Enhancing methacrylate-monolith-based downstream processes to champion plasmid DNA production

Increasing numbers of preclinical and clinical studies are utilizing pDNA (plasmid DNA) as the vector. In addition, there has been a growing trend towards larger and larger doses of pDNA utilized in human trials. The growing demand on pDNA manufacture leads to pressure to make more in less time. A key intervention has been the use of monoliths as stationary phases in liquid chromatography. Monolithic stationary phases offer fast separation to pDNA owing to their large pore size, making pDNA in the size range from 100 nm to over 300 nm easily accessible. However, the convective transport mechanism of monoliths does not guarantee plasmid purity. The recovery of pure pDNA hinges on a proper balance in the properties of the adsorbent phase, the mobile phase and the feedstock. The effects of pH and ionic strength of binding buffer, temperature of feedstock, active group density and the pore size of the stationary phase were considered as avenues to improve the recovery and purity of pDNA using a methacrylate-based monolithic adsorbent and Escherichia coli DH5α-pUC19 clarified lysate as feedstock. pDNA recovery was found to be critically dependent on the pH and ionic strength of the mobile phase. Up to a maximum of approx. 92% recovery was obtained under optimum conditions of pH and ionic strength. Increasing the feedstock temperature to 80°C increased the purity of pDNA owing to the extra thermal stability associated with pDNA over contaminants such as proteins. Results from toxicological studies of the plasmid samples using endotoxin standard (E. coli 0.55:B5 lipopolysaccharide) show that endotoxin level decreases with increasing salt concentration. It was obvious that large quantities of pure pDNA can be obtained with minimal extra effort simply by optimizing process parameters and conditions for pDNA purification.

Growth medium selection and its economic impact on plasmid DNA production

Current developments in gene medicine and vaccination studies are utilizing plasmid DNA (pDNA) as the vector. For this reason, there has been an increasing trend towards larger and larger doses of pDNA utilized in human trials: from 100-1000 μg in 2002 to 500-5000 μg in 2005. The increasing demand of pDNA has created the need to revolutionalize current production levels under optimum economy. In this work, different standard media (LB, TB and SOC) for culturing recombinant Escherichia coli DH5α harbouring pUC19 were compared to a medium optimised for pDNA production. Lab scale fermentations using the standard media showed that the highest pDNA volumetric and specific yields were for TB (11.4 μg/ml and 6.3 μg/mg dry cell mass respectively) and the lowest was for LB (2.8 μg/ml and 3.3 μg/mg dry cell mass respectively). A fourth medium, PDMR, designed by modifying a stoichiometrically-formulated medium with an optimised carbon source concentration and carbon to nitrogen ratio displayed pDNA volumetric and specific yields of 23.8 μg/ml and 11.2 μg/mg dry cell mass respectively. However, it is the economic advantages of the optimised medium that makes it so attractive. Keeping all variables constant except medium and using LB as a base scenario (100 medium cost [MC] units/mg pDNA), the optimised PDMR medium yielded pDNA at a cost of only 27 MC units/mg pDNA. These results show that greater amounts of pDNA can be obtained more economically with minimal extra effort simply by using a medium optimised for pDNA production.

Process engineering aspects of plasmid-based biopharmaceuticals production : tackling the threatening vaccine shortages to prevent global pandemics

Infectious diseases such as SARS, influenza and bird flu have the potential to cause global pandemics; a key intervention will be vaccination. Hence, it is imperative to have in place the capacity to create vaccines against new diseases in the shortest time possible. In 2004, The Institute of Medicine asserted that the world is tottering on the verge of a colossal influenza outbreak. The institute stated that, inadequate production system for influenza vaccines is a major obstruction in the preparation towards influenza outbreaks. Because of production issues, the vaccine industry is facing financial and technological bottlenecks: In October 2004, the FDA was caught off guard by the shortage of flu vaccine, caused by a contamination in a US-based plant (Chiron Corporation), one of the only two suppliers of US flu vaccine. Due to difficulties in production and long processing times, the bulk of the world's vaccine production comes from very small number of companies compared to the number of companies producing drugs. Conventional vaccines are made of attenuated or modified forms of viruses. Relatively high and continuous doses are administered when a non-viable vaccine is used and the overall protective immunity obtained is ephemeral. The safety concerns of viral vaccines have propelled interest in creating a viable replacement that would be more effective and safer to use.

History, status and future of Australia's native Sydney rock oyster industry

The Sydney rock oyster (Saccostrea glomerata) (SRO) is an oyster species that only occurs in estuaries along Australia's east coast. The SRO industry evolved from commercial gathering of oyster in the 1790s to a high production volume aquaculture industry in the 1970s. However, since the late 1970s the SRO industry has experienced a significant and continuous decline in production quantities and the industry's future commercial viably appears to be uncertain. The aim of this study was to review the history and the status of the SRO industry and to discuss the potential future prospects of this industry. This study summarised findings of the existing literature about the industry and defined development stages of the industry. Particular focus was put on the more recent development within the industry (1980s-present) which has not been covered adequately in the existing literature. The finding from this study revealed that major issues of the industry are linked to the management of prevailing diseases, the handling of water quality impairments from increasing coastal development, increasing competition from Australia's Pacific oyster (Crassostrea gigas) industry and the current socio-economic profile of the industry. The study also found that policy makers are currently confronted by the dilemma of saving a "dying art". Findings from this industry review may be vital for current and future fisheries managers and stakeholders as a basis for reviewing industry management and development strategies. This review may also be of interest for other aquaculture industries and fisheries who are dealing with similar challenges as the SRO industry.

Reframe QUT's evaluation framework : a case study of planning, policy and positioning leading to educational best practice

Every university in Australia has a set of policies that guide the institution in its educational practices, however, the policies are often developed in isolation to each other. Now imagine a space where policies are evidence-based, refined annually, cohesively interrelated, and meet stakeholders’ needs. Is this happenstance or the result of good planning? Culturally, Queensland University of Technology (QUT) is a risk-averse institution that takes pride in its financial solvency and is always keen to know “how are we going?” With a twenty-year history of annual reporting that assures the quality of course performance through multiple lines of evidence, QUT’s Learning and Teaching Unit went one step further and strategically aligned a suite of policies that take into consideration the needs of their stakeholders, collaborate with other areas across the institution and use multiple lines of evidence to inform curriculum decision-making. In QUT’s experience, strategic planning can lead to policy that is designed to meet stakeholders’ needs, not manage them; where decision-making is supported by evidence, not rhetoric; where all feedback is incorporated, not ignored; and where policies are cohesively interrelated, not isolated. While many may call this ‘policy nirvana’, QUT has positioned itself to demonstrate good educational practice through Reframe, its evaluation framework. In this case, best practice was achieved through the application of a theory of change and a design-led logic model that allows for transition to other institutions with different cultural specificity. The evaluation approach follows Seldin’s (2003) notion to offer depth and breadth to the evaluation framework along with Berk’s (2005) concept of multiple lines of evidence. In summary, this paper offers university executives, academics, planning and quality staff an opportunity to understand the critical steps that lead to strategic planning and design of evidence-based educational policy that positions a university for best practice in learning and teaching.

Endotoxin removal : a critical step in vaccine production

A major drawback to the immunological potency of conventional vaccines, resulting in reduced level of immune responses, tissue injury, shock and high cytotoxicity, thus making their applications contraindicated in immunodeficiency diseases, is the presence of high contaminant concentrations in vaccine titers. Vaccine contamination arises from the simultaneous occurrence of competitive pathways resulting in the formation of other bio-products during cellular metabolism aside the pathways necessary for the production of vaccine molecules. One of such vaccine contaminating molecules is endotoxins which are mainly lipopolysaccharides (LPS) complexes found in the membrane of bacterial cell wall. The structural dynamics of these molecules make their removal from vaccine titers problematic, thus making vaccine endotoxin removal a major research endeavour. This presentation will discuss a novel technique for reducing the endotoxin level of vaccines. The technique commences with the disentanglement of endotoxin-vaccine molecular bonding and then capturing the vaccine molecules on an affinity monolith to separate the vaccine molecules from the endotoxins.

Dancing as Knowledge Production/Dancer as Researcher in Practice-led Research

This chapter discusses research undertaken for a PhD in dance, highlighting the oscillating and ambivalent nature of practice-led research methods. Holding an unusual position within the field of practice-led research in dance, the researcher adopted the role of dancer within four creative processes led by four choreographers. Although instigating and leading this research, the author produced new knowledge of dancing practices whilst being directed creatively by the choreographers in the research environment. In line with many definitions of practice-led research, the methods used were emergent from the research arena and responsive to the requirements of the project. Deeply imbricated in the research environment, the researcher used her embodied self as a research tool, as the one who participated, discovered and recorded. This outsider/insider role of researcher/participant produced a series of meta-positions, leading to a mixed-modal outcome that encompassed live performance, a movement treatise and a written thesis.

Bio-hydrogen production from food waste through anaerobic fermentation

In order to protect our planet and ourselves from the adverse effects of excessive CO2 emissions and to prevent an imminent non-renewable fossil fuel shortage and energy crisis, there is a need to transform our current ‘fossil fuel dependent’ energy systems to new, clean, renewable energy sources. The world has recognized hydrogen as an energy carrier that complies with all the environmental quality and energy security, demands. This research aimed at producing hydrogen through anaerobic fermentation, using food waste as the substrate. Four food waste substrates were used: Rice, fish, vegetable and their mixture. Bio-hydrogen production was performed in lab scale reactors, using 250 mL serum bottles. The food waste was first mixed with the anaerobic sewage sludge and incubated at 37°C for 31 days (acclimatization). The anaerobic sewage sludge was then heat treated at 80°C for 15 min. The experiment was conducted at an initial pH of 5.5 and temperatures of 27, 35 and 55°C. The maximum cumulative hydrogen produced by rice, fish, vegetable and mixed food waste substrates were highest at 37°C (Rice =26.97±0.76 mL, fish = 89.70±1.25 mL, vegetable = 42.00±1.76 mL, mixed = 108.90±1.42 mL). A comparative study of acclimatized (the different food waste substrates were mixed with anaerobic sewage sludge and incubated at 37°C for 31days) and non-acclimatized food waste substrate (food waste that was not incubated with anaerobic sewage sludge) showed that acclimatized food waste substrate enhanced bio-hydrogen production by 90 - 100%.

Clinton, Bush, and Obama : changing policy and rhetoric in the United States Annual Trafficking in Persons Report

This chapter charts the political transitions in the anti-trafficking agenda and rhetoric of the U.S. Government across three Presidential administrations through a detailed examination of the annual Trafficking in Persons reports released by the Office to Monitor and Combat Trafficking in Persons between 2001 and 2012. We argue that the transitions in language and focus reflect key tensions that have dominated trafficking discourse throughout the Clinton, Bush and Obama Presidencies. These fissures include debate over law enforcement versus rights-based frameworks, competing approaches on victim protection and identification, and ongoing disputes about the relationship between prostitution and human trafficking.

ACE research vignette 039: The Ineffectiveness of Enterprise Policy : Is Policy Formulation to Blame?

This series of research vignettes is aimed at sharing current and interesting research findings from our team of international Entrepreneurship researchers. This vignette, written by Dr Norin Arshed, Professor Sara Carter and Professor Colin Mason examines whether the ineffectiveness of enterprise policy can be attributed to the formulation of the enterprise policy itself.

Practical linkages between cultural policy and education policy in promoting a creative workforce for youth in Tanzania

The broad aim of of this thesis is to contribute to understanding how the relationships between culture, employment and education can help Tanzania's young people secure jobs, and survive in the creative workforce so as to better their future. Based on a range of interviews and other data in Tanzania, the study considers how to integrate cultural expressions into arts education (education in art and education through art) as a tool for nurturing young people's creative talents for their future sustainable employment in Tanzania.

Illuminative evaluation as a method applied to Australian Government policy borrowing and implementation in higher education

Any government deciding to invoke widespread change in its higher education sector through implementation of new policies impacts on every institution and all staff and students, often in both the time taken up and the heightened emotions caused. The central phenomenon that this study addresses is the process and consequences of policy changes in higher education in Australia. The aim of this article is to record the research design through the perspective (evaluation research), theoretical framework (program evaluation) and methods (content analysis, descriptive statistical analysis and bibliometric analysis) applied to the investigation of the 2003 federal government higher education reform package. This approach allows both the intended and unintended consequences arising from the policy implementation of three national initiatives focused on learning and teaching in higher education in Australia to surface. As a result, this program evaluation, also known in some disciplines as policy implementation analysis, will demonstrate the applicability of illuminative evaluation as a methodology and reinforce how program evaluation will assist and advise future government reform and policy implementation, and will serve as a legacy for future evaluative research.Any government deciding to invoke widespread change in its higher education sector through implementation of new policies impacts on every institution and all staff and students, often in both the time taken up and the heightened emotions caused. The central phenomenon that this study addresses is the process and consequences of policy changes in higher education in Australia. The aim of this article is to record the research design through the perspective (evaluation research), theoretical framework (program evaluation) and methods (content analysis, descriptive statistical analysis and bibliometric analysis) applied to the investigation of the 2003 federal government higher education reform package. This approach allows both the intended and unintended consequences arising from the policy implementation of three national initiatives focused on learning and teaching in higher education in Australia to surface. As a result, this program evaluation, also known in some disciplines as policy implementation analysis, will demonstrate the applicability of illuminative evaluation as a methodology and reinforce how program evaluation will assist and advise future government reform and policy implementation, and will serve as a legacy for future evaluative research.

Energy use assessment of educational buildings: Toward a campus-wide sustainable energy policy

The purpose of this article is to assess the viability of blanket sustainability policies, such as Building Rating Systems in achieving energy efficiency in university campus buildings. We analyzed the energy consumption trends of 10 LEED-certified buildings and 14 non-LEED certified buildings at a major university in the US. Energy Use Intensity (EUI) of the LEED buildings was significantly higher (EUILEED= 331.20 kBtu/sf/yr) than non-LEED buildings (EUInon-LEED=222.70 kBtu/sf/yr); however, the median EUI values were comparable (EUILEED= 172.64 and EUInon-LEED= 178.16). Because the distributions of EUI values were non-symmetrical in this dataset, both measures can be used for energy comparisons—this was also evident when EUI computations exclude outliers, EUILEED=171.82 and EUInon-LEED=195.41. Additional analyses were conducted to further explore the impact of LEED certification on university campus buildings energy performance. No statistically significant differences were observed between certified and non-certified buildings through a range of robust comparison criteria. These findings were then leveraged to devise strategies to achieve sustainable energy policies for university campus buildings and to identify potential issues with portfolio level building energy performance comparisons.