Purinergic receptor mediated calcium signalling in urothelial cells

Non-neuronal ATP released from the urothelium in response to bladder stretch is a key modulator of bladder mechanosensation. Whilst non-neuronal ATP acts on the underlying bladder afferent nerves to facilitate sensation, there is also the potential for ATP to act in an autocrine manner, modulating urothelial cell function. The aim of this study was to systematically characterise the functional response of primary mouse urothelial cells (PMUCs) to ATP. PMUCs isolated from male mice (14-16 weeks) were used for live-cell fluorescent calcium imaging and qRT-PCR to determine the expression profile of P2X and P2Y receptors. The majority of PMUCs (74-92%) responded to ATP (1 μM-1 mM), as indicted by an increase in intracellular calcium (iCa2+). PMUCs exhibited dose-dependent responses to ATP (10 nM-1 mM) in both calcium containing (2 mM, EC50 = 3.49 ± 0.77 μM) or calcium free (0 mM, EC50 = 9.5 ± 1.5 μM) buffers. However, maximum iCa2+ responses to ATP were significantly attenuated upon repetitive applications in calcium containing but not in calcium free buffer. qRT-PCR revealed expression of P2X1-6, and P2Y1-2, P2Y4, P2Y6, P2Y11-14, but not P2X7 in PMUCs. These findings suggest the major component of ATP induced increases in iCa2+ are mediated via the liberation of calcium from intracellular stores, implicating functional P2Y receptors that are ubiquitously expressed on PMUCs.Russell Chess-Williams, Donna J. Sellers, Stuart M. Brierley, David Grundy, Luke Grund

Dendritic Targeting in the Leg Neuropil of Drosophila: The Role of Midline Signalling Molecules in Generating a Myotopic Map

During development of the Drosophila motor system, global guidance cues control and coordinate the targeting of both input and output elements of the neural system

Sampling the fish gill microbiome : a comparison of tissue biopsies and swabs

Funding Information: The research costs of this work were supported by the BBSRC EASTBIO DTP and Marine Alliance for Science and Technology Scotland (MASTS) small grants funding scheme. Acknowledgements The authors would like to thank Scottish Sea Farms (SSF) for the kind facilitation of fieldwork that provided material in this project, particularly the staff at the Loch Spelve facility, and the health team at SSF, particularly Dr. Ralph Bickerdike. Thanks are due as well to Professor Matt Holden and Kerry Pettigrew of the Infection Group within the Biomedical Sciences Research Complex, School of Medicine, University of St Andrews, for assistance within the laboratory, as well as Dr. David Bass at the Centre for Environment Fisheries and Aquaculture Science for helpful proofreading.Peer reviewedPublisher PD

α-Conotoxin Vc1.1 inhibits human dorsal root ganglion neuroexcitability and mouse colonic nociception via GABA\u3csub\u3eB\u3c/sub\u3e receptors

Objective α-Conotoxin Vc1.1 is a small disulfidebonded peptide from the venom of the marine cone snail Conus victoriae. Vc1.1 has antinociceptive actions in animal models of neuropathic pain, but its applicability to inhibiting human dorsal root ganglion (DRG) neuroexcitability and reducing chronic visceral pain (CVP) is unknown. Design We determined the inhibitory actions of Vc1.1 on human DRG neurons and on mouse colonic sensory afferents in healthy and chronic visceral hypersensitivity (CVH) states. In mice, visceral nociception was assessed by neuronal activation within the spinal cord in response to noxious colorectal distension (CRD). Quantitativereverse- transcription-PCR, single-cell-reversetranscription- PCR and immunohistochemistry determined ?-aminobutyric acid receptor B (GABABR) and voltagegated calcium channel (CaV2.2, CaV2.3) expression in human and mouse DRG neurons. Results Vc1.1 reduced the excitability of human DRG neurons, whereas a synthetic Vc1.1 analogue that is inactive at GABABR did not. Human DRG neurons expressed GABABR and its downstream effector channels CaV2.2 and CaV2.3. Mouse colonic DRG neurons exhibited high GABABR, CaV2.2 and CaV2.3 expression, with upregulation of the CaV2.2 exon-37a variant during CVH. Vc1.1 inhibited mouse colonic afferents ex vivo and nociceptive signalling of noxious CRD into the spinal cord in vivo, with greatest efficacy observed during CVH. A selective GABABR antagonist prevented Vc1.1-induced inhibition, whereas blocking both CaV2.2 and CaV2.3 caused inhibition comparable with Vc1.1 alone. Conclusions Vc1.1-mediated activation of GABABR is a novel mechanism for reducing the excitability of human DRG neurons. Vc1.1-induced activation of GABABR on the peripheral endings of colonic afferents reduces nociceptive signalling. The enhanced antinociceptive actions of Vc1.1 during CVH suggest it is a novel candidate for the treatment for CVP

Impact of the Mt. Pinatubo volcaniceruption on the lower ionosphere andatmospheric waves over Central Europe

The very strong volcanic eruption of Mt. Pinatubo in June 1991 directly affected the troposphere and lower and middle stratosphere. Here we look at its effects in the mesopause region as revealed by the radio wave absorption measurements in the lower ionosphere over Central Europe and inferred planetary and gravity wave activity. The gravity wave activity inferred from the nighttime LF radio wave absorption displays an evident enhancement for waves of periods of about 2-3 h coinciding with regional measurements of the optical depth of (volcanic) aerosols, while there is no detectable effect for short period waves (T < 1 h). There is no detectable effect in the planetary wave activity inferred from the daytime HF radio wave absorption. As for the absorption itself, the results on the impact of the Mt. Pinatubo eruption do not provide an observable effect

Growth or decline in the Church of England during the decade of Evangelism: did the Churchmanship of the Bishop matter?

The Decade of Evangelism occupied the attention of the Church of England throughout the 1990s. The present study employs the statistics routinely published by the Church of England in order to assess two matters: the extent to which these statistics suggest that the 43 individual dioceses finished the decade in a stronger or weaker position than they had entered it and the extent to which, according to these statistics, the performance of dioceses led by bishops shaped in the Evangelical tradition differed from the performance of dioceses led by bishops shaped in the Catholic tradition. The data demonstrated that the majority of dioceses were performing less effectively at the end of the decade than at the beginning, in terms of a range of membership statistics, and that the rate of decline varied considerably from one diocese to another. The only exception to the trend was provided by the diocese of London, which experienced some growth. The data also demonstrated that little depended on the churchmanship of the diocesan bishop in shaping diocesan outcomes on the performance indicators employed in the study

Real-time risk analysis for hybrid earthquake early warning systems

Earthquake Early Warning Systems (EEWS), based on real-time prediction of ground motion or structural response measures, may play a role in reducing vulnerability and/or exposition of buildings and lifelines. In fact, recently seismologists developed efficient methods for rapid estimation of event features by means of limited information of the P-waves. Then, when an event is occurring, probabilistic distributions of magnitude and source-to-site distance are available and the prediction of the ground motion at the site, conditioned to the seismic network measures, may be performed in analogy with the Probabilistic Seismic Hazard Analysis (PSHA). Consequently the structural performance may be obtained by the Probabilistic Seismic Demand Analysis (PSDA), and used for real-time risk management purposes. However, such prediction is performed in very uncertain conditions which have to be taken into proper account to limit false and missed alarms. In the present study, real-time risk analysis for early warning purposes is discussed. The magnitude estimation is performed via the Bayesian approach, while the earthquake localization is based on the Voronoi cells. To test the procedure it was applied, by simulation, to the EEWS under development in the Campanian region (southern Italy). The results lead to the conclusion that the PSHA, conditioned to the EEWS, correctly predicts the hazard at the site and that the false/missed alarm probabilities may be controlled by set up of an appropriate decisional rule and alarm threshold

Evaluating a multi-component intervention to reduce and break up office workers' sitting with sit-stand desks using the APEASE criteria

Objective: Sedentary workplace interventions have had success in reducing excessive sitting time in office workers, but barriers to implementation and uptake remain. This study formally assessed a theory-derived, sit-stand desk intervention using the APEASE (Acceptability, Practicability, Effectiveness, Affordability, Side-effects, Equity) criteria. / Methods: Thirteen adults (eight female, mean age 38 ± 10 years) from the treatment arm of a sedentary behaviour intervention participated in semi-structured interviews. Thematic codes were inductively assigned to data items followed by deductive charting using the APEASE criteria. / Results: The intervention was highly acceptable, practicable, safe to deploy, and helped workers reduce workplace sitting time, though individual preferences and workload mediated engagement. Affordability of sit-stand desks and Equity of access were potential barriers to uptake. / Conclusions: Through the lens of the APEASE criteria, this theory-derived, multi-component sit-stand desk intervention showed acceptability, practicability and effectiveness in reducing and breaking up sedentary time at work with minimal side effects. Using this approach with further tailoring and personalisation may help workers achieve greater reductions in workplace sitting, though affordability and equity should be considered further

Persistence in Linux-Based IoT Malware

The Internet of Things (IoT) is a rapidly growing collection of “smart” devices capable of communicating over the Internet. Being connected to the Internet brings new features and convenience, but it also poses new security threats, such as IoT malware. IoT malware has shown similar growth, making IoT devices highly vulnerable to remote compromise. However, most IoT malware variants do not exhibit the ability to gain persistence, as they typically lose control over the compromised device when the device is restarted. This paper investigates how persistence for various IoT devices can be implemented by attackers, such that they retain control even after the device has been rebooted. Having persistence would make it harder to remove IoT malware. We investigated methods that could be used by an attacker to gain persistence on a variety of IoT devices, and compiled the requirements and potential issues faced by these methods, in order to understand how best to combat this future threat. We successfully used these methods to gain persistence on four vulnerable IoT devices with differing designs, features and architectures. We also identified ways to counter them. This work highlights the enormous risk that persistence poses to potentially billions of IoT devices, and we hope our results and study will encourage manufacturers and developers to consider implementing our proposed countermeasures or create new techniques to combat this nascent threat