A potentiometric biosensor for rapid on-site disease diagnostics

Quantitative point-of-care (POC) devices are the next generation for serological disease diagnosis. Whilst pathogen serology is typically performed by centralized laboratories using Enzyme-Linked ImmunoSorbent Assay (ELISA), faster on-site diagnosis would infer improved disease management and treatment decisions. Using the model pathogen Bovine Herpes Virus-1 (BHV-1) this study employs an extended-gate field-effect transistor (FET) for direct potentiometric serological diagnosis. BHV-1 is a major viral pathogen of Bovine Respiratory Disease (BRD), the leading cause of economic loss ($2 billion annually in the US only) to the cattle and dairy industry. To demonstrate the sensor capabilities as a diagnostic tool, BHV-1 viral protein gE was expressed and immobilized on the sensor surface to serve as a capture antigen for a BHV-1-specific antibody (anti-gE), produced in cattle in response to viral infection. The gE-coated immunosensor was shown to be highly sensitive and selective to anti-gE present in commercially available anti-BHV-1 antiserum and in real serum samples from cattle with results being in excellent agreement with Surface Plasmon Resonance (SPR) and ELISA. The FET sensor is significantly faster than ELISA (<10 min), a crucial factor for successful disease intervention. This sensor technology is versatile, amenable to multiplexing, easily integrated to POC devices, and has the potential to impact a wide range of human and animal diseases.

Trait-Mediated Effects of Parasites on Invader-Native Interactions.

Parasites have a variety of behavioural effects on their hosts, which can in turn affect species with which the host interacts. Here we review how these trait-mediated indirect effects of parasites can alter the outcomes of invader-native interactions, illustrating with examples from the literature and with particular regard to the invader-native crustacean systems studied in our laboratories. Parasites may potentially inhibit or exacerbate invasions via their effects on host behaviour, in addition to their direct virulence effects on hosts. In several crustacean systems, we have found that parasites influence both host predation rates on intra- and inter-guild prey and host vulnerability to being preyed upon. These trait effects can theoretically alter invasion impact and patterns of coexistence, as they indirectly affect interactions between predators and prey with the potential for further ramifications to other species in the food web. The fitness consequences of parasite-induced trait-mediated effects are rarely considered in traditional parasitological contexts, but demand attention in the context of ecological communities. We can regard these trait effects as a form of cryptic virulence that only becomes apparent when hosts are examined in the context of the other species with which they interact.

Immunohistochemistry should undergo robust validation equivalent to that of molecular diagnostics

Immunohistochemistry (IHC) is a widely available and highly utilised tool in diagnostic histopathology and is used to guide treatment options as well as provide prognostic information. IHC is subjected to qualitative and subjective assessment, which has been criticised for a lack of stringency, while PCR-based molecular diagnostic validations by comparison are regarded as very rigorous. It is essential that IHC tests are validated through evidence-based procedures. With the move to ISO15189 (2012), not just of the accuracy, specificity and reproducibility of each test need to be determined and managed, but also the degree of uncertainty and the delivery of such tests. The recent update to ISO 15189 (2012) states that it is appropriate to consider the potential uncertainty of measurement of the value obtained in the laboratory and how that may impact on prognostic or predictive thresholds. In order to highlight the problems surrounding IHC validity, we reviewed the measurement of Ki67and p53 in the literature. Both of these biomarkers have been incorporated into clinical care by pathology laboratories worldwide. The variation seen appears excessive even when measuring centrally stained slides from the same cases. We therefore propose in this paper to establish the basis on which IHC laboratories can bring the same level of robust validation seen in the molecular pathology laboratories and the principles applied to all routine IHC tests.

Radioimmunoassay Quantitation of Neuropeptide Y (NPY) in Human Dental Pulps

Neuropeptide Y (NPY) is a 36 amino acid peptide that is abundantly expressed in both the central and peripheral nervous systems. NPY has previously been shown to be present in human dental pulp although its exact role in pulpal health and disease remains to be fully elucidated. In addition to serving a neurotransmitter role, NPY may also have a role in modulating the pulpal response to injury and inflammation. Indeed NPY is known to be a potent vasoconstrictor in a range of tissues. Recent work by our research group has demonstrated changes in sensory neuropeptide levels measured by radioimmunoassay (RIA) in healthy and carious teeth. In addition to elevated levels of sensory neuropeptides, it is also possible that the carious process is associated with increased levels of autonomic neuropeptides such as NPY. Objectives: The aim of the present study was to undertake a comprehensive quantitative RIA analysis of NPY expression in human dental pulps from carious and non-carious teeth. Methods: A total of 22 non-carious and 46 carious teeth were included in the study. NPY was measured in all samples using RIA. Briefly, the RIA system consisted of a total volume of 400 ul, comprising 100 ul anti-NPY antibody (Peninsula Laboratories), 200 ul human NPY synthetic standard or pulp sample, and 100 ul of 125I-labelled NPY as radioactive tracer. Results: The mean concentration of NPY in non-carious teeth was found to be 4.28 ng/g (4.34 SD) compared to 9.57 ng/g (9.39 SD) in carious teeth. Using ANOVA the difference in NPY levels between the non-carious group and the carious group was found to be statistically significant (p= 0.003). Conclusion: The significant increase in the levels of NPY in carious dental pulps reported in this study provides evidence for a role for NPY in the pulpal response to caries.

Atomic data for modelling fusion and astrophysical plasmas

Trends and focii of interest in atomic modelling and data are identified in connection with recent observations and experiments in fusion and astrophysics. In the fusion domain, spectral observations are included of core, beam penetrated and divertor plasma. The helium beam experiments at JET and the studies with very heavy species at ASDEX and JET are noted. In the astrophysics domain, illustrations are given from the SOHO and CHANDRA spacecraft which span from the solar upper atmosphere, through soft x-rays from comets to supernovae remnants. It is shown that non-Maxwellian, dynamic and possibly optically thick regimes must be considered. The generalized collisional-radiative model properly describes the collisional regime of most astrophysical and laboratory fusion plasmas and yields self-consistent derived data for spectral emission, power balance and ionization state studies. The tuning of this method to routine analysis of the spectral observations is described. A forward look is taken as to how such atomic modelling, and the atomic data which underpin it, ought to evolve to deal with the extended conditions and novel environments of the illustrations. It is noted that atomic physics influences most aspects of fusion and astrophysical plasma behaviour but the effectiveness of analysis depends on the quality of the bi-directional pathway from fundamental data production through atomic/plasma model development to the confrontation with experiment. The principal atomic data capability at JET, and other fusion and astrophysical laboratories, is supplied via the Atomic Data and Analysis Structure (ADAS) Project. The close ties between the various experiments and ADAS have helped in this path of communication.