Large-volume methacrylate monolith for plasmid purification : process engineering approach to synthesis and application

The extent of exothermicity associated with the construction of large-volume methacrylate monolithic columns has somewhat obstructed the realisation of large-scale rapid biomolecule purification especially for plasmid-based products which have proven to herald future trends in biotechnology. A novel synthesis technique via a heat expulsion mechanism was employed to prepare a 40 mL methacrylate monolith with a homogeneous radial pore structure along its thickness. Radial temperature gradient was recorded to be only 1.8 °C. Maximum radial temperature recorded at the centre of the monolith was 62.3 °C, which was only 2.3 °C higher than the actual polymerisation temperature. Pore characterisation of the monolithic polymer showed unimodal pore size distributions at different radial positions with an identical modal pore size of 400 nm. Chromatographic characterisation of the polymer after functionalisation with amino groups displayed a persistent dynamic binding capacity of 15.5 mg of plasmid DNA/mL. The maximum pressure drop recorded was only 0.12 MPa at a flow rate of 10 mL/min. The polymer demonstrated rapid separation ability by fractionating Escherichia coli DH5α-pUC19 clarified lysate in only 3 min after loading. The plasmid sample collected after the fast purification process was tested to be a homogeneous supercoiled plasmid with DNA electrophoresis and restriction analysis.

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.

Degradation of hydroxycinnamic acid mixtures in aqueous sucrose solutions by the Fenton process

The degradation efficiencies and behaviors of caffeic acid (CaA), p-coumaric acid (pCoA) and ferulic acid (FeA) in aqueous sucrose solutions containing the mixture of these hydroxycinnamic acids (HCAs) mixtures were studied by the Fenton oxidation process. Central composite design and multi-response surface methodology were used to evaluate and optimize the interactive effects of process parameters. Four quadratic polynomial models were developed for the degradation of each individual acid in the mixture and the total HCAs degraded. Sucrose was the most influential parameter that significantly affected the total amount of HCA degraded. Under the conditions studied there was < 0.01% loss of sucrose in all reactions. The optimal values of the process parameters for a 200 mg/L HCA mixture in water (pH 4.73, 25.15 °C) and sucrose solution (13 mass%, pH 5.39, 35.98 °C) were 77% and 57% respectively. Regression analysis showed goodness of fit between the experimental results and the predicted values. The degradation behavior of CaA differed from those of pCoA and FeA, where further CaA degradation is observed at increasing sucrose and decreasing solution pH. The differences (established using UV/Vis and ATR-FTIR spectroscopy) were because, unlike the other acids, CaA formed a complex with Fe(III) or with Fe(III) hydrogen-bonded to sucrose, and coprecipitated with lepidocrocite, an iron oxyhydroxide.

Process engineering approach to large-volume methacrylate monolith synthesis for plasmid purification

The construction of large?volume methacrylate monolithic columns for preparative-scale plasmid purification is obstructed by the enormous release of exotherms, thus introducing structural heterogeneity in the monolith pore system. A remarkable radial temperature gradient develops along the monolith thickness, reaching a terminal temperature that supersedes the maximum temperature required for the preparation of a structurally homogeneous monolith. A novel heat expulsion technique is employed to overcome the heat build-up during the synthesis process. The enormous heat build-up is perceived to encompass the heat associated with initiator decomposition and the heat released from free radical-monomer and monomer-monomer interactions. The heat resulting from the initiator decomposition was expelled along with some gaseous fumes before commencing polymerisation in a gradual addition fashion. Characteristics of a 50 mL monolith synthesized using this technique showed an improved uniformity in the pore structure radially along the length on the monolith. Chromatographic characterization of this adsorbent displayed a persistent binding capacity of 14.5 mg pDNA/mL of the adsorbent. The adsorbent was able to fractionate a clarified bacteria lysate in only 3 min (after loading) into RNA, protein and pDNA respectively. The pDNA fraction obtained was analyzed to be a homogeneous supercoiled pDNA.

Using DNA as a drug : challenges and opportunities for process engineers

The maturing of the biotechnology industry and a focus on productivity has seen a shift from discovery science to small-scale bench-top research to higher productivity, large scale production. Health companies are aggressively expanding their biopharmaceutical interests, an expansion which is facilitated by biochemical and bioprocess engineering. An area of continuous growth is vaccines. Vaccination will be a key intervention in the case of an influenza pandemic. The global manufacturing capacity for fast turn around vaccines is currently woefully inadequate at around 300 million shots. As the prevention of epidemics requires > 80 % vaccination, in theory the world should currently be aiming for the ability to produce around 5.3 billion vaccines. Presented is a production method for the creation of a fast turn around DNA vaccine. A DNA vaccine could have a production time scale of as little as two weeks. This process has been harnessed into a pilot scale production system for the creation of a pre-clinical grade malaria vaccine in a collaborative project with the Coppel Lab, Department of Microbiology, Monash University. In particular, improvements to the fermentation, chromatography and delivery stages will be discussed. Consideration will then be given as to how the fermentation stage affects the mid and downstream processing stages.

The applicability of the Precaution Adoption Process Model for understanding self-regulatory driving behaviour among older drivers

1. Introduction The success of self-regulation, in terms of enhancing older drivers’ safety and maintaining their mobility, depends largely upon older drivers’ awareness of the declines in their driving abilities. Therefore, interventions targeted at increasing older drivers’ safety should aim to enhance their awareness of their physical, sensory and cognitive limitations. Moreover, previous research suggests that driving behaviour change may occur through stages and that interventions and feedback may be perceived differently at each stage. 2. Study aims To further understand the process of driving self-regulation among older adults by exploring their perceptions and experiences of self-regulation, using the PAPM as a framework. To investigate the possible impact of feedback on their driving on their decision making process. 3. Methodology Research tool: Qualitative focus groups (n=5 sessions) Recruitment: Posters, media, newspaper advertisement and emails Inclusion criteria: Aged 70 or more, English-speaking, current drivers Participants: Convenience sample of 27 men and women aged 74 to 90 in the Sunshine Coast and Brisbane city, Queensland, Australia. 4. Analysis Thematic analysis was conducted following the process outlined by Braun and Clarke (2006) to identify, analyse and report themes within the data. Four main themes were identified.

Virtual business role-play : leveraging familiar environments to prime stakeholder memory during process elicitation

Business process models have traditionally been an effective way of examining business practices to identify areas for improvement. While common information gathering approaches are generally efficacious, they can be quite time consuming and have the risk of developing inaccuracies when information is forgotten or incorrectly interpreted by analysts. In this study, the potential of a role-playing approach for process elicitation and specification has been examined. This method allows stakeholders to enter a virtual world and role-play actions as they would in reality. As actions are completed, a model is automatically developed, removing the need for stakeholders to learn and understand a modelling grammar. Empirical data obtained in this study suggests that this approach may not only improve both the number of individual process task steps remembered and the correctness of task ordering, but also provide a reduction in the time required for stakeholders to model a process view.

On Merging Business Process Management and Geographic Information Systems : Modeling and Execution of Ecological Concerns in Processes

Business Process Management describes a holistic management approach for the systematic design, modeling, execution, validation, monitoring and improvement of organizational business processes. Traditionally, most attention within this community has been given to control-flow aspects, i.e., the ordering and sequencing of business activities, oftentimes in isolation with regards to the context in which these activities occur. In this paper, we propose an approach that allows executable process models to be integrated with Geographic Information Systems. This approach enables process models to take geospatial and other geographic aspects into account in an explicit manner both during the modeling phase and the execution phase. We contribute a structured modeling methodology, based on the well-known Business Process Model and Notation standard, which is formalized by means of a mapping to executable Colored Petri nets. We illustrate the feasibility of our approach by means of a sustainability-focused case example of a process with important ecological concerns.

‘It’s about finding a way’ : children, sites of opportunity, and building everyday peace in Colombia

The multiple forms of violence associated with protracted conflict disproportionately affect young people. Literature on conflict-affected children often focuses on the need to provide stability and security through institutions such as schools but rarely considers how young people themselves see these sites as part of their everyday lives. The enduring, pervasive, and complex nature of Colombia’s conflict means many young Colombians face the challenges of poverty, persistent social exclusion, and violence. Such conditions are exacerbated in ‘informal’ barrio communities such as los Altos de Cazucá, just south of the capital Bogotá. Drawing on field research in this community, particularly through interviews conducted with young people aged 10 to 17 this article explores how young people themselves understand the roles of the local school and ngo in their personal conceptualisations of the violence in their everyday lives. The evidence indicates that children use spaces available to them opportunistically and that these actions can and should be read as contributing to local, everyday forms of peacebuilding. The ways in which institutional spaces are understood and used by young people as ‘sites of opportunity’ challenges the assumed illegitimacy of young people’s voices and experiences in these environments.

A stakeholder process evaluation of a social marketing walking intervention targeting children

One quarter of Australian children are overweight or obese (ABS, 2010), putting them at increased risk of physical and psychological health problems (Reilly et al., 2003). Overweight and obesity in childhood tends to persist into adulthood and is associated with premature death and morbidity (Reilly & Kelly, 2011). Increases in Australian children’s weight have coincided with declines in active transportation, such as walking, to school (Salmon et al., 2005). To address this problem, the Victorian Health Promotion Foundation (VicHealth), which is an independent statutory authority which advises government and contributes to promoting good health in Victoria (VicHealth, 2014), developed the Walk to School program. Walk to School aims to encourage primary school children in Victoria to walk to and from school more often. Walking to school is a low cost and effective means of reducing excess weight (Rosenberg et al., 2006) that can be easily integrated into daily routine (Brophy et al., 2011). The purpose of this paper is to present the results of the stakeholder process evaluation of Walk to School 2013, which forms part of a broader outcome evaluation that is currently in field. Although there is an emphasis on outcome evaluation of programs, process evaluation can be equally important in determining program success (Saunders et al., 2005). Further, process evaluation to assess program delivery and utilization is explicitly recommended by two social marketing frameworks (see Lefebvre et al., 1988; Walsh et al., 1993).

Autonomous vehicle path planning for persistence monitoring under uncertainty using Gaussian based Markov decision process

One of the main challenges facing online and offline path planners is the uncertainty in the magnitude and direction of the environmental energy because it is dynamic, changeable with time, and hard to forecast. This thesis develops an artificial intelligence for a mobile robot to learn from historical or forecasted data of environmental energy available in the area of interest which will help for a persistence monitoring under uncertainty using the developed algorithm.

Process variability of pollutant build-up on urban road surfaces

Knowledge of the pollutant build-up process is a key requirement for developing stormwater pollution mitigation strategies. In this context, process variability is a concept which needs to be understood in-depth. Analysis of particulate build-up on three road surfaces in an urban catchment confirmed that particles <150µm and >150µm have characteristically different build-up patterns, and these patterns are consistent over different field conditions. Three theoretical build-up patterns were developed based on the size-fractionated particulate build-up patterns, and these patterns explain the variability in particle behavior and the variation in particle-bound pollutant load and composition over the antecedent dry period. Behavioral variability of particles <150µm was found to exert the most significant influence on the build-up process variability. As characterization of process variability is particularly important in stormwater quality modeling, it is recommended that the influence of behavioral variability of particles <150µm on pollutant build-up should be specifically addressed. This would eliminate model deficiencies in the replication of the build-up process and facilitate the accounting of the inherent process uncertainty, and thereby enhance the water quality predictions.

The challenge of developing a fleet driving risk assessment tool: What can be learned from the process?

One of the riskiest activities in the course of a person's work is driving. By developing and testing a new work driving risk assessment measurement tool for use by organisations this research will contribute to the safety of those who drive for work purposes. The research results highlighted limitations associated with current self-report measures and provided evidence that the work driving environment is extremely complex and involves constant interactions between humans, vehicles, the road environment, and the organisational context.

Culture in Business Process Management: How cultural values determine BPM success

There is consensus among practitioners and academics that culture is a critical factor that is able to determine success or failure of BPM initiatives. Yet, culture is a topic that seems difficult to grasp and manage. This may be the reason for the overall lack of guidance on how to address this topic in practice. We have conducted in-depth research for more than three years to examine why and how culture is relevant to BPM. In this chapter, we introduce a framework that explains the role of culture in BPM. We also present the relevant cultural values that compose a BPM culture, and we introduce a tool to examine the supportiveness of organizational cultures for BPM. Our research results provide the basis for further empirical analyses on the topic and support practitioners in the management of culture as an important factor in BPM initiatives.