Wegener's granulomatosis and autoantibodies to neutrophil antigens

We report five cases of Wegener's granulomatosis all of whom had clinical and histological evidence of disease activity at presentation and in whom autoantibodies to neutrophil antigens were detected. This test may prove useful for the diagnosis of this serious condition and help to monitor disease activity during treatment

Design of a high efficiency cyclone for collection of rare and low concentration airborne pathogens

Recent serious outbreaks of pathogens such as Chalara Fraxinea, (Ash Dieback) demonstrate the vulnerability of UK’s forests, woods, and trees. Early detection of threats is critical in the fight against such tree pests and pathogens. This requires a process for collecting and analysing the spores which is robust, flexible and can be deployed rapidly, but which also has sufficient sensitivity to detect the earliest precursors. The work presented here describes a newly-developed high-efficiency detection apparatus and is part of an ongoing BBRSC project to improve the UK’s biosecurity. Our automatic spore system is capable of collecting and analysing Chalara Fraxinea using a novel cyclonic pathogen collector. Cyclones are an effective way to separate small particles (spores) from large quantities of surrounding air and store them for analysis. Our system incorporates a purpose-designed high efficiency cyclone directly integrated with a low power custom impeller to maximise volumetric air sampling while minimising the power requirement. 3D printing was used extensively to validate theoretical models and the particle collection and retention capability of this series of prototypes was evaluated experimentally within our Aerosol test chamber. We found that we could collect greater than 90% of particulate in the target size range, thus providing a front-end to a detection platform. Our system is capable of operating autonomously and at low power, with a high sensitivity to ambient particles. The final system design incorporates the use of multiple cyclone storage vessels ensuring complete isolation of each sample, eradicating cross-contamination, and facilitating automated handling of the sample inside the same apparatus. COTS – commercial off the shelf components were incorporated into the cyclone to make a series of cost effective collection vessels. Early collection and detection of pathogens in-situ represents a considerable advance in surveillance and monitoring of pathogens strengthening UK biosecurity for the future

Development of an automated smart trap for wheat pathogens

National surveys show fungicide use on wheat continues to increase despite fluctuations in disease pressure, reaching a 30 year high in 2012 (Defra). Septoria tritici is the most significant foliar disease in UK wheat causing between £43M to £53M in yield losses annually; Yellow and brown rust are more sporadic but have caused significant losses during high disease years. In all cases control is by fungicide application costing £82M annually (GFK Kynetec 2013). Effective disease management relies on either prophylactic pesticide use or significant manual intervention and time consuming assessment of crop disease indicators by farmers and agronomists. Furthermore indications are that current levels of pesticide use could lead to increased risk of pesticide resistance, if this should occur it is estimated that wheat yields could reduce by up to 20%. To address this we have developed a prototype integrated and automated spore detection system, designed for unattended field application, to monitor and identify the presence of Septoria, brown and yellow rust. The prototype system incorporates novel cyclonic pathogen collection, on-board sample processing and isothermal DNA amplification chemistry (LAMP). We present the engineering design, optimisation and evaluation of our prototype system reporting on successfully completed laboratory testing and initial field trial results. This prototype will be the basis for the development of a commercially available system which, in addition to inoculum detection, will be capable of providing growers/agronomists with real-time information on inoculum moving into a crop enabling more effective timing and selection of fungicide application, and thus better control, increased yield, and improved environmental stewardship

Molecular analysis of intragenic recombination at the tryptophan synthetase locus in Neurospora crassa

Fifteen different classically generated and mapped mutations at the tryptophan synthetase locus in Neurospora crassa have been characterized to the level of the primary sequence of the gene. This sequence analysis has demonstrated that intragenic recombination is accurate to order mutations within one open reading frame. While classic genetic analysis correctly ordered the mutations, the position of mutations characterized by gene sequence analysis was more accurate. A leaky mutation was found to have a wild-type primary sequence. The presence of unique polymorphisms in the primary sequence of the trp-3 gene from strain 861 confirms that it has a unique history relative to the other strains studied. Most strains that were previously shown to be immunologically nonreactive with antibody preparations raised against tryptophan synthetase protein were shown to have nonsense mutations. This work defines 14 alleles of the N. crassa trp-3 gene.Citation: "Molecular analysis of intragenic recombination at the tryptophan synthetase locus in Neurospora crassa" (December 2013) A. Wiest D. Barchers M. Eaton R. Henderson R. Schnittker K. Mccluskey. Journal of Genetics, Indian Academy of Science. Volume 92 Issue 3. 523-528.Citation: Wiest, A., . . . & McCluskey, K. (2013). Molecular analysis of intragenic recombination at the tryptophan synthetase locus in Neurospora crassa. Journal of Genetics, 92(1), 523–528. https://doi.org/https://doi.org/10.1007/s12041-013-0305-

Protein droplet actuation on superhydrophobic surfaces: A new approach toward anti-biofouling electrowetting systems

Among Lab-on-a-chip techniques, digital microfluidics (DMF), allowing the precise actuation of discrete droplets, is a highly promising, flexible, biochemical assay platform for biomedical and bio-detection applications. However the durability of DMF systems remains a challenge due to biofouling of the droplet-actuating surface when high concentrations of biomolecules are employed. To address this issue, the use of superhydrophobic materials as the actuating surface in DMF devices is examined. The change in contact angle by electrowetting of deionised water and ovalbumin protein samples is characterised on different surfaces (hydrophobic and superhydrophobic). Ovalbumin droplets at 1 mg ml−1 concentration display better electrowetting reversibility on Neverwet®, a commercial superhydrophobic material, than on Cytop®, a typical DMF hydrophobic material. Biofouling rate, characterised by roll-off angle measurement of ovalbumin loaded droplets and further confirmed by measurements of the mean fluorescence intensity of labelled fibrinogen, appears greatly reduced on Neverwet®. Transportation of protein laden droplets (fibrinogen at concentration 0.1 mg ml−1 and ovalbumin at concentration 1 mg ml−1 and 10 mg ml−1) is successfully demonstrated using electrowetting actuation on both single-plate and parallel-plate configurations with performance comparable to that of DI water actuation. In addition, although droplet splitting requires further attention, merging and efficient mixing are demonstrated