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.

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.

Multi-action recognition via stochastic modelling of optical flow and gradients

In this paper we propose a novel approach to multi-action recognition that performs joint segmentation and classification. This approach models each action using a Gaussian mixture using robust low-dimensional action features. Segmentation is achieved by performing classification on overlapping temporal windows, which are then merged to produce the final result. This approach is considerably less complicated than previous methods which use dynamic programming or computationally expensive hidden Markov models (HMMs). Initial experiments on a stitched version of the KTH dataset show that the proposed approach achieves an accuracy of 78.3%, outperforming a recent HMM-based approach which obtained 71.2%.

Development and parameterization of a rain- and fire-driven model for exploring elephant effects in African savannas

We describe the development and parameterization of a grid-based model of African savanna vegetation processes. The model was developed with the objective of exploring elephant effects on the diversity of savanna species and structure, and in this formulation concentrates on the relative cover of grass and woody plants, the vertical structure of the woody plant community, and the distribution of these over space. Grid cells are linked by seed dispersal and fire, and environmental variability is included in the form of stochastic rainfall and fire events. The model was parameterized from an extensive review of the African savanna literature; when available, parameter values varied widely. The most plausible set of parameters produced long-term coexistence between woody plants and grass, with the tree-grass balance being more sensitive to changes in parameters influencing demographic processes and drought incidence and response, while less sensitive to fire regime. There was considerable diversity in the woody structure of savanna systems within the range of uncertainty in tree growth rate parameters. Thus, given the paucity of height growth data regarding woody plant species in southern African savannas, managers of natural areas should be cognizant of different tree species growth and damage response attributes when considering whether to act on perceived elephant threats to vegetation. © 2007 Springer Science+Business Media B.V.

Quantifying uncertainty in parameter estimates for stochastic models of collective cell spreading using approximate Bayesian computation

Wound healing and tumour growth involve collective cell spreading, which is driven by individual motility and proliferation events within a population of cells. Mathematical models are often used to interpret experimental data and to estimate the parameters so that predictions can be made. Existing methods for parameter estimation typically assume that these parameters are constants and often ignore any uncertainty in the estimated values. We use approximate Bayesian computation (ABC) to estimate the cell diffusivity, D, and the cell proliferation rate, λ, from a discrete model of collective cell spreading, and we quantify the uncertainty associated with these estimates using Bayesian inference. We use a detailed experimental data set describing the collective cell spreading of 3T3 fibroblast cells. The ABC analysis is conducted for different combinations of initial cell densities and experimental times in two separate scenarios: (i) where collective cell spreading is driven by cell motility alone, and (ii) where collective cell spreading is driven by combined cell motility and cell proliferation. We find that D can be estimated precisely, with a small coefficient of variation (CV) of 2–6%. Our results indicate that D appears to depend on the experimental time, which is a feature that has been previously overlooked. Assuming that the values of D are the same in both experimental scenarios, we use the information about D from the first experimental scenario to obtain reasonably precise estimates of λ, with a CV between 4 and 12%. Our estimates of D and λ are consistent with previously reported values; however, our method is based on a straightforward measurement of the position of the leading edge whereas previous approaches have involved expensive cell counting techniques. Additional insights gained using a fully Bayesian approach justify the computational cost, especially since it allows us to accommodate information from different experiments in a principled way.

No child’s play: Recognising the agency of former child soldiers in peace building processes

Engaging with the emerging discourse on children that recognises childhood as culturally specific and that children actively engage with their environment, this paper questions the dominant discourse’s view of children as passive recipients of socialisation. This paper argues that the discourse on children’s agency is a more useful framework for understanding the experiences of former child soldiers and that engaging meaningfully with this discourse will both improve life outcomes and reduce the risk of ongoing instability. This argument is made by an examination of the two discourses; examining their development and arguing for the usefulness of the agency discourse. This provides for an examination of children’s agency in education and skills training programs and of their political involvement (or marginalisation) in three conflicts: Colombia, Sierra Leone and Uganda. Recognising children as agents and engaging with how they navigate their lived experiences after involvement in conflict testifies to children’s resilience and their desire for change. Challenging the dominant discourse through the agency discourse allows for the acknowledgement of former child soldiers as both social and political agents in their own right and of their potential for contributing to stable and lasting peace.

Agent-based modelling and simulation of airplane boarding processes

Passenger flow simulations are an important tool for designing and managing airports. This thesis examines the different boarding strategies for the Boeing 777 and Airbus 380 aircraft in order to investigate their current performance and to determine minimum boarding times. The most optimal strategies have been discovered and new strategies that are more efficient are proposed. The methods presented offer reduced aircraft boarding times which plays an important role for reducing the overall aircraft Turn Time for an airline.

Alive SMC^2: Bayesian model selection for low-count time series models with intractable likelihoods

In this paper we present a new method for performing Bayesian parameter inference and model choice for low count time series models with intractable likelihoods. The method involves incorporating an alive particle filter within a sequential Monte Carlo (SMC) algorithm to create a novel pseudo-marginal algorithm, which we refer to as alive SMC^2. The advantages of this approach over competing approaches is that it is naturally adaptive, it does not involve between-model proposals required in reversible jump Markov chain Monte Carlo and does not rely on potentially rough approximations. The algorithm is demonstrated on Markov process and integer autoregressive moving average models applied to real biological datasets of hospital-acquired pathogen incidence, animal health time series and the cumulative number of poison disease cases in mule deer.

BP-XACML: An authorisation policy language for business processes

XACML has become the defacto standard for enterprise- wide, policy-based access control. It is a structured, extensible language that can express and enforce complex access control policies. There have been several efforts to extend XACML to support specific authorisation models, such as the OASIS RBAC profile to support Role Based Access Control. A number of proposals for authorisation models that support business processes and workflow systems have also appeared in the literature. However, there is no published work describing an extension to allow XACML to be used as a policy language with these models. This paper analyses the specific requirements of a policy language to express and enforce business process authorisation policies. It then introduces BP-XACML, a new profile that extends the RBAC profile for XACML so it can support business process authorisation policies. In particular, BP-XACML supports the notion of tasks, and constraints at the level of a task instance, which are important requirements in enforcing business process authorisation policies.

Fractional models in space for diffusive processes in heterogeneous media with applications in cell motility and electrical signal propagation

This work addresses fundamental issues in the mathematical modelling of the diffusive motion of particles in biological and physiological settings. New mathematical results are proved and implemented in computer models for the colonisation of the embryonic gut by neural cells and the propagation of electrical waves in the heart, offering new insights into the relationships between structure and function. In particular, the thesis focuses on the use of non-local differential operators of non-integer order to capture the main features of diffusion processes occurring in complex spatial structures characterised by high levels of heterogeneity.

Comparative exploration of narrative processes for better and poorer outcomes for depression

Background Contemporary psychotherapy research demonstrates that whilst most clients respond positively to psychological interventions, a small, but significant proportion of clients fail to experience the expected benefits of therapy. Although methodologies exist that enable the identification of successful and unsuccessful therapy, we have a limited understanding of the processes associated with these outcomes. Aim The current study sought to examine the relationship between therapeutic outcome and therapeutic language. Methodology: The therapeutic outcomes of 42 trainee-therapists who provided psychotherapy to 173 clients were tracked with the OQ-45.2 over a 5 year period with the view of identifying the client/ trainee-therapist dyads with the best and poorest outcomes. The 6 best outcome and 6 poorest outcome client/ trainee-therapist dyads were identified in order to examine the characteristics of therapeutic conversations associated with better and poorer therapy outcomes. Therapeutic conversations were analysed with the Narrative Process Coding System. Findings The best outcome client/ trainee-therapist dyads demonstrated significant increases in reflexive conversation over the course of psychotherapy. Implications Examining the practices of the best and poorest outcome client/ trainee-therapist dyads with objective measures of therapy outcome provides an important first step in understanding how therapeutic language may contribute to the greatest therapeutic improvement or deterioration.

"Social Stuff" and institutional micro-processes: Alcohol use by students in Australian university residential colleges

The literature on alcohol consumption among university and residential college students in Australia and comparable countries shows a high incidence of heavy and/or frequent drinking. In this article, we report the findings from a study on alcohol consumption among undergraduate university students living in residential colleges in Australia. The aim of the study was to examine residents’ alcohol use as part of a broader set of institutional practices in higher education that are constructed as central to the student experience. The data were collected through in-depth semistructured interviews with 29 students from seven residential colleges. We found that inclusion of alcohol in many students’ social and extracurricular activities while residing in college is associated with heavy and/or frequent drinking. We suggest that the use of alcohol among students is shaped by the colleges’ institutional micro-processes, leading to a tension between college managements’ aim to foster alcohol citizenship and students’ liberty to engage in frequent and/or heavy drinking.

Comparing the effect of inadequate documentation to the cost management and tendering processes in managing contractor and fixed lump sum contracts in Australia

Purpose of this paper This research aims to examine the effects of inadequate documentation to the cost management & tendering processes in Managing Contractor Contracts using Fixed Lump Sum as a benchmark. Design/methodology/approach A questionnaire survey was conducted with industry practitioners to solicit their views on documentation quality issues associated with the construction industry. This is followed by a series of semi-structured interviews with a purpose of validating survey findings. Findings and value The results showed that documentation quality remains a significant issue, contributing to the industries inefficiency and poor reputation. The level of satisfaction for individual attributes of documentation quality varies. Attributes that do appear to be affected by the choice of procurement method include coordination, build ability, efficiency, completeness and delivery time. Similarly the use and effectiveness of risk mitigation techniques appears to vary between the methods, based on a number of factors such as documentation completeness, early involvement, fast tracking etc. Originality/value of paper This research fills the gap of existing body of knowledge in terms of limited studies on the choice of a project procurement system has an influence on the documentation quality and the level of impact. Conclusions Ultimately research concludes that the entire project team including the client and designers should carefully consider the individual projects requirements and compare those to the trade-offs associated with documentation quality and the procurement method. While documentation quality is definitely an issue to be improved upon, by identifying the projects performance requirements a procurement method can be chosen to maximise the likelihood that those requirements will be met. This allows the aspects of documentation quality considered most important to the individual project to be managed appropriately.

A determinism and desire: Some neurological processes in perceiving the design object

The environment moderates behaviour using a subtle language of ‘affordances’ and ‘behaviour-settings’. Affordances are environmental offerings. They are objects that demand action; a cliff demands a leap and binoculars demand a peek. Behaviour-settings are ‘places;’ spaces encoded with expectations and meanings. Behaviour-settings work the opposite way to affordances; they demand inhibition; an introspective demeanour in a church or when under surveillance. Most affordances and behaviour-settings are designed, and as such, designers are effectively predicting brain reactions. • Affordances are nested within, and moderated by behaviour-settings. Both trigger automatic neural responses (excitation and inhibition). These, for the best part cancel each other out. This balancing enables object recognition and allows choice about what action should be taken (if any). But when excitation exceeds inhibition, instinctive action will automatically commence. In positive circumstances this may mean laughter or a smile. In negative circumstances, fleeing, screaming or other panic responses are likely. People with poor frontal function, due to immaturity (childhood or developmental disorders) or due to hypofrontality (schizophrenia, brain damage or dementia) have a reduced capacity to balance excitatory and inhibitory impulses. For these people, environmental behavioural demands increase with the decline of frontal brain function. • The world around us is not only encoded with symbols and sensory information. Opportunities and restrictions work on a much more primal level. Person/space interactions constantly take place at a molecular scale. Every space we enter has its own special dynamic, where individualism vies for supremacy between the opposing forces of affordance-related excitation and the inhibition intrinsic to behaviour-settings. And in this context, even a small change–the installation of a CCTV camera can turn a circus to a prison. • This paper draws on cutting-edge neurological theory to understand the psychological determinates of the everyday experience of the designed environment.