Functions of DNA damage machinery in the innate immune response to DNA virus infection.

DNA is potently immunostimulatory, and self-DNA is packaged in the nucleus or mitochondria allowing it to remain silent to cell-intrinsic sensors. However, damaged or mislocalised self-DNA is sensed by our innate immune systems, resulting in the production of type I interferons (IFNI), chemokines and inflammatory cytokines. During DNA virus infection the detection of viral DNA genomes by pattern recognition receptors (PRRs) is essential for the initiation of IFNI responses and host defence against these pathogens. It is intriguing that a number of molecular mechanisms have been found to be common to both of these DNA-induced stress responses and this has potentially important consequences for both sides of the host/pathogen arms race.This is the author accepted manuscript. The final version is available from Elsevier via http://dx.doi.org/10.1016/j.coviro.2015.08.001

ULTRASTRUCTURAL BASIS OF BIOCHEMICAL EFFECTS IN A SERIES OF LETHAL ALLELES IN THE MOUSE : Neonatal and Developmental Studies

The fine structure of newborn and fetal mouse liver and of newborn kidney cells homozygous for any of three albino alleles known to have multiple biochemical effects was investigated. Electron microscope studies of mutant cells revealed dilation and vesiculation of the rough endoplasmic reticulum in parenchymal liver cells, as well as dilation and other anomalies of the Golgi apparatus. These abnormalities were observed in all newborn mutants but never in littermate controls. Although they were most pronounced in liver parenchymal cells, they were found also to a lesser degree in kidney cells, but they were absent altogether in other cell types of the mutant newborn. Homozygous fetuses showed similar anomalies in the liver at 19 days of gestational age. In one of the alleles studied, mutant liver parenchymal cells were found to be abnormal as early as the 18th day of gestation. There appears to be a striking parallelism between the biochemical defects and those of the cellular membranes in homozygous mutant newborn and fetuses. Although the specific nature of the mutational effect on membrane structure remains unknown, the results are compatible with the assumption that a mutationally caused defect in a membrane component interferes with a mechanism vital in the integration of morphological and biochemical differentiation

Jumble Java Byte Code to Measure the Effectiveness of Unit Tests

Jumble is a byte code level mutation testing tool for Java which inter-operates with JUnit. It has been designed to operate in an industrial setting with large projects. Heuristics have been included to speed the checking of mutations, for example, noting which test fails for each mutation and running this first in subsequent mutation checks. Significant effort has been put into ensuring that it can test code which uses custom class loading and reflection. This requires careful attention to class path handling and coexistence with foreign class-loaders. Jumble is currently used on a continuous basis within an agile programming environment with approximately 370,000 lines of Java code under source control. This checks out project code every fifteen minutes and runs an incremental set of unit tests and mutation tests for modified classes. Jumble is being made available as open source

Modelling of flood hazard extent in data sparse areas: a case study of the Oti River basin, West Africa

Study region: Terrain and hydrological data are scarce in many African countries. The coarse spatial resolution of freely available Shuttle Radar Topographic Mission elevation data and the absence of flow gauges on flood-prone reaches, such as the Oti River studied here, make flood inundation modelling challenging in West Africa. Study focus: A flood modelling approach is developed here to simulate flood extent in data scarce regions. The methodology is based on a calibrated, distributed hydrological model for the whole basin to simulate the input discharges for a hydraulic model which is used to predict the flood extent for a 140 km reach of the Oti River. New hydrological insight for the region: Good hydrological model calibration (Nash Sutcliffe coefficient: 0.87) and validation (Nash Sutcliffe coefficient: 0.94) results demonstrate that even with coarse scale (5 km) input data, it is possible to simulate the discharge along this region's rivers, and importantly with a distributed model, derive model flows at any ungauged location within basin. With a lack of surveyed channel bathymetry, modelling the flood was only possible with a parametrized sub-grid hydraulic model. Flood model fit results relative to the observed 2007 flood extent and extensive sensitivity testing shows that this fit (64%) is likely to be as good as is possible for this region, given the coarseness of the terrain digital elevation model

⁹⁹ᵐTc SPECT imaging agent based on cFLFLFK for the detection of FPR1 in inflammation

Non-invasive imaging of the inflammatory process can provide a great deal of insight into a wide variety of diseases states, aiding diagnosis, evaluation and effective targeted treatment. During inflammation, blood borne leukocytes are recruited, through a series of activation and adhesion steps, to the site of injury or infection where they migrate across the blood vessel wall into the tissue. Thus, tracking leukocyte recruitment and accumulation provides a dynamic and localised read out of inflammatory events. Current leukocyte imaging techniques require ex vivo labelling of patient blood, involving laborious processing and potential risks to both patient and laboratory staff. Utilising high affinity ligands for leukocyte specific receptors may allow for injectable tracers that label leukocytes in situ, omitting potentially hazardous ex vivo handling. Formyl peptide receptors (FPRs) are a group of G-protein coupled receptors involved in the chemotaxis and inflammatory functioning of leukocytes. Highly expressed on leukocytes, and up regulated during inflammation, these receptors provide a potential target for imaging inflammatory events. Herein we present the synthesis and initial in vitro testing of a potential Single Photon Emission Computed Tomography (SPECT) leukocyte tracer. The FPR1 antagonist cFLFLFK-NH₂, which displays high affinity with little physiological effect, has been linked via a PEG motif to a ⁹⁹ᵐTc chelate. This tracer shows in vitro binding to human embryonic kidney cells expressing the FPR1 receptor, and functional in vitro tests reveal cFLFLFK-NH₂ compounds to have no effect on inflammatory cell functioning. Overall, these data show that ⁹⁹ᵐTc.cFLFLFK-NH₂ may be a useful tool for non-invasive imaging of leukocyte accumulation in inflammatory disease states

“Tell them what they want to hear and get back to work”: insights into the utility of current occupational health assessments from the perspectives of train drivers

Australian train drivers undergo periodic health assessments as part of a nationally standardised approach to reducing sudden incapacitation risk, given the demonstrated potential for occupational and public harm. These assessments occur pre-placement, then every 5 years to age 50, then every 2 years to age 60, and then every year. Despite some reported benefits to rail workforce health indicators since implementation, research suggests the assessments are not operating as effectively as they might. For example, the prevalence of obesity in drivers is higher than in the general population and continues to increase. To improve this, there is a need to understand the experiences of drivers undergoing workplace health assessments. The aims of this study were to examine train drivers’ perceptions and experiences of the assessments, understand how these experiences shape their engagement with the process, and to generate recommendations for improvement from a systems thinking perspective. A qualitative design was used, involving semi-structured interviews within five focus groups of train drivers (n = 29) held across four Australian rail organisations. Questions addressed drivers’ backgrounds, their understanding of the National Standard, experiences of and attitudes towards health assessments, lifestyle risk factors, and personal approach to health and wellbeing. Transcript data were subjected to thematic analysis. Five factors were identified: drivers’ unmet information needs, perceived low reliability and validity of assessment, need for psychological wellbeing assessment and support, maladaptive threat avoidance strategies, and focus on short-term outcomes and compliance. The global theme was reactive organisational culture. Findings suggest that driver engagement with health assessment can be improved by proactively addressing the identified factors in occupational health initiatives and preventative interventions to tackle the burgeoning problem of train driver health impairment.Anjum Naweed, Janine Chapman, Joshua Trig

Study of two-dimensional calculation of cratering

Time dependent, two-dimensional, coupled Eulerian-Lagrangian code designed for solution of cratering problem

18F-GE-180: a novel TSPO radiotracer compared to 11C-R-PK11195 in a preclinical model of stroke

PURPOSE: Neuroinflammation plays a critical role in various neuropathological conditions, and hence there is renewed interest in the translocator protein (TSPO) as a biomarker of microglial activation and macrophage infiltration in the brain. This is reflected in the large amount of research conducted seeking to replace the prototypical PET radiotracer 11C-R-PK11195 with a TSPO ligand with higher performance. Here we report the in vivo preclinical investigation of the novel TSPO tracer 18F-GE-180 in a rat model of stroke. METHODS: Focal cerebral ischaemia was induced in Wistar rats by 60-min occlusion of the middle cerebral artery (MCAO). Brain damage was assessed 24 h after MCAO by T2 MRI. Rats were scanned with 11C-R-PK11195 and 18F-GE-180 5 or 6 days after MCAO. Specificity of binding was confirmed by injection of unlabelled R-PK11195 or GE-180 20 min after injection of 18F-GE-180. In vivo data were confirmed by ex vivo immunohistochemistry for microglial (CD11b) and astrocytic biomarkers (GFAP). RESULTS: 18F-GE-180 uptake was 24 % higher in the core of the ischaemic lesion and 18 % lower in the contralateral healthy tissue than that of 11C-R-PK11195 uptake (1.5 ± 0.2-fold higher signal to noise ratio). We confirmed this finding using the simplified reference tissue model (BPND = 3.5 ± 0.4 and 2.4 ± 0.5 for 18F-GE-180 and 11C-R-PK11195, respectively, with R 1 = 1). Injection of unlabelled R-PK11195 or GE-180 20 min after injection of 18F-GE-180 significantly displaced 18F-GE-180 (69 ± 5 % and 63 ± 4 %, respectively). Specificity of the binding was also confirmed by in vitro autoradiography, and the location and presence of activated microglia and infiltrated macrophages were confirmed by immunohistochemistry. CONCLUSION: The in vivo binding characteristics of 18F-GE-180 demonstrate a better signal to noise ratio than 11C-R-PK11195 due to both a better signal in the lesion and lower nonspecific binding in healthy tissue. These results provide evidence that 18F-GE-180 is a strong candidate to replace 11C-R-PK11195