In vitro cultivation of adult human chondrocytes : importance of culture system, oxygen and zonal differences

Articular cartilage exhibits limited intrinsic regenerative capacity and focal tissue defects can lead to the development of osteoarthritis (OA), a painful and debilitating loss of cartilage tissue. In Australia, 1.4 million people are affected by OA and its prevalence is increasing in line with current demographics. As treatment options are limited, new therapeutic approaches are being investigated including biological resurfacing of joints with tissue-engineered cartilage. Despite some progress in the field, major challenges remain to be addressed for large scale clinical success. For example, large numbers of chondrogenic cells are required for cartilage formation, but chondrocytes lose their chondrogenic phenotype (dedifferentiate) during in vitro propagation. Additionally, the zonal organization of articular cartilage is critical for normal cartilage function, but development of zonal structure has been largely neglected in cartilage repair strategies. Therefore, we hypothesised that culture conditions for freshly isolated human articular chondrocytes from non-OA and OA sources can be improved by employing microcarrier cultures and a reduced oxygen environment and that oxygen is a critical factor in the maintenance of the zonal chondrocyte phenotype. Microcarriers have successfully been used to cultivate bovine chondrocytes, and offer a potential alternative for clinical expansion of human chondrocytes. We hypothesised that improved yields can be achieved by propagating human chondrocytes on microcarriers. We found that cells on microcarriers acquired a flattened, polygonal morphology and initially proliferated faster than monolayercultivated cells. However, microcarrier cultivation over four weeks did not improve growth rates or the chondrogenic potential of non-OA and OA human articular chondrocytes over conventional monolayer cultivation. Based on these observations, we aimed to optimise culture conditions by modifying oxygen tension, to more closely reflect the in vivo environment. We found that propagation at 5% oxygen tension (moderate hypoxia) did not improve proliferation or redifferentiation capacity of human osteoarthritic chondrocytes. Moderate hypoxia increased the expression of chondrogenic markers during redifferentiation. However, osteoarthritic chondrocytes cultivated on microcarriers exhibited lower expression levels of chondrogenic surface marker proteins and had at best equivalent redifferentiation capacities compared to monolayer-cultured cells. This suggests that monolayer culture with multiple passaging potentially selects for a subpopulation of cells with higher differentiation capacity, which are otherwise rare in osteoarthritic, aged cartilage. However, fibroblastic proteins were found to be highly expressed in all cultures of human osteoarthritic chondrocytes indicating the presence of a high proportion of dedifferentiated, senescent cells with a chondrocytic phenotype that was not rescued by moderate hypoxia. The different zones of cartilage support chondrocyte subpopulations, which exhibit characteristic protein expression and experience varying oxygen tensions. We, therefore, hypothesised that oxygen tension affects the zonal marker expression of human articular chondrocytes isolated from the different cartilage layers. We found that zonal chondrocytes maintained these phenotypic differences during in vitro cultivation. Low oxygen environments favoured the expression of the zonal marker proteoglycan 4 in superficial cells, most likely through the promotion of chondrogenesis. The putative zonal markers clusterin and cartilage intermediate layer protein were found to be expressed by all subpopulations of human osteoarthritic chondrocytes ex vivo and, thus, may not be reliable predictors of in vitro stratification using these clinically relevant cells. The findings in this thesis underline the importance of considering low oxygen conditions and zonal stratification when creating native-like cartilaginous constructs. We have not yet found the right cues to successfully cultivate clinically-relevant human osteoarthritic chondrocytes in vitro. A more thorough understanding of chondrocyte biology and the processes of chondrogenesis are required to ensure the clinical success of cartilage tissue engineering.

Market access of Papua New Guinea bananas (Musa spp.) with particular respect to banana fly (Bactrocera musae (Tryon)) (Diptera: Tephritidae)

International market access for fresh commodities is regulated by international accepted phytosanitary guidelines, the objectives of which are to reduce the biosecurity risk of plant pest and disease movement. Papua New Guinea (PNG) has identified banana as a potential export crop and to help meet international market access requirements, this thesis provides information for the development of a pest risk analysis (PRA) for PNG banana fruit. The PRA is a three step process which first identifies the pests associated with a particular commodity or pathway, then assesses the risk associated with those pests, and finally identifies risk management options for those pests if required. As the first step of the PRA process, I collated a definitive list on the organisms associated with the banana plant in PNG using formal literature, structured interviews with local experts, grey literature and unpublished file material held in PNG field research stations. I identified 112 organisms (invertebrates, vertebrate, pathogens and weeds) associated with banana in PNG, but only 14 of these were reported as commonly requiring management. For these 14 I present detailed information summaries on their known biology and pest impact. A major finding of the review was that of the 14 identified key pests, some research information occurs for 13. The single exception for which information was found to be lacking was Bactrocera musae (Tryon), the banana fly. The lack of information for this widely reported ‘major pest on PNG bananas’ would hinder the development of a PNG banana fruit PRA. For this reason the remainder of the thesis focused on this organism, particularly with respect to generation of information required by the PRA process. Utilising an existing, but previously unanalysed fruit fly trapping database for PNG, I carried out a Geographic Information System analysis of the distribution and abundance of banana in four major regions of PNG. This information is required for a PRA to determine if banana fruit grown in different parts of the country are at different risks from the fly. Results showed that the fly was widespread in all cropping regions and that temperature and rainfall were not significantly correlated with banana fly abundance. Abundance of the fly was significantly correlated (albeit weakly) with host availability. The same analysis was done with four other PNG pest fruit flies and their responses to the environmental factors differed to banana fly and each other. This implies that subsequent PRA analyses for other PNG fresh commodities will need to investigate the risk of each of these flies independently. To quantify the damage to banana fruit caused by banana fly in PNG, local surveys and one national survey of banana fruit infestation were carried out. Contrary to expectations, infestation was found to be very low, particularly in the widely grown commercial cultivar, Cavendish. Infestation of Cavendish fingers was only 0.41% in a structured, national survey of over 2 700 banana fingers. Follow up laboratory studies showed that fingers of Cavendish, and another commercial variety Lady-finger, are very poor hosts for B. musae, with very low host selection rates by female flies and very poor immature survival. An analysis of a recent (within last decade) incursion of B. musae into the Gazelle Peninsula of East New Britain Province, PNG, provided the final set of B. musae data. Surveys of the fly on the peninsular showed that establishment and spread of the fly in the novel environment was very rapid and thus the fly should be regarded as being of high biosecurity concern, at least in tropical areas. Supporting the earlier impact studies, however, banana fly has not become a significant banana fruit problem on the Gazelle, despite bananas being the primary starch staple of the region. The results of the research chapters are combined in the final Discussion in the form of a B. musae focused PRA for PNG banana fruit. Putting the thesis in a broader context, the Discussion also deals with the apparent discrepancy between high local abundance of banana fly and very low infestation rates. This discussion focuses on host utilisation patterns of specialist herbivores and suggests that local pest abundance, as determined by trapping or monitoring, need not be good surrogate for crop damage, despite this linkage being implicit in a number of international phytosanitary protocols.

Securing medical research data with a rights management system

We propose a digital rights management approach for sharing electronic health records for research purposes and argue advantages of the approach. We give an outline of our implementation, discuss challenges that we faced and future directions.

Demo abstract : FOS —- a new operating system for sensor networks

FOS, the Fleck Operating System, is a new operating system that implements cooperative threads—providing a simple and productive environment for applications programmers. This paper discusses sensor network operating systems in general and places this development in context.

System identification and control of an aerobot drive system

Fast thrust changes are important for authoritive control of VTOL micro air vehicles. Fixed-pitch rotors that alter thrust by varying rotor speed require high-bandwidth control systems to provide adequate performace. We develop a feedback compensator for a brushless hobby motor driving a custom rotor suitable for UAVs. The system plant is identified using step excitation experiments. The aerodynamic operating conditions of these rotors are unusual and so experiments are performed to characterise expected load disturbances. The plant and load models lead to a proportional controller design capable of significantly decreasing rise-time and propagation of disturbances, subject to bus voltage constraints.

Virtual fencing applications : Implementing and testing an automated cattle control system

Managing livestock movement in extensive systems has environmental and production benefits. Currently permanent wire fencing is used to control cattle; this is both expensive and inflexible. Cattle are known to respond to auditory and visual cues and we investigated whether these can be used to manipulate their behaviour. Twenty-five Belmont Red steers with a mean live weight of 270kg were each randomly assigned to one of five treatments. Treatments consisted of a combination of cues (audio, tactile and visual stimuli) and consequence (electrical stimulation). The treatments were electrical stimulation alone, audio plus electrical stimulation, vibration plus electrical stimulation, light plus electrical stimulation and electrified electric fence (6kV) plus electrical stimulation. Cue stimuli were administered for 3s followed immediately by electrical stimulation (consequence) of 1kV for 1s. The experiment tested the operational efficacy of an on-animal control or virtual fencing system. A collar-halter device was designed to carry the electronics, batteries and equipment providing the stimuli, including audio, vibration, light and electrical of a prototype virtual fencing device. Cattle were allowed to travel along a 40m alley to a group of peers and feed while their rate of travel and response to the stimuli were recorded. The prototype virtual fencing system was successful in modifying the behaviour of the cattle. The rate of travel of cattle along the alley demonstrated the large variability in behavioural response associated with tactile, visual and audible cues. The experiment demonstrated virtual fencing has potential for controlling cattle in extensive grazing systems. However, larger numbers of cattle need to be tested to derive a better understanding of the behavioural variance. Further controlled experimental work is also necessary to quantify the interaction between cues, consequences and cattle learning.

Low-Cost vision-based AUV guidance system for reef navigation

Ensuring the long term viability of reef environments requires essential monitoring of many aspects of these ecosystems. However, the sheer size of these unstructured environments (for example Australia’s Great Barrier Reef pose a number of challenges for current monitoring platforms which are typically remote operated and required significant resources and infrastructure. Therefore, a primary objective of the CSIRO robotic reef monitoring project is to develop and deploy a large number of AUV teams to perform broadscale reef surveying. In order to achieve this, the platforms must be cheap, even possibly disposable. This paper presents the results of a preliminary investigation into the performance of a low-cost sensor suite and associated processing techniques for vision and inertial-based navigation within a highly unstructured reef environment.

Low-cost flight control system for a small autonomous helicopter

In this paper we describe a low-cost flight control system for a small (60 class) helicopter which is part of a larger project to develop an autonomous flying vehicle. Our approach differs from that of others in not using an expensive inertial/GPS sensing system. The primary sensors for vehicle stabilization are a low-cost inertial sensor and a pair of CMOS cameras. We describe the architecture of our flight control system, the inertial and visual sensing subsystems and present some flight control results.

UAS Mission Path Planning System, (MPPS) using hybrid-game coupled to multi-objective optimizer

This paper presents the application of advanced optimization techniques to unmanned aerial system mission path planning system (MPPS) using multi-objective evolutionary algorithms (MOEAs). Two types of multi-objective optimizers are compared; the MOEA nondominated sorting genetic algorithm II and a hybrid-game strategy are implemented to produce a set of optimal collision-free trajectories in a three-dimensional environment. The resulting trajectories on a three-dimensional terrain are collision-free and are represented by using Bézier spline curves from start position to target and then target to start position or different positions with altitude constraints. The efficiency of the two optimization methods is compared in terms of computational cost and design quality. Numerical results show the benefits of adding a hybrid-game strategy to a MOEA and for a MPPS.

The legal framework of the Australian health system

This chapter is about the role of law in the creation and operation of Australian health systems. Accordingly, this chapter discusses how law regulates the way in which health services in Australia are funded, organised, regulated, managed, operated and governed. (The question of how health professionals are regulated is discussed in Chapter 15.) Although the focus of much of health law is on legal mechanisms for the resolution of disputes or disagreements between the state, health providers, professionals, patients and families and friends, and through dispute resolutions processes setting standards for practice, these are only some of the “jobs” that health law performs. In health systems where the state undertakes a significant role in regulating, funding, managing and providing health services, health law also performs an important constitutive function. Health law declares the values upon which the health system is based, shapes social processes to achieve public ends and provides a structure for the complex interactions that occur within a modern health system. Health law regulates decision-makers in health systems by establishing who has the power to participate in decisions and in what circumstances, establishing processes through which decisions are made and creating mechanisms for decision-makers to be held publicly accountable. It is this broader constitutive function of health law that is a primary focus of much of this chapter — how and why governments use their legislative powers to structure and shape the health system.

A decision support system for sustainable urban development : the integrated land use and transportation indexing model

Broad, early definitions of sustainable development have caused confusion and hesitation among local authorities and planning professionals. This confusion has arisen because loosely defined principles of sustainable development have been employed when setting policies and planning projects, and when gauging the efficiencies of these policies in the light of designated sustainability goals. The question of how this theory-rhetoric-practice gap can be filled is the main focus of this chapter. It examines the triple bottom line approach–one of the sustainability accounting approaches widely employed by governmental organisations–and the applicability of this approach to sustainable urban development. The chapter introduces the ‘Integrated Land Use and Transportation Indexing Model’ that incorporates triple bottom line considerations with environmental impact assessment techniques via a geographic, information systems-based decision support system. This model helps decision-makers in selecting policy options according to their economic, environmental and social impacts. Its main purpose is to provide valuable knowledge about the spatial dimensions of sustainable development, and to provide fine detail outputs on the possible impacts of urban development proposals on sustainability levels. In order to embrace sustainable urban development policy considerations, the model is sensitive to the relationship between urban form, travel patterns and socio-economic attributes. Finally, the model is useful in picturing the holistic state of urban settings in terms of their sustainability levels, and in assessing the degree of compatibility of selected scenarios with the desired sustainable urban future.

A planning support system for airport city development

Many airports around the world are diversifying their land use strategies to integrate non-aeronautical development. These airports embrace the “airport city” concept to develop a wide range of commercial and light industrial land uses to support airport revenues. The consequences of this changing urban form are profound for both airport and municipal planners alike and present numerous challenges with regard to integration of airport and regional planning. While several tools exist for regional planning and airport operational planning, no holistic airport landside and regional planning tool exist. What is required is a planning support system that can integrate the sometimes conflicting stakeholder interests into one common goal for the airport and the surrounding region. This paper presents a planning support system and evaluates its application to a case study involving Brisbane Airport and the South East Queensland region in Australia.