Highly potent delivery method of gp160 envelope vaccine combining lentivirus-like particles and DNA electrotransfer

Particulate antigen assemblies in the nanometer range and DNA plasmids are particularly interesting for designing vaccines. We hypothesised that a combination of these approaches could result in a new delivery method of gp160 envelope HIV-1 vaccine which could combine the potency of virus-like particles (VLPs) and the simplicity of use of DNA vaccines. Characterisation of lentivirus-like particles (lentiVLPs) by western blot, dynamic light scattering and electron microscopy revealed that their protein pattern, size and structure make them promising candidates for HIV-1 vaccines. Although all particles were similar with regard to size and distribution, they clearly differed in p24 capsid protein content suggesting that Rev may be required for particle maturation and Gag processing. In vivo, lentiVLP pseudotyping with the gp160 envelope or with a combination of gp160 and VSV-G envelopes did not influence the magnitude of the immune response but the combination of lentiVLPs with Alum adjuvant resulted in a more potent response. Interestingly, the strongest immune response was obtained when plasmids encoding lentiVLPs were co-delivered to mice muscles by electrotransfer, suggesting that lentiVLPs were efficiently produced in vivo or the packaging genes mediate an adjuvant effect. DNA electrotransfer of plasmids encoding lentivirus-like particles offers many advantages and appears therefore as a promising delivery method of HIV-1 vaccines. Keywords:VLP, Electroporation, Electrotransfer, HIV vaccine, DNA vaccine

Application of the direct Liapunov method to the problem of symmetric stability in the atmosphere

The problem of symmetric stability is examined within the context of the direct Liapunov method. The sufficient conditions for stability derived by Fjørtoft are shown to imply finite-amplitude, normed stability. This finite-amplitude stability theorem is then used to obtain rigorous upper bounds on the saturation amplitude of disturbances to symmetrically unstable flows.By employing a virial functional, the necessary conditions for instability implied by the stability theorem are shown to be in fact sufficient for instability. The results of Ooyama are improved upon insofar as a tight two-sided (upper and lower) estimate is obtained of the growth rate of (modal or nonmodal) symmetric instabilities.The case of moist adiabatic systems is also considered.

An assessment of a three-beam Doppler lidar wind profiling method for use in urban areas

Currently there are few observations of the urban wind field at heights other than rooftop level. Remote sensing instruments such as Doppler lidars provide wind speed data at many heights, which would be useful in determining wind loadings of tall buildings, and predicting local air quality. Studies comparing remote sensing with traditional anemometers carried out in flat, homogeneous terrain often use scan patterns which take several minutes. In an urban context the flow changes quickly in space and time, so faster scans are required to ensure little change in the flow over the scan period. We compare 3993 h of wind speed data collected using a three-beam Doppler lidar wind profiling method with data from a sonic anemometer (190 m). Both instruments are located in central London, UK; a highly built-up area. Based on wind profile measurements every 2 min, the uncertainty in the hourly mean wind speed due to the sampling frequency is 0.05–0.11 m s−1. The lidar tended to overestimate the wind speed by ≈0.5 m s−1 for wind speeds below 20 m s−1. Accuracy may be improved by increasing the scanning frequency of the lidar. This method is considered suitable for use in urban areas.

Representativity error for temperature and humidity using the Met Office high-resolution model

The observation-error covariance matrix used in data assimilation contains contributions from instrument errors, representativity errors and errors introduced by the approximated observation operator. Forward model errors arise when the observation operator does not correctly model the observations or when observations can resolve spatial scales that the model cannot. Previous work to estimate the observation-error covariance matrix for particular observing instruments has shown that it contains signifcant correlations. In particular, correlations for humidity data are more significant than those for temperature. However it is not known what proportion of these correlations can be attributed to the representativity errors. In this article we apply an existing method for calculating representativity error, previously applied to an idealised system, to NWP data. We calculate horizontal errors of representativity for temperature and humidity using data from the Met Office high-resolution UK variable resolution model. Our results show that errors of representativity are correlated and more significant for specific humidity than temperature. We also find that representativity error varies with height. This suggests that the assimilation scheme may be improved if these errors are explicitly included in a data assimilation scheme. This article is published with the permission of the Controller of HMSO and the Queen's Printer for Scotland.

Method of determining a geophysical-scale sea ice rheology from laboratory measurements

We present a methodology that allows a sea ice rheology, suitable for use in a General Circulation Model (GCM), to be determined from laboratory and tank experiments on sea ice when combined with a kinematic model of deformation. The laboratory experiments determine a material rheology for sea ice, and would investigate a nonlinear friction law of the form τ ∝ σ n⅔, instead of the more familiar Amonton's law, τ = μσn (τ is the shear stress, μ is the coefficient of friction and σ n is the normal stress). The modelling approach considers a representative region R containing ice floes (or floe aggregates), separated by flaws. The deformation of R is imposed and the motion of the floes determined using a kinematic model, which will be motivated from SAR observations. Deformation of the flaws is inferred from the floe motion and stress determined from the material rheology. The stress over R is then determined from the area-weighted contribution from flaws and floes

Integrated visual vocabulary in latent Dirichlet allocation-based scene classification for IKONOS image

Scene classification based on latent Dirichlet allocation (LDA) is a more general modeling method known as a bag of visual words, in which the construction of a visual vocabulary is a crucial quantization process to ensure success of the classification. A framework is developed using the following new aspects: Gaussian mixture clustering for the quantization process, the use of an integrated visual vocabulary (IVV), which is built as the union of all centroids obtained from the separate quantization process of each class, and the usage of some features, including edge orientation histogram, CIELab color moments, and gray-level co-occurrence matrix (GLCM). The experiments are conducted on IKONOS images with six semantic classes (tree, grassland, residential, commercial/industrial, road, and water). The results show that the use of an IVV increases the overall accuracy (OA) by 11 to 12% and 6% when it is implemented on the selected and all features, respectively. The selected features of CIELab color moments and GLCM provide a better OA than the implementation over CIELab color moment or GLCM as individuals. The latter increases the OA by only ∼2 to 3%. Moreover, the results show that the OA of LDA outperforms the OA of C4.5 and naive Bayes tree by ∼20%. © 2014 Society of Photo-Optical Instrumentation Engineers (SPIE) [DOI: 10.1117/1.JRS.8.083690]

Visual sensor fusion for active security in robotic industrial environments

This work presents a method of information fusion involving data captured by both a standard CCD camera and a ToF camera to be used in the detection of the proximity between a manipulator robot and a human. Both cameras are assumed to be located above the work area of an industrial robot. The fusion of colour images and time of light information makes it possible to know the 3D localization of objects with respect to a world coordinate system. At the same time this allows to know their colour information. Considering that ToF information given by the range camera contains innacuracies including distance error, border error, and pixel saturation, some corrections over the ToF information are proposed and developed to improve the results. The proposed fusion method uses the calibration parameters of both cameras to reproject 3D ToF points, expressed in a common coordinate system for both cameras and a robot arm, in 2D colour images. In addition to this, using the 3D information, the motion detection in a robot industrial environment is achieved, and the fusion of information is applied to the foreground objects previously detected. This combination of information results in a matrix that links colour and 3D information, giving the possibility of characterising the object by its colour in addition to its 3D localization. Further development of these methods will make it possible to identify objects and their position in the real world, and to use this information to prevent possible collisions between the robot and such objects.

The bioactivity of composite Fmoc-RGDS-collagen gels

The incorporation of small bioactive peptide motifs within robust hydrogels constitutes a facile procedure to chemically functionalise cell and tissue scaffolds. In this study, a novel approach to utilise Fmoc-linked peptide amphiphiles comprising the bio-functional cell-adhesion RGDS motif within biomimetic collagen gels was developed. The composite scaffolds thus created were shown to maintain the mechanical properties of the collagen gel while presenting additional bio-activity. In particular, these materials enhanced the adhesion and proliferation of viable human corneal stromal fibroblasts by 300% compared to nonfunctionalised gels. Furthermore, the incorporation of Fmoc-RGDS nanostructures within the collagen matrix significantly suppressed gel shrinkage resulting from the contractile action of encapsulated fibroblasts once activated by serum proteins. These mechanical and biological properties demonstrate that the incorporation of peptide amphiphiles provides a suitable and easy method to circumvent specific biomaterial limitations, such as cell-derived shrinkage, for improved performance in tissue engineering and regenerative medicine applications.

Visual sensor fusion for active security in robotic industrial environments

This work presents a method of information fusion involving data captured by both a standard charge-coupled device (CCD) camera and a time-of-flight (ToF) camera to be used in the detection of the proximity between a manipulator robot and a human. Both cameras are assumed to be located above the work area of an industrial robot. The fusion of colour images and time-of-flight information makes it possible to know the 3D localization of objects with respect to a world coordinate system. At the same time, this allows to know their colour information. Considering that ToF information given by the range camera contains innacuracies including distance error, border error, and pixel saturation, some corrections over the ToF information are proposed and developed to improve the results. The proposed fusion method uses the calibration parameters of both cameras to reproject 3D ToF points, expressed in a common coordinate system for both cameras and a robot arm, in 2D colour images. In addition to this, using the 3D information, the motion detection in a robot industrial environment is achieved, and the fusion of information is applied to the foreground objects previously detected. This combination of information results in a matrix that links colour and 3D information, giving the possibility of characterising the object by its colour in addition to its 3D localisation. Further development of these methods will make it possible to identify objects and their position in the real world and to use this information to prevent possible collisions between the robot and such objects.

Proposal of a floating mechanism based on magnetic levitation method toward EEG-driven robotic system

We discussed a floating mechanism based on quasi-magnetic levitation method that can be attached at the endpoint of a robot arm in order to construct a novel redundant robot arm for producing compliant motions. The floating mechanism can be composed of magnets and a constraint mechanism such that the repelling force of the magnets floats the endpoint part of the mechanism stable for the guided motions. The analytical and experimental results show that the proposed floating mechanism can produce stable floating motions with small inertia and viscosity. The results also show that the proposed mechanism can detect small force applied to the endpoint part because the friction force of the mechanism is very small.

A novel ensemble method for retrieving properties of warm cloud in 3-D using ground-based scanning radar and zenith radiances

We present a novel method for retrieving high-resolution, three-dimensional (3-D) nonprecipitating cloud fields in both overcast and broken-cloud situations. The method uses scanning cloud radar and multiwavelength zenith radiances to obtain gridded 3-D liquid water content (LWC) and effective radius (re) and 2-D column mean droplet number concentration (Nd). By using an adaption of the ensemble Kalman filter, radiances are used to constrain the optical properties of the clouds using a forward model that employs full 3-D radiative transfer while also providing full error statistics given the uncertainty in the observations. To evaluate the new method, we first perform retrievals using synthetic measurements from a challenging cumulus cloud field produced by a large-eddy simulation snapshot. Uncertainty due to measurement error in overhead clouds is estimated at 20% in LWC and 6% in re, but the true error can be greater due to uncertainties in the assumed droplet size distribution and radiative transfer. Over the entire domain, LWC and re are retrieved with average error 0.05–0.08 g m-3 and ~2 μm, respectively, depending on the number of radiance channels used. The method is then evaluated using real data from the Atmospheric Radiation Measurement program Mobile Facility at the Azores. Two case studies are considered, one stratocumulus and one cumulus. Where available, the liquid water path retrieved directly above the observation site was found to be in good agreement with independent values obtained from microwave radiometer measurements, with an error of 20 g m-2.

An optimized method for the extraction of bacterial mRNA from plant roots infected with Escherichia coli O157:H7

Analysis of microbial gene expression during host colonization provides valuable information on the nature of interaction, beneficial or pathogenic, and the adaptive processes involved. Isolation of bacterial mRNA for in planta analysis can be challenging where host nucleic acid may dominate the preparation, or inhibitory compounds affect downstream analysis, e.g., quantitative reverse transcriptase PCR (qPCR), microarray, or RNA-seq. The goal of this work was to optimize the isolation of bacterial mRNA of food-borne pathogens from living plants. Reported methods for recovery of phytopathogen-infected plant material, using hot phenol extraction and high concentration of bacterial inoculation or large amounts of infected tissues, were found to be inappropriate for plant roots inoculated with Escherichia coli O157:H7. The bacterial RNA yields were too low and increased plant material resulted in a dominance of plant RNA in the sample. To improve the yield of bacterial RNA and reduce the number of plants required, an optimized method was developed which combines bead beating with directed bacterial lysis using SDS and lysozyme. Inhibitory plant compounds, such as phenolics and polysaccharides, were counteracted with the addition of high-molecular-weight polyethylene glycol and hexadecyltrimethyl ammonium bromide. The new method increased the total yield of bacterial mRNA substantially and allowed assessment of gene expression by qPCR. This method can be applied to other bacterial species associated with plant roots, and also in the wider context of food safety.

Lattice Boltzmann method for simulating the viscous flow in large distensible blood vessels

A lattice Boltzmann method for simulating the viscous flow in large distensible blood vessels is presented by introducing a boundary condition for elastic and moving boundaries. The mass conservation for the boundary condition is tested in detail. The viscous flow in elastic vessels is simulated with a pressure-radius relationship similar to that of the Pulmonary blood vessels. The numerical results for steady flow agree with the analytical prediction to very high accuracy, and the simulation results for pulsatile flow are comparable with those of the aortic flows observed experimentally. The model is expected to find many applications for studying blood flows in large distensible arteries, especially in those suffering from atherosclerosis. stenosis. aneurysm, etc.

A novel transport assimilation method for the Atlantic meridional overturning circulation at 26°N

One of the prerequisites for achieving skill in decadal climate prediction is to initialize and predict the circulation in the Atlantic Ocean successfully. The RAPID array measures the Atlantic Meridional Overturning Circulation (MOC) at 26°N. Here we develop a method to include these observations in the Met Office Decadal Prediction System (DePreSys). The proposed method uses covariances of overturning transport anomalies at 26°N with ocean temperature and salinity anomalies throughout the ocean to create the density structure necessary to reproduce the observed transport anomaly. Assimilating transport alone in this way effectively reproduces the observed transport anomalies at 26°N and is better than using basin-wide temperature and salinity observations alone. However, when the transport observations are combined with in situ temperature and salinity observations in the analysis, the transport is not currently reproduced so well. The reasons for this are investigated using pseudo-observations in a twin experiment framework. Sensitivity experiments show that the MOC on monthly time-scales, at least in the HadCM3 model, is modulated by a mechanism where non-local density anomalies appear to be more important for transport variability at 26°N than local density gradients.