23 research outputs found

    An assessment of the causes of the errors in the 2015 UK General Election opinion polls

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    The opinion polls undertaken prior to the 2015 UK General Election under-estimated the Conservative lead over Labour by an average of 7 percentage points. This collective failure led politicians and commentators to question the validity and utility of political polling and raised concerns regarding a broader public loss of confidence in survey research. In this paper, we assess the likely causes of the 2015 polling errors. We begin by setting out a formal account of the statistical methodology and assumptions required for valid estimation of party vote shares using quota sampling. We then describe the current approach of polling organisations for estimating sampling variability and suggest a new method based on bootstrap re-sampling. Next, we use poll micro-data to assess the plausibility of different explanations of the polling errors. Our conclusion is that the primary cause of the polling errors in 2015 was unrepresentative sampling

    Cross-Interface Emulsification for Generating Size-Tunable Droplets

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    We report cross-interface emulsification (XiE), a simple method for the generation of monodisperse droplets of controllable volumes from picoliter to nanoliter. A device is set up in which a fused-silica capillary is vibrating across the surface of the continuous phase (mineral oil) in a reservoir, and the flow of the dispersed phase (aqueous solution) in the capillary is segmented into monodisperse droplets at the air/oil interface. We find that the volume of droplets is mainly dominated by the flow rate and vibrating frequency and not significantly influenced by other factors, such as the viscosity of the continuous phase and dispersed phase, the inner diameter of the capillary (20–100 μm), or the shape of the tip (tapered or flat). These features reflect high robustness, flexibility, and precision of XiE for on-demand volume control of droplets. The droplets automatically assemble into planar monolayer droplet arrays (PMDA) in flat-bottomed microwells of 96-well plates, offering excellent convenience for imaging of droplets. As a representative application, we carry out digital loop-mediated isothermal amplification using PMDAs with multivolume droplets for the absolute quantification of nucleic acids. Our results demonstrate that XiE is simple and controllable for the production of monodisperse size-tunable droplets, and it offers opportunities for common laboratories, even without microfabrication facilities, to perform digital quantification, single cell analysis, and other biochemical assays with high throughput

    Cross-Interface Emulsification for Generating Size-Tunable Droplets

    No full text
    We report cross-interface emulsification (XiE), a simple method for the generation of monodisperse droplets of controllable volumes from picoliter to nanoliter. A device is set up in which a fused-silica capillary is vibrating across the surface of the continuous phase (mineral oil) in a reservoir, and the flow of the dispersed phase (aqueous solution) in the capillary is segmented into monodisperse droplets at the air/oil interface. We find that the volume of droplets is mainly dominated by the flow rate and vibrating frequency and not significantly influenced by other factors, such as the viscosity of the continuous phase and dispersed phase, the inner diameter of the capillary (20–100 μm), or the shape of the tip (tapered or flat). These features reflect high robustness, flexibility, and precision of XiE for on-demand volume control of droplets. The droplets automatically assemble into planar monolayer droplet arrays (PMDA) in flat-bottomed microwells of 96-well plates, offering excellent convenience for imaging of droplets. As a representative application, we carry out digital loop-mediated isothermal amplification using PMDAs with multivolume droplets for the absolute quantification of nucleic acids. Our results demonstrate that XiE is simple and controllable for the production of monodisperse size-tunable droplets, and it offers opportunities for common laboratories, even without microfabrication facilities, to perform digital quantification, single cell analysis, and other biochemical assays with high throughput

    Heterogeneous Nanostructures Cause Anomalous Diffusion in Lipid Monolayers

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    The diffusion and mobility in biomembranes are crucial for various cell functions; however, the mechanisms involved in such processes remain ambiguous due to the complex membrane structures. Herein, we investigate how the heterogeneous nanostructures cause anomalous diffusion in dipalmitoylphosphatidylcholine (DPPC) monolayers. By identifying the existence of condensed nanodomains and clarifying their impact, our findings renew the understanding of the hydrodynamic description and the statistical feature of the diffusion in the monolayers. We find a universal characteristic of the multistage mean square displacement (MSD) with an intermediate crossover, signifying two membrane viscosities at different scales: the short-time scale describes the local fluidity and is independent of the nominal DPPC density, and the long-time scale represents the global continuous phase taking into account nanodomains and increases with DPPC density. The constant short-time viscosity reflects a dynamic equilibrium between the continuous fluid phase and the condensed nanodomains in the molecular scale. Notably, we observe an “anomalous yet Brownian” phenomenon exhibiting an unusual double-peaked displacement probability distribution (DPD), which is attributed to the net dipolar repulsive force from the heterogeneous nanodomains around the microdomains. The findings provide physical insights into the transport of membrane inclusions that underpin various biological functions and drug deliveries

    PKC and downstream NF-κB are required for IL-6-induced proliferation in CD45<sup>+</sup> myeloma cells.

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    <p>CD45<sup>+</sup> U266 cells (A) or CD45<sup>+</sup> CD138<sup>+</sup> primary cells (B) were incubated with or without PKC inhibitor Ro31-8220 (1 μM), or NF-κB inhibitor BAY11-7082 (5 μM) for 1 hour and stimulated with IL-6 (10 ng/ml). PKC phosphorylation level was measured after 10 minutes of stimulation with IL-6, and the IκB phosphorylation level was analyzed after 60 minutes stimulation. Western blotting was performed by using specific antibodies. The representative blots of three independent experiments are shown. BrdU incorporation was used to detect the DNA synthesis by IL-6 after 72 hours. DMSO is used as a control. Data are shown as mean ± SD of triplicate cultures and are from one experiment representative of three performed. ** <i>p</i> < 0.01 vs DMSO control in the absence of IL-6; ## <i>p</i> < 0.01 vs DMSO control in the presence of IL-6 by a one-way ANOVA with HSD test.</p

    Probing Non-Gaussianity in Confined Diffusion of Nanoparticles

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    Confined diffusion is ubiquitous in nature. Ever since the “anomalous yet Brownian” motion was observed, the non-Gaussianity in confined diffusion has been unveiled as an important issue. In this Letter, we experimentally investigate the characteristics and source of non-Gaussian behavior in confined diffusion of nanoparticles suspended in polymer solutions. A time-varied and size-dependent non-Gaussianity is reported based on the non-Gaussian parameter and displacement probability distribution, especially when the nanoparticle’s size is smaller than the typical polymer mesh size. This non-Gaussianity does not vanish even at the long-time Brownian stage. By inspecting the displacement autocorrelation, we observe that the nanoparticle–structure interaction, indicated by the anticorrelation, is limited in the short-time stage and makes little contribution to the non-Gaussianity in the long-time stage. The main source of the non-Gaussianity can therefore be attributed to hopping diffusion that results in an exponential probability distribution with the large displacements, which may also explain certain processes dominated by rare events in the biological environment

    Model of CD45 isoforms-mediated IL-6 signaling in multiple myeloma cells.

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    <p>Engagement of IL-6R with IL-6 leads to complex formation of IL-6R, gp130, Lyn as well as CD45RO/RB in raft microdomains. In response to IL-6 stimulation, CD45RO/RB moves into lipid rafts to induce dephosphorylation of the negative regulatory of Tyr507, phosphorylation of Tyr396, and subsequent conformation change and Lyn activation [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0119780#pone.0119780.ref017" target="_blank">17</a>]. We confirmed our hypothesis that lipid rafts-targeted CD45RO/RB facilitates IL-6-induced STAT3 and Lyn/PKC/NF-κB activation in rafts microdomains, while raft-excluded CD45RA remains outside of lipid rafts after IL-6 stimulation and negatively regulates ERK-activation.</p

    Effects of Exogenously expressed CD45RO, RB, or RA-EGFP on phosphorylation of STAT3, MAPK, PKC and IκB.

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    <p>CD45<sup>-</sup> U266 cells were transfected with untagged (mock) or CD45RO-EGFP, CD45RB-EGFP or CD45RA-EGFP expression plasmids, and either treated or untreated with IL-6 (10 ng/ml) at different time points. The representative blots are from three independent experiments and separate blotting using antibodies to P-STAT3, STAT3 (A), P-ERK, ERK (B), P-PKC, PKC (C) and P-IκB, IκB (D) are shown. The densities of protein bands were determined by densitometry and the data represent a change from the control mock density. * <i>p</i> < 0.05 vs mock control in the presence of IL-6; ** <i>p</i> < 0.01 vs mock control in the presence of IL-6 by a one-way ANOVA with HSD test.</p
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