Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy

Whilst there is a large body of evidence looking at the design of cationic liposomes as transfection agents, correlates of formulation to function remain elusive. In this research, we investigate if lipid packaging can give further insights into transfection efficacy. DNA lipoplexes composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method. Each of the formulations was prepared by hydration in dH2O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. In addition, Langmuir monolayer studies were performed to investigate any possible correlation between lipid packaging and liposome attributes. Using PBS, rather than dH2O, to prepare the lipoplexes increased the size of vesicles in most of formulations and resulted in variation in transfection efficacies. However, one combination of lipids (DSPE:DOTAP) could not form liposomes in PBS, whilst the DSPE:DSTAP combination could not form liposomes in either aqueous media. Monolayer studies demonstrated saturated lipid combinations offered dramatically closer molecular packing compared to the other combinations which could suggest why this lipid combination could not form vesicles. Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent

Rethinking the law and politics of democratic police accountability

This paper evaluates the work and impact of a number of Police and Crime Commissioners (PCCs) in England and Wales and attempts to refocus public discourse and scrutiny on their Police and Crime Plans as a key prism through which their performance should be measured. Drawing upon the literature published by various PCCs, the Stevens Commission, the Home Affairs Committee and numerous academics, the paper will argue that a major reform of democratic police accountability in England and Wales is needed. Due to the often voluminous and piecemeal nature of the documents published on the PCCs’ websites, the textual analysis is limited to the Police and Crime Plans for Greater Manchester, the West Midlands and the London Metropolitan are

Early-life course factors and oral health among young Norwegian adults

Objective Using a national sample of young Norwegian adults, we examined whether unpleasant experience with dental care during childhood is associated with tooth loss and oral health–related quality of life in adulthood after accounting for early- and later-life socio-behavioural circumstances and dental avoidance behaviour. Methods 2433 individuals aged 25-35 years participated in an electronic survey. Oral quality of life was measured using the oral impact of daily performance (OIDP) inventory. Generalized linear models and negative binomial regression models were used to estimate the association of early unpleasant experiences with dental care and tooth loss and OIDP scores. Incidence rate ratio (IRR) and 95% confidence intervals (CI) were used to estimate the relative differences in prevalence of tooth loss and OIDP scores. Results Adjusting for early-life characteristics only, the prevalence of tooth loss was 1.42 (IRR = 1.42, 95% CI: 1.24-1.64) and 1.96 (IRR = 1.96, 95% CI: 1.70-2.26) times higher among individuals who reported unpleasant experiences a few times or several times, than in individuals who did not report unpleasant experiences with dental care in childhood. Adjusting further for educational level, smoking and tooth brushing attenuated the relative differences (IRR = 1.40, 95% CI: 1.22-1.62 and IRR = 1.88, 95% CI: 1.62-2.17, respectively). Lastly, when adjusting for dental avoidance behaviour, the prevalence of tooth loss was 1.29 (IRR = 1.29, 95% CI: 1.11-1.50) and 1.58 (IRR = 1.58, 95% CI: 1.32-1.88) times higher among individuals who reported unpleasant experiences a few times or several times than in those who did not. Corresponding associations of early unpleasant experience with OIDP were (IRR = 1.41 95% CI: 1.22-1.63) and (IRR = 1.69, 95% CI: 1.42-2.01) when adjusting for early-life characteristics, and (IRR = 1.39, 95% CI: 1.20-1.60) and (IRR = 1.51, 95% CI: 1.27-1.80) when adjusting for education, smoking and tooth brushing. When adjusting for dental avoidance behaviour, the association of early unpleasant experience with OIDP became nonsignificant. Conclusion Unpleasant dental care experiences during childhood are associated with poor oral health in adulthood, independent of later-life socio-behavioural characteristics including negative dental care seeking. This highlights the importance of tailoring regular contacts with dental healthcare services in childhood to build confidence in children and thus has implications for healthcare policy.publishedVersio

Lead levels in fur of rats treated with inorganic lead measured by inductively coupled argon plasma mass spectrometry

The aim of this study was to investigate the relationship between continuous lead exposure and the concentration of this metal in fur. The two main questions we wanted to answer were: 1) Are the fur lead concentrations different according to exposure level? 2) Is the kinetics of lead concentration linear in different compartments

Phase transitions in biological membranes

Native membranes of biological cells display melting transitions of their lipids at a temperature of 10-20 degrees below body temperature. Such transitions can be observed in various bacterial cells, in nerves, in cancer cells, but also in lung surfactant. It seems as if the presence of transitions slightly below physiological temperature is a generic property of most cells. They are important because they influence many physical properties of the membranes. At the transition temperature, membranes display a larger permeability that is accompanied by ion-channel-like phenomena even in the complete absence of proteins. Membranes are softer, which implies that phenomena such as endocytosis and exocytosis are facilitated. Mechanical signal propagation phenomena related to nerve pulses are strongly enhanced. The position of transitions can be affected by changes in temperature, pressure, pH and salt concentration or by the presence of anesthetics. Thus, even at physiological temperature, these transitions are of relevance. There position and thereby the physical properties of the membrane can be controlled by changes in the intensive thermodynamic variables. Here, we review some of the experimental findings and the thermodynamics that describes the control of the membrane function.Comment: 23 pages, 15 figure

Computational optical imaging with a photonic lantern

[EN] The thin and flexible nature of optical fibres often makes them the ideal technology to view biological processes in-vivo, but current microendoscopic approaches are limited in spatial resolution. Here, we demonstrate a route to high resolution microendoscopy using a multicore fibre (MCF) with an adiabatic multimode-to-single-mode "photonic lantern" transition formed at the distal end by tapering. We show that distinct multimode patterns of light can be projected from the output of the lantern by individually exciting the single-mode MCF cores, and that these patterns are highly stable to fibre movement. This capability is then exploited to demonstrate a form of single-pixel imaging, where a single pixel detector is used to detect the fraction of light transmitted through the object for each multimode pattern. A custom computational imaging algorithm we call SARA-COIL is used to reconstruct the object using only the pre-measured multimode patterns themselves and the detector signals.This work was funded through the "Proteus" Engineering and Physical Sciences Research Council (EPSRC) Interdisciplinary Research Collaboration (IRC) (EP/K03197X/1), by the Science and Technology Facilities Council (STFC) through STFC-CLASP grants ST/K006509/1 and ST/K006460/1, STFC Consortium grants ST/N000625/1 and ST/N000544/1. S.L. acknowledges support from the National Natural Science Foundation of China under Grant no. 61705073. DBP acknowledges support from the Royal Academy of Engineering, and the European Research Council (PhotUntangle, 804626). The authors thank Philip Emanuel for the use of his confocal image of A549 cells and Eckhardt Optics for their image of the USAF 1951 target. The authors sincerely thank the anonymous reviewers of this paper for their detailed and considered feedback which helped us to improve the quality of this paper significantly.Choudhury, D.; Mcnicholl, DK.; Repetti, A.; Gris-Sánchez, I.; Li, S.; Phillips, DB.; Whyte, G.... (2020). Computational optical imaging with a photonic lantern. Nature Communications. 11(1):1-9. https://doi.org/10.1038/s41467-020-18818-6S19111Wood, H. A. C., Harrington, K., Birks, T. A., Knight, J. C. & Stone, J. M. High-resolution air-clad imaging fibers. Opt. Lett. 43, 5311–5314 (2018).Akram, A. R. et al. In situ identification of Gram-negative bacteria in human lungs using a topical fluorescent peptide targeting lipid A. Sci. Transl. Med. 10, eaal0033 (2018).Shin, J., Bosworth, B. T. & Foster, M. A. Compressive fluorescence imaging using a multi-core fiber and spatially dependent scattering. Opt. Lett. 42, 109–112 (2017).Papadopoulos, I. N., Farahi, S., Moser, C. & Psaltis, D. Focusing and scanning light through a multimode optical fiber using digital phase conjugation. Opt. Express 20, 10583–10590 (2012).Čižmár, T. & Dholakia, K. Exploiting multimode waveguides for pure fibre-based imaging. Nat. Commun. 3, 1027 (2012).Plöschner, M., Tyc, T. & Čižmár, T. Seeing through chaos in multimode fibres. Nat. Photon. 9, 529–535 (2015).Birks, T. A., Gris-Sánchez, I., Yerolatsitis, S., Leon-Saval, S. G. & Thomson, R. R. The photonic lantern. Adv. Opt. Photon. 7, 107–167 (2015).Birks, T. A., Mangan, B. J., Díez, A., Cruz, J. L. & Murphy, D. F. Photonic lantern’ spectral filters in multi-core fiber. Opt. Express 20, 13996–14008 (2012).Edgar, M. P., Gibson, G. M. & Padgett, M. J. Principles and prospects for single-pixel imaging. Nat. Photon. 13, 13–20 (2019).Mahalati, R. N., Yu, Gu. R. & Kahn, J. M. Resolution limits for imaging through multi-mode fiber. Opt. Express 21, 1656–1668 (2013).Amitonova, L. V. & de Boer, J. F. Compressive imaging through a multimode fiber. Opt. Lett. 43, 5427–5430 (2018).Mallat, S. A Wavelet Tour of Signal Processing 2nd edn (Academic Press, Burlington, MA, 2009).Lustig, M., Donoho, D. & Pauly, J. M. Sparse MRI: the application of compressed sensing for rapid MR imaging. Magn. Reson. Med. 58, 1182–1195 (2007).Davies, M., Puy, G., Vandergheynst, P. & Wiaux, Y. A compressed sensing framework for magnetic resonance fingerprinting. SIAM J. Imaging Sci. 7, 2623–2656 (2014).Wiaux, Y., Puy, G., Scaife, A. M. M. & Vandergheynst, P. Compressed sensing imaging techniques in radio interferometry. Mon. Not. R. Astron. Soc. 395, 1733–1742 (2009).Carrillo, R. E., McEwen, J. D. & Wiaux, Y. Sparsity averaging reweighted analysis (SARA): a novel algorithm for radio-interferometric imaging. Mon. Not. R. Astron. Soc. 426, 1223–1234 (2012).Katz, O., Bromberg, Y. & Silberberg, Y. Compressive ghost imaging. Appl. Phys. Lett. 95, 131110 (2009).Sun, B., Welsh, S. S., Edgar, M. P., Shapiro, J. H. & Padgett, M. J. Normalized ghost imaging. Opt. Express 20, 16892–16901 (2012).Kim, M., Park, C., Rodriguez, C., Park, Y. & Cho, Y.-H. Superresolution imaging with optical fluctuation using speckle patterns illumination. Sci. Rep. 5, 16525 (2015).Combettes, P. L. & Pesquet, J. -C. in Fixed-Point Algorithms for Inverse Problems in Science and Engineering (Springer, New York, 2011).Komodakis, N. & Pesquet, J.-C. Playing with duality: an overview of recent primal dual approaches for solving large-scale optimization problems. IEEE Signal Proc. Mag. 32, 31–54 (2015).Chandrasekharan, H. K. et al. Multiplexed single-mode wavelength-to-time mapping of multimode light. Nat. Commun. 8, 14080 (2017).Wadsworth, W. J. et al. Very high numerical aperture fibers. Photon. Technol. Lett. 16, 843–845 (2004).Pesquet, J.-C. & Repetti, A. A class of randomized primal-dual algorithms for distributed optimization. J. Nonlinear Convex Anal. 16, 2353–2490 (2015).Chambolle, A., Ehrhardt, M. J., Richtárik, P. & Schönlieb, C.-B. Stochastic primal-dual hybrid gradient algorithm with arbitrary sampling and imaging applications. SIAM J. Optim. 28, 2783–2808 (2018).Bolte, J., Sabach, S. & Teboulle, M. Proximal alternating linearized minimization for nonconvex and nonsmooth problems. Math. Program. 146, 459–494 (2014).Chouzenoux, E., Pesquet, J.-C. & Repetti, A. A block coordinate variable metric forward-backward algorithm. J. Glob. Optim. 66, 457–485 (2016).Flusberg, B. A. et al. Fiber-optic fluorescence imaging. Nat. Methods 2, 941–950 (2005).Tsvirkun, V. et al. Bending-induced inter-core group delays in multicore fibers. Opt. Express 25, 31863–31875 (2017).Candès, E. J., Wakin, M. B. & Boyd, S. Enhancing sparsity by reweighted l1 minimization. J. Fourier Anal. Appl. 14, 877–905 (2008).Condat, L. A primal–dual splitting method for convex optimization involving lipschitzian, proximable and linear composite terms. J. Optim. Theory Appl. 158, 460–479 (2013).Vu, B. C. A splitting algorithm for dual monotone inclusions involving cocoercive operators. Adv. Comp. Math. 38, 667–681 (2013)

Anomalous Diffusion Induced by Cristae Geometry in the Inner Mitochondrial Membrane

Diffusion of inner membrane proteins is a prerequisite for correct functionality of mitochondria. The complicated structure of tubular, vesicular or flat cristae and their small connections to the inner boundary membrane impose constraints on the mobility of proteins making their diffusion a very complicated process. Therefore we investigate the molecular transport along the main mitochondrial axis using highly accurate computational methods. Diffusion is modeled on a curvilinear surface reproducing the shape of mitochondrial inner membrane (IM). Monte Carlo simulations are carried out for topologies resembling both tubular and lamellar cristae, for a range of physiologically viable crista sizes and densities. Geometrical confinement induces up to several-fold reduction in apparent mobility. IM surface curvature per se generates transient anomalous diffusion (TAD), while finite and stable values of projected diffusion coefficients are recovered in a quasi-normal regime for short- and long-time limits. In both these cases, a simple area-scaling law is found sufficient to explain limiting diffusion coefficients for permeable cristae junctions, while asymmetric reduction of the junction permeability leads to strong but predictable variations in molecular motion rate. A geometry-based model is given as an illustration for the time-dependence of diffusivity when IM has tubular topology. Implications for experimental observations of diffusion along mitochondria using methods of optical microscopy are drawn out: a non-homogenous power law is proposed as a suitable approach to TAD. The data demonstrate that if not taken into account appropriately, geometrical effects lead to significant misinterpretation of molecular mobility measurements in cellular curvilinear membranes

Bayesian Model Selection Applied to the Analysis of Fluorescence Correlation Spectroscopy Data of Fluorescent Proteins in Vitro and in Vivo

Fluorescence correlation spectroscopy (FCS) is a powerful technique to investigate molecular dynamics with single molecule sensitivity. In particular, in the life sciences it has found widespread application using fluorescent proteins as molecularly specific labels. However, FCS data analysis and interpretation using fluorescent proteins remains challenging due to typically low signal-to-noise ratio of FCS data and correlated noise in autocorrelated data sets. As a result, naive fitting procedures that ignore these important issues typically provide similarly good fits for multiple competing models without clear distinction of which model is preferred given the signal-to-noise ratio present in the data. Recently, we introduced a Bayesian model selection procedure to overcome this issue with FCS data analysis. The method accounts for the highly correlated noise that is present in FCS data sets and additionally penalizes model complexity to prevent over interpretation of FCS data. Here, we apply this procedure to evaluate FCS data from fluorescent proteins assayed in vitro and in vivo. Consistent with previous work, we demonstrate that model selection is strongly dependent on the signal-to-noise ratio of the measurement, namely, excitation intensity and measurement time, and is sensitive to saturation artifacts. Under fixed, low intensity excitation conditions, physical transport models can unambiguously be identified. However, at excitation intensities that are considered moderate in many studies, unwanted artifacts are introduced that result in nonphysical models to be preferred. We also determined the appropriate fitting models of a GFP tagged secreted signaling protein, Wnt3, in live zebrafish embryos, which is necessary for the investigation of Wnt3 expression and secretion in development. Bayes model selection therefore provides a robust procedure to determine appropriate transport and photophysical models for fluorescent proteins when appropriate models are provided, to help detect and eliminate experimental artifacts in solution, cells, and in living organisms.National Science Foundation (U.S.). Physics of Living Systems ProgramNational Institute of Mental Health (U.S.) (Award U01MH106011