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.

Spherical image-based visual servo and structure estimation

This paper presents a formulation of image-based visual servoing (IBVS) for a spherical camera where coordinates are parameterized in terms of colatitude and longitude: IBVSSph. The image Jacobian is derived and simulation results are presented for canonical rotational, translational as well as general motion. Problems with large rotations that affect the planar perspective form of IBVS are not present on the sphere, whereas the desirable robustness properties of IBVS are shown to be retained. We also describe a structure from motion (SfM) system based on camera-centric spherical coordinates and show how a recursive estimator can be used to recover structure. The spherical formulations for IBVS and SfM are particularly suitable for platforms, such as aerial and underwater robots, that move in SE(3).

Speech recognition using ad-hoc microphone arrays

While close talking microphones give the best signal quality and produce the highest accuracy from current Automatic Speech Recognition (ASR) systems, the speech signal enhanced by microphone array has been shown to be an effective alternative in a noisy environment. The use of microphone arrays in contrast to close talking microphones alleviates the feeling of discomfort and distraction to the user. For this reason, microphone arrays are popular and have been used in a wide range of applications such as teleconferencing, hearing aids, speaker tracking, and as the front-end to speech recognition systems. With advances in sensor and sensor network technology, there is considerable potential for applications that employ ad-hoc networks of microphone-equipped devices collaboratively as a virtual microphone array. By allowing such devices to be distributed throughout the users’ environment, the microphone positions are no longer constrained to traditional fixed geometrical arrangements. This flexibility in the means of data acquisition allows different audio scenes to be captured to give a complete picture of the working environment. In such ad-hoc deployment of microphone sensors, however, the lack of information about the location of devices and active speakers poses technical challenges for array signal processing algorithms which must be addressed to allow deployment in real-world applications. While not an ad-hoc sensor network, conditions approaching this have in effect been imposed in recent National Institute of Standards and Technology (NIST) ASR evaluations on distant microphone recordings of meetings. The NIST evaluation data comes from multiple sites, each with different and often loosely specified distant microphone configurations. This research investigates how microphone array methods can be applied for ad-hoc microphone arrays. A particular focus is on devising methods that are robust to unknown microphone placements in order to improve the overall speech quality and recognition performance provided by the beamforming algorithms. In ad-hoc situations, microphone positions and likely source locations are not known and beamforming must be achieved blindly. There are two general approaches that can be employed to blindly estimate the steering vector for beamforming. The first is direct estimation without regard to the microphone and source locations. An alternative approach is instead to first determine the unknown microphone positions through array calibration methods and then to use the traditional geometrical formulation for the steering vector. Following these two major approaches investigated in this thesis, a novel clustered approach which includes clustering the microphones and selecting the clusters based on their proximity to the speaker is proposed. Novel experiments are conducted to demonstrate that the proposed method to automatically select clusters of microphones (ie, a subarray), closely located both to each other and to the desired speech source, may in fact provide a more robust speech enhancement and recognition than the full array could.

Franchise nations : the future of nations?

In this paper, I extend the notion of franchise nations, borrowed from Neal Stephenson’s cyberpunk novel Snow Crash (1993), in order to employ it as a device for thinking about the future of the nation. I argue the concept to be particularly well suited for such contemplation because of its sound grounding in the historical intermesh of economic, political and cultural motivations intrinsic to the concept as well as lived experience of the nation. I illustrate this very briefly by casting (mainland) China as the master franchisor and the overseas Chinese as franchisees. Specifically, I discuss the media events concerning China that took place during 2008, such as the protests and counter-protests that occurred at various legs of the Olympic Torch Relay, the Sichuan earthquake of 12 May and the opening ceremony of the Beijing Olympics on 8 August, and reactions to these happenings from overseas Chinese located variously in Australia, Canada and the United States. I argue that employing the notion of franchise nations lays bare the commercial and political instrumentalism behind the promotion and courtship of diasporas by home nations but, crucially, also aids in the understanding of the reciprocal processes by which franchisees are fashioned out of these communities. Finally, I suggest that, aside from China, franchise nations may also be a useful approach for thinking about how nations like India and Singapore are expanded, exported and explained into the future.

Theory of radar imaging using time-frequency distribution

The concept of radar was developed for the estimation of the distance (range) and velocity of a target from a receiver. The distance measurement is obtained by measuring the time taken for the transmitted signal to propagate to the target and return to the receiver. The target's velocity is determined by measuring the Doppler induced frequency shift of the returned signal caused by the rate of change of the time- delay from the target. As researchers further developed conventional radar systems it become apparent that additional information was contained in the backscattered signal and that this information could in fact be used to describe the shape of the target itself. It is due to the fact that a target can be considered to be a collection of individual point scatterers, each of which has its own velocity and time- delay. DelayDoppler parameter estimation of each of these point scatterers thus corresponds to a mapping of the target's range and cross range, thus producing an image of the target. Much research has been done in this area since the early radar imaging work of the 1960s. At present there are two main categories into which radar imaging falls. The first of these is related to the case where the backscattered signal is considered to be deterministic. The second is related to the case where the backscattered signal is of a stochastic nature. In both cases the information which describes the target's scattering function is extracted by the use of the ambiguity function, a function which correlates the backscattered signal in time and frequency with the transmitted signal. In practical situations, it is often necessary to have the transmitter and the receiver of the radar system sited at different locations. The problem in these situations is 'that a reference signal must then be present in order to calculate the ambiguity function. This causes an additional problem in that detailed phase information about the transmitted signal is then required at the receiver. It is this latter problem which has led to the investigation of radar imaging using time- frequency distributions. As will be shown in this thesis, the phase information about the transmitted signal can be extracted from the backscattered signal using time- frequency distributions. The principle aim of this thesis was in the development, and subsequent discussion into the theory of radar imaging, using time- frequency distributions. Consideration is first given to the case where the target is diffuse, ie. where the backscattered signal has temporal stationarity and a spatially white power spectral density. The complementary situation is also investigated, ie. where the target is no longer diffuse, but some degree of correlation exists between the time- frequency points. Computer simulations are presented to demonstrate the concepts and theories developed in the thesis. For the proposed radar system to be practically realisable, both the time- frequency distributions and the associated algorithms developed must be able to be implemented in a timely manner. For this reason an optical architecture is proposed. This architecture is specifically designed to obtain the required time and frequency resolution when using laser radar imaging. The complex light amplitude distributions produced by this architecture have been computer simulated using an optical compiler.

An ecology of centre-based child care

The studies in the thesis were derived from a program of research focused on centre-based child care in Australia. The studies constituted an ecological analysis as they examined proximal and distal factors which have the potential to affect children's developmental opportunities (Bronfenbrenner, 1979). The project was conducted in thirty-two child care centres located in south-east Queensland. Participants in the research included staff members at the centres, families using the centres and their children. The first study described the personal and professional characteristics of one hundred and forty-four child care workers, as well as their job satisfaction and job commitment. Factors impinging on the stability of care afforded to children were examined, specifically child care workers' intentions to leave their current position and actual staff turnover at a twelve month follow-up. This is an ecosystem analysis (Bronfenbrenner & Crouter, 1983), as it examined the world of work for carers; a setting not directly involving the developing child, but which has implications for children's experiences. Staff job satisfaction was focused on working with children and other adults, including parents and colleagues. Involvement with children was reported as being the most rewarding aspect of the work. This intrinsic satisfaction was enough to sustain caregivers' efforts to maintain their employment in child care programs. It was found that, while improving working conditions may help to reduce turnover, it is likely that moderate turnover rates will remain as child care staff work in relatively small centres and they leave in order to improve career prospects. Departure from a child care job appeared to be as much about improving career opportunities or changing personal circumstances, as it was about poor wages and working conditions. In the second study, factors that influence maternal satisfaction with child care arrangements were examined. The focus included examination of the nature and qualities of parental interaction with staff. This was a mesosystem analysis (Bronfenbrenner & Crouter, 1983), as it considered the links between family and child care settings. Two hundred and twenty-two questionnaires were returned from mothers whose children were enrolled in the participating centres. It was found that maternal satisfaction with child care encompassed the domains of child-centred and parent-centred satisfaction. The nature and range of responses in the quantitative and qualitative data indicated that these parents were genuinely satisfied with their children's care. In the prediction of maternal satisfaction with child care, single parents, mothers with high role satisfaction, and mothers who were satisfied with the frequency of staff contact and degree of supportive communication had higher levels of satisfaction with their child care arrangements. The third study described the structural and process variations within child care programs and examined program differences for compliance with regulations and differences by profit status of the centre, as a microsystem analysis (Bronfenbrenner, 1979). Observations were made in eighty-three programs which served children from two to five years. The results of the study affirmed beliefs that nonprofit centres are superior in the quality of care provided, although this was not to a level which meant that the care in for-profit centres was inadequate. Regulation of structural features of child care programs, per se, did not guarantee higher quality child care as measured by global or process indicators. The final study represented an integration of a range of influences in child care and family settings which may impact on development. Features of child care programs which predict children's social and cognitive development, while taking into account child and family characteristics, were identified. Results were consistent with other research findings which show that child and family characteristics and child care quality predict children's development. Child care quality was more important to the prediction of social development, while family factors appeared to be more predictive of cognitive/language development. An influential variable predictive of development was the period of time which the child had been in the centre. This highlighted the importance of the stability of child care arrangements. Child care quality features which had most influence were global ratings of the qualities of the program environment. However, results need to be interpreted cautiously as the explained variance in the predictive models developed was low. The results of these studies are discussed in terms of the implications for practice and future research. Considerations for an expanded view of ecological approaches to child care research are outlined. Issues discussed include the need to generate child care research which is relevant to social policy development, the implications of market driven policies for child care services, professionalism and professionalisation of child care work, and the need to reconceptualise child care research when the goal is to develop greater theoretical understanding about child care environments and developmental processes.

Art in early childhood : the discourse of 'proper' teaching

This thesis is a problematisation of the teaching of art to young children. To problematise a domain of social endeavour, is, in Michel Foucault's terms, to ask how we come to believe that "something ... can and must be thought" (Foucault, 1985:7). The aim is to document what counts (i.e., what is sayable, thinkable, feelable) as proper art teaching in Queensland at this point ofhistorical time. In this sense, the thesis is a departure from more recognisable research on 'more effective' teaching, including critical studies of art teaching and early childhood teaching. It treats 'good teaching' as an effect of moral training made possible through disciplinary discourses organised around certain epistemic rules at a particular place and time. There are four key tasks accomplished within the thesis. The first is to describe an event which is not easily resolved by means of orthodox theories or explanations, either liberal-humanist or critical ones. The second is to indicate how poststructuralist understandings of the self and social practice enable fresh engagements with uneasy pedagogical moments. What follows this discussion is the documentation of an empirical investigation that was made into texts generated by early childhood teachers, artists and parents about what constitutes 'good practice' in art teaching. Twenty-two participants produced text to tell and re-tell the meaning of 'proper' art education, from different subject positions. Rather than attempting to capture 'typical' representations of art education in the early years, a pool of 'exemplary' teachers, artists and parents were chosen, using "purposeful sampling", and from this pool, three videos were filmed and later discussed by the audience of participants. The fourth aspect of the thesis involves developing a means of analysing these texts in such a way as to allow a 're-description' of the field of art teaching by attempting to foreground the epistemic rules through which such teacher-generated texts come to count as true ie, as propriety in art pedagogy. This analysis drew on Donna Haraway's (1995) understanding of 'ironic' categorisation to hold the tensions within the propositions inside the categories of analysis rather than setting these up as discursive oppositions. The analysis is therefore ironic in the sense that Richard Rorty (1989) understands the term to apply to social scientific research. Three 'ironic' categories were argued to inform the discursive construction of 'proper' art teaching. It is argued that a teacher should (a) Teach without teaching; (b) Manufacture the natural; and (c) Train for creativity. These ironic categories work to undo modernist assumptions about theory/practice gaps and finding a 'balance' between oppositional binary terms. They were produced through a discourse theoretical reading of the texts generated by the participants in the study, texts that these same individuals use as a means of discipline and self-training as they work to teach properly. In arguing the usefulness of such approaches to empirical data analysis, the thesis challenges early childhood research in arts education, in relation to its capacity to deal with ambiguity and to acknowledge contradiction in the work of teachers and in their explanations for what they do. It works as a challenge at a range of levels - at the level of theorising, of method and of analysis. In opening up thinking about normalised categories, and questioning traditional Western philosophy and the grand narratives of early childhood art pedagogy, it makes a space for re-thinking art pedagogy as "a game oftruth and error" (Foucault, 1985). In doing so, it opens up a space for thinking how art education might be otherwise.

The mechanism of maturation of insulin-like growth factor-1

This study, to elucidate the role of des(1-3)IGF-I in the maturation of IGF-I,used two strategies. The first was to detect the presence of enzymes in tissues, which would act on IGF-I to produce des(1-3)IGF-I, and the second was to detect the potential products of such enzymic activity, namely Gly-Pro-Glu(GPE), Gly-Pro(GP) and des(l- 3)IGF-I. No neutral tripeptidyl peptidase (TPP II), which would release the tripeptide GPE from IGF-I, was detected in brain, urine nor in red or white blood cells. The TPPlike activity which was detected, was attributed to a combined action of a dipeptidyl peptidase (DPP N) and an aminopeptidase (AP A). A true TPP II was, however, detected in platelets. Two purified TPP II enzymes were investigated but they did not release GPE from IGF-I under a variety of conditions. Consequently, TPP II seemed unlikely to participate in the formation of des(1-3)IGF-I. In contrast, an acidic tripeptidyl peptidase activity (TPP I) was detected in brain and colostrum, the former with a pH optimum of 4.5 and the latter 3.8. It seems likely that such an enzyme would participate in the formation of des( 1-3 )IGF-I in these tissues in vitro, ie. that des(1-3)IGF-I may have been produced as an artifact in the isolation of IGF-I from brain and colostrum in acidic conditions. This contrasts with suggestions of an in vivo role for des(1-3)IGF-I, as reported by others. The activity of a dipeptidyl peptidase N (DPP N) from urine, which should release the dipeptide GP from IGF-I, was assessed under a variety of conditions and with a variety of additives and potential enzyme stimulants, but there was no release of GP. The DPP N also exhibited a transferase activity with synthetic substrates in the presence of dipeptides, at lower concentrations than previously reported for other acceptors or other proteolytic enzymes. In addition, a low concentration of a product,possibly the tetrapeptide Gly-Pro-Gly-Leu, was detected with the action of the enzyme on IGF-I in the presence of the dipeptide Gly-Leu. As part of attempts to detect tissue production of des(1-3)IGF-I, a monoclonal antibody (MAb ), directed towards the GPE- end ofiGF-I was produced by immunisation with a 10-mer covalently attached to a carrier protein. By the use of indirect ELISA and inhibitor studies, the MAb was shown to selectively recognise peptides with anNterminal GPE- sequence, and applied to the indirect detection of des(1-3)IGF-I. The concentration of GPE in brain, measured by mass spectrometry ( MS), was low, and the concentration of total IGF-I (measured by ELISA with a commercial polyclonal antibody [P Ab]) was 40 times higher at 50 nmol/kg. This also, was not consistent with the action of a tripeptidyl peptidase in brain that converted all IGF-I to des(1-3)IGF-I plus GPE. Contrasting ELISA results, using the MAb prepared in this study, suggest an even higher concentration of intact IGF-I of 150 nmollkg. This would argue against the presence of any des( 1-3 )IGF-I in brain, but in turn, this indicates either the presence of other substances containing a GPE amino-terminus or other cross reacting epitope. Although the results of the specificity studies reported in Chapter 5 would make this latter possibility seem unlikely, it cannot be completely excluded. No GP was detected in brain by MS. No GPE was detected in colostrum by capillary electrophoresis (CE) but the interference from extraneous substances reduced the detectability of GPE by CE and this approach would require further, prior, purification and concentration steps. A molecule, with a migration time equal to that of the peptide GP, was detected in colostrum by CE, but the concentration (~ 10 11mo/L) was much higher than the IGF-I concentration measured by radio-immunoassay using a PAb (80 nmol/L) or using a Mab (300-400 nmolL). A DPP IV enzyme was detected in colostrum and this could account for the GP, derived from substrates other than IGF-1. Based on the differential results of the two antibody assays, there was no indication of the presence of des(1-3)IGF-I in brain or colostrum. In the absence of any enzyme activity directed towards the amino terminus of IGF-I and the absence any potential products, IGF-I, therefore, does not appear to "mature" via des(1-3)IGF-I in the brain, nor in the neutral colostrum. In spite of these results which indicate the absence of an enzymic attack on IGF-I and the absence of the expected products in tissues, the possibility that the conversion of IGF-I may occur in neutral conditions in limited amounts, cannot be ruled out. It remains possible that in the extracellular environment of the membrane, a complex interaction of IGF-I, binding protein, aminopeptidase(s) and receptor, produces des(1- 3)IGF-I as a transient product which is bound to the receptor and internalised.

Whitening Race: Essays In Social And Cultural Criticism In Australia

Aileen Moreton-Robinson has brought together scholars from a range of disciplines: philosophy, cultural and gender studies, education, social work, sociology and literary studies. All engage critically with the location of the social and discursive construction of whiteness.

Struggles Over Difference: Curriculum, texts and pedagogy in the Asia-Pacific

Struggles over Difference addresses education, schools, textbooks, and pedagogies in various countries of the Asia-Pacific, offering critical curriculum studies and policy analyses of national and regional educational systems. These systems face challenges linked to new economic formations, cultural globalization, and emergent regional and international geopolitical instabilities and conflicts. Contributors offer insights on how official knowledge, text, discourse and discipline should be shaped; who should shape it; through which institutional agencies it should be administered: and social and cultural practices through which this should occur.

From Civil Strife to Civil Society: civil and military responsibilities in disrupted states

"The 1990s saw the United Nations, the militaries of key member states, and NGOs increasingly entangled in the complex affairs of disrupted states. Whether as deliverers of humanitarian assistance or as agents of political, social, and civic reconstruction, whether in Somalia, Bosnia, Kosovo, or East Timor, these actors have had to learn ways of interacting with each other in order to optimize the benefits for the populations they seek to assist. Yet the challenges have proved daunting. Civil and military actors have different organizational cultures and standard operating procedures and are confronted with the need to work together to perform tasks to which different actors may attach quite different priorities."--BOOK JACKET.