Investigation of molecular mechanisms regulating biomineralization of pearl oyster Pinctada maxima

Biomineralization is a process encompassing all mineral containing tissues produced within an organism. The most dynamic example of this process is the formation of the mollusk shell, comprising a variety of crystal phases and microstructures. The organic component incorporated within the shell is said to dictate this remarkable architecture. Subsequently, for the past decade considerable research have been undertaken to identify and characterize the protein components involved in biomineralization. Despite these efforts the general understanding of the process remains ambiguous. This study employs a novel molecular approach to further the elucidation of the shell biomineralization. A microarray platform has been custom generated (PmaxArray 1.0) from the pearl oyster Pinctada maxima. PmaxArray 1.0 consists of 4992 expressed sequence tags (ESTs) originating from the mantle, an organ involved in shell formation. This microarray has been used as the primary tool for three separate investigations in an effort to associate transcriptional gene expression from P. maxima to the process of shell biomineralization. The first investigation analyzes the spatial expression of ESTs throughout the mantle organ. The mantle was dissected into five discrete regions and each analyzed for gene expression with PmaxArray 1.0. Over 2000 ESTs were differentially expressed among the tissue sections, identifying five major expression regions. Three of these regions have been proposed to have shell formation functions belonging to nacre, prismatic calcite and periostracum. The spatial gene expression map was confirmed by in situ hybridization, localizing a subset of ESTs from each expression region to the same mantle area. Comparative sequence analysis of ESTs expressed in the proposed shell formation regions with the BLAST tool, revealed a number of the transcripts were novel while others showed significant sequence similarities to previously characterized shell formation genes. The second investigation correlates temporal EST expression during P. maxima larval ontogeny with transitions in shell mineralization during the same period. A timeline documenting the morphologicat microstructural and mineralogical shell characteristics of P. maxima throughout larval ontogeny has been established. Three different shell types were noted based on the physical characters and termed, prodissoconch I, prodissoconch 11 and dissoconch. PmaxArray 1.0 analyzed ESTs expression of animals throughout the larval development of P. maxima, noting up-regulation of 359 ESTs in association with the shell transitions from prodissoconch 1 to prodissoconch 11 to dissoconch. Comparative sequence analysis of these ESTs indicates a number of the transcripts are novel as well as showing significant sequence similarities between ESTs and known shell matrix associated genes and proteins. These ESTs are discussed in relation to the shell characters associated with their temporal expression. The third investigation uses PmaxArray 1.0 to analyze gene expression in the mantle tissue of P. maxima specimens exposed to sub-lethal concentrations of a shell-deforming toxin, tributyltin (TBT). The shell specific effects of TBT are used in this investigation to interpret differential expression of ESTs with respect to shell formation functions. A lethal and sublethal TBT concentration range was established for P. maxima, noting a concentration of 50 ng L- 1 TBT as sub-lethal over a 21 day period. Mantle tissue from P. maxima animals treated with 50 ng L- 1 TBT was assessed for differential EST expression with untreated control animals. A total of 102 ESTs were identified as differentially expressed in association with TBT exposure, comparative sequence identities included an up-regulation of immunity and detoxification related genes and down-regulation of several shell matrix genes. A number of transcripts encoding novel peptides were additionally identified. The potential actions of these genes are discussed with reference to TBT toxicity and shell biomineralization. This thesis has used a microarray platform to analyze gene expression in spatial, temporal and toxicity investigations, revealing the involvement of numerous gene transcripts in specific shell formation functions. Investigation of thousands of transcripts simultaneously has provided a holistic interpretation of the organic components regulating shell biomineralization.

Action learning of online marketing via a global contest

Introduction: The Google Online Marketing Challenge is a global competition in which student teams run advertising campaigns for small and medium-sized businesses (SMEs) using AdWords, Google’s text-based advertisements. In 2008, its inaugural year, over 8,000 students and 300 instructors from 47 countries representing over 200 schools participated. The Challenge ran in undergraduate and graduate classes in disciplines such as marketing, tourism, advertising, communication and information systems. Combining advertising and education, the Challenge gives student hands-on experience in the increasingly important field of online marketing, engages them with local businesses and motivates them through the thrill of a global competition. Student teams receive US$200 in AdWords credits, Google’s premier advertising product that offers cost-per-click advertisements. The teams then recruit and work with a local business to devise an effective online marketing campaign. Students first outline a strategy, run a series of campaigns, and provide their business with recommendations to improve their online marketing. Teams submit two written reports for judging by 14 academics in eight countries. In addition, Google AdWords experts judge teams on their campaign statistics such as success metrics and account management. Rather than a marketing simulation against a computer or hypothetical marketing plans for hypothetical businesses, the Challenges has student teams develop and manage real online advertising campaigns for their clients and compete against peers globally.

Wear mechanisms and scale effects in two-body abrasion

The ‘particle size effect’ and its manifestation in abrasion still attracts considerable debate as to its origins and the ranking of its likely causes. Experiments have been conducted to study the important contribution that the formation of wear debris can have on the progression of wear. The experiments consist of unlubricated (dry) pin-on-disk tests with silicon carbide coated paper of varying particle size, with different pin material, diameter and loads. It has been observed that the influence of debris formation on wear rate is more pronounced for fine abrasives and soft-wearing materials. Consequently, it is proposed that the particle size effect can be explained in terms of geometrical scaling and the evolution of third-body effects with diminishing particle diameter.

Reciprocity : where art meets the community : action research in response to artistic encounters and relationships

This practice-led research project examines some of the factors and issues facing artists working in the public domain who wish to engage with the community as audience. Using the methodology of action research, the three major creative projects in this study use art as a socio-political tool with the aim of providing an effective vehicle for broadening awareness, understanding forms of social protest and increasing tolerance for diversity. The three projects: Floodline November 7, 2004, Look in, Look out, and The Urban Terrorist Project, dealt with issues of marginalisation of communities, audiences and graffiti artists respectively. The artist/researcher is outlined as both creator and collaborator in the work. Processes included ephemeral elements, such as temporary installation and performance, as well as interactive elements that encouraged direct audience involvement as part of the work. In addition to the roles of creator and collaborator, both of which included audience as well as artist, the presence of an outside entity was evident. Whether local, legal authorities or prevailing attitudes, outside entities had an unavoidable impact on the processes and outcomes of the work. Each project elicited a range of responses from their respective audiences; however, the overarching concept of reciprocity was seen to be the crucial factor in conception, artistic methods and outcomes.

Malnutrition and pressure ulcer risk in adults in Australian health care facilities

Aims To determine the effect of nutritional status on the presence and severity of pressure ulcers in statewide? public healthcare facilities, in Queensland, Australia. Research Methods A multicentre, cross sectional audit of nutritional status of a convenience sample of subjects was carried out as part of a large audit of pressure ulcers in a sample of state based public healthcare facilities in 2002 and 2003. Dietitians in 20 hospitals and six residential aged care facilities conducted single day nutritional status audits of 2208 acute and 839 aged care subjects using the Subjective Global Assessment. The effect of nutritional status on the presence, highest stage and number of pressure ulcers was determined by logistic regression in a model controlling for age, gender, medical specialty and facility location. The potential clustering effect of facility was accounted for in the model using an analysis of correlated data approach. Results Subjects with malnutrition had an adjusted odds risk of 2.6 (95% CI 1.8-3.5, p<0.001) of having a pressure ulcer in acute facilities and 2.0 (95% CI 1.5-2.7, p<0.001) for residential aged care facilities. There was also increased odds risk of having a pressure ulcer, having a higher stage pressure ulcer and a higher number of pressure ulcers with increased severity of malnutrition. Conclusion Malnutrition was associated with at least twice the odds risk of having a pressure ulcer of in public healthcare facilities in Queensland. Action must be taken to identify, prevent and treat malnutrition, especially in patients at risk of pressure ulcer.

Discovery and characterization of IGFBP-mediated endocytosis in the human retinal pigment epithelial cell line ARPE-19

Insulin-like growth factor binding proteins (IGFBPs) are prime regulators of IGF-action in numerous cell types including the retinal pigment epithelium (RPE). The RPE performs several functions essential for vision, including growth factor secretion and waste removal via a phagocytic process mediated in part by vitronectin (Vn). In the course of studying the effects of IGFBPs on IGF-mediated VEGF secretion and Vn-mediated phagocytosis in the RPE cell line ARPE-19, we have discovered that these cells avidly ingest synthetic microspheres (2.0 μm diameter) coated with IGFBPs. Given the novelty of this finding and the established role for endocytosis in mediating IGFBP actions in other cell types, we have explored the potential role of candidate cell surface receptors. Moreover, we have examined the role of key IGFBP structural motifs, by comparing responses to three members of the IGFBP family (IGFBP-3, IGFBP-4 and IGFBP-5) which display overlapping variations in primary structure and glycosylation status. Coating of microspheres (FluoSpheres®, sulfate modified polystyrene filled with a fluorophore) was conducted at 37 °C for 1 h using 20 μg/mL of test protein, followed by extensive washing. Binding of proteins was confirmed using a microBCA assay. The negative control consisted of microspheres treated with 0.1% bovine serum albumin (BSA), and all test samples were post-treated with BSA in an effort to coat any remaining free protein binding sites, which might otherwise encourage non-specific interactions with the cell surface. Serum-starved cultures of ARPE-19 cells were incubated with microspheres for 24 h, using a ratio of approximately 100 microspheres per cell. Uptake of microspheres was quantified using a fluorometer and was confirmed visually by confocal fluorescence microscopy. The ARPE-19 cells displayed little affinity for BSA-treated microspheres, but avidly ingested large quantities of those pre-treated with Vn (ANOVA; p < 0.001). Strong responses were also observed towards recombinant formulations of non-glycosylated IGFBP-3, glycosylated IGFBP-3 and glycosylated IGFBP-5 (all p < 0.001), while glycosylated IGFBP-4 induced a relatively minor response (p < 0.05). The response to IGFBP-3 was unaffected in the presence of excess soluble IGFBP-3, IGF-I or Vn. Likewise, soluble IGFBP-3 did not induce uptake of BSA-treated microspheres. Antibodies to either the transferrin receptor or type 1 IGF-receptor displayed slight inhibitory effects on responses to IGFBPs and Vn. Heparin abolished responses to Vn, IGFBP-5 and non-glycosylated IGFBP-3, but only partially inhibited the response to glycosylated IGFBP-3. Our results demonstrate for the first time IGFBP-mediated endocytosis in ARPE-19 cells and suggest roles for the IGFBP-heparin-binding domain and glycosylation status. These findings have important implications for understanding the mechanisms of IGFBP actions on the RPE, and in particular suggest a role for IGFBP-endocytosis.

Realistic yet humanitarian? The comprehensive plan of action and refugee policy in Southeast Asia

The 1989 Comprehensive Plan of Action (CPA) has recently been described as a successful example of how to manage large protracted refugee flows. However, this article revisits the circumstances surrounding the CPA used to resolve the prolonged Indo-Chinese refugee crisis to highlight that part of its development was linked to the fact that Southeast Asian states refused to engage with proposed solutions, which did not include repatriation for the majority of the Indo-Chinese asylum seekers who were deemed to be ‘non-genuine’1 ( UNGA, 1989a) refugees. This resulted in the CPA often forcibly repatriating ‘non-genuine’ refugees, particularly near the end of its program. This article reviews the CPA in order to assess whether its practices and results should be repeated.

Theoretical investigation of mechanisms of formation and interaction of nanoparticles

In this thesis an investigation into theoretical models for formation and interaction of nanoparticles is presented. The work presented includes a literature review of current models followed by a series of five chapters of original research. This thesis has been submitted in partial fulfilment of the requirements for the degree of doctor of philosophy by publication and therefore each of the five chapters consist of a peer-reviewed journal article. The thesis is then concluded with a discussion of what has been achieved during the PhD candidature, the potential applications for this research and ways in which the research could be extended in the future. In this thesis we explore stochastic models pertaining to the interaction and evolution mechanisms of nanoparticles. In particular, we explore in depth the stochastic evaporation of molecules due to thermal activation and its ultimate effect on nanoparticles sizes and concentrations. Secondly, we analyse the thermal vibrations of nanoparticles suspended in a fluid and subject to standing oscillating drag forces (as would occur in a standing sound wave) and finally on lattice surfaces in the presence of high heat gradients. We have described in this thesis a number of new models for the description of multicompartment networks joined by a multiple, stochastically evaporating, links. The primary motivation for this work is in the description of thermal fragmentation in which multiple molecules holding parts of a carbonaceous nanoparticle may evaporate. Ultimately, these models predict the rate at which the network or aggregate fragments into smaller networks/aggregates and with what aggregate size distribution. The models are highly analytic and describe the fragmentation of a link holding multiple bonds using Markov processes that best describe different physical situations and these processes have been analysed using a number of mathematical methods. The fragmentation of the network/aggregate is then predicted using combinatorial arguments. Whilst there is some scepticism in the scientific community pertaining to the proposed mechanism of thermal fragmentation,we have presented compelling evidence in this thesis supporting the currently proposed mechanism and shown that our models can accurately match experimental results. This was achieved using a realistic simulation of the fragmentation of the fractal carbonaceous aggregate structure using our models. Furthermore, in this thesis a method of manipulation using acoustic standing waves is investigated. In our investigation we analysed the effect of frequency and particle size on the ability for the particle to be manipulated by means of a standing acoustic wave. In our results, we report the existence of a critical frequency for a particular particle size. This frequency is inversely proportional to the Stokes time of the particle in the fluid. We also find that for large frequencies the subtle Brownian motion of even larger particles plays a significant role in the efficacy of the manipulation. This is due to the decreasing size of the boundary layer between acoustic nodes. Our model utilises a multiple time scale approach to calculating the long term effects of the standing acoustic field on the particles that are interacting with the sound. These effects are then combined with the effects of Brownian motion in order to obtain a complete mathematical description of the particle dynamics in such acoustic fields. Finally, in this thesis, we develop a numerical routine for the description of "thermal tweezers". Currently, the technique of thermal tweezers is predominantly theoretical however there has been a handful of successful experiments which demonstrate the effect it practise. Thermal tweezers is the name given to the way in which particles can be easily manipulated on a lattice surface by careful selection of a heat distribution over the surface. Typically, the theoretical simulations of the effect can be rather time consuming with supercomputer facilities processing data over days or even weeks. Our alternative numerical method for the simulation of particle distributions pertaining to the thermal tweezers effect use the Fokker-Planck equation to derive a quick numerical method for the calculation of the effective diffusion constant as a result of the lattice and the temperature. We then use this diffusion constant and solve the diffusion equation numerically using the finite volume method. This saves the algorithm from calculating many individual particle trajectories since it is describes the flow of the probability distribution of particles in a continuous manner. The alternative method that is outlined in this thesis can produce a larger quantity of accurate results on a household PC in a matter of hours which is much better than was previously achieveable.

Investigation of mechanisms governing emission of odorants

The literature identifies several models that describe inter-phase mass transfer, key to the emission process. While the emission process is complex and these models may be more or less successful at predicting mass transfer rates, they identify three key variables for a system involving a liquid and an air phase in contact with it: • A concentration (or partial pressure) gradient driving force; • The fluid dynamic characteristics within the liquid and air phases, and • The chemical properties of the individual components within the system. In three applied research projects conducted prior to this study, samples collected with two well-known sampling devices resulted in very different odour emission rates. It was not possible to adequately explain the differences observed. It appeared likely, however, that the sample collection device might have artefact effects on the emission of odorants, i.e. the sampling device appeared to have altered the mass transfer process. This raised the obvious question: Where two different emission rates are reported for a single source (differing only in the selection of sampling device), and a credible explanation for the difference in emission rate cannot be provided, which emission rate is correct? This research project aimed to identify the factors that determine odour emission rates, the impact that the characteristics of a sampling device may exert on the key mass transfer variables, and ultimately, the impact of the sampling device on the emission rate itself. To meet these objectives, a series of targeted reviews, and laboratory and field investigations, were conducted. Two widely-used, representative devices were chosen to investigate the influence of various parameters on the emission process. These investigations provided insight into the odour emission process generally, and the influence of the sampling device specifically.

Mechanisms of change in negative thinking and urinary monoamines in depressed patients during acute treatment with group cognitive behavior therapy and antidepressant medication

This naturalistic study investigated the mechanisms of change in measures of negative thinking and in 24-h urinary metabolites of noradrenaline (norepinephrine), dopamine and serotonin in a sample of 43 depressed hospital patients attending an eight-session group cognitive behavior therapy program. Most participants (91%) were taking antidepressant medication throughout the therapy period according to their treating Psychiatrists' prescriptions. The sample was divided into outcome categories (19 Responders and 24 Non-responders) on the basis of a clinically reliable change index [Jacobson, N.S., & Truax, P., 1991. Clinical significance: a statistical approach to defining meaningful change in psychotherapy research. Journal of Consulting and Clinical Psychology, 59, 12–19.] applied to the Beck Depression Inventory scores at the end of the therapy. Results of repeated measures analysis of variance [ANOVA] analyses of variance indicated that all measures of negative thinking improved significantly during therapy, and significantly more so in the Responders as expected. The treatment had a significant impact on urinary adrenaline and metadrenaline excretion however, these changes occurred in both Responders and Non-responders. Acute treatment did not significantly influence the six other monoamine metabolites. In summary, changes in urinary monoamine levels during combined treatment for depression were not associated with self-reported changes in mood symptoms.

Venture creation and resource processes : using Bricolage sustainability ventures

Principal Topic: Resource decisions are critical to the venture creation process, which has important subsequent impacts on venture creation and performance (Boeker, 1989). Most entrepreneurs however, suffer substantial resource constraints in venture creation and during venture growth (Shepherd et al., 2000). Little is known about how high potential, sustainability ventures (the ventures of interest in this research), despite resource constraints, achieve continued venture persistence and venture success. One promising theory that explicitly links to resource constraints is a concept developed by Levi Strauss (1967) termed bricolage. Bricolage aligns with notions of resourcefulness: using what's on hand, through making do, and recombining resources for new or novel purposes (Baker & Nelson 2005). To the best of our knowledge, previous studies have not systematically investigated internal and external constraints, their combinations, and subsequent bricolage patterns. The majority of bricolage literature focuses on external environmental constraints (e.g. Wieck 1989; Baker & Nelson 2005), thereby paying less attention to in evaluating internal constraints (e.g. skills and capabilities) or constraint combinations. In this paper we focus on ventures that typically face resource-poor environments. High potential, nascent and young sustainability ventures are often created and developed with resource constraints and in some cases, have greater resource requirements owing to higher levels of technical sophistication of their products (Rothaermel & Deeds 2006). These ventures usually have high aspirations and potential for growth who ''seeks to meet the needs and aspirations without compromising the ability to meet those of the future'' (Brundtland Commission 1983). High potential ventures are increasingly attributed with a central role in the development of innovation, and employment in developed economies (Acs 2008). Further, increasing awareness of environmental and sustainability issues has fostered demand for business processes that reduce detrimental environmental impacts of global development (Dean & McMullen 2007) and more environmentally sensitive products and services: representing an opportunity for the development of ventures that seek to satisfy this demand through entrepreneurial action. These ventures may choose to ''make do'' with existing resources in developing resource combinations that produce the least impact on the environment. The continuous conflict between the greater requirements for resources and limited resource availability in high potential sustainable ventures, with the added complexity of balancing this with an uncompromising focus on using ''what's on hand'' to lessen environment impacts may make bricolage behaviours critical for these ventures. Research into bricolage behaviour is however, the exception rather than the rule (Cunha 2005). More research is therefore needed to further develop and extend this emerging concept, especially in the context of sustainability ventures who are committed to personal and social goals of resourcefulness. To date, however, bricolage has not been studied specifically among high potential sustainable ventures. This research seeks to develop an in depth understanding of the impact of internal and external constraints and their combinations on the mechanisms employed in bricolage behaviours in differing dynamic environments. The following research question was developed to investigate this: How do internal, external resource constraints (or their combinations) impact bricolage resource decisions in high potential sustainability ventures? ---------- Methodology/Key Propositions: 6 case studies will be developed utilizing survey data from the Comprehensive Australian Study of Entrepreneurial Emergence (CAUSEE) large-scale longitudinal study of new venture start-ups in Australia. Prior to commencing case studies, 6 scoping interviews were conducted with key stakeholders including industry members, established businesses and government to ensure practical relevance in case development. The venture is considered the unit of analysis with the key informant being the entrepreneur and other management team members where appropriate. Triangulation techniques are used in this research including semi-structured interviews, survey data, onsite visits and secondary documentation website analysis, resumes, and business plans. These 6 sustainability ventures have been selected based on different environmental dynamism conditions including a traditionally mature market (building industry) and a more dynamic, evolving industry (renewable energy/solar ventures). In evaluating multidisciplinary literature, we expect the following external constraints are critical including: technology constraints (seen through lock-in of incumbents existing technology), institutional regulation and standards, access to markets, knowledge and training to nascent and young venture bricolage processes. The case studies will investigate internal constraints including resource fungability, resource combination capabilities, translating complex science/engineering knowledge into salient, valuable market propositions, i.e. appropriate market outcomes, and leveraging relationships may further influence bricolage decisions. ---------- Results and Implications: Intended ventures have been identified within the CAUSEE sample and have agreed to participate and secondary data collection for triangulation purposes has already commenced. Data collection of the case studies commenced 27th of May 2009. Analysis is expected to be completed finalised by 25th September 2009. This paper will report on the pattern of resource constraints and its impact on bricolage behaviours: its subsequent impact on resource deployment within venture creation and venture growth. As such, this research extends the theory of bricolage through the systematic analysis of constraints on resource management processes in sustainability ventures. For practice, this research may assist in providing a better understanding of the resource requirements and processes needed for continued venture persistence and growth in sustainability ventures. In these times of economic uncertainty, a better understanding of the influence on constraints and bricolage: the interplay of behaviours, processes and outcomes may enable greater venture continuance and success.