Synthesis and characterisation of hybrid gold/polymer nanoparticles for bioassay application

A bioassay technique, based on surface-enhanced Raman scattering (SERS) tagged gold nanoparticles encapsulated with a biotin functionalised polymer, has been demonstrated through the spectroscopic detection of a streptavidin binding event. A methodical series of steps preceded these results: synthesis of nanoparticles which were found to give a reproducible SERS signal; design and synthesis of polymers with RAFT-functional end groups able to encapsulate the gold nanoparticle. The polymer also enabled the attachment of a biotin molecule functionalised so that it could be attached to the hybrid nanoparticle through a modular process. Finally, the demonstrations of a positive bioassay for this model construct using streptavidin/biotin binding. The synthesis of silver and gold nanoparticles was performed by using tri-sodium citrate as the reducing agent. The shape of the silver nanoparticles was quite difficult to control. Gold nanoparticles were able to be prepared in more regular shapes (spherical) and therefore gave a more consistent and reproducible SERS signal. The synthesis of gold nanoparticles with a diameter of 30 nm was the most reproducible and these were also stable over the longest periods of time. From the SERS results the optimal size of gold nanoparticles was found to be approximately 30 nm. Obtaining a consistent SERS signal with nanoparticles smaller than this was particularly difficult. Nanoparticles more than 50 nm in diameter were too large to remain suspended for longer than a day or two and formed a precipitate, rendering the solutions useless for our desired application. Gold nanoparticles dispersed in water were able to be stabilised by the addition of as-synthesised polymers dissolved in a water miscible solvent. Polymer stabilised AuNPs could not be formed from polymers synthesised by conventional free radical polymerization, i.e. polymers that did not possess a sulphur containing end-group. This indicated that the sulphur-containing functionality present within the polymers was essential for the self assembly process to occur. Polymer stabilization of the gold colloid was evidenced by a range of techniques including, visible spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis and Raman spectroscopy. After treatment of the hybrid nanoparticles with a series of SERS tags, focussing on 2-quinolinethiol the SERS signals were found to have comparable signal intensity to the citrate stabilised gold nanoparticles. This finding illustrates that the stabilization process does not interfere with the ability of gold nanoparticles to act as substrates for the SERS effect. Incorporation of a biotin moiety into the hybrid nanoparticles was achieved through a =click‘ reaction between an alkyne-functionalised polymer and an azido-functionalised biotin analogue. This functionalized biotin was prepared through a 4-step synthesis from biotin. Upon exposure of the surface-bound streptavidin to biotin-functionalised polymer hybrid gold nanoparticles, then washing, a SERS signal was obtained from the 2-quinolinethiol which was attached to the gold nanoparticles (positive assay). After exposure to functionalised polymer hybrid gold nanoparticles without biotin present then washing a SERS signal was not obtained as the nanoparticles did not bind to the streptavidin (negative assay). These results illustrate the applicability of the use of SERS active functional-polymer encapsulated gold nanoparticles for bioassay application.

Towards automated and in-situ, near-real time 3-D reconstruction of coral reef environments

Coral reefs are biologically complex ecosystems that support a wide variety of marine organisms. These are fragile communities under enormous threat from natural and human-based influences. Properly assessing and measuring the growth and health of reefs is essential to understanding impacts of ocean acidification, coastal urbanisation and global warming. In this paper, we present an innovative 3-D reconstruction technique based on visual imagery as a non-intrusive, repeatable, in situ method for estimating physical parameters, such as surface area and volume for efficient assessment of long-term variability. The reconstruction algorithms are presented, and benchmarked using an existing data set. We validate the technique underwater, utilising a commercial-off-the-shelf camera and a piece of staghorn coral, Acropora cervicornis. The resulting reconstruction is compared with a laser scan of the coral piece for assessment and validation. The comparison shows that 77% of the pixels in the reconstruction are within 0.3 mm of the ground truth laser scan. Reconstruction results from an unknown video camera are also presented as a segue to future applications of this research.

A qualitative study of older adults’ responses to sitting-time questions : do we get the information we want?

Background: In the last decade, there has been increasing interest in the health effects of sedentary behavior, which is often assessed using self-report sitting-time questions. The aim of this qualitative study was to document older adults’ understanding of sitting-time questions from the International Physical Activity (PA) Questionnaire (IPAQ) and the PA Scale for the Elderly (PASE). Methods: Australian community-dwelling adults aged 65+ years answered the IPAQ and PASE sitting questions in face-to-face semi-structured interviews. IPAQ uses one open-ended question to assess sitting on a weekday in the last 7 days 'at work, at home, while doing coursework and during leisure time'; PASE uses a three-part closed question about daily leisure-time sitting in the last 7 days. Participants expressed their thoughts out loud while answering each question. They were then probed about their responses. Interviews were recorded, transcribed and coded into themes. Results: Mean age of the 28 male and 27 female participants was 73 years (range 65-89). The most frequently reported activity was watching TV. For both questionnaires, many participants had difficulties understanding what activities to report. Some had difficulty understanding what activities should be classified as ‘leisure-time sitting’. Some assumed they were being asked to only report activities provided as examples. Most reported activities they normally do, rather than those performed on a day in the previous week. Participants used a variety of strategies to select ‘a day’ for which they reported their sitting activities and to calculate sitting time on that day. Therefore, many different ways of estimating sitting time were used. Participants had particular difficulty reporting their daily sitting-time when their schedules were not consistent across days. Some participants declared the IPAQ sitting question too difficult to answer. Conclusion: The accuracy of older adults’ self-reported sitting time is questionable given the challenges they have in answering sitting-time questions. Their responses to sitting-time questions may be more accurate if our recommendations for clarifying the sitting domains, providing examples relevant to older adults and suggesting strategies for formulating responses are incorporated. Future quantitative studies should include objective criterion measures to assess validity and reliability of these questions.

Building the foundations : skill acquisition in children

Introduction: Why we need to base childrens’ sport and physical education on the principles of dynamical systems theory and ecological psychology As the childhood years are crucial for developing many physical skills as well as establishing the groundwork leading to lifelong participation in sport and physical activities, (Orlick & Botterill, 1977, p. 11) it is essential to examine current practice to make sure it is meeting the needs of children. In recent papers (e.g. Renshaw, Davids, Chow & Shuttleworth, in press; Renshaw, Davids, Chow & Hammond, in review; Chow et al., 2009) we have highlighted that a guiding theoretical framework is needed to provide a principled approach to teaching and coaching and that the approach must be evidence- based and focused on mechanism and not just on operational issues such as practice, competition and programme management (Lyle, 2002). There is a need to demonstrate how nonlinear pedagogy underpins teaching and coaching practice for children given that some of the current approaches underpinning children’s sport and P.E. may not be leading to optimal results. For example, little time is spent undertaking physical activities (Tinning, 2006) and much of this practice is not representative of the competition demands of the performance environment (Kirk & McPhail, 2002; Renshaw et al., 2008). Proponents of a non- linear pedagogy advocate the design of practice by applying key concepts such as the mutuality of the performer and environment, the tight coupling of perception and action, and the emergence of movement solutions due to self organisation under constraints (see Renshaw, et al., in press). As skills are shaped by the unique interacting individual, task and environmental constraints in these learning environments, small changes to individual structural (e.g. factors such as height or limb length) or functional constraints (e.g. factors such as motivation, perceptual skills, strength that can be acquired), task rules, equipment, or environmental constraints can lead to dramatic changes in movement patterns adopted by learners to solve performance problems. The aim of this chapter is to provide real life examples for teachers and coaches who wish to adopt the ideas of non- linear pedagogy in their practice. Specifically, I will provide examples related to specific issues related to individual constraints in children and in particular the unique challenges facing coaches when individual constraints are changing due to growth and development. Part two focuses on understanding how cultural environmental constraints impact on children’s sport. This is an area that has received very little attention but plays a very important part in the long- term development of sporting expertise. Finally, I will look at how coaches can manipulate task constraints to create effective learning environments for young children.

Optimal control of distributed generators and capacitors by hybrid DPSO

In this paper, a comprehensive planning methodology is proposed that can minimize the line loss, maximize the reliability and improve the voltage profile in a distribution network. The injected active and reactive power of Distributed Generators (DG) and the installed capacitor sizes at different buses and for different load levels are optimally controlled. The tap setting of HV/MV transformer along with the line and transformer upgrading is also included in the objective function. A hybrid optimization method, called Hybrid Discrete Particle Swarm Optimization (HDPSO), is introduced to solve this nonlinear and discrete optimization problem. The proposed HDPSO approach is a developed version of DPSO in which the diversity of the optimizing variables is increased using the genetic algorithm operators to avoid trapping in local minima. The objective function is composed of the investment cost of DGs, capacitors, distribution lines and HV/MV transformer, the line loss, and the reliability. All of these elements are converted into genuine dollars. Given this, a single-objective optimization method is sufficient. The bus voltage and the line current as constraints are satisfied during the optimization procedure. The IEEE 18-bus test system is modified and employed to evaluate the proposed algorithm. The results illustrate the unavoidable need for optimal control on the DG active and reactive power and capacitors in distribution networks.

Time variation effects of weather conditions in rural MUSA-MIMO-OFDM channels

This paper presents channel measurements and weather data collection experiments conducted in a rural environment for an innovative Multi-User-Single-Antenna (MUSA) MIMO-OFDM technology, proposed for rural areas. MUSA MIMO-OFDM uplink channels are established by placing six user terminals (UT) around one access point (AP). Generated terrain profiles and relative received power plots are presented based on the experimental data. According to the relative received signal, MUSA-MIMO-OFDM uplink channels experience temporal fading. Moreover, the correlation between the relative received power and weather variables are presented. Results show that all weather variables exhibit a negative average correlation with received power. Wind speed records the highest average negative correlation coefficient of -0.35. Local maxima of negative correlation, ranging from 0.49 to 0.78, between the weather variables and relative received signals were registered between 5-6 a.m. The highest measured correlation (-0.78) of this time of the day was exhibited by wind speed. These results show the extend of time variation effects experienced by MUSA-MIMO-OFDM channels deployed in rural environments.

Path planning, guidance and control for a UAV forced landing

A forced landing is an unscheduled event in flight requiring an emergency landing, and is most commonly attributed to engine failure, failure of avionics or adverse weather. Since the ability to conduct a successful forced landing is the primary indicator for safety in the aviation industry, automating this capability for unmanned aerial vehicles (UAVs) will help facilitate their integration into, and subsequent routine operations over civilian airspace. Currently, there is no commercial system available to perform this task; however, a team at the Australian Research Centre for Aerospace Automation (ARCAA) is working towards developing such an automated forced landing system. This system, codenamed Flight Guardian, will operate onboard the aircraft and use machine vision for site identification, artificial intelligence for data assessment and evaluation, and path planning, guidance and control techniques to actualize the landing. This thesis focuses on research specific to the third category, and presents the design, testing and evaluation of a Trajectory Generation and Guidance System (TGGS) that navigates the aircraft to land at a chosen site, following an engine failure. Firstly, two algorithms are developed that adapts manned aircraft forced landing techniques to suit the UAV planning problem. Algorithm 1 allows the UAV to select a route (from a library) based on a fixed glide range and the ambient wind conditions, while Algorithm 2 uses a series of adjustable waypoints to cater for changing winds. A comparison of both algorithms in over 200 simulated forced landings found that using Algorithm 2, twice as many landings were within the designated area, with an average lateral miss distance of 200 m at the aimpoint. These results present a baseline for further refinements to the planning algorithms. A significant contribution is seen in the design of the 3-D Dubins Curves planning algorithm, which extends the elementary concepts underlying 2-D Dubins paths to account for powerless flight in three dimensions. This has also resulted in the development of new methods in testing for path traversability, in losing excess altitude, and in the actual path formation to ensure aircraft stability. Simulations using this algorithm have demonstrated lateral and vertical miss distances of under 20 m at the approach point, in wind speeds of up to 9 m/s. This is greater than a tenfold improvement on Algorithm 2 and emulates the performance of manned, powered aircraft. The lateral guidance algorithm originally developed by Park, Deyst, and How (2007) is enhanced to include wind information in the guidance logic. A simple assumption is also made that reduces the complexity of the algorithm in following a circular path, yet without sacrificing performance. Finally, a specific method of supplying the correct turning direction is also used. Simulations have shown that this new algorithm, named the Enhanced Nonlinear Guidance (ENG) algorithm, performs much better in changing winds, with cross-track errors at the approach point within 2 m, compared to over 10 m using Park's algorithm. A fourth contribution is made in designing the Flight Path Following Guidance (FPFG) algorithm, which uses path angle calculations and the MacCready theory to determine the optimal speed to fly in winds. This algorithm also uses proportional integral- derivative (PID) gain schedules to finely tune the tracking accuracies, and has demonstrated in simulation vertical miss distances of under 2 m in changing winds. A fifth contribution is made in designing the Modified Proportional Navigation (MPN) algorithm, which uses principles from proportional navigation and the ENG algorithm, as well as methods specifically its own, to calculate the required pitch to fly. This algorithm is robust to wind changes, and is easily adaptable to any aircraft type. Tracking accuracies obtained with this algorithm are also comparable to those obtained using the FPFG algorithm. For all three preceding guidance algorithms, a novel method utilising the geometric and time relationship between aircraft and path is also employed to ensure that the aircraft is still able to track the desired path to completion in strong winds, while remaining stabilised. Finally, a derived contribution is made in modifying the 3-D Dubins Curves algorithm to suit helicopter flight dynamics. This modification allows a helicopter to autonomously track both stationary and moving targets in flight, and is highly advantageous for applications such as traffic surveillance, police pursuit, security or payload delivery. Each of these achievements serves to enhance the on-board autonomy and safety of a UAV, which in turn will help facilitate the integration of UAVs into civilian airspace for a wider appreciation of the good that they can provide. The automated UAV forced landing planning and guidance strategies presented in this thesis will allow the progression of this technology from the design and developmental stages, through to a prototype system that can demonstrate its effectiveness to the UAV research and operations community.

Practical stability of approximating discrete-time filters with respect to model mismatch using relative entropy concepts

This paper establishes practical stability results for an important range of approximate discrete-time filtering problems involving mismatch between the true system and the approximating filter model. Using local consistency assumption, the practical stability established is in the sense of an asymptotic bound on the amount of bias introduced by the model approximation. Significantly, these practical stability results do not require the approximating model to be of the same model type as the true system. Our analysis applies to a wide range of estimation problems and justifies the common practice of approximating intractable infinite dimensional nonlinear filters by simpler computationally tractable filters.

Optimal online prediction in adversarial environments

In many prediction problems, including those that arise in computer security and computational finance, the process generating the data is best modelled as an adversary with whom the predictor competes. Even decision problems that are not inherently adversarial can be usefully modeled in this way, since the assumptions are sufficiently weak that effective prediction strategies for adversarial settings are very widely applicable.

Optimal online prediction in adversarial environments

In many prediction problems, including those that arise in computer security and computational finance, the process generating the data is best modelled as an adversary with whom the predictor competes. Even decision problems that are not inherently adversarial can be usefully modeled in this way, since the assumptions are sufficiently weak that effective prediction strategies for adversarial settings are very widely applicable.

Optimal strategies and minimax lower bounds for online convex games

A number of learning problems can be cast as an Online Convex Game: on each round, a learner makes a prediction x from a convex set, the environment plays a loss function f, and the learner’s long-term goal is to minimize regret. Algorithms have been proposed by Zinkevich, when f is assumed to be convex, and Hazan et al., when f is assumed to be strongly convex, that have provably low regret. We consider these two settings and analyze such games from a minimax perspective, proving minimax strategies and lower bounds in each case. These results prove that the existing algorithms are essentially optimal.

An analysis of the nature and effectiveness of corporate governance in smaller listed Australian companies

The objective of this thesis is to investigate the corporate governance attributes of smaller listed Australian firms. This study is motivated by evidence that these firms are associated with more regulatory concerns, the introduction of ASX Corporate Governance Recommendations in 2004, and a paucity of research to guide regulators and stakeholders of smaller firms. While there is an extensive body of literature examining the effectiveness of corporate governance, the literature principally focuses on larger companies, resulting in a deficiency in the understanding of the nature and effectiveness of corporate governance in smaller firms. Based on a review of agency theory literature, a theoretical model is developed that posits that agency costs are mitigated by internal governance mechanisms and transparency. The model includes external governance factors but in many smaller firms these factors are potentially absent, increasing the reliance on the internal governance mechanisms of the firm. Based on the model, the observed greater regulatory intervention in smaller companies may be due to sub-optimal internal governance practices. Accordingly, this study addresses four broad research questions (RQs). First, what is the extent and nature of the ASX Recommendations that have been adopted by smaller firms (RQ1)? Second, what firm characteristics explain differences in the recommendations adopted by smaller listed firms (RQ2), and third, what firm characteristics explain changes in the governance of smaller firms over time (RQ3)? Fourth, how effective are the corporate governance attributes of smaller firms (RQ4)? Six hypotheses are developed to address the RQs. The first two hypotheses explore the extent and nature of corporate governance, while the remaining hypotheses evaluate its effectiveness. A time-series, cross-sectional approach is used to evaluate the effectiveness of governance. Three models, based on individual governance attributes, an index of six items derived from the literature, and an index based on the full list of ASX Recommendations, are developed and tested using a sample of 298 smaller firms with annual observations over a five-year period (2002-2006) before and after the introduction of the ASX Recommendations in 2004. With respect to (RQ1) the results reveal that the overall adoption of the recommendations increased from 66 per cent in 2004 to 74 per cent in 2006. Interestingly, the adoption rate for recommendations regarding the structure of the board and formation of committees is significantly lower than the rates for other categories of recommendations. With respect to (RQ2) the results reveal that variations in rates of adoption are explained by key firm differences including, firm size, profitability, board size, audit quality, and ownership dispersion, while the results for (RQ3) were inconclusive. With respect to (RQ4), the results provide support for the association between better governance and superior accounting-based performance. In particular, the results highlight the importance of the independence of both the board and audit committee chairs, and of greater accounting-based expertise on the audit committee. In contrast, while there is little evidence that a majority independent board is associated with superior outcomes, there is evidence linking board independence with adverse audit opinion outcomes. These results suggest that board and chair independence are substitutes; in the presence of an independent chair a majority independent board may be an unnecessary and costly investment for smaller firms. The findings make several important contributions. First, the findings contribute to the literature by providing evidence on the extent, nature and effectiveness of governance in smaller firms. The findings also contribute to the policy debate regarding future development of Australia’s corporate governance code. The findings regarding board and chair independence, and audit committee characteristics, suggest that policy-makers could consider providing additional guidance for smaller companies. In general, the findings offer support for the “if not, why not?” approach of the ASX, rather than a prescriptive rules-based approach.

Practical stability with respect to model mismatch of approximate discrete-time output feedback control

This paper establishes a practical stability result for discrete-time output feedback control involving mismatch between the exact system to be stabilised and the approximating system used to design the controller. The practical stability is in the sense of an asymptotic bound on the amount of error bias introduced by the model approximation, and is established using local consistency properties of the systems. Importantly, the practical stability established here does not require the approximating system to be of the same model type as the exact system. Examples are presented to illustrate the nature of our practical stability result.

Measurement of total body water (TBW) and total energy expenditure (TEE) using stable isotopes

Understanding the relationship between diet, physical activity and health in humans requires accurate measurement of body composition and daily energy expenditure. Stable isotopes provide a means of measuring total body water and daily energy expenditure under free-living conditions. While the use of isotope ratio mass spectrometry (IRMS) for the analysis of 2H (Deuterium) and 18O (Oxygen-18) is well established in the field of human energy metabolism research, numerous questions remain regarding the factors which influence analytical and measurement error using this methodology. This thesis was comprised of four studies with the following emphases. The aim of Study 1 was to determine the analytical and measurement error of the IRMS with regard to sample handling under certain conditions. Study 2 involved the comparison of TEE (Total daily energy expenditure) using two commonly employed equations. Further, saliva and urine samples, collected at different times, were used to determine if clinically significant differences would occur. Study 3 was undertaken to determine the appropriate collection times for TBW estimates and derived body composition values. Finally, Study 4, a single case study to investigate if TEE measures are affected when the human condition changes due to altered exercise and water intake. The aim of Study 1 was to validate laboratory approaches to measure isotopic enrichment to ensure accurate (to international standards), precise (reproducibility of three replicate samples) and linear (isotope ratio was constant over the expected concentration range) results. This established the machine variability for the IRMS equipment in use at Queensland University for both TBW and TEE. Using either 0.4mL or 0.5mL sample volumes for both oxygen-18 and deuterium were statistically acceptable (p>0.05) and showed a within analytical variance of 5.8 Delta VSOW units for deuterium, 0.41 Delta VSOW units for oxygen-18. This variance was used as “within analytical noise” to determine sample deviations. It was also found that there was no influence of equilibration time on oxygen-18 or deuterium values when comparing the minimum (oxygen-18: 24hr; deuterium: 3 days) and maximum (oxygen-18: and deuterium: 14 days) equilibration times. With regard to preparation using the vacuum line, any order of preparation is suitable as the TEE values fall within 8% of each other regardless of preparation order. An 8% variation is acceptable for the TEE values due to biological and technical errors (Schoeller, 1988). However, for the automated line, deuterium must be assessed first followed by oxygen-18 as the automated machine line does not evacuate tubes but merely refills them with an injection of gas for a predetermined time. Any fractionation (which may occur for both isotopes), would cause a slight elevation in the values and hence a lower TEE. The purpose of the second and third study was to investigate the use of IRMS to measure the TEE and TBW of and to validate the current IRMS practices in use with regard to sample collection times of urine and saliva, the use of two TEE equations from different research centers and the body composition values derived from these TEE and TBW values. Following the collection of a fasting baseline urine and saliva sample, 10 people (8 women, 2 men) were dosed with a doubly labeled water does comprised of 1.25g 10% oxygen-18 and 0.1 g 100% deuterium/kg body weight. The samples were collected hourly for 12 hrs on the first day and then morning, midday, and evening samples were collected for the next 14 days. The samples were analyzed using an isotope ratio mass spectrometer. For the TBW, time to equilibration was determined using three commonly employed data analysis approaches. Isotopic equilibration was reached in 90% of the sample by hour 6, and in 100% of the sample by hour 7. With regard to the TBW estimations, the optimal time for urine collection was found to be between hours 4 and 10 as to where there was no significant difference between values. In contrast, statistically significant differences in TBW estimations were found between hours 1-3 and from 11-12 when compared with hours 4-10. Most of the individuals in this study were in equilibrium after 7 hours. The TEE equations of Prof Dale Scholler (Chicago, USA, IAEA) and Prof K.Westerterp were compared with that of Prof. Andrew Coward (Dunn Nutrition Centre). When comparing values derived from samples collected in the morning and evening there was no effect of time or equation on resulting TEE values. The fourth study was a pilot study (n=1) to test the variability in TEE as a result of manipulations in fluid consumption and level of physical activity; the magnitude of change which may be expected in a sedentary adult. Physical activity levels were manipulated by increasing the number of steps per day to mimic the increases that may result when a sedentary individual commences an activity program. The study was comprised of three sub-studies completed on the same individual over a period of 8 months. There were no significant changes in TBW across all studies, even though the elimination rates changed with the supplemented water intake and additional physical activity. The extra activity may not have sufficiently strenuous enough and the water intake high enough to cause a significant change in the TBW and hence the CO2 production and TEE values. The TEE values measured show good agreement based on the estimated values calculated on an RMR of 1455 kcal/day, a DIT of 10% of TEE and activity based on measured steps. The covariance values tracked when plotting the residuals were found to be representative of “well-behaved” data and are indicative of the analytical accuracy. The ratio and product plots were found to reflect the water turnover and CO2 production and thus could, with further investigation, be employed to identify the changes in physical activity.