applying them to test images with known properties. The parameters in preprocessing of the scanning electron microscope (SEM) images used in image processing have been tested and optimized. The fibre orientation distribution is estimated using fast Fourier transform (FFT) and Hough transform (HT) methods. The results obtained using these two methods are in good agreement. The HT method is more demanding in computational time compared with the Fourier transform (FT) method. However, the advantage of the HT method is that the actual orientation of the lines can be concluded directly from the result of the transform without the need for any further computation. The distribution of the length of the straight fibre segments of the fabrics is evaluated by the HT method. The effect of curl of the fibres on the result of this evaluation is shown.

Development and characterization of self-assembling nanoparticles using a bio-inspired amphipathic peptide for gene delivery

The design of a non-viral gene delivery vehicle capable of delivering and releasing a functional nucleic acid cargo intracellularly remains a formidable challenge. For systemic gene therapy to be successful a delivery vehicle is required that protects the nucleic acid cargo from enzymatic degradation, extravasates from the vasculature, traverses the cell membrane, disrupts the endosomal vesicles and unloads the cargo at its destination site, namely the nucleus for the purposes of gene delivery. This manuscript reports the extensive investigation of a novel amphipathic peptide composed of repeating RALA units capable of overcoming the biological barriers to gene delivery both in vitro and in vivo. Our data demonstrates the spontaneous self-assembly of cationic DNA-loaded nanoparticles when the peptide is complexed with pDNA. Nanoparticles were < 100 nm, were stable in the presence of serum and were fusogenic in nature, with increased peptide α-helicity at a lower pH. Nanoparticles proved to be non-cytotoxic, readily traversed the plasma membrane of both cancer and fibroblast cell lines and elicited reporter-gene expression following intravenous delivery in vivo. The results of this study indicate that RALA presents an exciting delivery platform for the systemic delivery of nucleic acid therapeutics.

Development and validation of Fluid Structure Interaction Methods for an Oscillating Wave Surge Converter

The term fatigue loads on the Oyster Oscillating Wave Surge Converter (OWSC) is used to describe hydrostatic loads due to water surface elevation with quasi-static changes of state. Therefore a procedure to implement hydrostatic pressure distributions into finite element analysis of the structure is desired. Currently available experimental methods enable one to measure time variant water surface elevation at discrete locations either on or around the body of the scale model during tank tests. This paper discusses the development of a finite element analysis procedure to implement time variant, spatially distributed hydrostatic pressure derived from discretely measured water surface elevation. The developed method can process differently resolved (temporal and spatial) input data and approximate the elevation over the flap faces with user defined properties. The structural loads, namely the forces and moments on the body can then be investigated by post processing the numerical results. This method offers the possibility to process surface elevation or hydrostatic pressure data from computational fluid dynamics simulations and can thus be seen as a first step to a fluid-structure interaction model.

Knowledge translation in men’s health research: development and delivery of content for use online

Background: Men can be hard to reach with face-to-face health-related information, while increasingly, research shows that they are seeking health information from online sources. Recognizing this trend, there is merit in developing innovative online knowledge translation (KT) strategies capable of translating research on men’s health into engaging health promotion materials. While the concept of KT has become a new mantra for researchers wishing to bridge the gap between research evidence and improved health outcomes, little is written about the process, necessary skills, and best practices by which researchers can develop online knowledge translation.
Objective: Our aim was to illustrate some of the processes and challenges involved in, and potential value of, developing research knowledge online to promote men’s health.

Methods: We present experiences of KT across two case studies of men’s health. First, we describe a study that uses interactive Web apps to translate knowledge relating to Canadian men’s depression. Through a range of mechanisms, study findings were repackaged with the explicit aim of raising awareness and reducing the stigma associated with men’s depression and/or help-seeking. Second, we describe an educational resource for teenage men about unintended pregnancy, developed for delivery in the formal Relationship and Sexuality Education school curricula of Ireland, Northern Ireland (United Kingdom), and South Australia. The intervention is based around a Web-based interactive film drama entitled “If I Were Jack”.

Results: For each case study, we describe the KT process and strategies that aided development of credible and well-received online content focused on men’s health promotion. In both case studies, the original research generated the inspiration for the interactive online content and the core development strategy was working with a multidisciplinary team to develop this material through arts-based approaches. In both cases also, there is an acknowledgment of the need for gender and culturally sensitive information. Both aimed to engage men by disrupting stereotypes about men, while simultaneously addressing men through authentic voices and faces. Finally, in both case studies we draw attention to the need to think beyond placement of content online to delivery to target audiences from the outset.

Conclusions: The case studies highlight some of the new skills required by academics in the emerging paradigm of translational research and contribute to the nascent literature on KT. Our approach to online KT was to go beyond dissemination and diffusion to actively repackage research knowledge through arts-based approaches (videos and film scripts) as health promotion tools, with optimal appeal, to target male audiences. Our findings highlight the importance of developing a multidisciplinary team to inform the design of content, the importance of adaptation to context, both in terms of the national implementation context and consideration of gender-specific needs, and an integrated implementation and evaluation framework in all KT work.

Novel photocatalytic reactor development for removal of hydrocarbons from water

Hydrocarbons contamination of the marine environment generated by the offshore oil and gas industry is generated from a number of sources including oil contaminated drill cuttings and produced waters. The removal of hydrocarbons from both these sources is one of the most significant challenges facing this sector as it moves towards zero emissions. The application of a number of techniques which have been used to successfully destroy hydrocarbons in produced water and waste water effluents has previously been reported. This paper reports the application of semiconductor photocatalysis as a final polishing step for the removal of hydrocarbons from two waste effluent sources. Two reactor concepts were considered: a simple flat plate immobilised film unit, and a new rotating drum photocatalytic reactor. Both units proved to be effective in removing residual hydrocarbons from the effluent with the drum reactor reducing the hydrocarbon content by 90% under 10 minutes.