Inactivation properties of sodium channel Nav1.8 maintain action potential amplitude in small DRG neurons in the context of depolarization

<p>Abstract</p> <p>Background</p> <p>Small neurons of the dorsal root ganglion (DRG) express five of the nine known voltage-gated sodium channels. Each channel has unique biophysical characteristics which determine how it contributes to the generation of action potentials (AP). To better understand how AP amplitude is maintained in nociceptive DRG neurons and their centrally projecting axons, which are subjected to depolarization within the dorsal horn, we investigated the dependence of AP amplitude on membrane potential, and how that dependence is altered by the presence or absence of sodium channel Na_v1.8.</p> <p>Results</p> <p>In small neurons cultured from wild type (WT) adult mouse DRG, AP amplitude decreases as the membrane potential is depolarized from -90 mV to -30 mV. The decrease in amplitude is best fit by two Boltzmann equations, having V_1/2values of -73 and -37 mV. These values are similar to the V_1/2values for steady-state fast inactivation of tetrodotoxin-sensitive (TTX-s) sodium channels, and the tetrodotoxin-resistant (TTX-r) Na_v1.8 sodium channel, respectively. Addition of TTX eliminates the more hyperpolarized V_1/2component and leads to increasing AP amplitude for holding potentials of -90 to -60 mV. This increase is substantially reduced by the addition of potassium channel blockers. In neurons from Na_v1.8(-/-) mice, the voltage-dependent decrease in AP amplitude is characterized by a single Boltzmann equation with a V_1/2value of -55 mV, suggesting a shift in the steady-state fast inactivation properties of TTX-s sodium channels. Transfection of Na_v1.8(-/-) DRG neurons with DNA encoding Na_v1.8 results in a membrane potential-dependent decrease in AP amplitude that recapitulates WT properties.</p> <p>Conclusion</p> <p>We conclude that the presence of Na_v1.8 allows AP amplitude to be maintained in DRG neurons and their centrally projecting axons even when depolarized within the dorsal horn.</p

Consequences of Biomarker Analysis on the Cost-Effectiveness of Cetuximab in Combination with FOLFIRI as a First-Line Treatment of Metastatic Colorectal Cancer: Personalised Medicine at Work

Background Therapies may be more efficacious when targeting a patient subpopulation with specific attributes, thereby enhancing the cost-effectiveness of treatment. In the CRYSTAL study, patients with metastatic colorectal cancer (mCRC) were treated with cetuximab plus FOLFIRI or FOLFIRI alone until disease progression, unacceptable toxic effects or withdrawal of consent. Objective To determine if stratified use of cetuximab based on genetic biomarker detection improves cost-effectiveness. Methods We used individual patient data from CRYSTAL to compare the cost-effectiveness, cost per life-year (LY) and cost per quality-adjusted LY (QALY) gained of cetuximab plus FOLFIRI versus FOLFIRI alone in three cohorts of patients with mCRC: all randomised patients (intent-to-treat; ITT), tumours with no detectable mutations in codons 12 and 13 of exon 2 of the KRAS protein (‘KRAS wt’) and no detectable mutations in exons 2, 3 and 4 of KRAS and exons 2, 3 and 4 of NRAS (‘RAS wt’). Survival analysis was conducted using RStudio, and a cost-utility model was modified to allow comparison of the three cohorts. Results The deterministic base-case ICER (cost per QALY gained) was £130,929 in the ITT, £72,053 in the KRAS wt and £44,185 in the RAS wt cohorts for cetuximab plus FOLFIRI compared with FOLFIRI alone. At a £50,000 willingness-to-pay threshold, cetuximab plus FOLFIRI has a 2.8, 20 and 63% probability of being cost-effective for the ITT, KRAS wt and RAS wt cohorts, respectively, versus FOLFIRI alone. Conclusion Screening for mutations in both KRAS and NRAS may provide the most cost-effective approach to patient selection

Interweaving in hybrid methodologies

The paper will consider instances of the interweaving of theory and practice within drawing research, in order to suggest potential approaches to the development of hybrid methodologies in fine art practice-led research. The paper is written from the position of two current supervisors and creative research collaborators: Deborah Harty and Phil Sawdon (aka humhyphenhum from 2007), who historically were supervisee/supervisor. The paper will make reference to Harty's experience as a Ph.D. researcher undertaking practice-led research within a fine art context (completed 2010) and supervised by Sawdon. A discussion of Harty's hybrid methodology: action theoria, will provide an instance of the interweaving of theory and practice. Action theoria incorporates the cyclical and iterative process of action research – intention; action; review – with a process of theoria – the dialogue of both practice and theory's relationship to a given subject matter. Following this, the paper will discuss the interweaving of action theoria into humhyphenhum's collaborative research methodology: meaningful play. This interwoven methodology evolved during collaborative practice-led research projects from 2005 to the present. The paper will make reference to several of humhyphenhum's projects as a means to identify the interweaving of theory and practice within collaborative research. As current supervisors (2015), the paper will conclude with a discussion of how reflection on these experiences has informed our position as supervisors. We will consider, for example, how this has impacted on our ability, as individual supervisors, to offer insights into the interweaving of theory and practice, without defaulting to the position of compelling our supervisees to adopt our methodology

Probing Qubit Memory Errors at the Part-per-Million Level

Robust qubit memory is essential for quantum computing, both for near-term devices operating without error correction, and for the long-term goal of a fault-tolerant processor. We directly measure the memory error $\epsilon_m$ for a $^{43}$Ca$^+$ trapped-ion qubit in the small-error regime and find $\epsilon_m<10^{-4}$ for storage times t\lesssim50\,\mbox{ms}. This exceeds gate or measurement times by three orders of magnitude. Using randomized benchmarking, at t=1\,\mbox{ms} we measure $\epsilon_m=1.2(7)\times10^{-6}$, around ten times smaller than that extrapolated from the $T_{2}^{\ast}$ time, and limited by instability of the atomic clock reference used to benchmark the qubit.Comment: 8 pages, 5 figure

Timing of ergogenic aids and micronutrients on muscle and exercise performance

The timing of macronutrient ingestion in relation to exercise is a purported strategy to augment muscle accretion, muscle and athletic performance, and recovery. To date, the majority of macronutrient nutrient timing research has focused on carbohydrate and protein intake. However, emerging research suggests that the strategic ingestion of various ergogenic aids and micronutrients may also have beneficial effects. Therefore, the purpose of this narrative review is to critically evaluate and summarize the available literature examining the timing of ergogenic aids (caffeine, creatine, nitrates, sodium bicarbonate, beta-alanine) and micronutrients (iron, calcium) on muscle adaptations and exercise performance. In summary, preliminary data is available to indicate the timing of caffeine, nitrates, and creatine monohydrate may impact outcomes such as exercise performance, strength gains and other exercise training adaptations. Furthermore, data is available to suggest that timing the administration of beta-alanine and sodium bicarbonate may help to minimize known untoward adverse events while maintaining potential ergogenic outcomes. Finally, limited data indicates that timed ingestion of calcium and iron may help with the uptake and metabolism of these nutrients. While encouraging, much more research is needed to better understand how timed administration of these nutrients and others may impact performance, health, or other exercise training outcomes

Risk Assessment and the Effects of Refuge Availability on the Defensive Behaviors of the Southern Unstriped Scorpion (Vaejovis carolinianus)

Selection should favor individuals that acquire, process, and act on relevant environmental signals to avoid predation. Studies have found that scorpions control their use of venom: both when it is released and the total volume expelled. However, this research has not included how a scorpion’s awareness of environmental features influences these decisions. The current study tested 18 Vaejovis carolinianus scorpions (nine females and nine males) by placing them in circular arenas supplied with varying numbers (zero, two, or four) of square refuges and by tracking their movements overnight. The following morning, defensive behaviors were elicited by prodding scorpions on the chelae, prosoma, and metasoma once per second over 90 s. We recorded stings, venom use, chelae pinches, and flee duration. We found strong evidence that, across all behaviors measured, V. carolinianus perceived prods to the prosoma as more threatening than prods to the other locations. We found that stinging was a common behavior and became more dominant as the threat persisted. Though tenuous, we found evidence that scorpions’ defensive behaviors changed based on the number of refuges and that these differences may be sex specific. Our findings suggest that V. carolinianus can assess risk and features of the local environment and, therefore, alter their defensive strategies accordingly

Temperate Grassland Yields and Nitrogen Uptake Are Influenced by Fertilizer Nitrogen Source

This research was supported under the National Development Plan through the Research Stimulus Fund administered by the Department of Agriculture, Food and the Marine (Grants RSF10-/RD/SC/716 and RSF11S138) and from the Department of Agriculture and Rural Development for Northern Ireland and by the Teagasc Walsh Fellowship Scheme.peer reviewedIn temperate grasslands, N source influences greenhouse gas emissions. Nitrification and urea hydrolysis inhibitors can reduce these losses. The objective of this study was to evaluate the impact of N source, urease inhibitors, and nitrification inhibitors on temperate grassland yields and N uptake. Experiments were conducted at three locations over 2 years (6 site-years) on the island of Ireland, covering a range of soils and climatic conditions. Results showed that calcium ammonium nitrate (CAN), urea+N-(n-butyl) thiophosphoric triamide (NBPT), urea+NBPT+dicyandiamide (DCD), and urea had equal annual dry matter yield. Urea+DCD had lower dry matter yield than CAN for 3 site-years. Calcium ammonium nitrate and urea+NBPT consistently had the same N uptake, urea+DCD had lower N uptake than CAN in 4 of 6 site-years, urea had lower N uptake than CAN in 2 site-years, and urea+NBPT+DCD had lower N uptake than CAN in 1 site-year. Urea+NBPT is a cost-effective alternative to CAN, which is consistently equal in terms of yield and N uptake in temperate grassland.Teagasc Walsh Fellowship ProgrammeDepartment of Agriculture and Rural Development for Northern IrelandDepartment of Agriculture, Food and the Marin

Mitochondrial Activity and Cyr1 are Key Regulators of Ras1 Activation of C. albicans Virulence Pathways

Candida albicans is both a major fungal pathogen and a member of the commensal human microflora. The morphological switch from yeast to hyphal growth is associated with disease and many environmental factors are known to influence the yeast-to-hyphae switch. The Ras1-Cyr1-PKA pathway is a major regulator of C. albicans morphogenesis as well as biofilm formation and white-opaque switching. Previous studies have shown that hyphal growth is strongly repressed by mitochondrial inhibitors. Here, we show that mitochondrial inhibitors strongly decreased Ras1 GTP-binding and activity in C. albicans and similar effects were observed in other Candida species. Consistent with there being a connection between respiratory activity and GTP-Ras1 binding, mutants lacking complex I or complex IV grew as yeast in hypha-inducing conditions, had lower levels of GTP-Ras1, and Ras1 GTP-binding was unaffected by respiratory inhibitors. Mitochondria-perturbing agents decreased intracellular ATP concentrations and metabolomics analyses of cells grown with different respiratory inhibitors found consistent perturbation of pyruvate metabolism and the TCA cycle, changes in redox state, increased catabolism of lipids, and decreased sterol content which suggested increased AMP kinase activity. Biochemical and genetic experiments provide strong evidence for a model in which the activation of Ras1 is controlled by ATP levels in an AMP kinase independent manner. The Ras1 GTPase activating protein, Ira2, but not the Ras1 guanine nucleotide exchange factor, Cdc25, was required for the reduction of Ras1-GTP in response to inhibitor-mediated reduction of ATP levels. Furthermore, Cyr1, a well-characterized Ras1 effector, participated in the control of Ras1-GTP binding in response to decreased mitochondrial activity suggesting a revised model for Ras1 and Cyr1 signaling in which Cyr1 and Ras1 influence each other and, together with Ira2, seem to form a master-regulatory complex necessary to integrate different environmental and intracellular signals, including metabolic status, to decide the fate of cellular morphology

Influence of Maternal Protein Restriction in Primiparous Heifers During Mid- and/or Late-gestation on Dam and Suckling Calf Performance

Nutrient status in gestating beef cows has been shown to impact performance of the dam and offspring; however, most research has focused on energy or a total diet restriction and a single period of gestation. The objective of this study was to evaluate the effects of maternal metabolizable protein (MP) restriction in primiparous heifers during mid- and/or late gestation on dam and suckling calf performance through weaning

Background-free detection of trapped ions

We demonstrate a Doppler cooling and detection scheme for ions with low-lying D levels which almost entirely suppresses scattered laser light background, while retaining a high fluorescence signal and efficient cooling. We cool a single ion with a laser on the 2S1/2 to 2P1/2 transition as usual, but repump via the 2P3/2 level. By filtering out light on the cooling transition and detecting only the fluorescence from the 2P_3/2 to 2S1/2 decays, we suppress the scattered laser light background count rate to 1 per second while maintaining a signal of 29000 per second with moderate saturation of the cooling transition. This scheme will be particularly useful for experiments where ions are trapped in close proximity to surfaces, such as the trap electrodes in microfabricated ion traps, which leads to high background scatter from the cooling beam