Development of a Rep-inducible, BBTV-based expression system in banana

Banana bunchy top is regarded as the most important viral disease of banana, causing significant yield losses worldwide. The disease is caused by Banana bunchy top virus (BBTV), which is a circular ssDNA virus belonging to the genus Babuvirus in the family Nanoviridae. There are currently few effective control strategies for this and other ssDNA viruses. “In Plant Activation” (InPAct) is a novel technology being developed at QUT for ssDNA virus-activated suicide gene expression. The technology exploits the rolling circle replication mechanism of ssDNA viruses and is based on a unique “split” gene design such that suicide gene expression is only activated in the presence of the viral Rep. This PhD project aimed to develop a BBTV-based InPAct system as a suicide gene strategy to control BBTV. The BBTV-based InPAct vector design requires a BBTV intergenic region (IR) to be embedded within an intron in the gene expression cassette. To ensure that the BBTV IR would not interfere with intron splicing, a TEST vector was initially generated that contained the entire BBTV IR embedded within an intron in a β-glucuronidase (GUS) expression vector. Transient GUS assays in banana embryogenic cell suspensions indicated that cryptic intron splice sites were present within the IR. Transcript analysis revealed two cryptic intron splice sites in the Domain III sequence of the CR-M within the IR. Removal of the CR-M from the TEST vector resulted in an enhancement of GUS expression suggesting that the cryptic intron splice sites had been removed. An InPAct GUS vector was subsequently generated that contained the modified BBTV IR, with the CR-M (minus Domain III) repositioned within the InPAct cassette. Using transient histochemical and fluorometric GUS assays in banana embryogenic cells, the InPAct GUS vector was shown to be activated in the presence of the BBTV Rep. However, the presence of both BBTV Rep and Clink was shown to have a deleterious effect on GUS expression suggesting that these proteins were cytotoxic at the levels expressed. Analysis of replication of the InPAct vectors by Southern hybridisation revealed low levels of InPAct cassette-based episomal DNA released from the vector through the nicking/ligation activity of BBTV Rep. However, Rep-mediated episomal replicons, indicative of rolling circle replication of the released circularised cassettes, were not observed. The inability of the InPAct cassette to be replicated was further investigated. To examine whether the absence of Domain III of the CR-M was responsible, a suite of modified BBTV-based InPAct GUS vectors was constructed that contained the CR-M with the inclusion of Domain III, the CR-M with the inclusion of Domain III and additional upstream IR sequence, or no CR-M. Analysis of replication by Southern hybridisation revealed that neither the presence of Domain III, nor the entire CR-M, had an effect on replication levels. Since the InPAct cassette was significantly larger than the native BBTV genomic components (approximately 1 kb), the effect of InPAct cassette size on replication was also investigated. A suite of size variant BBTV-based vectors was constructed that increased the size of a replication competent cassette to 1.1 kbp through to 2.1 kbp.. Analysis of replication by Southern hybridisation revealed that an increase in vector size above approximately 1.5 - 1.7 kbp resulted in a decrease in replication. Following the demonstration of Rep-mediated release, circularisation and expression from the InPAct GUS vector, an InPAct vector was generated in which the uidA reporter gene was replaced with the ribonuclease-encoding suicide gene, barnase. Initially, a TEST vector was generated to assess the cytotoxicity of Barnase on banana cells. Although transient assays revealed a Barnase-induced cytotoxic effect in banana cells, the expression levels were sub-optimal. An InPAct BARNASE vector was generated and tested for BBTV Rep-activated Barnase expression using transient assays in banana embryogenic cells. High levels of background expression from the InPAct BARNASE vector made it difficult to accurately assess Rep-activated Barnase expression. Analysis of replication by Southern hybridisation revealed low levels of InPAct cassette-based episomal DNA released from the vector but no Rep-mediated episomal replicons indicative of rolling circle replication of the released circularised cassettes were again observed. Despite the inability of the InPAct vectors to replicate to enable high level gene expression, the InPAct BARNASE vector was assessed in planta for BBTV Rep-mediated activation of Barnase expression. Eleven lines of transgenic InPAct BARNASE banana plants were generated by Agrobacterium-mediated transformation and were challenged with viruliferous Pentalonia nigronervosa. At least one clonal plant in each line developed bunchy top symptoms and infection was confirmed by PCR. No localised lesions were observed on any plants, nor was there any localised GUS expression in the one InPAct GUS line challenged with viruliferous aphids. The results presented in this thesis are the first study towards the development of a BBTV-based InPAct system as a Rep-activatable suicide gene expression system to control BBTV. Although further optimisation of the vectors is necessary, the preliminary results suggest that this approach has the potential to be an effective control strategy for BBTV. The use of iterons within the InPAct vectors that are recognised by Reps from different ssDNA plant viruses may provide a broad-spectrum resistance strategy against multiple ssDNA plant viruses. Further, this technology holds great promise as a platform technology for the molecular farming of high-value proteins in vitro or in vivo through expression of the ssDNA virus Rep protein.

Solutions for the flow of a micropolar fluid in a porous channel

How various additives can increase some cardio-vascular diseases and effects of transport for albumin and glucose through permeable membranes are some important studies in biomechanics. The rolling phenomena of the leucocytes gives rise to an inflammatory reaction along a vascular wall. Initiated by Eringen [5], a micropolar fluid is a satisfactory model for flows of fluids which contain micro-constituents which can undergo rotation.

Social networking : investigating the features of Facebook application

Research on social networking sites like Facebook is emerging but sparse. The exploratory study investigates the value users derive from self-described ‘cool’ Facebook applications, and explores the features that either encourage or discourage users to recommend application to their friends. Thus the concepts of value and cool are explored in a social networking setting. Our qualitative data shows that consumers derive a combination of functional value along with either social or emotional value from the applications. Female Facebook users indicated self-expression as important, while mates then to use Facebook application to socially compete. Three broad categories emerged for application features; symmetrical features can both encourage or discourage recommendation, asymmetrical features one encourage or discourage but not both, and polar features where different levels of the same feature encourage or discourage. Recommending or not recommending an application tends to be the result of a combination of features rather than one feature in isolation.

Pram and stroller related injury in Queensland children under 5 years of age

An estimated 200 Queensland children under 5 years of age are injured every year in incidents involving prams or strollers. The majority of injuries are due to falls from or falls with the pram or stroller Nineteen children were identified as having been caught in the pram or stroller mechanism (13 sustained finger injuries). Stairs and escalators were a factor in nearly 10 percent of pram or stroller fall injuries, with children being tipped out of the pram or stroller, or rolling down the stairs in the device. Roll away injuries accounted for eight percent of all pram or stroller fall injuries (some also involving stairs) Roll away injuries could be prevented by a default brake system similar to airport trolleys. Pram or stroller failure was identified in 2% of injuries

Intelligent prognostics of machinery health utilising suspended condition monitoring data

The ability to forecast machinery failure is vital to reducing maintenance costs, operation downtime and safety hazards. Recent advances in condition monitoring technologies have given rise to a number of prognostic models for forecasting machinery health based on condition data. Although these models have aided the advancement of the discipline, they have made only a limited contribution to developing an effective machinery health prognostic system. The literature review indicates that there is not yet a prognostic model that directly models and fully utilises suspended condition histories (which are very common in practice since organisations rarely allow their assets to run to failure); that effectively integrates population characteristics into prognostics for longer-range prediction in a probabilistic sense; which deduces the non-linear relationship between measured condition data and actual asset health; and which involves minimal assumptions and requirements. This work presents a novel approach to addressing the above-mentioned challenges. The proposed model consists of a feed-forward neural network, the training targets of which are asset survival probabilities estimated using a variation of the Kaplan-Meier estimator and a degradation-based failure probability density estimator. The adapted Kaplan-Meier estimator is able to model the actual survival status of individual failed units and estimate the survival probability of individual suspended units. The degradation-based failure probability density estimator, on the other hand, extracts population characteristics and computes conditional reliability from available condition histories instead of from reliability data. The estimated survival probability and the relevant condition histories are respectively presented as “training target” and “training input” to the neural network. The trained network is capable of estimating the future survival curve of a unit when a series of condition indices are inputted. Although the concept proposed may be applied to the prognosis of various machine components, rolling element bearings were chosen as the research object because rolling element bearing failure is one of the foremost causes of machinery breakdowns. Computer simulated and industry case study data were used to compare the prognostic performance of the proposed model and four control models, namely: two feed-forward neural networks with the same training function and structure as the proposed model, but neglected suspended histories; a time series prediction recurrent neural network; and a traditional Weibull distribution model. The results support the assertion that the proposed model performs better than the other four models and that it produces adaptive prediction outputs with useful representation of survival probabilities. This work presents a compelling concept for non-parametric data-driven prognosis, and for utilising available asset condition information more fully and accurately. It demonstrates that machinery health can indeed be forecasted. The proposed prognostic technique, together with ongoing advances in sensors and data-fusion techniques, and increasingly comprehensive databases of asset condition data, holds the promise for increased asset availability, maintenance cost effectiveness, operational safety and – ultimately – organisation competitiveness.

Recombinant protein production using a Tobacco yellow dwarf virus-based episomal expression vector : control of Rep activity

Over the past decade, plants have been used as expression hosts for the production of pharmaceutically important and commercially valuable proteins. Plants offer many advantages over other expression systems such as lower production costs, rapid scale up of production, similar post-translational modification as animals and the low likelihood of contamination with animal pathogens, microbial toxins or oncogenic sequences. However, improving recombinant protein yield remains one of the greatest challenges to molecular farming. In-Plant Activation (InPAct) is a newly developed technology that offers activatable and high-level expression of heterologous proteins in plants. InPAct vectors contain the geminivirus cis elements essential for rolling circle replication (RCR) and are arranged such that the gene of interest is only expressed in the presence of the cognate viral replication-associated protein (Rep). The expression of Rep in planta may be controlled by a tissue-specific, developmentally regulated or chemically inducible promoter such that heterologous protein accumulation can be spatially and temporally controlled. One of the challenges for the successful exploitation of InPAct technology is the control of Rep expression as even very low levels of this protein can reduce transformation efficiency, cause abnormal phenotypes and premature activation of the InPAct vector in regenerated plants. Tight regulation over transgene expression is also essential if expressing cytotoxic products. Unfortunately, many tissue-specific and inducible promoters are unsuitable for controlling expression of Rep due to low basal activity in the absence of inducer or in tissues other than the target tissue. This PhD aimed to control Rep activity through the production of single chain variable fragments (scFvs) specific to the motif III of Tobacco yellow dwarf virus (TbYDV) Rep. Due to the important role played by the conserved motif III in the RCR, it was postulated that such scFvs can be used to neutralise the activity of the low amount of Rep expressed from a “leaky” inducible promoter, thus preventing activation of the TbYDV-based InPAct vector until intentional induction. Such scFvs could also offer the potential to confer partial or complete resistance to TbYDV, and possibly heterologous viruses as motif III is conserved between geminiviruses. Studies were first undertaken to determine the levels of TbYDV Rep and TbYDV replication-associated protein A (RepA) required for optimal transgene expression from a TbYDV-based InPAct vector. Transient assays in a non-regenerable Nicotiana tabacum (NT-1) cell line were undertaken using a TbYDV-based InPAct vector containing the uidA reporter gene (encoding GUS) in combination with TbYDV Rep and RepA under the control of promoters with high (CaMV 35S) or low (Banana bunchy top virus DNA-R, BT1) activity. The replication enhancer protein of Tomato leaf curl begomovirus (ToLCV), REn, was also used in some co-bombardment experiments to examine whether RepA could be substituted by a replication enhancer from another geminivirus genus. GUS expression was observed both quantitatively and qualitatively by fluorometric and histochemical assays, respectively. GUS expression from the TbYDV-based InPAct vector was found to be greater when Rep was expected to be expressed at low levels (BT1 promoter) rather than high levels (35S promoter). GUS expression was further enhanced when Rep and RepA were co-bombarded with a low ratio of Rep to RepA. Substituting TbYDV RepA with ToLCV REn also enhanced GUS expression but more importantly highest GUS expression was observed when cells were co-transformed with expression vectors directing low levels of Rep and high levels of RepA irrespective of the level of REn. In this case, GUS expression was approximately 74-fold higher than that from a non-replicating vector. The use of different terminators, namely CaMV 35S and Nos terminators, in InPAct vectors was found to influence GUS expression. In the presence of Rep, GUS expression was greater using pInPActGUS-Nos rather than pInPActGUS-35S. The only instance of GUS expression being greater from vectors containing the 35S terminator was when comparing expression from cells transformed with Rep, RepA and REnexpressing vectors and either non-replicating vectors, p35SGS-Nos or p35SGS-35S. This difference was most likely caused by an interaction of viral replication proteins with each other and the terminators. These results indicated that (i) the level of replication associated proteins is critical to high transgene expression, (ii) the choice of terminator within the InPAct vector may affect expression levels and (iii) very low levels of Rep can activate InPAct vectors hence controlling its activity is critical. Prior to generating recombinant scFvs, a recombinant TbYDV Rep was produced in E. coli to act as a control to enable the screening for Rep-specific antibodies. A bacterial expression vector was constructed to express recombinant TbYDV Rep with an Nterminal His-tag (N-His-Rep). Despite investigating several purification techniques including Ni-NTA, anion exchange, hydrophobic interaction and size exclusion chromatography, N-His-Rep could only be partially purified using a Ni-NTA column under native conditions. Although it was not certain that this recombinant N-His-Rep had the same conformation as the native TbYDV Rep and was functional, results from an electromobility shift assay (EMSA) showed that N-His-Rep was able to interact with the TbYDV LIR and was, therefore, possibly functional. Two hybridoma cell lines from mice, immunised with a synthetic peptide containing the TbYDV Rep motif III amino acid sequence, were generated by GenScript (USA). Monoclonal antibodies secreted by the two hybridoma cell lines were first screened against denatured N-His-Rep in Western analysis. After demonstrating their ability to bind N-His-Rep, two scFvs (scFv1 and scFv2) were generated using a PCR-based approach. Whereas the variable heavy chain (VH) from both cell lines could be amplified, only the variable light chain (VL) from cell line 2 was amplified. As a result, scFv1 contained VH and VL from cell line 1, whereas scFv2 contained VH from cell line 2 and VL from cell line 1. Both scFvs were first expressed in E. coli in order to evaluate their affinity to the recombinant TbYDV N-His-Rep. The preliminary results demonstrated that both scFvs were able to bind to the denatured N-His-Rep. However, EMSAs revealed that only scFv2 was able to bind to native N-His-Rep and prevent it from interacting with the TbYDV LIR. Each scFv was cloned into plant expression vectors and co-bombarded into NT-1 cells with the TbYDV-based InPAct GUS expression vector and pBT1-Rep to examine whether the scFvs could prevent Rep from mediating RCR. Although it was expected that the addition of the scFvs would result in decreased GUS expression, GUS expression was found to slightly increase. This increase was even more pronounced when the scFvs were targeted to the cell nucleus by the inclusion of the Simian virus 40 large T antigen (SV40) nuclear localisation signal (NLS). It was postulated that the scFvs were binding to a proportion of Rep, leaving a small amount available to mediate RCR. The outcomes of this project provide evidence that very high levels of recombinant protein can theoretically be expressed using InPAct vectors with judicious selection and control of viral replication proteins. However, the question of whether the scFvs generated in this project have sufficient affinity for TbYDV Rep to prevent its activity in a stably transformed plant remains unknown. It may be that other scFvs with different combinations of VH and VL may have greater affinity for TbYDV Rep. Such scFvs, when expressed at high levels in planta, might also confer resistance to TbYDV and possibly heterologous geminiviruses.

Mathematical modelling of muscle recruitment and function in the lumbar spine

Low back pain is an increasing problem in industrialised countries and although it is a major socio-economic problem in terms of medical costs and lost productivity, relatively little is known about the processes underlying the development of the condition. This is in part due to the complex interactions between bone, muscle, nerves and other soft tissues of the spine, and the fact that direct observation and/or measurement of the human spine is not possible using non-invasive techniques. Biomechanical models have been used extensively to estimate the forces and moments experienced by the spine. These models provide a means of estimating the internal parameters which can not be measured directly. However, application of most of the models currently available is restricted to tasks resembling those for which the model was designed due to the simplified representation of the anatomy. The aim of this research was to develop a biomechanical model to investigate the changes in forces and moments which are induced by muscle injury. In order to accurately simulate muscle injuries a detailed quasi-static three dimensional model representing the anatomy of the lumbar spine was developed. This model includes the nine major force generating muscles of the region (erector spinae, comprising the longissimus thoracis and iliocostalis lumborum; multifidus; quadratus lumborum; latissimus dorsi; transverse abdominis; internal oblique and external oblique), as well as the thoracolumbar fascia through which the transverse abdominis and parts of the internal oblique and latissimus dorsi muscles attach to the spine. The muscles included in the model have been represented using 170 muscle fascicles each having their own force generating characteristics and lines of action. Particular attention has been paid to ensuring the muscle lines of action are anatomically realistic, particularly for muscles which have broad attachments (e.g. internal and external obliques), muscles which attach to the spine via the thoracolumbar fascia (e.g. transverse abdominis), and muscles whose paths are altered by bony constraints such as the rib cage (e.g. iliocostalis lumborum pars thoracis and parts of the longissimus thoracis pars thoracis). In this endeavour, a separate sub-model which accounts for the shape of the torso by modelling it as a series of ellipses has been developed to model the lines of action of the oblique muscles. Likewise, a separate sub-model of the thoracolumbar fascia has also been developed which accounts for the middle and posterior layers of the fascia, and ensures that the line of action of the posterior layer is related to the size and shape of the erector spinae muscle. Published muscle activation data are used to enable the model to predict the maximum forces and moments that may be generated by the muscles. These predictions are validated against published experimental studies reporting maximum isometric moments for a variety of exertions. The model performs well for fiexion, extension and lateral bend exertions, but underpredicts the axial twist moments that may be developed. This discrepancy is most likely the result of differences between the experimental methodology and the modelled task. The application of the model is illustrated using examples of muscle injuries created by surgical procedures. The three examples used represent a posterior surgical approach to the spine, an anterior approach to the spine and uni-lateral total hip replacement surgery. Although the three examples simulate different muscle injuries, all demonstrate the production of significant asymmetrical moments and/or reduced joint compression following surgical intervention. This result has implications for patient rehabilitation and the potential for further injury to the spine. The development and application of the model has highlighted a number of areas where current knowledge is deficient. These include muscle activation levels for tasks in postures other than upright standing, changes in spinal kinematics following surgical procedures such as spinal fusion or fixation, and a general lack of understanding of how the body adjusts to muscle injuries with respect to muscle activation patterns and levels, rate of recovery from temporary injuries and compensatory actions by other muscles. Thus the comprehensive and innovative anatomical model which has been developed not only provides a tool to predict the forces and moments experienced by the intervertebral joints of the spine, but also highlights areas where further clinical research is required.

Best basis-based intelligent machine fault diagnosis

The wavelet packet transform decomposes a signal into a set of bases for time–frequency analysis. This decomposition creates an opportunity for implementing distributed data mining where features are extracted from different wavelet packet bases and served as feature vectors for applications. This paper presents a novel approach for integrated machine fault diagnosis based on localised wavelet packet bases of vibration signals. The best basis is firstly determined according to its classification capability. Data mining is then applied to extract features and local decisions are drawn using Bayesian inference. A final conclusion is reached using a weighted average method in data fusion. A case study on rolling element bearing diagnosis shows that this approach can greatly improve the accuracy ofdiagno sis.

Flexible high-power multi DC-DC converters for train systems

This thesis reports on the investigations, simulations and analyses of novel power electronics topologies and control strategies. The research is financed by an Australian Research Council (ARC) Linkage (07-09) grant. Therefore, in addition to developing original research and contributing to the available knowledge of power electronics, it also contributes to the design of a DC-DC converter for specific application to the auxiliary power supply in electric trains. Specifically, in this regard, it contributes to the design of a 7.5 kW DC-DC converter for the industrial partner (Schaffler and Associates Ltd) who supported this project. As the thesis is formatted as a ‘thesis by publication’, the contents are organized around published papers. The research has resulted in eleven papers, including seven peer reviewed and published conference papers, one published journal paper, two journal papers accepted for publication and one submitted journal paper (provisionally accepted subject to few changes). In this research, several novel DC-DC converter topologies are introduced, analysed, and tested. The similarity of all of the topologies devised lies in their ‘current circulating’ switching state, which allows them to store some energy in the inductor, as extra inductor current. The stored energy may be applied to enhance the performance of the converter in the occurrence of load current or input voltage disturbances. In addition, when there is an alternating load current, the ability to store energy allows the converter to perform satisfactorily despite frequently and highly varying load current. In this research, the capability of current storage has been utilised to design topologies for specific applications, and the enhancement of the performance of the considered applications has been illustrated. The simplest DC-DC converter topology, which has a ‘current circulating’ switching state, is the Positive Buck-Boost (PBB) converter (also known as the non-inverting Buck-Boost converter). Usually, the topology of the PBB converter is operating as a Buck or a Boost converter in applications with widely varying input voltage or output reference voltage. For example, in electric railways (the application of our industrial partner), the overhead line voltage alternates from 1000VDC to 500VDC and the required regulated voltage is 600VDC. In the course of this research, our industrial partner (Schaffler and Associates Ltd) industrialized a PBB converter–the ‘Mudo converter’–operating at 7.5 kW. Programming the onboard DSP and testing the PBB converter in experimental and nominal power and voltage was part of this research program. In the earlier stages of this research, the advantages and drawbacks of utilization of the ‘current circulating’ switching state in the positive Buck-Boost converter were investigated. In brief, the advantages were found to be robustness against input voltage and current load disturbances, and the drawback was extra conduction and switching loss. Although the robustness against disturbances is desirable for many applications, the price of energy loss must be minimized to attract attention to the utilization of the PBB converter. In further stages of this research, two novel control strategies for different applications were devised to minimise the extra energy loss while the advantages of the positive Buck-Boost converter were fully utilized. The first strategy is Smart Load Controller (SLC) for applications with pre-knowledge or predictability of input voltage and/or load current disturbances. A convenient example of these applications is electric/hybrid cars where a master controller commands all changes in loads and voltage sources. Therefore, the master controller has a pre-knowledge of the load and input voltage disturbances so it can apply the SLC strategy to utilize robustness of the PBB converter. Another strategy aiming to minimise energy loss and maximise the robustness in the face of disturbance is developed to cover applications with unexpected disturbances. This strategy is named Dynamic Hysteresis Band (DHB), and is used to manipulate the hysteresis band height after occurrence of disturbance to reduce dynamics of the output voltage. When no disturbance has occurred, the PBB converter works with minimum inductor current and minimum energy loss. New topologies based on the PBB converter have been introduced to address input voltage disturbances for different onboard applications. The research shows that the performance of applications of symmetrical/asymmetrical multi-level diode-clamped inverters, DC-networks, and linear-assisted RF amplifiers may be enhanced by the utilization of topologies based on the PBB converter. Multi-level diode-clamped inverters have the problem of DC-link voltage balancing when the power factor of their load closes to unity. This research has shown that this problem may be solved with a suitable multi-output DC-DC converter supplying DClink capacitors. Furthermore, the multi-level diode-clamped inverters supplied with asymmetrical DC-link voltages may improve the quality of load voltage and reduce the level of Electromagnetic Interference (EMI). Mathematical analyses and experiments on supplying symmetrical and asymmetrical multi-level inverters by specifically designed multi-output DC-DC converters have been reported in two journal papers. Another application in which the system performance can be improved by utilization of the ‘current circulating’ switching state is linear-assisted RF amplifiers in communicational receivers. The concept of ‘linear-assisted’ is to divide the signal into two frequency domains: low frequency, which should be amplified by a switching circuit; and the high frequency domain, which should be amplified by a linear amplifier. The objective is to minimize the overall power loss. This research suggests using the current storage capacity of a PBB based converter to increase its bandwidth, and to increase the domain of the switching converter. The PBB converter addresses the industrial demand for a DC-DC converter for the application of auxiliary power supply of a typical electric train. However, after testing the industrial prototype of the PBB converter, there were some voltage and current spikes because of switching. To attenuate this problem without significantly increasing the switching loss, the idea of Active Gate Signalling (AGS) is presented. AGS suggests a smart gate driver that selectively controls the switching process to reduce voltage/current spikes, without unacceptable reduction in the efficiency of switching.

A new multilevel converter configuration for high power and high quality applications

The Queensland University of Technology (QUT) allows the presentation of theses for the Degree of Doctor of Philosophy in the format of published or submitted papers, where such papers have been published, accepted or submitted during the period of candidature. This thesis is composed of ten published /submitted papers and book chapters of which nine have been published and one is under review. This project is financially supported by an Australian Research Council (ARC) Discovery Grant with the aim of investigating multilevel topologies for high quality and high power applications, with specific emphasis on renewable energy systems. The rapid evolution of renewable energy within the last several years has resulted in the design of efficient power converters suitable for medium and high-power applications such as wind turbine and photovoltaic (PV) systems. Today, the industrial trend is moving away from heavy and bulky passive components to power converter systems that use more and more semiconductor elements controlled by powerful processor systems. However, it is hard to connect the traditional converters to the high and medium voltage grids, as a single power switch cannot stand at high voltage. For these reasons, a new family of multilevel inverters has appeared as a solution for working with higher voltage levels. Besides this important feature, multilevel converters have the capability to generate stepped waveforms. Consequently, in comparison with conventional two-level inverters, they present lower switching losses, lower voltage stress across loads, lower electromagnetic interference (EMI) and higher quality output waveforms. These properties enable the connection of renewable energy sources directly to the grid without using expensive, bulky, heavy line transformers. Additionally, they minimize the size of the passive filter and increase the durability of electrical devices. However, multilevel converters have only been utilised in very particular applications, mainly due to the structural limitations, high cost and complexity of the multilevel converter system and control. New developments in the fields of power semiconductor switches and processors will favor the multilevel converters for many other fields of application. The main application for the multilevel converter presented in this work is the front-end power converter in renewable energy systems. Diode-clamped and cascade converters are the most common type of multilevel converters widely used in different renewable energy system applications. However, some drawbacks – such as capacitor voltage imbalance, number of components, and complexity of the control system – still exist, and these are investigated in the framework of this thesis. Various simulations using software simulation tools are undertaken and are used to study different cases. The feasibility of the developments is underlined with a series of experimental results. This thesis is divided into two main sections. The first section focuses on solving the capacitor voltage imbalance for a wide range of applications, and on decreasing the complexity of the control strategy on the inverter side. The idea of using sharing switches at the output structure of the DC-DC front-end converters is proposed to balance the series DC link capacitors. A new family of multioutput DC-DC converters is proposed for renewable energy systems connected to the DC link voltage of diode-clamped converters. The main objective of this type of converter is the sharing of the total output voltage into several series voltage levels using sharing switches. This solves the problems associated with capacitor voltage imbalance in diode-clamped multilevel converters. These converters adjust the variable and unregulated DC voltage generated by renewable energy systems (such as PV) to the desirable series multiple voltage levels at the inverter DC side. A multi-output boost (MOB) converter, with one inductor and series output voltage, is presented. This converter is suitable for renewable energy systems based on diode-clamped converters because it boosts the low output voltage and provides the series capacitor at the output side. A simple control strategy using cross voltage control with internal current loop is presented to obtain the desired voltage levels at the output voltage. The proposed topology and control strategy are validated by simulation and hardware results. Using the idea of voltage sharing switches, the circuit structure of different topologies of multi-output DC-DC converters – or multi-output voltage sharing (MOVS) converters – have been proposed. In order to verify the feasibility of this topology and its application, steady state and dynamic analyses have been carried out. Simulation and experiments using the proposed control strategy have verified the mathematical analysis. The second part of this thesis addresses the second problem of multilevel converters: the need to improve their quality with minimum cost and complexity. This is related to utilising asymmetrical multilevel topologies instead of conventional multilevel converters; this can increase the quality of output waveforms with a minimum number of components. It also allows for a reduction in the cost and complexity of systems while maintaining the same output quality, or for an increase in the quality while maintaining the same cost and complexity. Therefore, the asymmetrical configuration for two common types of multilevel converters – diode-clamped and cascade converters – is investigated. Also, as well as addressing the maximisation of the output voltage resolution, some technical issues – such as adjacent switching vectors – should be taken into account in asymmetrical multilevel configurations to keep the total harmonic distortion (THD) and switching losses to a minimum. Thus, the asymmetrical diode-clamped converter is proposed. An appropriate asymmetrical DC link arrangement is presented for four-level diode-clamped converters by keeping adjacent switching vectors. In this way, five-level inverter performance is achieved for the same level of complexity of the four-level inverter. Dealing with the capacitor voltage imbalance problem in asymmetrical diodeclamped converters has inspired the proposal for two different DC-DC topologies with a suitable control strategy. A Triple-Output Boost (TOB) converter and a Boost 3-Output Voltage Sharing (Boost-3OVS) converter connected to the four-level diode-clamped converter are proposed to arrange the proposed asymmetrical DC link for the high modulation indices and unity power factor. Cascade converters have shown their abilities and strengths in medium and high power applications. Using asymmetrical H-bridge inverters, more voltage levels can be generated in output voltage with the same number of components as the symmetrical converters. The concept of cascading multilevel H-bridge cells is used to propose a fifteen-level cascade inverter using a four-level H-bridge symmetrical diode-clamped converter, cascaded with classical two-level Hbridge inverters. A DC voltage ratio of cells is presented to obtain maximum voltage levels on output voltage, with adjacent switching vectors between all possible voltage levels; this can minimize the switching losses. This structure can save five isolated DC sources and twelve switches in comparison to conventional cascade converters with series two-level H bridge inverters. To increase the quality in presented hybrid topology with minimum number of components, a new cascade inverter is verified by cascading an asymmetrical four-level H-bridge diode-clamped inverter. An inverter with nineteen-level performance was achieved. This synthesizes more voltage levels with lower voltage and current THD, rather than using a symmetrical diode-clamped inverter with the same configuration and equivalent number of power components. Two different predictive current control methods for the switching states selection are proposed to minimise either losses or THD of voltage in hybrid converters. High voltage spikes at switching time in experimental results and investigation of a diode-clamped inverter structure raised another problem associated with high-level high voltage multilevel converters. Power switching components with fast switching, combined with hard switched-converters, produce high di/dt during turn off time. Thus, stray inductance of interconnections becomes an important issue and raises overvoltage and EMI issues correlated to the number of components. Planar busbar is a good candidate to reduce interconnection inductance in high power inverters compared with cables. The effect of different transient current loops on busbar physical structure of the high-voltage highlevel diode-clamped converters is highlighted. Design considerations of proper planar busbar are also presented to optimise the overall design of diode-clamped converters.

From the cathedral to the classroom : the emergence of new discourses on religious education

In 2009, Religious Education is a designated key learning area in Catholic schools in the Archdiocese of Brisbane and, indeed, across Australia. Over the years, though, different conceptualisations of the nature and purpose of religious education have led to the construction of different approaches to the classroom teaching of religion. By investigating the development of religious education policy in the Archdiocese of Brisbane from 1984 to 2003, the study seeks to trace the emergence of new discourses on religious education. The study understands religious education to refer to a lifelong process that occurs through a variety of forms (Moran, 1989). In Catholic schools, it refers both to co-curricula activities, such as retreats and school liturgies, and the classroom teaching of religion. It is the policy framework for the classroom teaching of religion that this study explores. The research was undertaken using a policy case study approach to gain a detailed understanding of how new conceptualisations of religious education emerged at a particular site of policy production, in this case, the Archdiocese of Brisbane. The study draws upon Yeatman’s (1998) description of policy as occurring “when social actors think about what they are doing and why in relation to different and alternative possible futures” (p. 19) and views policy as consisting of more than texts themselves. Policy texts result from struggles over meaning (Taylor, 2004) in which specific discourses are mobilised to support particular views. The study has a particular interest in the analysis of Brisbane religious education policy texts, the discursive practices that surrounded them, and the contexts in which they arose. Policy texts are conceptualised in the study as representing “temporary settlements” (Gale, 1999). Such settlements are asymmetrical, temporary and dependent on context: asymmetrical in that dominant actors are favoured; temporary because dominant actors are always under challenge by other actors in the policy arena; and context - dependent because new situations require new settlements. To investigate the official policy documents, the study used Critical Discourse Analysis (hereafter referred to as CDA) as a research tool that affords the opportunity for researchers to map and chart the emergence of new discourses within the policy arena. As developed by Fairclough (2001), CDA is a three-dimensional application of critical analysis to language. In the Brisbane religious education arena, policy texts formed a genre chain (Fairclough, 2004; Taylor, 2004) which was a focus of the study. There are two features of texts that form genre chains: texts are systematically linked to one another; and, systematic relations of recontextualisation exist between the texts. Fairclough’s (2005) concepts of “imaginary space” and “frameworks for action” (p. 65) within the policy arena were applied to the Brisbane policy arena to investigate the relationship between policy statements and subsequent guidelines documents. Five key findings emerged from the study. First, application of CDA to policy documents revealed that a fundamental reconceptualisation of the nature and purpose of classroom religious education in Catholic schools occurred in the Brisbane policy arena over the last twenty-five years. Second, a disjuncture existed between catechetical discourses that continued to shape religious education policy statements, and educational discourses that increasingly shaped guidelines documents. Third, recontextualisation between policy documents was evident and dependent on the particular context in which religious education occurred. Fourth, at subsequent links in the chain, actors created their own “imaginary space”, thereby altering orders of discourse within the policy arena, with different actors being either foregrounded or marginalised. Fifth, intertextuality was more evident in the later links in the genre chain (i.e. 1994 policy statement and 1997 guidelines document) than in earlier documents. On the basis of the findings of the study, six recommendations are made. First, the institutional Church should carefully consider the contribution that the Catholic school can make to the overall pastoral mission of the diocese in twenty-first century Australia. Second, policymakers should articulate a nuanced understanding of the relationship between catechesis and education with regard to the religion classroom. Third, there should be greater awareness of the connections among policies relating to Catholic schools – especially the connection between enrolment policy and religious education policy. Fourth, there should be greater consistency between policy documents. Fifth, policy documents should be helpful for those to whom they are directed (i.e. Catholic schools, teachers). Sixth, “imaginary space” (Fairclough, 2005) in policy documents needs to be constructed in a way that allows for multiple “frameworks for action” (Fairclough, 2005) through recontextualisation. The findings of this study are significant in a number of ways. For religious educators, the study highlights the need to develop a shared understanding of the nature and purpose of classroom religious education. It argues that this understanding must take into account the multifaith nature of Australian society and the changing social composition of Catholic schools themselves. Greater recognition should be given to the contribution that religious studies courses such as Study of Religion make to the overall religious development of a person. In view of the social composition of Catholic schools, there is also an issue of ecclesiological significance concerning the conceptualisation of the relationship between the institutional Catholic Church and Catholic schools. Finally, the study is of significance because of its application of CDA to religious education policy documents. Use of CDA reveals the foregrounding, marginalising, or excluding of various actors in the policy arena.