Development of a 3D culture system to study the skeletal metastasis of prostate cancer

In the cancer research field, most in vitro studies still rely on two-dimensional (2D) cultures. However, the trend is rapidly shifting towards using a three-dimensional (3D) culture system. This is because 3D models better recapitulate the microenvironment of cells, and therefore, yield cellular and molecular responses that more accurately describe the pathophysiology of cancer. By adopting technology platforms established by the tissue engineering discipline, it is now possible to grow cancer cells in extracellular matrix (ECM)-like environments and dictate the biophysical and biochemical properties of the matrix. In addition, 3D models can be modified to recapitulate different stages of cancer progression for instance from the initial development of tumor to metastasis. Inevitably, to recapitulate a heterotypic condition, comprising more than one cell type, it requires a more complex 3D model. To date, 3D models that are available for studying the prostate cancer (CaP)-bone interactions are still lacking. Therefore, the aim of this study is to establish a co-culture model that allows investigation of direct and indirect CaP-bone interactions. Prior to that, 3D polyethylene glycol (PEG)-based hydrogel cultures for CaP cells were first developed and growth conditions were optimised. Characterization of the 3D hydrogel cultures show that LNCaP cells form a multicellular mass that resembles avascular tumor. In comparison to 2D cultures, besides the difference in cell morphology, the response of LNCaP cells to the androgen analogue (R1881) stimulation is different compared to the cells in 2D cultures. This discrepancy between 2D and 3D cultures is likely associated with the cell-cell contact, density and ligand-receptor interactions. Following the 3D monoculture study, a 3D direct co-culture model of CaP cells and the human tissue engineered bone (hTEBC) construct was developed. Interactions between the CaP cells and human osteoblasts (hOBs) resulted in elevation of Matrix Metalloproteinase 9 (MMP9) for PC-3 cells and Prostate Specific Antigen (PSA) for LNCaP cells. To further investigate the paracrine interaction of CaP cells and (hOBs), a 3D indirect co-culture model was developed, where LNCaP cells embedded within PEG hydrogels were co-cultured with hTEBC. It was found that the cellular changes observed reflect the early event of CaP colonizing the bone site. In the absence of androgens, interestingly, up-regulation of PSA and other kallikreins is also detected in the co-culture compared to the LNCaP monoculture. This non androgenic stimulation could be triggered by the soluble factors secreted by the hOB such as Interleukin-6. There are also decrease in alkaline phosphatase (ALP) activity and down-regulation of genes of the hOB when co-cultured with LNCaP cells that have not been previously described. These genes include transforming growth factor β1 (TGFβ1), osteocalcin and Vimentin. However, no changes to epithelial markers (e.g E-cadherin, Cytokeratin 8) were observed in both cell types from the co-culture. Some of these intriguing changes observed in the co-cultures that had not been previously described have enriched the basic knowledge of the CaP cell-bone interaction. From this study, we have shown evidence of the feasibility and versatility of our established 3D models. These models can be adapted to test various hypotheses for studies pertaining to underlying mechanisms of bone metastasis and could provide a vehicle for anticancer drug screening purposes in the future.

Development of a transformation system for sorghum (Sorghum bicolor L.)

Sorghum (Sorghum bicolor (L.) Moench) is the world’s fifth major cereal crop and holds importance as a construction material, food and fodder source. More recently, the potential of this plant as a biofuel source has been noted. Despite its agronomic importance, the use of sorghum production is being constrained by both biotic and abiotic factors. These challenges could be addressed by the use of genetic engineering strategies to complement conventional breeding techniques. However, sorghum is one of the most recalcitrant crops for genetic modification with the lack of an efficient tissue culture system being amongst the chief reasons. Therefore, the aim of this study was to develop an efficient tissue culture system for establishing regenerable embryogenic cell lines, micropropagation and acclimatisation for Sorghum bicolor and use this to optimise parameters for genetic transformation via Agrobacterium-mediated transformation and microprojectile bombardment. Using five different sorghum cultivars, SA281, 296B, SC49, Wray and Rio, numerous parameters were investigated in an attempt to establish an efficient and reproducible tissue culture and transformation system. Using immature embryos (IEs) as explants, regenerable embryogenic cell lines (ECLs) could only be established from cultivars SA281 and 296B. Large amounts of phenolics were produced from IEs of cultivars, SC49, Wary and Rio, and these compounds severely hindered callus formation and development. Cultivar SA281 also produced phenolics during regeneration. Attempts to suppress the production of these compounds in cultivars SA281 and SC49 using activated charcoal, PVP, ascorbic acid, citric acid and liquid filter paper bridge methods were either ineffective or had a detrimental effect on embryogenic callus formation, development and regeneration. Immature embryos sourced during summer were found to be far more responsive in vitro than those sourced during winter. In an attempt to overcome this problem, IEs were sourced from sorghum grown under summer conditions in either a temperature controlled glasshouse or a growth chamber. However, the performance of these explants was still inferior to that of natural summer-sourced explants. Leaf whorls, mature embryos, shoot tips and leaf primordia were found to be unsuitable as explants for establishing ECLs in sorghum cultivars SA281 and 296B. Using the florets of immature inflorescences (IFs) as explants, however, ECLs were established and regenerated for these cultivars, as well as for cultivar Tx430, using callus induction media, SCIM, and regeneration media, VWRM. The best in vitro responses, from the largest possible sized IFs, were obtained using plants at the FL-2 stage (where the last fully opened leaf was two leaves away from the flag leaf). Immature inflorescences could be stored at 25oC for up to three days without affecting their in vitro responses. Compared to IEs, the IFs were more robust in tissue culture and showed responses which were season and growth condition independent. A micropropagation protocol for sorghum was developed in this study. The optimum plant growth regulator (PGR) combination for the micropropagation of in vitro regenerated plantlets was found to be 1.0 mg/L BAP in combination with 0.5 mg/L NAA. With this protocol, cultivars 296B and SA281 produced an average of 57 and 13 off-shoots per plantlet, respectively. The plantlets were successfully acclimatised and developed into phenotypically normal plants that set seeds. A simplified acclimatisation protocol for in vitro regenerated plantlets was also developed. This protocol involved deflasking in vitro plantlets with at least 2 fully-opened healthy leaves and at least 3 roots longer than 1.5 cm, washing the media from the roots with running tap water, planting in 100 mm pots and placing in plastic trays covered with a clear plastic bag in a plant growth chamber. After seven days, the corners of the plastic cover were opened and the bags were completely removed after 10 days. All plantlets were successfully acclimatised regardless of whether 1:1 perlite:potting mix, potting mix, UC mix or vermiculite were used as potting substrates. Parameters were optimised for Agrobacterium-mediated transformation (AMT) of cultivars SA281, 296B and Tx430. The optimal conditions were the use of Agrobacterium strain LBA4404 at an inoculum density of 0.5 OD600nm, heat shock at 43oC for 3 min, use of the surfactant Pluronic F-68 (0.02% w/v) in the inoculation media with a pH of 5.2 and a 3 day co-cultivation period in dark at 22oC. Using these parameters, high frequencies of transient GFP expression was observed in IEs precultured on callus initiation media for 1-7 days as well as in four weeks old IE- and IF-derived callus. Cultivar SA281 appeared very sensitive to Agrobacterium since all tissue turned necrotic within two weeks post-exposure. For cultivar 296B, GFP expression was observed up to 20 days post co-cultivation but no stably transformed plants were regenerated. Using cultivar Tx430, GFP was expressed for up to 50 days post co-cultivation. Although no stably transformed plants of this cultivar were regenerated, this was most likely due to the use of unsuitable regeneration media. Parameters were optimised for transformation by particle bombardment (PB) of cultivars SA281, 296B and Tx430. The optimal conditions were use of 3-7 days old IEs and 4 weeks old IF callus, 4 hour pre- and post-bombardment osmoticum treatment, use of 0.6 µm gold microparticles, helium pressure of 1500 kPa and target distance of 15 cm. Using these parameters for PB, transient GFP expression was observed for up to 14, 30 and 50 days for cultivars SA281, 296B and Tx430, respectively. Further, the use of PB resulted in less tissue necrosis compared to AMT for the respective cultivars. Despite the presence of transient GFP expression, no stably transformed plants were regenerated. The establishment of regenerable ECLs and the optimization of AMT and PB parameters in this study provides a platform for future efforts to develop an efficient transformation protocol for sorghum. The development of GM sorghum will be an important step towards improving its agronomic properties as well as its exploitation for biofuel production.

In vitro breeding strategies in the development of Australian native plants

Plant tissue culture is a technique that exploits the ability of many plant cells to revert to a meristematic state. Although originally developed for botanical research, plant tissue culture has now evolved into important commercial practices and has become a significant research tool in agriculture, horticulture and in many other areas of plant sciences. Plant tissue culture is the sterile culture of plant cells, tissues, or organs under aseptic conditions leading to cell multiplication or regeneration or organs and whole plants. The steps required to develop reliable systems for plant regeneration and their application in plant biotechnology are reviewed in countless books. Some of the major landmarks in the evolution of in vitro techniques are summarised in Table 5.1. In this chapter the current applications of this technology to agriculture, horticulture, forestry and plant breeding are briefly described with specific examples from Australian plants when applicable.

Development of a gas nanosensor node powered by solar cells

This work focuses on the development of a stand-alone gas nanosensor node, powered by solar energy to track concentration of polluted gases such as NO2, N2O, and NH3. Gas sensor networks have been widely developed over recent years, but the rise of nanotechnology is allowing the creation of a new range of gas sensors [1] with higher performance, smaller size and an inexpensive manufacturing process. This work has created a gas nanosensor node prototype to evaluate future field performance of this new generation of sensors. The sensor node has four main parts: (i) solar cells; (ii) control electronics; (iii) gas sensor and sensor board interface [2-4]; and (iv) data transmission. The station is remotely monitored through wired (ethernet cable) or wireless connection (radio transmitter) [5, 6] in order to evaluate, in real time, the performance of the solar cells and sensor node under different weather conditions. The energy source of the node is a module of polycrystalline silicon solar cells with 410cm2 of active surface. The prototype is equipped with a Resistance-To-Period circuit [2-4] to measure the wide range of resistances (KΩ to GΩ) from the sensor in a simple and accurate way. The system shows high performance on (i) managing the energy from the solar panel, (ii) powering the system load and (iii) recharging the battery. The results show that the prototype is suitable to work with any kind of resistive gas nanosensor and provide useful data for future nanosensor networks.

Development of a data mining-based analysis framework for multi-attribute construction project information

Data mining techniques extract repeated and useful patterns from a large data set that in turn are utilized to predict the outcome of future events. The main purpose of the research presented in this paper is to investigate data mining strategies and develop an efficient framework for multi-attribute project information analysis to predict the performance of construction projects. The research team first reviewed existing data mining algorithms, applied them to systematically analyze a large project data set collected by the survey, and finally proposed a data-mining-based decision support framework for project performance prediction. To evaluate the potential of the framework, a case study was conducted using data collected from 139 capital projects and analyzed the relationship between use of information technology and project cost performance. The study results showed that the proposed framework has potential to promote fast, easy to use, interpretable, and accurate project data analysis.

Comparing: knowledge-based urban development of Vancouver, Melbourne, Manchester and Boston

The research reported in this paper introduces a knowledge-based urban development assessment framework, which is constructed in order to evaluate and assist in the (re)formulation of local and regional policy frameworks and applications necessary in knowledge city transformations. The paper also reports the findings of an application of this framework in a comparative study of Boston, Vancouver, Melbourne and Manchester. The paper with its assessment framework: demonstrates an innovative way of examining the knowledge-based development capacity of cities by scrutinising their economic, socio-cultural, enviro-urban and institutional development mechanisms and capabilities; presents some of the generic indicators used to evaluate knowledge-based development performance of cities; reveals how a city can benchmark its development level against that of other cities, and; provides insights for achieving a more sustainable and knowledge-based development.

Towards the development of a gas sensor system for monitoring pollutant gases in the low troposphere using small unmanned aerial vehicles

The world is facing problems due to the effects of increased atmospheric pollution, climate change and global warming. Innovative technologies to identify, quantify and assess fluxes exchange of the pollutant gases between the Earth’s surface and atmosphere are required. This paper proposes the development of a gas sensor system for a small UAV to monitor pollutant gases, collect data and geo-locate where the sample was taken. The prototype has two principal systems: a light portable gas sensor and an optional electric–solar powered UAV. The prototype will be suitable to: operate in the lower troposphere (100-500m); collect samples; stamp time and geo-locate each sample. One of the limitations of a small UAV is the limited power available therefore a small and low power consumption payload is designed and built for this research. The specific gases targeted in this research are NO2, mostly produce by traffic, and NH3 from farming, with concentrations above 0.05 ppm and 35 ppm respectively which are harmful to human health. The developed prototype will be a useful tool for scientists to analyse the behaviour and tendencies of pollutant gases producing more realistic models of them.

Threat appraisal for harm from falls: Insights for development of education-based intervention

Objective: This study aims to describe how patients perceive the threat of falls in hospitals, to identify patient characteristics that are associated with greater or lesser perceptions of the threat of falls, and to examine whether there is a discord between the risk that patients perceive in general and the risk that they perceive for themselves personally. Method: A cross-sectional survey amongst geriatric rehabilitation inpatients in Brisbane, Australia, was implemented. The first component of the survey dealt with the ‘general’ nature of in-hospital falls and falls related risks while the second component of the survey was directed at identifying whether the patient held the same belief for themselves. Results: A total of 21 out of 125 participants (17%) indicated that they felt that they were at risk of falling during their hospitalisation and 28 (22%) felt that they would injure themselves if they were to fall. Self-perceived risk of falls was associated with decreasing age and lower cognitive function (Functional Independence Measure Cognitive score). A majority of patients felt that falls most commonly occur in the bathroom [n=67 (54%)] and that if they were to fall, they would fall in the bathroom [n=56 (45%)]. Discussion: Patients generally do not think they are at risk of falling while in hospital and this may contribute to poor adherence to falls prevention strategies. It is possible that raising patient perception of the risk of falls and injury from falls in hospitals may help improve adherence to falls prevention strategies in this setting.

An action research approach for the professional development of Vietnamese nurse educators

Nurse education in Viet Nam is undergoing substantial reform. In order to facilitate the change, in 2007 the Viet Nam Nurses Association formed a collaborative partnership with the School of Nursing and Midwifery at an Australia university. This collaboration gave rise to the Viet Nam Nursing Capacity Building Project under the leadership of Professor Genevieve Gray, funded by the Atlantic Philanthropies. The new four year competency based nursing curriculum frame is expected to be implemented in September 2011 following approval by the Viet Nam Ministry of Education. The focus of this paper is the Teaching Fellowships Program, an initiative of the Viet Nam Nursing Capacity Building Project, developed to help meet the challenges associated with leading and dealing with the curriculum change. The paper explores the development of the program and justifies an action research approach, illuminates key issues, and briefly refers to changes to the next fellowship program.

Communications materials on methods of development of revenue and cost estimates to non-technical audiences

This CDROM includes PDFs of presentations on the following topics: "TXDOT Revenue and Expenditure Trends;" "Examine Highway Fund Diversions, & Benchmark Texas Vehicle Registration Fees;" "Evaluation of the JACK Model;" "Future highway construction cost trends;" "Fuel Efficiency Trends and Revenue Impact"