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.

Airway cellularity, lipid laden macrophages and microbiology of gastric juice and airways in children with reflux oesophagitis

Background: Gastroesophageal reflux disease (GORD) can cause respiratory disease in children from recurrent aspiration of gastric contents. GORD can be defined in several ways and one of the most common method is presence of reflux oesophagitis. In children with GORD and respiratory disease, airway neutrophilia has been described. However, there are no prospective studies that have examined airway cellularity in children with GORD but without respiratory disease. The aims of the study were to compare (1) BAL cellularity and lipid laden macrophage index (LLMI) and, (2) microbiology of BAL and gastric juices of children with GORD (G+) to those without (G-). Methods: In 150 children aged <14-years, gastric aspirates and bronchoscopic airway lavage (BAL) were obtained during elective flexible upper endoscopy. GORD was defined as presence of reflux oesophagitis on distal oesophageal biopsies. Results: BAL neutrophil% in G- group (n = 63) was marginally but significantly higher than that in the G+ group (n = 77), (median of 7.5 and 5 respectively, p = 0.002). Lipid laden macrophage index (LLMI), BAL percentages of lymphocyte, eosinophil and macrophage were similar between groups. Viral studies were negative in all, bacterial cultures positive in 20.7% of BALs and in 5.3% of gastric aspirates. BAL cultures did not reflect gastric aspirate cultures in all but one child. Conclusion: In children without respiratory disease, GORD defined by presence of reflux oesophagitis, is not associated with BAL cellular profile or LLMI abnormality. Abnormal microbiology of the airways, when present, is not related to reflux oesophagitis and does not reflect that of gastric juices. © 2005 Chang et al; licensee BioMed Central Ltd.

Culture independent analysis of greyback canegrub-associated microflora and microbial community comparison using subtractive hybridisation

Greyback canegrubs cost the Australian sugarcane industry around $13 million per annum in damage and control. A novel and cost effective biocontrol bacterium could play an important role in the integrated pest management program currently in place to reduce damage and control associated costs. During the course of this project, terminal restriction fragment length polymorphism (TRFLP), 16-S rDNA cloning, suppressive subtractive hybridisation (SSH) and entomopathogen-specific PCR screening were used to investigate the little studied canegrub-associated microflora in an attempt to discover novel pathogens from putatively-diseased specimens. Microflora associated with these soil-dwelling insects was found to be both highly diverse and divergent between individual specimens. Dominant members detected in live specimens were predominantly from taxa of known insect symbionts while dominant sequences amplified from dead grubs were homologous to putativelysaprophytic bacteria and bacteria able to grow during refrigeration. A number of entomopathogenic bacteria were identified such as Photorhabdus luminescens and Pseudomonas fluorescens. Dead canegrubs prior to decomposition need to be analysed if these bacteria are to be isolated. Novel strategies to enrich putative pathogen-associated sequences (SSH and PCR screening) were shown to be promising approaches for pathogen discovery and the investigation of canegrubsassociated microflora. However, due to inter- and intra-grub-associated community diversity, dead grub decomposition and PCR-specific methodological limitations (PCR bias, primer specificity, BLAST database restrictions, 16-S gene copy number and heterogeneity), recommendations have been made to improve the efficiency of such techniques. Improved specimen collection procedures and utilisation of emerging high-throughput sequencing technologies may be required to examine these complex communities in more detail. This is the first study to perform a whole-grub analysis and comparison of greyback canegrub-associated microbial communities. This work also describes the development of a novel V3-PCR based SSH technique. This was the first SSH technique to use V3-PCR products as a starting material and specifically compare bacterial species present in a complex community.

Co-creative media : theorising digital storytelling as a platform for researching and developing participatory culture

This paper considers the question, ‘what is co-creative media, and why is it a useful idea in social media research’? The term ‘co-creative media’ is now used by Creative Industries researchers at QUT to describe their digital storytelling practices. Digital storytelling is a set of collaborative digital media production techniques that have been used to facilitate social participation in numerous Australian and international contexts. Digital storytelling has been adapted by Creative Industries researchers at QUT as a platform for researching the potential of vernacular creativity in a variety of contexts, including social inclusion of marginalized and disadvantaged groups; inclusion in public histories of narratives that might be overlooked; and articulation of voices that otherwise remain silent in the formulation of social and economic development strategies. The adaption of digital storytelling to different contexts has been shaped by the reflexive, recursive, and pragmatic requirements of action research. Amongst other things, this activity draws attention to the agency of researchers in facilitating these kinds of participatory media processes and outcomes. This discussion serves to problematise concepts of participatory media by introducing the term ‘co-creative media’ and differentiating these from other social media production practices.

Developing a supportive culture for teaching and learning : a university, faculty and school perspective

In the early 1990's the University of Salford was typical of most pre-1992 Universities in that whilst students provided much of it's income, little attention was paid to pedagogy. As Warren Piper (1994) observed, University teachers were professional in their subject areas but generally did not seek to acquire a pedagogy of HE. This was the case in Alsford. Courses were efficiently run but only a minority of staff were engaged in actively considering learning and teaching issues. Instead staff time was spent on research and commercial activity.----- In the mid-1990's the teaching environment began to change significantly. As well as Dearing, the advent of QAA and teaching quality reviews, Salford was already experiencing changes in the characteristics of its student body. Wideing access was on our agenda before it was so predominant nationally. With increasing numbers and heterogeneity of students as well as these external factors, new challenges were facing the University and teaching domain.----- This paper describes how a culture which values teaching, learning and pedagogic inquiry is being created in the university. It then focuses on parts of this process specific to the Faculty of Business and Informatics, namely the Faculty's Learning and Teaching Research Network and the establishment of the Centre for Construction Education in the School of Construction and Property Management.----- The Faculty of Business and Informatics' Learning and Teaching Research Network aims to raise the profile, quality and volume of pedagogic research across the five schools in the faculty. The initiative is targeted at all academics regardless of previous research experience. We hope to grow and nurture research potential where it exists and to acknowledge and use the existing expertise of subject-based researchers in collaborative ventures. We work on the principle that people are deliged to share what they know but need appreciation and feedback for doing so. A further ain is to surface and celebrate the significant amount of tacit knowledge in the area of pedagogy evidenced by the strength of student and employer feedback in many areas of the faculty's teaching.----- The Faculty embraces generic and core management expertise but also includes applied management disciplines in information systems and construction and property management where internationally leading research activities and networked centres of excellence have been established. Drawing from this experience, and within the context of the Faculty network, a Centre for Construction Education is being established with key international external partners to develop a sustainable business model of an enterprising pedagogic centre that can undertake useful research to underpin teaching in the Faculty whilst offering sustainable business services to allow it to benefit from pump-priming grant funding.----- Internal and external networking are important elements in our plans and ongoing work. Key to this are our links with the LTSN subject centres (BEST and CEBE) and the LTSN generic centre. The paper discusses networking as a concept and gives examples of practices which have proved useful in this context.----- The academic influences on our approach are also examined. Dixon’s (2000) work examining how a range of companies succeed through internal knowledge sharing has provided a range of transferable practices. We also examine the notion of dialogue in this context, defined by Ballantyne (1999) as ‘The interactive human process of reasoning together which comes into being through interactions based on spontaneity or need and is enabled by trust’ Social constructionist principles of Practical Authorship (Shotter, 1993, Pavlica, Holman and Thorpe, 1998)) have also proved useful in developing our perspective on learning and knowledge creation within our community of practice.