Preparation and characterization of poly(lactic-co-glycolic acid) microparticles containing DNA molecules encoding a malaria vaccine candidate

Background A novel ultrasonic atomization approach for the formulation of biodegradable poly(lactic-co-glycolic acid) (PLGA) microparticles of a malaria DNA vaccine is presented. A 40 kHz ultrasonic atomization device was used to create the microparticles from a feedstock containing 5 volumes of 0.5% w/v PLGA in acetone and 1 volume of condensed DNA which was fed at a flow rate of 18ml h-1. The plasmid DNA vectors encoding a malaria protein were condensed with a cationic polymer before atomization. Results High levels of gene expression in vitro were observed in COS-7 cells transfected with condensed DNA at a nitrogen to phosphate (N/P) ratio of 10. At this N/P ratio, the condensed DNA exhibited a monodispersed nanoparticle size (Z-average diameter of 60.8 nm) and a highly positive zeta potential of 38.8mV. The microparticle formulations of malaria DNA vaccine were quality assessed and it was shown that themicroparticles displayed high encapsulation efficiencies between 82-96% and a narrow size distribution in the range of 0.8-1.9 μm. In vitro release profile revealed that approximately 82% of the DNA was released within 30 days via a predominantly diffusion controlledmass transfer system. Conclusions This ultrasonic atomization technique showed excellent particle size reproducibility and displayed potential as an industrially viable approach for the formulation of controlled release particles.

Towards the design of a scalable and commercially viable technique for plasmid purification using a methacrylate monolithic stationary phase

A monolithic stationary phase was prepared via free radical co-polymerization of ethylene glycol dimethacrylate (EDMA) and glycidyl methacrylate (GMA) with pore diameter tailored specifically for plasmid binding, retention and elution. The polymer was functionalized. with 2-chloro-N,N-diethylethylamine hydrochloride (DEAE-Cl) for anion-exchange purification of plasmid DNA (pDNA) from clarified lysate obtained from E. coli DH5α-pUC19 culture in a ribonuclease/ protease-free environment. Characterization of the monolithic resin showed a porous material, with 68% of the pores existing in the matrix having diameters above 300 nm. The final product isolated from a single-stage 5 min anion-exchange purification was a pure and homogeneous supercoiled (SC) pDNA with no gDNA, RNA and protein contamination as confirmed by ethidium bromide agarose gel electrophoresis (EtBr-AGE), enzyme restriction analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This non-toxic technique is cGMP compatible and highly scalable for production of pDNA on a commercial level.

Enhancing methacrylate-monolith-based downstream processes to champion plasmid DNA production

Increasing numbers of preclinical and clinical studies are utilizing pDNA (plasmid DNA) as the vector. In addition, there has been a growing trend towards larger and larger doses of pDNA utilized in human trials. The growing demand on pDNA manufacture leads to pressure to make more in less time. A key intervention has been the use of monoliths as stationary phases in liquid chromatography. Monolithic stationary phases offer fast separation to pDNA owing to their large pore size, making pDNA in the size range from 100 nm to over 300 nm easily accessible. However, the convective transport mechanism of monoliths does not guarantee plasmid purity. The recovery of pure pDNA hinges on a proper balance in the properties of the adsorbent phase, the mobile phase and the feedstock. The effects of pH and ionic strength of binding buffer, temperature of feedstock, active group density and the pore size of the stationary phase were considered as avenues to improve the recovery and purity of pDNA using a methacrylate-based monolithic adsorbent and Escherichia coli DH5α-pUC19 clarified lysate as feedstock. pDNA recovery was found to be critically dependent on the pH and ionic strength of the mobile phase. Up to a maximum of approx. 92% recovery was obtained under optimum conditions of pH and ionic strength. Increasing the feedstock temperature to 80°C increased the purity of pDNA owing to the extra thermal stability associated with pDNA over contaminants such as proteins. Results from toxicological studies of the plasmid samples using endotoxin standard (E. coli 0.55:B5 lipopolysaccharide) show that endotoxin level decreases with increasing salt concentration. It was obvious that large quantities of pure pDNA can be obtained with minimal extra effort simply by optimizing process parameters and conditions for pDNA purification.

Finite volume methods for simulating anomalous transport

In this thesis a new approach for solving a certain class of anomalous diffusion equations was developed. The theory and algorithms arising from this work will pave the way for more efficient and more accurate solutions of these equations, with applications to science, health and industry. The method of finite volumes was applied to discretise the spatial derivatives, and this was shown to outperform existing methods in several key respects. The stability and convergence of the new method were rigorously established.

Application of anomaly technique in wind turbine bearing fault detection

Bearing faults are the most common cause of wind turbine failures. Unavailability and maintenance cost of wind turbines are becoming critically important, with their fast growing in electric networks. Early fault detection can reduce outage time and costs. This paper proposes Anomaly Detection (AD) machine learning algorithms for fault diagnosis of wind turbine bearings. The application of this method on a real data set was conducted and is presented in this paper. For validation and comparison purposes, a set of baseline results are produced using the popular one-class SVM methods to examine the ability of the proposed technique in detecting incipient faults.

Fine-grained plant classification using convolutional neural networks for feature extraction

We present an overview of the QUT plant classification system submitted to LifeCLEF 2014. This system uses generic features extracted from a convolutional neural network previously used to perform general object classification. We examine the effectiveness of these features to perform plant classification when used in combination with an extremely randomised forest. Using this system, with minimal tuning, we obtained relatively good results with a score of 0:249 on the test set of LifeCLEF 2014.

Instructional objectives to core-gameplay : a serious game design technique

This paper explores a gap within the serious game design research. That gap is the ambiguity surrounding the process of aligning the instructional objectives of serious games with their core-gameplay i.e. the moment-to-moment activity that is the core of player interaction. A core-gameplay focused design framework is proposed that can work alongside existing, more broadly focused serious games design frameworks. The framework utilises an inquiry-based approach that allows the serious game designer to use key questions as a means to clearly outline instructional objectives with the core-gameplay. The use of this design framework is considered in the context of a small section of gameplay from an educational game currently in development. This demonstration of the framework brings shows how instructional objectives can be embedded into a serious games core-gameplay.

World architecture 1900-2000 : A critical mosaic : Southeast Asia and Oceania [Volume 10]

"World Architecture records the major architectural contributions made in all regions of the world to the development of human culture. Grouped into 10 geographical regions and representing five twenty-year-periods, the buildings have been selected by approximately 80 eminent international architectural critics. Each volume contains 100 buildings from one particular region, each object accompanied by an analytical text as well as by drawings and photographs. Introduction essays by the general editor, Kenneth Frampton, and the editor(s) of each volume complete the survey. The series comprises 10 volumes. The books are handsome, linen-bound and stitched, generously formatted (21,5 x 28,5 cm/8,4 x 11 inches) and contain approx. 300 pages and 400 colour prints each. This unique project gives the most precise and authoritative description of 1000 of the century's most notable buildings. Countries: Brunei, Burma, Cambodia, Indonesia, Laos, Malaysia, The Philippines, Singapore, Thailand, Vietnam and Australia, New Zealand, Papua New Guinea, and Oceania."

Stability and convergence of a new finite volume method for a two-sided space-fractional diffusion equation

In this paper, we consider a two-sided space-fractional diffusion equation with variable coefficients on a finite domain. Firstly, based on the nodal basis functions, we present a new fractional finite volume method for the two-sided space-fractional diffusion equation and derive the implicit scheme and solve it in matrix form. Secondly, we prove the stability and convergence of the implicit fractional finite volume method and conclude that the method is unconditionally stable and convergent. Finally, some numerical examples are given to show the effectiveness of the new numerical method, and the results are in excellent agreement with theoretical analysis.

Microscale consolidation analysis of relaxation behavior of single living chondrocytes subjected to varying strain-rates

Besides the elastic stiffness, the relaxation behavior of single living cells is also of interest of various researchers when studying cell mechanics. It is hypothesized that the relaxation response of the cells is governed by both intrinsic viscoelasticity of the solid phase and fluid-solid interactions mechanisms. There are a number of mechanical models have been developed to investigate the relaxation behavior of single cells. However, there is lack of model enable to accurately capture both of the mechanisms. Therefore, in this study, the porohyperelastic (PHE) model, which is an extension of the consolidation theory, combined with inverse Finite Element Analysis (FEA) technique was used at the first time to investigate the relaxation response of living chondrocytes. This model was also utilized to study the dependence of relaxation behavior of the cells on strain-rates. The stress-relaxation experiments under the various strain-rates were conducted with the Atomic Force Microscopy (AFM). The results have demonstrated that the PHE model could effectively capture the stress-relaxation behavior of the living chondrocytes, especially at intermediate to high strain-rates. Although this model gave some errors at lower strain-rates, its performance was acceptable. Therefore, the PHE model is properly a promising model for single cell mechanics studies. Moreover, it has been found that the hydraulic permeability of living chondrocytes reduced with decreasing of strain-rates. It might be due to the intracellular fluid volume fraction and the fluid pore pressure gradients of chondrocytes were higher when higher strain-rates applied.

A technique for analysing GOOSE packets when testing relays in an IEC 61850-8-1 environment

While the implementation of the IEC 61850 standard has significantly enhanced the performance of communications in electrical substations, it has also increased the complexity of the system. Subsequently, these added elaborations have introduced new challenges in relation to the skills and tools required for the design, test and maintenance of 61850-compatible substations. This paper describes a practical experience of testing a protection relay using a non-conventional test equipment; in addition, it proposes a third party software technique to reveal the contents of the packets transferred on the substation network. Using this approach, the standard objects can be linked and interpreted to what the end-users normally see in the IED and test equipment proprietary software programs.

Fault detection of wind turbine drivetrain utilizing power-speed characteristics

Wind energy, being the fastest growing renewable energy source in the present world, requires a large number of wind turbines to transform wind energy into electricity. One factor driving the cost of this energy is the reliable operation of these turbines. Therefore, it is a growing requirement within the wind farm community, to monitor the operation of the wind turbines on a continuous basis so that a possible fault can be detected ahead of time. As the wind turbine operates in an environment of constantly changing wind speed, it is a challenging task to design a fault detection technique which can accommodate the stochastic operational behavior of the turbines. Addressing this issue, this paper proposes a novel fault detection criterion which is robust against operational uncertainty, as well as having the ability to quantify severity level specifically of the drivetrain abnormality within an operating wind turbine. A benchmark model of wind turbine has been utilized to simulate drivetrain fault condition and effectiveness of the proposed technique has been tested accordingly. From the simulation result it can be concluded that the proposed criterion exhibits consistent performance for drivetrain faults for varying wind speed and has linear relationship with the fault severity level.

Wide-area damping control for inter-area oscillations using inverse filtering technique

In this study, a non-linear excitation controller using inverse filtering is proposed to damp inter-area oscillations. The proposed controller is based on determining generator flux value for the next sampling time which is obtained by maximising reduction rate of kinetic energy of the system after the fault. The desired flux for the next time interval is obtained using wide-area measurements and the equivalent area rotor angles and velocities are predicted using a non-linear Kalman filter. A supplementary control input for the excitation system, using inverse filtering approach, to track the desired flux is implemented. The inverse filtering approach ensures that the non-linearity introduced because of saturation is well compensated. The efficacy of the proposed controller with and without communication time delay is evaluated on different IEEE benchmark systems including Kundur's two area, Western System Coordinating Council three-area and 16-machine, 68-bus test systems.

From ImageNet to mining: Adapting visual object detection with minimal supervision

This paper presents visual detection and classification of light vehicles and personnel on a mine site.We capitalise on the rapid advances of ConvNet based object recognition but highlight that a naive black box approach results in a significant number of false positives. In particular, the lack of domain specific training data and the unique landscape in a mine site causes a high rate of errors. We exploit the abundance of background-only images to train a k-means classifier to complement the ConvNet. Furthermore, localisation of objects of interest and a reduction in computation is enabled through region proposals. Our system is tested on over 10km of real mine site data and we were able to detect both light vehicles and personnel. We show that the introduction of our background model can reduce the false positive rate by an order of magnitude.